Continental J. Food Science and Technology



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mso-hansi-font-family:"Times New Roman"; mso-bidi-font-family:"Times New Roman"; color:#4F81BD; mso-ansi-language:EN-US; mso-fareast-language:EN-US; font-weight:bold; font-style:italic;} span.msoIns {mso-style-type:export-only; mso-style-name:""; text-decoration:underline; text-underline:single; color:teal;} span.msoDel {mso-style-type:export-only; mso-style-name:""; text-decoration:line-through; color:red;} .MsoChpDefault {mso-style-type:export-only; mso-default-props:yes; font-size:10.0pt; mso-ansi-font-size:10.0pt; mso-bidi-font-size:10.0pt; mso-ascii-font-family:Calibri; mso-fareast-font-family:Calibri; mso-hansi-font-family:Calibri; mso-bidi-font-family:Arial;} @page Section1 {size:612.0pt 792.0pt; margin:72.0pt 90.0pt 72.0pt 90.0pt; mso-header-margin:36.0pt; mso-footer-margin:36.0pt; mso-paper-source:0;} div.Section1 {page:Section1;} @page Section2 {size:792.0pt 612.0pt; mso-page-orientation:landscape; margin:90.0pt 72.0pt 90.0pt 72.0pt; mso-header-margin:36.0pt; mso-footer-margin:36.0pt; mso-paper-source:0;} div.Section2 {page:Section2;} @page Section3 {size:612.0pt 792.0pt; margin:72.0pt 90.0pt 72.0pt 90.0pt; mso-header-margin:36.0pt; mso-footer-margin:36.0pt; mso-paper-source:0;} div.Section3 {page:Section3;} @page Section4 {size:792.0pt 612.0pt; mso-page-orientation:landscape; margin:89.85pt 72.0pt 89.85pt 72.0pt; mso-header-margin:36.0pt; mso-footer-margin:36.0pt; mso-paper-source:0;} div.Section4 {page:Section4;} @page Section5 {size:612.0pt 792.0pt; margin:72.0pt 90.0pt 72.0pt 90.0pt; mso-header-margin:36.0pt; mso-footer-margin:36.0pt; mso-paper-source:0;} div.Section5 {page:Section5;} /* List Definitions */ @list l0	{mso-list-id:339241392; mso-list-type:hybrid; mso-list-template-ids:-129468198 1074331663 1074331673 1074331675 1074331663 1074331673 1074331675 1074331663 1074331673 1074331675;} @list l0:level1 {mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l0:level2 {mso-level-number-format:alpha-lower; mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l0:level3 {mso-level-number-format:roman-lower; mso-level-tab-stop:none; mso-level-number-position:right; text-indent:-9.0pt;} @list l0:level4 {mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l0:level5 {mso-level-number-format:alpha-lower; mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l0:level6 {mso-level-number-format:roman-lower; mso-level-tab-stop:none; mso-level-number-position:right; text-indent:-9.0pt;} @list l0:level7 {mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l0:level8 {mso-level-number-format:alpha-lower; mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l0:level9 {mso-level-number-format:roman-lower; mso-level-tab-stop:none; mso-level-number-position:right; text-indent:-9.0pt;} @list l1	{mso-list-id:499778286; mso-list-type:hybrid; mso-list-template-ids:-1558688584 1074331657 1074331651 1074331653 1074331649 1074331651 1074331653 1074331649 1074331651 1074331653;} @list l1:level1 {mso-level-number-format:bullet; mso-level-text:; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:75.55pt; text-indent:-18.0pt; font-family:Wingdings; mso-bidi-font-family:Wingdings;} @list l1:level2 {mso-level-number-format:bullet; mso-level-text:o; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:111.55pt; text-indent:-18.0pt; font-family:"Courier New";} @list l1:level3 {mso-level-number-format:bullet; mso-level-text:; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:147.55pt; text-indent:-18.0pt; font-family:Wingdings; mso-bidi-font-family:Wingdings;} @list l1:level4 {mso-level-number-format:bullet; mso-level-text:; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:183.55pt; text-indent:-18.0pt; font-family:Symbol; mso-bidi-font-family:Symbol;} @list l1:level5 {mso-level-number-format:bullet; mso-level-text:o; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:219.55pt; text-indent:-18.0pt; font-family:"Courier New";} @list l1:level6 {mso-level-number-format:bullet; mso-level-text:; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:255.55pt; text-indent:-18.0pt; font-family:Wingdings; mso-bidi-font-family:Wingdings;} @list l1:level7 {mso-level-number-format:bullet; mso-level-text:; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:291.55pt; text-indent:-18.0pt; font-family:Symbol; mso-bidi-font-family:Symbol;} @list l1:level8 {mso-level-number-format:bullet; mso-level-text:o; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:327.55pt; text-indent:-18.0pt; font-family:"Courier New";} @list l1:level9 {mso-level-number-format:bullet; mso-level-text:; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:363.55pt; text-indent:-18.0pt; font-family:Wingdings; mso-bidi-font-family:Wingdings;} @list l2	{mso-list-id:530534171; mso-list-type:hybrid; mso-list-template-ids:-120584192 1074331671 1074331673 1074331675 1074331663 1074331673 1074331675 1074331663 1074331673 1074331675;} @list l2:level1 {mso-level-number-format:alpha-lower; mso-level-text:"%1\)";	mso-level-tab-stop:none;	mso-level-number-position:left;	text-indent:-18.0pt;} @list l2:level2	{mso-level-number-format:alpha-lower;	mso-level-tab-stop:none;	mso-level-number-position:left;	text-indent:-18.0pt;} @list l2:level3	{mso-level-number-format:roman-lower;	mso-level-tab-stop:none;	mso-level-number-position:right;	text-indent:-9.0pt;} @list l2:level4	{mso-level-tab-stop:none;	mso-level-number-position:left;	text-indent:-18.0pt;} @list l2:level5	{mso-level-number-format:alpha-lower;	mso-level-tab-stop:none;	mso-level-number-position:left;	text-indent:-18.0pt;} @list l2:level6	{mso-level-number-format:roman-lower;	mso-level-tab-stop:none;	mso-level-number-position:right;	text-indent:-9.0pt;} @list l2:level7	{mso-level-tab-stop:none;	mso-level-number-position:left;	text-indent:-18.0pt;} @list l2:level8	{mso-level-number-format:alpha-lower;	mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l2:level9 {mso-level-number-format:roman-lower; mso-level-tab-stop:none; mso-level-number-position:right; text-indent:-9.0pt;} @list l3	{mso-list-id:621303726; mso-list-type:hybrid; mso-list-template-ids:-1867973640 -611810844 67698713 67698715 67698703 67698713 67698715 67698703 67698713 67698715;} @list l3:level1 {mso-level-tab-stop:54.0pt; mso-level-number-position:left; margin-left:54.0pt; text-indent:-18.0pt; mso-ansi-font-weight:normal; vertical-align:super;} @list l4	{mso-list-id:679550830; mso-list-type:hybrid; mso-list-template-ids:648962962 1074331663 1074331673 1074331675 1074331663 1074331673 1074331675 1074331663 1074331673 1074331675;} @list l4:level1 {mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l4:level2 {mso-level-number-format:alpha-lower; mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l4:level3 {mso-level-number-format:roman-lower; mso-level-tab-stop:none; mso-level-number-position:right; text-indent:-9.0pt;} @list l4:level4 {mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l4:level5 {mso-level-number-format:alpha-lower; mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l4:level6 {mso-level-number-format:roman-lower; mso-level-tab-stop:none; mso-level-number-position:right; text-indent:-9.0pt;} @list l4:level7 {mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l4:level8 {mso-level-number-format:alpha-lower; mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l4:level9 {mso-level-number-format:roman-lower; mso-level-tab-stop:none; mso-level-number-position:right; text-indent:-9.0pt;} @list l5	{mso-list-id:814109284; mso-list-template-ids:398486370;} @list l5:level1 {mso-level-start-at:3; mso-level-text:%1; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:24.0pt; text-indent:-24.0pt;} @list l5:level2 {mso-level-start-at:3; mso-level-text:"%1\.%2"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:24.0pt; text-indent:-24.0pt;} @list l5:level3 {mso-level-start-at:7; mso-level-text:"%1\.%2\.%3"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:36.0pt; text-indent:-36.0pt;} @list l5:level4 {mso-level-text:"%1\.%2\.%3\.%4"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:36.0pt; text-indent:-36.0pt;} @list l5:level5 {mso-level-text:"%1\.%2\.%3\.%4\.%5"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:54.0pt; text-indent:-54.0pt;} @list l5:level6 {mso-level-text:"%1\.%2\.%3\.%4\.%5\.%6"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:54.0pt; text-indent:-54.0pt;} @list l5:level7 {mso-level-text:"%1\.%2\.%3\.%4\.%5\.%6\.%7"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:72.0pt; text-indent:-72.0pt;} @list l5:level8 {mso-level-text:"%1\.%2\.%3\.%4\.%5\.%6\.%7\.%8"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:72.0pt; text-indent:-72.0pt;} @list l5:level9 {mso-level-text:"%1\.%2\.%3\.%4\.%5\.%6\.%7\.%8\.%9"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:90.0pt; text-indent:-90.0pt;} @list l6	{mso-list-id:862283899; mso-list-template-ids:-367209190;} @list l6:level1 {mso-level-start-at:3; mso-level-text:%1; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:24.0pt; text-indent:-24.0pt;} @list l6:level2 {mso-level-start-at:3; mso-level-text:"%1\.%2"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:24.0pt; text-indent:-24.0pt;} @list l6:level3 {mso-level-start-at:8; mso-level-text:"%1\.%2\.%3"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:36.0pt; text-indent:-36.0pt;} @list l6:level4 {mso-level-text:"%1\.%2\.%3\.%4"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:36.0pt; text-indent:-36.0pt;} @list l6:level5 {mso-level-text:"%1\.%2\.%3\.%4\.%5"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:54.0pt; text-indent:-54.0pt;} @list l6:level6 {mso-level-text:"%1\.%2\.%3\.%4\.%5\.%6"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:54.0pt; text-indent:-54.0pt;} @list l6:level7 {mso-level-text:"%1\.%2\.%3\.%4\.%5\.%6\.%7"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:72.0pt; text-indent:-72.0pt;} @list l6:level8 {mso-level-text:"%1\.%2\.%3\.%4\.%5\.%6\.%7\.%8"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:72.0pt; text-indent:-72.0pt;} @list l6:level9 {mso-level-text:"%1\.%2\.%3\.%4\.%5\.%6\.%7\.%8\.%9"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:90.0pt; text-indent:-90.0pt;} @list l7	{mso-list-id:905650051; mso-list-type:hybrid; mso-list-template-ids:706616340 1074331663 1074331673 1074331675 1074331663 1074331673 1074331675 1074331663 1074331673 1074331675;} @list l7:level1 {mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l7:level2 {mso-level-number-format:alpha-lower; mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l7:level3 {mso-level-number-format:roman-lower; mso-level-tab-stop:none; mso-level-number-position:right; text-indent:-9.0pt;} @list l7:level4 {mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l7:level5 {mso-level-number-format:alpha-lower; mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l7:level6 {mso-level-number-format:roman-lower; mso-level-tab-stop:none; mso-level-number-position:right; text-indent:-9.0pt;} @list l7:level7 {mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l7:level8 {mso-level-number-format:alpha-lower; mso-level-tab-stop:none; mso-level-number-position:left; text-indent:-18.0pt;} @list l7:level9 {mso-level-number-format:roman-lower; mso-level-tab-stop:none; mso-level-number-position:right; text-indent:-9.0pt;} @list l8	{mso-list-id:914586124; mso-list-template-ids:-773927140;} @list l8:level1 {mso-level-start-at:3; mso-level-text:%1; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:24.0pt; text-indent:-24.0pt; mso-ansi-font-weight:bold; mso-bidi-font-weight:bold;} @list l8:level2 {mso-level-start-at:3; mso-level-text:"%1\.%2"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:24.0pt; text-indent:-24.0pt; mso-ansi-font-weight:bold; mso-bidi-font-weight:bold;} @list l8:level3 {mso-level-start-at:6; mso-level-text:"%1\.%2\.%3"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:36.0pt; text-indent:-36.0pt; mso-ansi-font-weight:bold; mso-bidi-font-weight:bold;} @list l8:level4 {mso-level-text:"%1\.%2\.%3\.%4"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:36.0pt; text-indent:-36.0pt; mso-ansi-font-weight:bold; mso-bidi-font-weight:bold;} @list l8:level5 {mso-level-text:"%1\.%2\.%3\.%4\.%5"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:54.0pt; text-indent:-54.0pt; mso-ansi-font-weight:bold; mso-bidi-font-weight:bold;} @list l8:level6 {mso-level-text:"%1\.%2\.%3\.%4\.%5\.%6"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:54.0pt; text-indent:-54.0pt; mso-ansi-font-weight:bold; mso-bidi-font-weight:bold;} @list l8:level7 {mso-level-text:"%1\.%2\.%3\.%4\.%5\.%6\.%7"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:72.0pt; text-indent:-72.0pt; mso-ansi-font-weight:bold; mso-bidi-font-weight:bold;} @list l8:level8 {mso-level-text:"%1\.%2\.%3\.%4\.%5\.%6\.%7\.%8"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:72.0pt; text-indent:-72.0pt; mso-ansi-font-weight:bold; mso-bidi-font-weight:bold;} @list l8:level9 {mso-level-text:"%1\.%2\.%3\.%4\.%5\.%6\.%7\.%8\.%9"; mso-level-tab-stop:none; mso-level-number-position:left; margin-left:90.0pt; text-indent:-90.0pt; mso-ansi-font-weight:bold; mso-bidi-font-weight:bold;} @list l9	{mso-list-id:961881085; mso-list-template-ids:-1250937094;} @list l9:level1 {mso-level-start-at:3; mso-level-text:%1; mso-level-tab-stop:24.0pt; mso-level-number-position:left; margin-left:24.0pt; text-indent:-24.0pt; mso-ansi-font-weight:bold; mso-bidi-font-weight:bold;} @list l9:level2 {mso-level-start-at:3; mso-level-text:"%1\.%2"; mso-level-tab-stop:24.0pt; mso-level-number-position:left; margin-left:24.0pt; text-indent:-24.0pt; mso-ansi-font-weight:bold; mso-bidi-font-weight:bold;} @list l9:level3 {mso-level-start-at:5; mso-level-text:"%1\.%2\.%3"; mso-level-tab-stop:36.0pt; mso-level-number-position:left; margin-left:36.0pt; text-indent:-36.0pt; mso-ansi-font-weight:bold; mso-bidi-font-weight:bold;} @list l9:level4 {mso-level-text:"%1\.%2\.%3\.%4"; mso-level-tab-stop:36.0pt; mso-level-number-position:left; margin-left:36.0pt; text-indent:-36.0pt; mso-ansi-font-weight:bold; mso-bidi-font-weight:bold;} @list l9:level5 {mso-level-text:"%1\.%2\.%3\.%4\.%5"; mso-level-tab-stop:54.0pt; mso-level-number-position:left; margin-left:54.0pt; text-indent:-54.0pt; mso-ansi-font-weight:bold; mso-bidi-font-weight:bold;} @list l9:level6 {mso-level-text:"%1\.%2\.%3\.%4\.%5\.%6"; mso-level-tab-stop:54.0pt; mso-level-number-position:left; margin-left:54.0pt; text-indent:-54.0pt; mso-ansi-font-weight:bold; mso-bidi-font-weight:bold;} @list l9:level7 {mso-level-text:"%1\.%2\.%3\.%4\.%5\.%6\.%7"; mso-level-tab-stop:72.0pt; mso-level-number-position:left; margin-left:72.0pt; text-indent:-72.0pt; mso-ansi-font-weight:bold; mso-bidi-font-weight:bold;} @list l9:level8 {mso-level-text:"%1\.%2\.%3\.%4\.%5\.%6\.%7\.%8"; mso-level-tab-stop:72.0pt; mso-level-number-position:left; margin-left:72.0pt; text-indent:-72.0pt; mso-ansi-font-weight:bold; mso-bidi-font-weight:bold;} @list l9:level9 {mso-level-text:"%1\.%2\.%3\.%4\.%5\.%6\.%7\.%8\.%9"; mso-level-tab-stop:90.0pt; mso-level-number-position:left; margin-left:90.0pt; text-indent:-90.0pt; mso-ansi-font-weight:bold; mso-bidi-font-weight:bold;} @list l10 {mso-list-id:978732129; mso-list-type:hybrid; mso-list-template-ids:-241154292 1074331663 1074331673 1074331675 1074331663 1074331673 1074331675 1074331663 1074331673 1074331675;} @list l10:level1 {mso-level-tab-stop:none; mso-level-number-position:left; 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Food Science and Technology 4: 1 - 6, 2010                                      ISSN: 2141 – 422X

©Wilolud Journals, 2010                                                                                  http://www.wiloludjournal.com



PROXIMATE AND MINERAL COMPOSITION IN SOME FRESHWATER FISHES IN UPPER RIVER BENUE, YOLA, NIGERIA.



Onyia,L.U1., Milam, C2.; Manu, J.M3. and Allison,D.S.4

1Department of Fisheries; 2Department of Chemistry; 3Department of Chemical Engineering; 4Department of Food Science and Technology, Federal University of Technology, P.M.B.2076, Yola, Adamawa State.



ABSTRACT

The study showed that freshwater fishes are good sources of protein, lipid and micronutrients. The proximate composition of B. filamelosus, T. niloticus, B. niloticus, C. gariepinus and M, rume revealed encouraging high crude protein contents 33.5%, 34.4%, 37.1%, 38.5% and 38.6% respectively. There was significant difference(p>0.05) in percentage crude protein, fibre content, ether extract, ash content, NFE energy values among the fishes sampled. The mineral elements and heavy metals in fish tissues were in the following order-K>Na>Fe>Zn>Cu>Ni>Pb, but Cadmium was not detected in all the fishes sampled. Levels of heavy metals are below WHO limits for human consumption.

The aim of this work is to determine the micronutrients and proximate composition of some freshwater species from arid zone of Nigeria.



KEYWORDS: Proximate composition, mineral elements, heavy metal, freshwater fishes.



INTRODUCTION

Fish is widely accepted because of its high palatability, low cholesterol and tender flesh (Eyo, 2000). It is the cheapest source of animal protein and other essential nutrients required in human diet (Sadiku and Oladimeji, 1991). Fish may be the sole accessible and/or affordable source of animal protein for poor households in urban or semi-urban areas (Bene and Heck, 2005).The nature and quality of nutrients in most animals depend largely on their food type. More so, the feeding habit of an individual fish species greatly affects its body nutrient composition (Lagler et.al., 1977).

<p style="text-align: justify;">

<p style="text-align: justify;">The measurement of some proximate profiles such as protein content, lipids, ash content, nitrogen free extract and crude fibre is often necessary to ensure that they meet the dietary requirements and commercial specifications (Watchman, 2000, Anon, 2000).The study of micro-nutrients present in living organisms is of biological importance because many of such micro-nutrients take part in some metabolic processes and are known to be indispensable to all living things(Shul’man,1974). Fishes contain small amount of these micro-nutrients, some of which are essential nutrients, being components of many enzymes system and metabolic mechanisms that contribute to the growth of the fish. The most important micro-nutrients in form of mineral salts include calcium, potassium, phosphorus, iron, chlorine, while many others are needed in trace amounts. The deficiency in these principal nutritional mineral elements induces a lot of malfunctioning, as it reduces productivity and causes diseases, such as inability of blood to clot, osteoporosis, anaemia, etc (Shul’man,1974 and Mills,1980). The deficiencies of these micro-nutrients, vitamins and protein result in the death of 60% of children under age 5 annually in Africa (Bene and Heck, 2005). The study of these micro-nutrients in fish will reveal the quantity available to fish consumers and prevent the resultant mortality rate due to their deficiencies.

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<p style="text-align: justify;">Furthermore, one major pollution sources that pose serious health risk and environmental concern is heavy metals commonly found in fish and aquatic environment (Onyia, et. al., 2007a and Milam and Onyia, 2007b). However, aquatic organisms require these mineral elements at moderate levels, but when they exceed metabolic demand or requirement they accumulate in tissues of organisms such as fish. Fish can only metabolize heavy metals to a lesser extent because most of them are non-biodegradable (Lenntech, 2006). Therefore, considering the various health risk and nutritional benefits associated with fish consumption, it has become important that, micro-nutrients and proximate composition of fish and their

<p style="text-align: center;">Onyia,L.U et al.,: Continental J. Food Science and Technology 4: 1 - 6, 2010

<p style="text-align: justify;">

<p style="text-align: justify;">

<p style="text-align: justify;">health status be examined in order to establish the safety level of table-sized fish species before consumption. The aim of this work is to determine the micro-nutrients and proximate composition of some freshwater species from arid zone of Nigeria.

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<p style="text-align: justify;">MATERIALS AND METHODS

<p style="text-align: justify;">The fishes were collected from the fish-landing site at Vininkilang within Girei Local Government Area, Adamawa State Nigeria in May 2007. This site is very close Upper Benue River. The fishes were cut, weighed and dried to constant weight (70-800C). The dried samples were ground with mortar and pestle into fine powder and stored in labeled polythene bags until required for analysis.

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<p style="text-align: justify;">Proximate Analysis.

<p style="text-align: justify;">The methods for analyses were the standard procedures of AOAC (1990). Moisture content was determined by oven drying(at 105ºC) overnight, ash by incineration of 2g of each sample in a muffle furnace (Lenton Furnaces, England) at 600ºc for 2hours, protein( N x 6.25) by the Micro-Kjeldahl method, crude lipid was extracted with n-hexane in a soxhlet extractor, crude fibre by acid-base digestion using 1.25% H2S04(w/v) and 1.25% NaoH(w;v) solution, while available nitrogen free extract was calculated by difference. The energy value of the sample was estimated (Kcal; 100g) by multiplying the percentage of crude protein, crude fibre and NFE by the factors of 16.7, 37.7 and 16.7 respectively (Vadivel and Janardhanan, 2004). All proximate components were analyzed in triplicate and reported as mean on %dry weight basis.

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<p style="text-align: justify;">Mineral Analysis

<p style="text-align: justify;">Mineral analysis was done after triple acid digestion according to the method described by Hassan and Umar (2004). Iron, copper, magnesium, calcium, zinc, lead, cadmium and nickel were analyzed by atomic absorption spectrophotometer (Alpha 4 model), while flame photometer (Corning 400 UK) was used for Sodium and Potassium analysis. All determinations were carried out in triplicate and reported as mean mineral content in mg/100g dry matter (DM). Let us know the statistical methods used

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<p style="text-align: justify;">Data analysis

<p style="text-align: justify;">Data collected from each parameter was subjected to computation and analysis of variance (ANOVA) test followed by the least significant difference (LSD).

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<p style="text-align: justify;">ResultS

<p style="text-align: justify;">The mean percentage proximate composition (i. e. protein, fibre, ether extract, ash, moisture content, NFE and dry matter) of the analyzed samples is shown in Table 1. The food values analyzed showed varied values of their presence in the body tissue of the fishes analyzed; with the percentage dry matter, NFE and crude protein recording higher value in that order; followed by moisture content, ash content, ether extract and crude fibre. The fish samples  presented a relatively higher and lower amount of proximate concentrations in the order of ''M. rume> C. gariepinus>B. niloticus> T. niloticus> B. filamelosus. The energy values of the fish samples analyzed showed a relatively higher values in fatty fishes except M. rume''(349.49 Kcal/100g). The lean fishes had B. niloticus (370.12Kcal/100g) and T. niloticus (357.12Kcal/100g). The rest fatty fishes- C.gariepinus and B. filamelosus- recorded 393.26Kcal/100g and 363.7Kcal/100g.

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<p style="text-align: justify;">The result of the means of mineral elements in the body tissue of fish samples analyzed was given in Table2. The variations observed in the values observed in the values obtained showed significant difference. All the mineral elements investigated were represented in all species analyzed except cadmium. The most abundant mineral elements in the fish samples analyzed were Potassium, Sodium, Iron, Zinc and Copper, while Nickel and Lead were in trace amount. Cadmium was not detected in all the fish sampled. T. niloticus, C. gariepinus, B. filamelosus, M. rume and B. niloticus presented a relatively higher amount of mineral content in that order with which they are listed.

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<p style="text-align: center;">Onyia,L.U et al.,: Continental J. Food Science and Technology 4: 1 - 6, 2010

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<p style="text-align: justify;">Discussion

<p style="text-align: justify;">Fish is a source of animal protein and the declining livestock production (Cattle and Poultry) in Nigeria has increased the relative substitution value of fish for meat. Fish is widely accepted because of its high palatability, low cholesterol and tender flesh (Eyo, 2001). However, fewer numbers of consumers eat fish because of its nutritional value. It is therefore necessary to make information available to consumers and fishery workers on the nutritional contribution of some fish species in their daily diets (Adewoye et. al., 2003; Barminas, et. al 1998).

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<p style="text-align: justify;">Concentrations of Potassium (K+) were observed to have appreciably dominated other elements analyzed in all the fish samples examined. This tends to disagree with the work done by Fawole,'' et. al''., (2007) at Asa reservoir, in Ilorin, Kwara State Nigeria; where the dominant element in the fishes sampled was sodium. It could be inferred from the high concentration of potassium (K+) in the tissues of the fish species that the water body from which the fishes were collected is rich in potassium (K+). This must have allowed an active movement of this ion across the gill structure, which in turn may depend on the concentration in the external medium and that the richness in potassium (K+) concentrations would boost the osmoregulatory activities in the organisms(Bently,1971). The concentration of sodium (Na+) in the fish samples examined ranked second among the mineral elements analyzed. The variations recorded in the concentration of mineral in fish muscles examined could be as a result of the rate in which they are available in the water body and the ability of the fish to absorb these inorganic elements from their diet and the environment where they live (Adewoye and Omotosho, 1997).

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<p style="text-align: justify;">Other elements (such as iron, copper, nickel, zinc, lead and cadmium) composition of the fish samples recorded variations in their concentrations both within and between the selected species sampled. This variation in concentrations of the mineral elements in sampled fish tissues agree with the work of Windom ''et. al.'', (1987) which stated that such variation was due to the chemical forms of the elements and their concentrations in the environment. The concentrations of the mineral elements in the fish tissues are in the following order (i.e. Fe>Zn>Cu.>Ni.> Pb). Cadmium was not detected in any of the fish tissues. This report is in agreement with the one obtained by Ako and Salihu (2004). The levels of most of these mineral elements present are in trace amount and are still below World Health Organization limits for human consumption.

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<p style="text-align: justify;">The proximate composition of B. filamelosus, T. niloticus, B. niloticus, C. gariepinus and M. rume reveal encouraging high crude protein contents of 33.5%, 34.4%, 37.1%, 38.5% and 38.6% respectively(Table 1).The relatively high to moderate percentage crude protein may be attributed to the fact that fishes are good source of pure protein, but the differences observed, in values obtained could also be as a result of fish consumption or absorption capability and conversion potentials of essential nutrients from their diets or their local environment into such biochemical attributes needed the organisms body(Burgress, 1975; and Adewoye and Omotosho, 1997). The fatty fishes recorded high ether extract (9.1% and 7.7%). Fish contain essential fatty acids that are useful to human body. The ether extract level in the fish tissues could have been due to the influence of food (Reinitz, 1983; Degani Dosoretz, 1988). The percentage ash in B. filamelosus, M. rume and T. niloticus was within the stipulated levels of 10-15 % (Emokpae and Ajayi, 1989). B. niloticus and C. gariepinus had percentage ash level of 5%. This result was far lower (for C. gariepinus) than the result obtained from Lake Geriyo (Edward, 2007). The percentage ash content in the fishes analyzed is an indication of ample mineral content in fish. Percentage moisture in fish muscles was within the acceptable level in all the samples without any significant difference (P>0.05) could be due to the stable water levels in the environmental location where the fish were collected. There is significant difference in the energy value (kcal/100g) of the fish samples.

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<p style="text-align: justify;">The study therefore, showed that freshwater fishes are good sources of minerals. It could be inferred that the mineral elemental levels of each species is a function of the availability preferential accumulation. However, it was revealed from the study that, micro-nutrients were not low, which could be due to the fact that the body needs of the fish are met and the concentrations in the water body is high. It therefore becomes necessary to equally consider the mineral status of the fish and the persistent food safety of the

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<p style="text-align: justify;">fish prior to consumption in addition to the prevailing choice for fish as a high protein source. This work has unveiled the importance of freshwater fishes as good sources of protein and micro-nutrients. Since nutritional value of freshwater fishes examined are now known, consumers can now know what benefits to derive when these fish species are eaten.

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<p style="text-align: justify;">REFERENCES

<p style="text-align: justify;">Adewoye, S.O. and Omotosho, J. S.(1997). Nutrient Composition of some freshwater Fishes in Nigeria. BioSci. Res. Commun.11 (4)333-336.

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<p style="text-align: justify;">Adewoye, S. O., Fawole, O. O. and Omotosho J. S.(2003). Concentrations of selected Elements in some freshwater fishes in Nigeria. Science Focus Vol.4: Pp10108.←Rewrite thus Science Focus. 4:106-108

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<p style="text-align: justify;">Ako, P. A. and Salihu, S. O.(2004). Studies on some major and trace metals in smoked and over –dried fish. Journal of Applied Sciences and Environmental Management. 8.(2): 5-9.

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<p style="text-align: justify;">Anon,(2000). Communication from the Commission to the Council and Parliament Conf.(2000) 724 Brussade European Commission, 20Pp.

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<p style="text-align: justify;">AOAC (Association of Official Analytical Chemist)(1990). Official Method of analysis of AOAC.14th Ed. W. Horwitz(Editor). Allengton, Washington D. C.

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<p style="text-align: justify;">Barminas, J. T., Charles, M. and D. Emmanuel (1998). Mineral composition of non-Conventional leafy vegetables. Plant Food for Human Nutrition53:29-36.Kluwer Academic Publishers, the Netherlands.

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<p style="text-align: justify;">Bene, C. and S. Heck(2005). Fish and food security in Africa. NAGA World Fish Centre Quarterly.Vol. 28, No.3 and 4:8-13.

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<p style="text-align: justify;">Bentley, P. J. (1971). Endocrine and Osmoregulation, Springer-Verdag Heidelberg. Pp.220-230.

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<p style="text-align: justify;">Burgress, G. H.O.(1975). Increasing the direct consumption of fish. In: W W Pirie(Edu).Food Protein Sources. International Biological Programme 4. Cambridge,Pp.187- 200.

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<p style="text-align: justify;">Edward, V.(2006). A comparative study of body composition of the African catfish (Clarias gariepinus) obtained from Lake Gerio and Gesse-Daddo fish farm, Yola Adamawa State, Nigeria. B. Tech. Project 32Pp.

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<p style="text-align: justify;">Emokpae, A. O. and A.Ajayi (1989). A study of the quality of fish meat. NIOMR 1989 Annual Report: 66-69.

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<p style="text-align: justify;">Eyo, A. A.(2001). Fish Processing Technology in the tropics. University of Ilorin, Nigeria. 430Pp.

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<p style="text-align: justify;">Fawole, O.O., Ogundiran, M. A., Ayandiran, T. A. and Olagunju, O. F.(2007).Proximate And Mineral Composition in some freshwater fishes in Nigeria. Internet Journal of Food Safety, Vol.9, 2007. Pp. 52-55.

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<p style="text-align: justify;">Hassan, L. G. and Umar, K. J.(2004). Proximate and mineral compositions of seeds and Pulp of African Locust Beans(Parkia biglobosa L.). Nigerian Journal of Basic and Applied Sciences13:15-27.

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<p style="text-align: justify;">Lagler, K. F., Bardach, J. E. and Miller R. R.(1977). Lethology, the study of fishes. Wiley, New York. Pp156-163.

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<p style="text-align: justify;">Lenntech (2006). Lenntech Water treatment and air purification holding B. V.Retrieved June2006, from http/www/lenntech com/ feedback 2htm.

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<p style="text-align: center;">Onyia,L.U et al.,: Continental J. Food Science and Technology 4: 1 - 6, 2010

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<p style="text-align: justify;">Mills, C. F.(1980).The mineral nutrient of livestock(Underwood, E. J. 1981 Ed.) Commonwealth Bureaux Pg9.

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<p style="text-align: justify;">Milam, Charles and L. U. Onyia(2007). Comparative Investigation of heavy metals in pelagic and benthic fishes from Upper River Benue, Yola Adamawa State. ''Journal of Physical Sciences, Vol. 3, No. 4, 2007 ''

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<p style="text-align: justify;">Onyia, L. U. and Milam, C. and Ese, D.S.(2007).Investigation of heavy metals in four Commercial fishes along Upper River Benue, Yola.International Journal of Physical Sciences, Vol. 2, No. 2, 2007.

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<p style="text-align: justify;">Sadiku, S. O. E. and A. A. Oladimeji (1991). Relationship of proximate composition of Lates niloticus (L). Synodontis schall RES. COMM. 3(1) 29-40.

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<p style="text-align: justify;">Shul’man, G.E.(1974). Life cycle of fish physiology and Biochemistry, Halted Press a Division of John Wiley& Sons Inc. N. Y.(Ist Ed.) Pp101-104.

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<p style="text-align: justify;">Vadivel, V. and Janardhanan, K.(2000b). Chemical Composition of the Underutilized Legume Cassia hirsute L. Plant Foods for Human Nutrition 55:369-381.

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<p style="text-align: justify;">Watchman, J. J.(2000). Composition and Quality of fish, Edinburgh, Torry Research Station.

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<p style="text-align: justify;">Windom, H., Stein, D., Scheldon, R. and Smith, J. R.(1987). Composition of trace metal Concentrations in muscle of a benthopelagic fish. (Cory phaenoides armatus) from the Atlantic and Pacific oceans. Deepsea Research 34(2)213-220.undefined

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<p style="text-align: justify;">Table1: The percentage means proximate composition in the body of fish species from Upper Benue River.

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<p style="text-align: center;">Onyia,L.U et al.,: Continental J. Food Science and Technology 4: 1 - 6, 2010

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<p style="text-align: justify;">Table2: Means of mineral elements in the body tissue of from Upper Benue River (mg/g).

<p style="text-align: justify;"> <p style="text-align: justify;"> Key:

<p style="text-align: justify;"> BN- B. niloticus, MR- M. rume,   BF-   B, filamelosus,  TL- T. niloticus,  CL-C.gariepinus, ND-Not detected.

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<p style="text-align: justify;">Received for Publication: 03/04/2010

<p style="text-align: justify;">Accepted for Publication: 30/04/2010

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<p style="text-align: justify;">Corresponding Author:

Onyia,L.U

Department of Fisheries; Federal University of Technology, P.M.B.2076, Yola, Adamawa State.

<p style="text-align: justify;">Email [mailto:uchelucky2005@yahoo.com uchelucky2005@yahoo.com]

<p style="text-align: justify;">Continental J. Food Science and Technology 4: 7 – 13, 2010                                  ISSN: 2141 – 422X

<p style="text-align: justify;">©Wilolud Journals, 2010                                                                                  http://www.wiloludjournal.com

EFFECT OF ADDED DEFATTED BENISEED ON THE  QUALITY OF ACHA BASED BISCUITS

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<p style="text-align: center;">1,2Ayo, J.A; 1Ikuomola, D.S; 1Esan,Y.O; 2Onuoha. O. G;  2Ayo, V.A   and 2Ekele. V

<p style="text-align: center;">Dept. Food Science and Technology,

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<p style="text-align: center;">1Joseph Ayo Babalola University, Ikeji-Arakeji, PMB 5006, Ilesa. Osun State. Nigeria

<p style="text-align: center;">2The Federal Polytechnic, PMB 0231, Bauchi. Bauchi State. Nigeria

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<p style="margin: 0cm 43.2pt 0.0001pt;">ABSTRACT

<p style="margin: 0cm 43.2pt 0.0001pt; text-align: justify;">The acha (Digitaria exile staff) grain was washed (tap water), sun dried, milled and sieved (400mn sieve aperture) to produce acha flour. The beniseed(Sesamum indicum) seeds were sorted, washed, sun dried, milled, defatted (using ethanol), dried and sieved (400mn sieve aperture) to produce beniseed flour. The defatted beniseed flour was substituted into the acha flour at 0-20% to produce beniseed-acha composite flour. This was mixed with fat, sugar, salt, baking powder to produce batter. The batter was rolled (on a steel table), cut into shapes (using biscuit cutter), placed on greased trays and baked at 160oC for 20 minutes. The effect of added defatted beniseed on  the physical, chemical and sensory quality of acha–beniseed composite biscuit were evaluated. The protein, fat, moisture, ash, breaking strength and spread ratio increased from 10.5 to 12.9, 1.80 to 2.15, 1.0 to 1.6, 1.20 to 3.20%, 1.49 to 1.67kg and 6.0 to 7.25, respectively with increase in the percentage of added defatted beniseed. However, the average means of the sensory properties decreased from 8.30 to 4.30, 7.20 to 5.80, 8.50 to 6.30, 8.30 to 4.20, and 7.90 to 3.70 for color, texture, appearance, flavor and general acceptability, respectively at p ≤ 0.05 level of acceptance. The addition of defatted beniseed above 10% has a significant negative effect on the acceptances of the product. There was an increase of 12 and 16.4% of protein and fat content respectively, at the maximum acceptance level (10%) of the added beniseed flour.

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<p style="margin: 0cm 43.2pt 0.0001pt;">KEYWORDS:  Effect, Defatted Beniseed, Quality, Acha Biscuits

 

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INTRODUCTION

<p style="text-align: justify;">The nutritional value of biscuits varies with the type of cereal used. Biscuit is known to generally contain fat (18.5%), carbohydrate (78.23%), ash (1.0%), and protein (7.1%) and salt (0.85%) (Okeagu, 2001). The dependence on the use of wheat flour is one major constraint in biscuit production. The high demand of wheat flour and the in ability of the country to meet this demand calls for research into alternative local sources of flour baking example cassava, acha, millet, sorghum, etc.

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<p style="text-align: justify;">Nigeria is finding itself more and more caught up in “wheat trap” in which most of its foods are made from wheat a foreign cereal. (Whitey, 1971). Most of the common local cereal grains including acha, though having similar structure and composition were left in a state of under development and inadequate processing. Due to lack of attention, acha is still ergonomically primitive because of its very tiny seeds size resulting in difficulty in dehulling.

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<p style="text-align: justify;">Today, plant proteins play significant roles in human nutrition, particularly in developing countries where average protein intake is less than the required. Due to inadequate supplies of animal proteins, there has been a constant search for new protein sources, for use as both functional food ingredients and nutritional supplements, (Obizoba, et al., 1994). Plant protein products are gaining increased interest as ingredients in food systems throughout many parts of the world. The final success of utilizing plant proteins as additives depends greatly upon the favorable characteristics that they impart on foods. Beniseed flour contains substantial quantity of tryptophan and methionine, an important essential amino acids needed for the

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<p style="text-align: center;">Ayo, J.A et al.,:Continental J. Food Science and Technology 4: 7 – 13, 2010

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<p style="text-align: justify;">healthy living (Eleazar et al., 2003). The development of defatted beniseed flour would provide industry with new high protein food ingredients for products formulation and protein fortification (Uwala, 2002).

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<p style="text-align: justify;">Acha has been researched in several areas (Bolaji et al., 1993; Funanbial, 1998; Irving and Jideani, 1997; Jideani and Akingbala, 1993; Jideani et al., 1994; Lasekan 1998; Ayo and Nkama, 2003; Ayo and Nkama, 2004; Ayo and Nkama, 2006; Ayo et al., 2008a; Ayo et al., 2008b; Ayo et al., 2007a; Ayo et al., 2007b).

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<p style="text-align: justify;">The relatively low quantity of acha protein has called for supplementation particularly with inexpensive stable pulse and legumes such as soybean. Research has shown that nutrients composition or proximate composition of food is not enough to determine nutrient bio-availability (Davies and Austine 1982) hence the use of animal feed. The biological utilization of a protein is primarily dependent on its digestibility (Hooda and Jood, 2005).

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<p style="text-align: justify;">Acha, contains high water absorption capacity that gives it capacity to be utilized in baked foods. It also contain pentosans(Lasekan and Lasekan 1998, Ayo 1998) which gives it the ability to form gel in the presence of oxidizing agents at room temperature with high residual protein coupled with high levels of sulphur and hydrophobic amino acid residues which makes it useful in baking (Obizoba et al, 1994). The recent finding of the unique property of acha flour in relatively lowering the blood glucose level (Ayo et al., 2007) and the subsequent reducing the diabetic populace has made the same an attractive research focus in the recent times.

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<p style="text-align: justify;">This work is aimed at improving the nutrient intake of populace particularly the diabetics, by supplementing the underutilized benniseed grains into acha flour to produce acha-beniseed composite biscuits.

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MATERIALS AND METHODS

<p style="text-align: justify;">The acha and beniseed grains were purchased from the Bauchi Central Market. The acha grain was dried (APV Cabinet Drier, 45oC), milled (Attrition Mill Yamaha Japan), and sieved (350nm aperture) to produce acha flour. The beniseed grains was sorted, washed, dried (APV Cabinet Drier, 45oC), milled and defatted using ethanol and dried (APV Cabinet Drier, 45oC), and sieved (350nm aperture) to produce benniseed flour. The defatted beniseed was supplemented (at 0.5, 10, 15, and 20%) into the acha flour to produce composite flour. Fat and sugar was fluffed into sucrose sugar and added to different proportion of composite flour added and mixed with condiments (salt, baking powder, egg and milk) to form batter. The batter was rolled (on a steel table), cut into shape (using biscuit cutter), placed on greased trays and baked at 160oC for 20 minutes.

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<p style="text-align: justify;">Physical Analysis

<p style="text-align: justify;">The Spread Ratio was determined by measuring the length and height of three rows and column, respectively of five well formed biscuits. The spread ratio is calculated as diameter divided by height (Gomez et al., 1997).

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<p style="text-align: justify;">The breaking strength was determined by adapting Okaka and Isieh’s (1997) method. Biscuit of known thickness (0.4cm) was placed centrally between two parallel metal bars (3 cm apart). Weights were added on the biscuit until the biscuit snapped. The least weight that caused the breaking of the biscuit was regarded as the break strength of the biscuit.

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<p style="text-align: justify;">Chemical Analysis

<p style="text-align: justify;">The protein, fat, ash and moisture content of the samples were determined. The carbohydrate content was obtained by difference of values above from hundred (AOAC, 1990).

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<p style="text-align: justify;">Sensory Evaluation of beneseed-acha composite biscuits

<p style="text-align: justify;">The sensory evaluation of the samples was carried out for consumer acceptance and preference using 20judges (students from Department of Food Science and Technology and students from Department of

<p style="text-align: center;">Ayo, J.A et al.,:Continental J. Food Science and Technology 4: 7 – 13, 2010

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<p style="text-align: justify;">Mechanical Engineering of Federal Polytechnic, Bauchi), randomly selected using a Nine point Hedonic scale (1 and 9 representing “extremely dislike” and “extremely like”, respectively).  The mean scores were differentiated using Analysis of Variance Methods.  The qualities assessed include colour, texture, taste, flavour and general acceptance.  Coded samples of the same size and temperature (29oC) were served on a coloured (white) plate of the same size to judges in each panel cupboard under the florescent light.  Only one sensory attribute was tasted at one sitting.

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RESULTS AND DISCUSSION

<p style="text-align: justify;">Effect of added defatted beniseed flour on physiochemical quality of acha – beniseed composite biscuit.

<p style="text-align: justify;">The effect of added beniseed flour on the physiochemical quality of acha based biscuit is summarized on Table 1.

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<p style="text-align: justify;">The moisture content of the acha–beniseed biscuit increased from 1.0 to 1.6% with the increase in the defatted beniseed flour concentrate (0-20%). This could be due to the increase in protein content. Proteins particularly from cereals have been noted to have high affinity for moisture and will always absorb the same if available (Jideani and Akingbade 1993; Alabo 2006)

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<p style="text-align: justify;">The protein content increased from 10.5 to 12.9% with increase in the percentage (0-20) addition of defatted beniseed flour. This could be due to the addition of the defatted beniseed, a legumes which are noted to be a good source of protein (Olayanju et al., 2006; Eneche, 2005). They contain powerful antioxidants called lignin which are also anti-carcinogenic and contain phytosterols which block cholesterol production. Their protein has a good balance of amino acids with a chemical score of 62%, and a net protein utilization of 54% (Alobo, 2007). These characteristics give sesame the potential of being a source of protein supplementation in cereal based foods.

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<p style="text-align: justify;">The fat content increased from 1.8 to 2.15% with increase in the percentage (0 – 20%) of defatted beniseed flour. The increase was not significant due to the defatting of the added beniseed. Similar non significant increase has been noted in the use of defatted legumes such as pigeon, and groundnut (Olayanju et al., 2006; Ayo, 2003).

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<p style="text-align: justify;">The ash content increased from 1.2 to 3.2% with increase in the percentage (0 – 20%) of defatted beniseed flour. The increase in the ash content could make the product a good source of minerals as observed in similar works (De Lemen et al., 2003; Addo, 2005). The seeds are rich in manganese, copper and calcium (90mg per table spoon for unhulled seeds, 10mg for hulled seeds and contain vitamin B, (thiamine and vitamin E (tocophenol) (Sosanya, 2007).

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<p style="text-align: justify;">The carbohydrate content decreased from 35.59 to 80.15%.as the percentage of defatted beniseed flour increased (0 – 20%). The decrease could be due to the low  carbohydrate content of the added defatted beniseed flour as has been observed in similar research works using legumes (Kent 1984; Ayo 1998).

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<p style="text-align: justify;">Effect on the Physical quality

<p style="text-align: justify;">The effect of added beniseed flour on the breaking strength of acha–beniseed biscuit increased from 1.49 to 1.67kg with increase in defatted beniseed flour concentrate(0 – 20%). This could be due to low quantity of carbohydrate content and poor quality of protein in term of functionality in beniseed and acha flour which has been observed to be the important factors in the formation of bonds that determine the strength of food (Fennema 1995).

<p style="text-align: justify;">

<p style="text-align: justify;">The spread ratio of acha–beniseed biscuit increased from 6.0 to 7.25 with added defatted benniseed flour. The increase in spread ratio is an indication of poor cohesions of the net work of the protein and carbohydrates which are the principal nutrients in the product (Gomez et al, 1997; Ayo and Nkama, 2003). The poor cohesion could allow the outflow of some ingredients such as sugar that could melt at the high temperature of baking hence increasing the spreadibility of the material.

<p style="margin-left: 72pt; text-align: justify;">

<p style="text-align: center;">Ayo, J.A et al.,:Continental J. Food Science and Technology 4: 7 – 13, 2010

<p style="text-align: justify;">

<p style="text-align: justify;">

<p style="text-align: justify;">Effect of added defated beneseed flour on the sensory quality of acha based biscuit.

<p style="text-align: justify;">The addition of defatted beniseed flour decreased the mean score of colour and texture from 8.30 to 4.30  and 7.20 to 5.80, respectively with increased percentages (0 – 20%) flour of beniseed as shown in Table 2. The decrease in the means score of the texture could be due to the same reason adduced to that of break strength.

<p style="text-align: justify;">

<p style="text-align: justify;">The addition of defatted beniseed flour increased the mean score of flavor from 4.20 to 8.30 with increase in the percentage (0-20%) of added defatted beniseed flour. This could be due to the possible by products flavor (fufury, amyl compounds) of protein in the beniseed and the acha carbohydrate which is acceptable in most baked products like biscuits (Fennema, 1985). However, the addition of defatted beniseed flour decreased the mean core of taste from 7.70 to 3.70 with increase in the percentages (0 – 20%) of defatted beniseed flour. This could be due to the bitter taste of some inherent compounds in beniseed flour particular at high temperature (Iwe, 2000).

The addition of defatted beniseed flour decreased the acceptability of the product on mean score that ranges from 7.90 to 3.70. The addition of defatted beniseed flour is acceptable up to 10% fortification with beniseed flour in acha – beniseed composite biscuit with a corresponding 11.9% protein.

CONCLUSION

<p style="text-align: justify;">The addition of defatted beniseed flour in the production of beniseed-acha composite biscuits has been found to be acceptable up to 10% which has a corresponding 125% (10.5-11.90%) and 16.3% (1.80-2.10%) of protein and fat respectively.

<p style="text-align: justify;">

<p style="text-align: justify;">The economic impact of using beniseed-acha composite flour in manufacture of biscuits lies in the reduced need for wheat for wheat being imported. In developing countries like Nigeria, where there is ban on wheat importation, utilization of acha and beniseed for biscuit production should be encouraged as the biscuits are a good source of protein and energy. Biscuit consumption is high in Nigeria; therefore, acha-beniseed biscuits will serve as vehicle for increasing intake of protein and calories in Nigerian diets. The production and acceptability of acha-beniseed composite biscuits will to a great extent provide variety of food for the diabetics as recent findings has recommended acha for such populace

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<p style="margin-left: 36pt; text-indent: -36pt;">De lumen B.O, Thompson, S, Odegaard J.W.(1993) sulphur Amino-Acid rich proteins in Acha (Digitana

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<p style="margin-left: 36pt; text-align: justify; text-indent: -36pt;">the Department of Food Science and Technology, Federal Polytechnic, .Bauchi) Pp. 15.

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<p style="text-align: center;">Ayo, J.A et al.,:Continental J. Food Science and Technology 4: 7 – 13, 2010

<p style="margin-left: 72pt;">Table1: Physiochemical Quality of Acha Beneseed Composite Biscuit

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<p style="margin-left: 72pt; text-indent: 36pt; line-height: 200%;">Ayo, J.A et al.,:Continental J. Food Science and Technology 4: 7 – 13, 2010''' '''

<p style="margin-left: 72pt; text-indent: 36pt; line-height: 200%;">'''Table 2: Sensory  Quality of Acha – Beniseed  Composite Biscuit '''

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<p style="margin-left: 36pt; text-indent: 36pt; line-height: 150%;">Score with the same subscript are not significantly different as 5% level of a acceptance

<p style="text-align: justify;">Received for Publication: 03/04/2010

<p style="text-align: justify;">Accepted for Publication: 30/04/2010

<p style="text-align: justify;">

<p style="text-align: justify;">Corresponding Author:

Ayo, J.A

Joseph Ayo Babalola University, Ikeji-Arakeji, PMB 5006, Ilesa. Osun State. Nigeria, Email: [mailto:jeromeayo@yahoo.com jeromeayo@yahoo.com]

<p style="text-align: justify;">Continental J. Food Science and Technology 4: 14 – 23, 2010                                ISSN: 2141 – 422X

<p style="text-align: justify;">©Wilolud Journals, 2010                                                                                  http://www.wiloludjournal.com

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NUTRITIONAL AND ANTINUTRITIONAL VALUES OF BOJER (Argyreia nervosa) SEEDS

<p style="margin-left: 108pt;">

<p style="margin-left: 108pt;">                   Fowomola, M. A.

<p style="text-align: center;">Science Technology Department, School of Applied Sciences and Technology, Federal Polytechnic, Offa, KwaraState.E-mail: moshoodfowomola@yahoo.com

<p style="text-align: center; page-break-after: avoid;"> =ABSTRACT= <p style="margin: 0cm 43.2pt 0.0001pt; text-align: justify;">The proximate composition, mineral, amino acid, vitamin and antinutrient contents of Bojer (Argyreia nervosa) seeds were investigated. The results of the proximate analysis show that bojer seed is very rich in crude protein (22.47 ±1.12%), lipid (13.67 ±0.68%), crude fibre (15.72 ± 0.79%), ash (3.68 ± 0.18%), carbohydrate (44.46 ± 2.22%) and energy content (390.75 ± 19.48 KJ/100g). The results also indicated that bojer seed. containsNa(23.11mg/100),Mg(4.96mg/100),Fe(16.22mg/100),Zn(1.24mg/100),Ca(5.77mg/100),Cu(0.14 mg/100),K(30.25 mg/100) and Mn(33.29 mg/100). Amino acid analysis revealed that bojer seed contains mainly aspartate (4.49 g/ 100g protein), lysine (4.48 g/ 100g protein), leucine (5.18 g/ 100g protein), glutamate (7.66 g/ 100g protein) and histidine (0.65 g/ 100g protein). The vitamin  analysis showed that bojer contained  1764.72±2.12 (IU) vitamin A, 0.30±0.03 mg/100g vitamin E, 0.082±0.02 mg/100g Vitamin K, 0.038±0.03 mg/100g Vitamin B1,  0.04±0 mg/100g, Vitamin B2, 0.12±0 mg/100g, Vitamin B6, 0.125±0.01 mg/100g, Vitamin B12 and  4.66±0.02 mg/100g Vitamin C.  Results of antinutrient analysis show that bojer seed contains alkaloid (2.01±0.10 mg/100g), tannins (1.21±0.06 mg/100g), phytate (1.52 ± 0.08 mg/100g), cyanogenic glycoside (0.043 ± 0.01 mg/100g), and saponin (0.63 ± 0.03 mg/100g).Trypsin inhibitor activity was found to be (11.84 ± .59 TIU/mg protein). The results showed that bojer seed is not only rich in caloric level but also low in concentrations of antinutritional factors. = = =KEYWORDS antinutritional, argyreia nervosa, bojer, trypsin inhibitor.= <p style="text-align: justify;"> =INTRODUCTION= <p style="text-align: justify;">Protein-energy malnutrition (PEM), remains the most common serious nutritional problem in almost all countries of Asia, Africa, Latin America and the Near East. This may be attributed to war, political instability and poverty. In addition, high rates of population growth and current patterns of population dispersion threaten world health and welfare. For instant, in Nigeria where the  population is over 120,000,000, most people can not afford to buy conventional protein sources (such as meat, fish, egg, chicken and milk) as a result of the poor economic. The search for alternative sources of foods which will be readily available to all is inevitable.

<p style="text-align: justify;">

<p style="text-align: justify;">Several workers, such as Umoh et. al., (1980) and Mba (1980) had highlighted a good number of lesser known animal foods such as clam, periwinkle and land snail, whose consumption was limited to a small population due to lack of adequate information about their nutritional potentials. In addition, Aletor and Aladetimi (1989) provided information on the nutritional potentials of some locally available under-utilized leguminous plant seeds. Furthermore, Fowomola and Akidahunsi (2007) reported on the nutritional quality of sandbox tree, an under-utilized plant in Nigeria.

<p style="text-align: justify;">'' ''

<p style="text-align: justify;"> Argyreia nervosa, otherwise known as Bojer, Hawaiian baby wood rose, Elephant creeper and woolly Morning Glory belongs to the family ''Convolvulaceae. ''It  originated from the Indian subcontinent  and it introduced to numerous areas worldwide, including Hawaii, Africa and Caribbean. Presently, the plant is cultivated in all the tropical regions as an ornamental tree and as a narcotic (amazingnature,2005).This plant  has large heart-shaped foliage (hairy from the bottom and 30cm across), violet coloured funnel-shaped flowers and round fruits that contain one to four seeds. The vine can reach 10 meters in height and the plant can be propagated vegetatively (by cutting), by seeds in humid soil at temperatures of 20-25o C.

Fowomola, M. A.: Continental J. Food Science and Technology 4: 14 – 23, 2010

<p style="text-align: justify;">

<p style="text-align: justify;">

<p style="text-align: justify;">The seeds of A. nervosa have hallucinogenic properties and are used as “cheap buzz”, an alternative to alcohol in Hawaii, Haiti and Puerto Rico. Three to eight seeds may produce psychedelic effects comparative to Lysergic acid diethylamide (LSD). The experience usually reported include extreme lassitude, changes in visual and auditory perception, emotional disturbances, nausea, papillary dilation, tremor, increase in blood pressure and body temperature .In addition, the seed can cause uterine contractions ,which may lead to miscarriage in a pregnant woman if she consumes it .However, with appropriate dosage and supervision, the effects can be used to aid child birth (amazingnature, 2005). The leaves and roots of A    .nervosa which are not psychoactive are used as antiseptic and anti-inflammatory drugs in India.

<p style="text-align: justify;">

<p style="text-align: justify;">Phytochemical analysis has revealed that the seeds contain 0.3 to 1 % ergot-alkaloids by weight. These include Ergine (d-Lysergic acid amide) and its derivatives such as ergometrine. The hallucigenic or psychedelic effects experienced after consumption of A.nervosa seeds are usually attributed to Ergine (d-Lysergic acid amide) and its derivatives (amazingnature, 2005). The “furry” outer skin of the seeds contains cyanogenic glycosides, the ingestion of which is the likely cause of nausea reported by those who have eaten the seeds. The main body of the plant contains small amount of strychnine, a potent toxin, but its presence is negligible in the seeds (amazingnature, 2005). However there is scarcity of information on the nutritional and antinutritional factors in bojer (argyreia nervosa) seeds which can serve as a guide for the possible utilisation of bojer seeds by animal/ human feed manufacturers, public health authorities and other food regulatory bodies. Therefore, the present work is aimed to provide this information.

<p style="text-align: justify;"> =MATERIALS AND METHODS= =Materials= <p style="text-align: justify;">Collection of Samples

<p style="text-align: justify;">Bojer seeds were collected from a mechanical workshop, Tipper Garage,  Offa in Kwara State, Nigeria, identified and authenticated by a botanist at Department of Science Technology, School of Applied Sciences and Technology, Federal Polytechnic Offa Kwara State, Nigeria.

<p style="text-align: justify;">

<p style="text-align: justify;">All Analytical grade reagents used for chemical analysis were supplied by BDH Ltd., Poole, UK.

<p style="text-align: justify;">

<p style="text-align: justify;">Methods

<p style="text-align: justify;">Sample Treatment

<p style="text-align: justify;">The seeds were hulled from their coats, air – dried in the laboratory for two weeks, ground to a mesh size of 1 mm (using  Laboratory pestle and  Mortal ) and kept in air–tight sterilised containers for analysis.

<p style="text-align: justify;">

<p style="text-align: justify;"> Proximate Analysis

<p style="text-align: justify;"> Crude protein, Lipid, Ash, Crude fibre, Carbohydrate and energy contents of Bojer seed were determined by using the methods described by Horwitz (1980), Pearson (1973), Foster and Leslie (1971), Norman and Waddington (1989) and Okoye (1992), respectively.

<p style="text-align: justify;">

<p style="text-align: justify;">Determination of minerals

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Preparation of sample

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Sample (2g) was ashed at 5000C to a constant weight. 15ml of 20% (v/v) nitric acid solution was added to the crucible to break up the ash. This was boiled and filtered through No.43 Whatman filter paper. The residue and the paper were washed 3 times with deionised water into a standard 100ml volumetric flask and then diluted to 100ml with    deionised water.

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Atomic absorption spectrophotometer (AAS) (Pye Unicam Sp 9 AAS) was used for the determination of Calcium,Copper,Manganese, Zinc, Magnesium and Iron as described by Perkin-Elmer(1982) while Sodium and Potassium were determined by using flame spectrophotometer.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="text-indent: 36pt;">Fowomola, M. A.: Continental J. Food Science and Technology 4: 14 – 23, 2010

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Amino acid profile

<p style="text-align: justify;">The amino acid profile in the sample was determined using method described by Spackman ''et. al'',. (1958). The sample was dried to constant weight, defatted, hydrolyzed, evaporated in a rotary evaporator and then loaded into the Technicon Sequential Multisample amino acid analyzer (TSM) for analysis.

<p style="text-align: justify;">

<p style="text-align: justify;">Determination of Vitamins

Vitamins were determined using the methods described by Association of Vitamin Chemists (1987).

<p style="text-align: justify;">

<p style="text-align: justify;">Determination of Antinutrients

<p style="text-align: justify;">The contents of cyanide, tannin, alkaloid, oxalate, saponin and trypsin inhibitor activity in bojer seed were determined according to the methods described by Harborne (1998), Makkar ''et. al., (1993), Harbone (1973), Ukpabi and Ejidoh (1989), Fenwick and Oakenful (1983) and Kakade et. al''., (1974) respectively.

<p style="text-align: justify;">

Statistical analysis: Results of Proximate analysis, Vitamins and antinutrients were presented as mean ±SD of three determinations.

<p style="text-align: justify;">

<p style="text-align: justify;">RESULTS

<p style="text-align: justify;">Table 1 shows the proximate composition and energy content of bojer seed. The results indicated that bojer seed is very rich in Carbohydrate (44.46± 2.22 %) while the protein content is 22.47±1.12%, Lipid 13.67±0.68% and crude fibre15.72 ± 0.79%. The energy content is 390.75±19.48KJ/100g. Table 2 depicts the concentrations of mineral assayed in bojer seed. The results showed that bojer seed contains 23.11(mg/100) Na, 3.4(mg/100) K, 5.77 (mg/100)Ca,4.96(mg/100) Mg, 16.22(mg/100) Fe and 1.24 (mg/100) Zn,0.14(mg/100) Cu and  33.29(mg/100) Mn. The amino acid profile of bojer seed is as presented in Table 3. The results revealed  that bojer seed is very rich in glutamate (7.66g/100g of protein) while cysteine (0.78g/100g of protein) has the lowest value. Among the essential amino acids, arginine has the highest value (5.97g/100g of protein). This is followed by leucine which has the value of 5.18 g/100g of protein. Table 4 shows the vitamins contents (mg\100g) of bojer (argyreia nervosa) seeds. The results  showed that bojer contained  1764.72±2.12 (IU) vitamin A, 0.30±0.03 mg/100g vitamin E, 0.082±0.02 mg/100g Vitamin K, 0.038±0.03 mg/100g Vitamin B1,  0.04±0 mg/100g, Vitamin B2, 0.12±0 mg/100g, Vitamin B6, 0.125±0.01 mg/100g, Vitamin B12 and  4.66±0.02 mg/100g Vitamin C.  Table 5 depicts the antinutrients contents of bojer seed. The results revealed that trypsin inhibitor has the highest value (11.84±0.59TIU/mg) followed by alkaloid  (2.01±0.01mg/100g) while cyanide has the lowest value (0.043±0.01mg/100g).

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<p style="text-align: justify;">DISCUSSION

<p style="text-align: justify;">The results of proximate analysis showed that bojer seed is a nutritionally promising seed because of its high levels of oil, protein and carbohydrate. It is more nutritious when compared with sweet yellow maize(Cortéz,, and Wild-Altamirano,. 1972). High level of crude fibre in bojer seed is of benefit to mankind. It has been reported that fruits containing high level of crude fibre are able to control glycaemia by reducing the absorption of glucose in the intestine (Anderson et al., 1988). In addition, insoluble fibre has been found to be helpful in the treatment and prevention of constipation, haemorrhoids, diverticulosis, high blood cholesterol levels, the risk of coronary heart disease and colon cancer (Slavin, 1990).

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify;">

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify;">The results of the amino acid analysis showed that bojer seed is very rich in some essential amino acids that are needed for normal functioning of the body. The amino acid requirements for infants and 10–12-year-old children have been reported to be 31 mg/kg/day for isoleucine, 64 mg/kg/day for lysine, 27 mg/kg/day for sulfur amino acids, 37 mg/kg/day for threonine and 14 mg/kg/day for tryptophan (Pineda et. al.,1981). There are also evidences that histidine is an essential amino acid for adults and its dietary requirement in both normal and uremic men has been reported to be between 8 and 12 mg/kg/day (Giordano et. al., 1972, Bergstrom et. al., 1970 and Kopple et. al., 1975).

<p style="text-indent: 36pt;">Fowomola, M. A.: Continental J. Food Science and Technology 4: 14 – 23, 2010 = =

=The results of mineral assay showed that bojer seed is very rich in sodium and manganese. Sodium plays a vital role in active transport across the cell membrane and is also required for maintenance of osmotic balance (Ann, 2010) while manganese serves as a coenzyme for Superoxide Dismutase (Jefferson = = = =et.al.,2009). The presence of antioxidant vitamins such as vitamin C, E and A suggests that bojer seed could be used as an alternative source of these vitamins. Antioxidant vitamins have been reported to reduce oxidative processes which are known to be vital in the initiation of arthrosclerosis Steinberg, ''et. al.'' (1989).= <p style="text-align: justify;">The results of antinutrient analysis showed that bojer seed contained some secondary metabolites which may cause problems if consume raw. However, they are very low in concentration when compared with those of maize (Hassan et. al., 2005).These included tannins which are aromatic compounds containing phenolic groups. They interact with salivary proteins and glycoproteins in the mouth and render the tissues astringent to taste (Howes, 1953). Astringency gives tannin the medicinal value in preventing diarrhoea and dysentery and for controlling haemorrhage (Sollman, 1957; Jones, 1965). Furthermore, tannins protect plant against dehydration, rotting, damage by animals and pathogens. When they are polymerised, insoluble protective barrier is formed which prevents microbial attack (Stumpf and Conn, 1981). Therefore they can be applied to wounds as protective coating. Bichel and Bach (1968) reported that the symptoms of continued intake of tannin include gastritis as well as irritation and edema of the intestine. Glick and Joslyn (1970) also reported that feeding 0.5% of tannic acid decreased nitrogen retention and caused 5% mortality in rats. Tannins are known  to bind strongly to proteins in vitro and form a compound called tannin-protein complex (T-PC). The complexes are quite resistant to degradation by digestive enzymes, thereby interfering with nitrogen balance in vivo (Feeny,1976; Mcleod, 1974; Rhoades, and Cates, 1976).

<p style="text-align: justify;">

<p style="text-align: justify;">In addition, the work of Chung et.al., (1998) revealed that tannins have anticarcinogenic and antimutagenic potentials, which is important in protecting cellular oxidative damage, including lipid peroxidation in human.

<p style="text-align: justify;">

<p style="text-align: justify;">Savage et.al., (1964) reported that phytate depressed the growth of chicks fed with phytate – casein diet by forming complex with zinc thereby making the later unavailable. Omosaiye and Cheryan (1979) also reported that phytate formed complex with protein by the actions of cations, usually calcium, zinc or magnesium which act as a bridge between the negatively charged protein carboxyl groups and former. The report of Chen et.al., (1934), shown that the minimum lethal dose of hydrogen cyanide taken by man through mouth to be between 0.5mg and 3.5mg/kg body weight. The symptoms of hydrogen cyanide including peripheral numbness, light-headedness, mental confusion, stupor, cyanosis and convulsion were reported by Halstrom and Moiler (1945). Oxalate forms complex with calcium thereby making it unavailable when fed to animals and more so high oxalate diets can increase the risk of renal calcium absorption (Osagie and Eka, 1998). Saponins have characteristic bitter taste, foaming properties and haemolytic effect on red blood cells (Osagie and Eka, 1998).Some alkaloids for example potato alkaloid (solanine) cause gastrointestinal upsets and neurological disorders, especially when taken in excess of 20mg/100g sample (Osagie and Eka, 1998). Trypsin inhibitors are low molecular weight proteins which form complexes with trypsin thereby reducing its proteolytic activity which in turn reduces the availability of amino  acids which eventually lead to reduced growth and pancreatic enlargement ( Leiner, 1989).

<p style="text-align: justify;">

<p style="text-align: justify;">CONCLUSION

<p style="text-align: justify;">The present study has shown that bojer seeds are not only rich in caloric level but also low in concentrations of antinutritional factors and could serve as a guide for the possible utilisation of bojer seeds by animal feed manufacturers, public health authorities and other food regulatory bodies.

<p style="margin-left: 6pt; text-align: justify; text-indent: -1.5pt;">

<p style="margin-left: 6pt; text-align: justify; text-indent: -1.5pt;">ACKNOWLEDGEMENTS

<p style="margin-left: 6pt; text-align: justify; text-indent: -1.5pt;">The assistance rendered by my departmental Staff and the supports from my family are gratefully acknowledged.

<p style="text-align: justify;">

<p style="text-align: justify;">

<p style="text-align: justify;">

<p style="margin-left: 36pt; text-indent: 36pt;">Fowomola, M. A.: Continental J. Food Science and Technology 4: 14 – 23, 2010

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<p style="text-align: justify;"> Table1. Proximate composition of bojer seeds

<p style="text-align: justify;"> <p style="text-align: justify;">Each value is a mean of three determinations. [mean ± SD]

<p style="text-align: justify;">

<p style="text-align: justify;">Table2. some mineral contents of bojer  seeds

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<p style="text-align: justify;">Table 3. Amino acid profile (g/100g Dry Weight) of bojer seeds <p style="text-align: justify;">

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<p style="margin-left: 36pt; text-indent: 36pt;">Fowomola, M. A.: Continental J. Food Science and Technology 4: 14 – 23, 2010

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<p style="text-align: justify;">Table 4. Vitamins contents (mg\100g Dry Weight) of bojer seeds <p style="margin-right: 72pt; text-align: justify;"> Each value is a mean of three determinations [mean ± SD].

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<p style="text-align: justify;"> <p style="text-align: justify;">Table 5. Antinutrients content (mg/100g) of  bojer  seeds

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<p style="margin-left: -72pt; text-align: justify;">                               undefinedEach value is a mean of three determinations [mean ± SD].

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Stumpf, P.K. and Conn, E.E. (1981). Secondary plant products. In the Biochemistry of plant. (Conn. Edition). Academic press. New York. pp.33-40.

<p style="margin-left: 36pt; text-align: justify; text-indent: -36pt;">

<p style="margin-left: 36pt; text-align: justify; text-indent: -36pt;">

<p style="margin-left: 36pt; text-indent: 36pt;">Fowomola, M. A.: Continental J. Food Science and Technology 4: 14 – 23, 2010

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<p style="margin-left: 36pt; text-align: justify; text-indent: -36pt;">

<p style="margin-left: 36pt; text-align: justify; text-indent: -36pt;">Savage J.E., Yohe J.M., Pickett, E.E. and O’Dell B.L. (1964). Zinc metabolism in the growing chicks.

<p style="margin-left: 36pt; text-align: justify; text-indent: -36pt;">Tissue concentration and effect of phytate on absorption. ''PoultryScience. '' 43. pp. 420 – 426.

<p style="margin-left: 36pt; text-align: justify; text-indent: -36pt;">

<p style="margin-left: 36pt; text-align: justify; text-indent: -36pt;">Slavin, J. (1990) "Dietary Fiber: Mechanisms or Magic in Disease Prevention?" Nutrition Today.

<p style="margin-left: 36pt; text-align: justify; text-indent: -36pt;">Nov/Dec. 1990. Tropilab inc.(2005).http.WWW.trobilab\hura-cre.html Udoessien, E.I. and Ifon, E.T.

<p style="margin-left: 36pt; text-align: justify; text-indent: -36pt;">(1984). Chemical evaluation of some antinutritional constituents in four species of yam. Tropical Science

<p style="margin-left: 36pt; text-align: justify; text-indent: -36pt;">32. pp. 115 – 119.

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<p style="margin-left: 4.5pt; text-align: justify;">Ukpabi, U.J. and Ejidoh. J.I. (1989): Effect of deep oil frying on the oxalate content and the degree of itching of cocoyams (Xanthosoma and colocasia SPP). Technical paper presented at the 5th Annual Conference of the Agricultural Society of Nigeria. Federal University of Technology, Owerri Nigeria. 3rd – 6th September 1989. pp.13 - 14.

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<p style="margin-left: 4.5pt; text-align: justify;">Umoh, I.B. and Bassir, O. (1977). Lesserknown sources of protein in some Nigeria peasant diets. Food Chemistry. 2. pp. 315-321.

<p style="margin-left: 4.5pt; text-align: justify;">

<p style="margin-left: 4.5pt; text-align: justify;">Umoh, I.B., Ayalogy, E.O. and Bassir, O. (1980). Evaluations of the nutritive value of some lesser know protein sources in Nigeria peasant diets. Ecology, Food and Nutrition. 2. pp. 81-86.

<p style="margin-left: 4.5pt; text-align: justify;">

<p style="margin-left: 4.5pt; text-align: justify;">Wheeler; E.L. and Ferrel, R.E. (1971): A method for phytic acid determination in wheat fractions''. Cereal Chemistry''. 48. pp. 312 – 320.

<p style="margin-left: 4.5pt; text-align: justify;">

<p style="margin-left: 4.5pt; text-align: justify;">Wink, M (1993): Quinolizidine alkaloids methods in plants biochemistry 8. Academic Press Ltd. pp.112-120.

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<p style="text-align: justify;">Received for Publication: 03/04/2010

<p style="text-align: justify;">Accepted for Publication: 30/04/2010

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<p style="text-align: justify;">Continental J. Food Science and Technology 4: 24 – 37, 2010                                ISSN: 2141 – 422X

<p style="text-align: justify;">©Wilolud Journals, 2010                                                                                  http://www.wiloludjournal.com

<p style="text-align: center;">CRYOGENIC GRINDING OF SPICES IS A NOVEL APPROACH WHEREAS AMBIENT GRINDING NEEDS IMPROVEMENT

<p style="text-align: center;">

<p style="text-align: center;">Murlidhar Meghwal, T K Goswami <p style="border: medium none ; padding: 0cm; text-align: center;">Agricultural and Food Engineering Department (AgFE), Indian Institute of Technology (IIT) Kharagpur, India <p style="text-align: center;">

<p style="margin: 0cm 43.2pt 0.0001pt;">ABSTRACT

<p style="margin: 0cm 43.2pt 0.0001pt; text-align: justify;">Study on ambient and cryogenic grinding was performed to test the novelty of cryogenic grinding and pin point the drawbacks of ambient grinding. Comparative study had shown that ambient grinding need more power (8.92%) and specific energy (14.5%) than cryogenic grinding. Particle size analysis had shown that cryogenic grinding produced coarser particles. Comparative study of energy law constant shows that ambient is more power consumptive. The higher amount of volatile oil (2.15 ml/100 g) content was found in cryogenic grinding and also powder of freshness and lower whiteness (40%) and higher yellowness (14%) indices found for cryogenic grinding.

<p style="margin: 0cm 43.2pt 0.0001pt; text-align: justify;">

<p style="margin: 0cm 43.2pt 0.0001pt;">KEYWORDS: Ambient, cryogenic, grinding, volatile oil, mill, particle, diameter, power, specific energy.

<p style="margin: 0cm 0cm 0.0001pt; line-height: normal;">INTRODUCTION

<p style="text-align: justify;">Grinding is an important unit operation in which the size of the particle is reduced and their surface area is increased. When increasing surface area of particles, it means the availability of constituents (such as oil inside the cells, fragrance and flavouring components) that are available in the material is increases. Power consumption in grinding, size of the commented particles and increase in the surface area depends on the initial size, shape and strength of the particle or material; the kind of grinder or mill used for this unit operation and the fixing of operating parameter to running the grinder or mill such as temperature, size of sieve, number of rotor ribs, etc (Das, 2005).

<p style="text-align: justify;">

<p style="text-align: justify;">Grinding is the most power consuming operation because only 1% of the energy imparted into the material is utilized loosening the bond between particles, whereas almost 99% of input energy is dissipated as heat, rising the temperature of the ground product etc. In spice grinding temperature rises to the extent of 42 - 93 (Singh and Goswami, 1997) and this causes the loss of volatile oil and flavouring constituents; for high oil bearing material, oil comes out from oil bearing material during grinding, which makes ground product gummy, sticky and results in chocking of sieves through which the product passes (Singh and Goswami, 1997).

<p style="text-align: justify;">

<p style="text-align: justify;">Thermal damage is one of the main limitations of the conventional grinding process, so it is especially important to perform the grinding under controlled temperatures conditions. Calculation of temperature and its effect on thermal damage to the material undergoing grinding was carried out by Malkin and Guo (2007) and suggest that if we can reduce the temperature of two rubbing surfaces, we can obtain better product.

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<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">The fundamental principle of cryogenic grinding is similar to that of  conventional grinding  methods  for materials, but the compositions are very complex, containing aromatics  of  high  volatility,  oils and  fats, which  are  easily oxidized. Using liquid  nitrogen  or  liquid  air  as  the cryogen,  all of thermo-sensitive herbal medicines, spices and important food commodity can be ground  below  their brittle temperature. The colour and other properties of the products of  cryogenic grinding  will  not  be  changed  and  their  flavour and  nutritional value  will  not  be  lost (Shimo et al.,1991). The usefulness of cryogenic grinding can be summed up as (1) the conventional or ambient grinding of spices results in inferior quality of the product having several operational problems such dust formation. (2) the application of cryogenic

<p style="text-align: center;">Murlidhar Meghwal, T K Goswami: Continental J. Food Science and Technology 4: 24 – 37, 2010

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">technology for grinding of spices has been scientifically proved to be suitable technique with less loss of volatile oil content, improved colour and grinding operation. (3) the research information and data generated on properties of spices and cryogenic grinding would help to understand grinding phenomena and develop efficient grinding system. Cryogenic grinding produced Gelucire 44/14 in a powder form and did not change its physical properties, emulsification capacities and dissolution performances of the formulation (Chambin et al., 2004). The normal grinding produces poor quality of powder that does not conform to the international quality standard; as a result either fetches lower prices or not accepted by the importer countries. The temperature rise of the product can be minimized to some extent by circulating cold air or water around the grinder. But this technique is not sufficient enough to significantly reduce the temperature rise of the product. The loss of volatile can be significantly reduced by the cryogenic grinding technique using liquid nitrogen or liquid carbon dioxide that provides the refrigeration needed to pre-cool the spices and maintain the desired low temperature by absorbing heat generation during grinding operation. The extremely low temperature in the grinder solidifies the oil so that the spices become brittle, they crumble easily permitting grinding to a finer and more consistent size. The high quality ground product would have domestic as well as international market. There is need for modelling of grinding process (Stepien, 2009). Limited research information is available on the cryogenic grinding of spices. The present study was undertaken with the objectives of comparative study on power and specific energy requirement for ambient and cryogenic grinding of black pepper; study of different energy law constants; effect of ambient and cryogenic grinding on particle size; volatile oil content and colour retention.

<p style="text-align: justify;">

<p style="text-align: justify;">MATERIALS AND METHODS

<p style="text-align: justify;">Sample preparation

<p style="text-align: justify;">For the present investigation, black pepper was collected from Indian Institute of Spices Research (IISR), Calicut, Kerala, India during May 2009. The pepper were cleaned manually to separate out the stones, dirt, dust, broken, foreign, unwanted matters and immature seeds from the main sample of black pepper. The initial moisture content (mc) and mc after grinding of the black pepper on dry basis (db), was determined by oven drying method at 72oC for 24 h until a constant weight was obtained (Ranganna, 1995).

<p style="margin-left: 72pt; text-align: justify; text-indent: 36pt;">                                                                                                (1)

<p style="margin-right: -16.55pt; text-align: justify;">

<p style="margin-right: -16.55pt; text-align: justify;">where, Q is amount of water to be added in ml, Wi is initial mass of sample (g), Mi is initial mc (% db), Mf is final mc (% db). The pepper was kept in sealed and moisture resistant flexible polyethylene bags.

<p style="margin-right: -16.55pt; text-align: justify;">

<p style="text-align: justify;">Ambient and Cryogenic Grinding

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">Rotor mill

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">Rotor mill (Model Pulverisette 14, Fritsch, Germany) is one of the types among the several types of size reduction devices were used for ambient and cryogenic grinding. In this device the size reduction of particle takes place by impacts of the rotating ribs and attrition of the particle on sieve and mill’s stationery surfaces. The peripheral speed of the rotors ranges between 70 (15000 RPM) to 90 (20000 RPM) m s-1. The major components of the mill are rotor (88.5 mm dia.) having 8 to 12 number of fixed ribs, sieve rings of different opening sizes (0.08, 0.12, 0.2, 0.5 and 1.0 mm) are available and 1 and 0.5 mm sieves are used for present study. The speed of rotor could be controlled through an in built mechanism.

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">Experimental procedure

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">The experiment for ambient grinding was carried out at room temperature. First of all cleaned and sieved sample was made ready for grinding, grinder was switched on, sample was fed manually into the feed hopper slowly to avoid chocking of sieve and product was obtained at collector pan from an outlet. Similarly, in cryogenic grinding Liquid nitrogen (LN2) was fed along with the sample and ground product was obtained at collector pan from an outlet. A temperature indicator, 600 to -200oC (Testo, Germany) was inserted into the outlet of powder to record the temperature of the product.

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">

<p style="text-align: center;">Murlidhar Meghwal, T K Goswami: Continental J. Food Science and Technology 4: 24 – 37, 2010

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">Size reduction theory in relation to black pepper grinding

<p style="text-align: justify;">Black pepper is a seed and it can’t be used as such. It has to be ground for consumption purposes. Grinding also helps in separation of various ingredients of black pepper. The size reduction theory of black pepper involves particle size measurement, particle size analysis, power consumption in grinding (Geankoplis, 2004) etc. Different laws which explain energy requirement in grinding are described in the following sub sections:

<p style="text-align: justify;">

<p style="text-align: justify;">Power Requirement in Ambient and Cryogenic Grinding

<p style="text-align: justify;">In size reduction mechanical actions are required to reduce the particles into smaller ones. There is need of energy to fracture and creating new surfaces. Approximate calculations give actual efficiencies of about 0.1 to 2% (Geankoplis, 2004). Theories were derived depending upon the assumption that the energy E required to produce a change dD in a particle of size D in a power function of D.

<p style="text-align: justify;">

<p style="text-align: justify;">                                                                                                                               (2)

<p style="text-align: justify;">

<p style="text-align: justify;">where, D is the diameter of particle in mm, n and C are constants.

<p style="text-align: justify;">

<p style="text-align: justify;">A single phage wattmeter (range 0 - 750 W, least count 5 W) was connected with the machine to measure the power consumed and ultimately to measure the energy required in grinding. Mill was run empty and by using stop watch, number of revolutions (m) completed by the energy meter in time ‘tm’ (s) was recorded. Similarly, the total number of revolutions completed (n) to grind the whole sample in time ‘tn’ (s) was also recorded. Power under load was measured at each set of experiments. The circular panel of energy meter was divided into 10 large divisions, each large division had 10 intermediate divisions and each intermediate division was divided into 2 small divisions. So, disc had 200 small divisions. Thus, 1 kWh and 60 revolutions will correspond to 12,000 divisions.

<p style="margin-left: 36pt; text-align: justify; text-indent: 36pt;">

<p style="margin-left: 36pt; text-align: justify; text-indent: 36pt;">    1division =  ''kWh ''

<p style="margin-left: 36pt; text-align: justify; text-indent: 36pt;">Power consumption = [ ] Х Х kWs

<p style="text-align: justify;">                                                                   = 0.3 [  -  ] kW                                                            (3)

<p style="text-align: justify;">

<p style="text-align: justify;">Specific Energy Consumption

<p style="text-align: justify;">It is the amount of energy required to grind the unit amount of material in unit time. It may be expressed in kW kg-1 or kJ kg-1. The following formula was used to calculate the specific energy consumed in grinding (Singh & Goswami, 1997).

<p style="text-align: justify;">

<p style="text-align: justify;">                      (4)

<p style="text-align: justify;">

<p style="text-align: justify;">Energy constants

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">a). Rittenger’s law constants (Kr) :

<p style="text-align: justify;">Rittinger proposed a law which states that the work in crushing is proportional to the new surface created. From equation (2) it turns to be-

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">                                               E= Kr[ ]                                                                (5)

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">

<p style="text-align: center;">Murlidhar Meghwal, T K Goswami: Continental J. Food Science and Technology 4: 24 – 37, 2010

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">where, DF is the diameter of feed, DP is the diameter of product, E is the amount of work required to reduce a unit mass of feed from DF to DP and Kr is a constant. This law implies that the same amount of energy is required to reduce a material from 100 to 50 mm as is required to reduce the same material from 50 to 33.33 mm. It has been found experimentally that this law has some validity in grinding fine powders (Geankoplis, 2004).

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">

<p style="text-align: justify;">

<p style="text-align: justify;">b). Kick’s law constants (Kk):

<p style="text-align: justify;">Kick’s law states that energy required to reduce a material in size was directly proportional to the size-reduction ratio. This implies that n = 1 in eq (2) and this can mathematical be expressed as follows:

<p style="text-align: justify;">

<p style="text-align: justify;">                                         E = C ln

<p style="text-align: justify;">                                         E =  KKln[ ]                                                             (6)

<p style="text-align: justify;">

<p style="text-align: justify;">where, Kk is a constant. This law implies that the same amount of energy is required to reduce a material from 100 to 50 mm as is needed to reduce the same material from 50 to 25 mm (Singh & Sahay, 2004 & Geankoplis, 2004).

<p style="text-align: justify;">

<p style="text-align: justify;">c). Bond’s law constants (Kb):

<p style="text-align: justify;">This law states that the work required using a large-size feed is proportional to the squire root of the surface/volume ratio of the product. This corresponds to n = 1.5 and mathematically this can be expressed as given below (Singh & Sahay, 2004 & Geankoplis, 2004).

<p style="text-align: justify;">

<p style="text-align: justify;">                                                                 E = Kb

<p style="margin: 0cm 0cm 0.0001pt 72pt; text-align: justify; line-height: normal;">             E =   =Kb [ -  ]                                                                               (7)

<p style="text-align: justify;">

<p style="text-align: justify;">d). Rosin Rammler Sperling Bennet (RRSB) constant

<p style="margin-left: 72pt; text-align: justify; text-indent: 36pt;"> Φ = ]                                                                                                 (8)

<p style="margin-left: 72pt; text-align: justify; text-indent: 36pt;">

<p style="text-align: justify;">where,

<p style="text-align: justify;">       Φ =                                (9)

<p style="text-align: justify;">D is the sieve size (m), M and l are constants

<p style="text-align: justify;">

<p style="text-align: justify;">Particle size analysis

<p style="text-align: justify;">Black pepper seed were ground by using 1 and 0.5 mm size sieve. Particle size analysis of ground spice powder were carried out by using shaker setup and then calculating different diameter. Mass Mean Diameter (MMD): It can be defined as the ratio of mass fraction in individual increment to the total mass fraction together (McCabe et al., 2000).

<p style="margin-left: 36pt; text-align: justify; text-indent: 36pt;">

<p style="margin-left: 36pt; text-align: justify; text-indent: 36pt;">MMD μm =                                                                                               (10)

<p style="text-align: center;">Murlidhar Meghwal, T K Goswami: Continental J. Food Science and Technology 4: 24 – 37, 2010

<p style="text-align: justify;">

<p style="text-align: justify;">

<p style="text-align: justify;">Volume Surface Mean Diameter (VSMD) or Sauter Mean Diameter (Ds): It can be defined as the 6 times the ratio of volume of 1 kg ground particle (which are assumed to be spherical) and its surface area (Das, 2005). The value of Ds can be estimated from the Eq. (11).

<p style="margin-left: 36pt; text-align: justify; text-indent: 36pt;">

<p style="margin-left: 36pt; text-align: justify; text-indent: 36pt;"> Dsμm = 1/                                                                                             (11)

<p style="text-align: justify;">The arithmetic mean diameter (AMD) usually termed as the mean diameter, is the arithmetic average particle diameter of the distribution. The value of the arithmetic mean is sensitive to the quantities of particulate matter at the extreme lower and upper ends of the distribution. It can be mathematically be expressed as Eq. (12)

<p style="margin-left: 72pt; text-align: justify;">

<p style="margin-left: 72pt; text-align: justify;">AMD μm = /                                                                                           (12)

<p style="text-align: justify;">

<p style="text-align: justify;">Volume Mean Diameter (VMD): It is the ratio of the volume of total mass based on average diameter of total particle to the sum of the volume of each fraction.

<p style="margin-left: 72pt; text-align: justify;">

<p style="margin-left: 72pt; text-align: justify;">VMD μm =  [1/3 ]^0.33                                                                     (13)

<p style="text-align: justify;">'' ''

<p style="text-align: justify;">Dmode: The mode represents the value that occurs most frequently in a distribution. In particle size distributions, the mode is the particle diameter that occurs most frequently. Dmedian: It is the sieve size in mm or μm through 50% ground material passes out. D80: It the sieve size in mm or μm through which 80% ground material passes. It also helps to know about the extent of fineness obtained by grinding (McCabe et al., 2000), (Fig. 1).

<p style="text-align: justify;">

<p style="text-align: justify;">

<p style="text-align: center;">Fig. 1 Showing Dmode, Dmedian and D80

<p style="text-align: center;">Murlidhar Meghwal, T K Goswami: Continental J. Food Science and Technology 4: 24 – 37, 2010

<p style="text-align: justify;">

<p style="text-align: justify;">

<p style="text-align: justify;">Initial Surface Area (Ai, m2kg-1) of black pepper seed before grinding can be estimated using the Eq. (14)

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">                          Ai =                                                                                                                         (14)

<p style="margin: 0cm 0cm 0.0001pt; line-height: normal;">

<p style="margin: 0cm 0cm 0.0001pt; line-height: normal;">where, Diis diameter of pepper before grinding in m, ψiis spherecity of pepper before grinding.

<p style="text-align: justify;">Total Surface Area (At, m2 kg-1)undefinedcreated by Grinding can be obtained using Eq. (15)

<p style="margin-bottom: 0.0001pt; text-align: justify; text-indent: 36pt; line-height: normal;">'' ''

<p style="margin-bottom: 0.0001pt; text-align: justify; text-indent: 36pt; line-height: normal;">At=                                                                                                                        (15)

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">where, Ds is sauter mean diameter in m, Ψo is spherecity of pepper after grinding (Das, 2005).

<p style="margin-bottom: 0.0001pt; text-align: justify; text-indent: 36pt; line-height: normal;">

<p style="text-align: justify;">Fineness Modulus (FM): It is an empirical factor obtained by adding the total percentages of a sample of the aggregate retained on each of a specified series of sieves and dividing the sum by 100. This concept helps to describe particle-size distributions by an index number. Many agencies use fineness modulus variation as a convenient means of keeping quality history data on uniformity of particle-size distribution of aggregate production, delivery, and use. Some agencies require that aggregates be processed to remain within upper and lower limits of fineness modulus. Such requirements are more frequently applicable to fine particles.

<p style="text-align: justify;">

<p style="text-align: justify;">                       FM =                                                                                         (16)

<p style="text-align: justify;">

<p style="text-align: justify;">Quality attributes of spice powder

<p style="text-align: justify;">Volatile oil

<p style="text-align: justify;">Interest in the production of high-quality spice products has encouraged scientists to investigate the effects of cryogenic milling/grinding on spice quality. Ambient and cryogenically milled spices were studied for volatiles oil content retention. Oil was extracted from ambient and cryogenic ground samples by steam distillation method (Masango, 2005; Li et al., 2009).

<p style="text-align: justify;">

<p style="text-align: justify;">Colour

<p style="text-align: justify;">The colour is an important quality attributes to accept or reject the spices because it has direct appealing effect in the mind of consumer. For the colour determination purpose chromameter (CR-400/410, Konica Minolta, Tokyo, Japan) was used, which gave L, a and b value, where L value varies between 0 and 100. A perfectly white body has L=100 and a black body has L = 0. A positive value of ‘a’ indicates the redness and negative value greenness. A positive value of ‘b’ indicates yellowness and negative value of b shows blueness.

<p style="text-align: justify;">

<p style="text-align: justify;">In standard colour chart, at centre a and b zero and they shows gray colour (Das, 2005). For whiteness index (WI) determinations following relations are used.

<p style="text-align: justify;">

<p style="text-align: justify;">Yellowness (Y) = L2/100                                                                                                                              (17)

<p style="text-align: justify;">Z = 1.18103L (L/100 – b/70)                                                                                                                                  (18)

<p style="text-align: justify;">WI = 3.388Z-3Y                                                                                                                                         (19)

<p style="text-align: justify;">YI = 142.86b/L                                                                                                                                           (20)

<p style="text-align: justify;">

<p style="text-align: center;">Murlidhar Meghwal, T K Goswami: Continental J. Food Science and Technology 4: 24 – 37, 2010

<p style="text-align: justify;">

<p style="text-align: justify;">

<p style="text-align: justify;">Aroma and fragrance

<p style="text-align: justify;"> Aroma and fragrance was just observed by human smelling sense but for better understanding and to know the change in chemical constitute we have to go for Gas chromatic analysis.

<p style="text-align: justify;">

<p style="text-align: justify;">Liquid nitrogen (LN2) requirement

<p style="text-align: justify;">LN2 was obtained from Cryogenic Engineering Centre (IIT Kharagpur) @ Rs. 15/ litre. The amount of LN2undefinedrequired for grinding was estimated by filling the known amount of LN2 is Dewar before grinding and then measuring the left LN2 after grinding in Dewar.

RESULTS AND DISCUSSION

<p style="text-align: justify;">Ambient and Cryogenic Grinding

<p style="text-align: justify;">Power consumption (P)

<p style="text-align: justify;">Power is the time rate at which work is done or energy is transferred. In calculus terms, power is the derivative of work with respect to time. Ambient and cryogenic grinding was carried out by using rotor speed mill with 0.5 mm and 1 mm sieve opening size. Data such as material taken for grinding and results obtained like product, time taken for grinding, rps, feed rate, power consumption and specific energy are tabulated in Table 1. It is clear from Table 1 that ambient grinding needs more power for grinding compare to cryogenic grinding. It is because during cryogenic grinding material become more brittle. On the contrary, in ambient grinding oil will come out of the cells and make the material sticky in nature and material sticks on the grinding surface which require more amount of power to grind the material.

<p style="text-align: justify;">

<p style="text-align: justify;">Table 1 Sieve size, moisture content, grinding time, grinding temperature, feed rate, revolution, power consumption, and specific energy <p style="text-align: justify;">AG = Ambient grinding; CG = Cryogenic grinding

The data are expressed as mean and the coefﬁcient of variation was <5%.

Fig. 2 shows variations in specific energy requirement with variations in grinding temperature.

<p style="text-align: center;">

<p style="text-align: center;">

<p style="text-align: center;">

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<p style="text-align: center;">Murlidhar Meghwal, T K Goswami: Continental J. Food Science and Technology 4: 24 – 37, 2010

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<p style="text-align: center;">Fig. 2 Variation in power requirement with varying temperature

0.5 mm opening sieve                                       1 mm opening sieve

<p style="text-align: center;">

<p style="text-align: center;">Eq. (21) and (22) shows the variation in power consumption with different temperature while grinding with 1 and 0.5 mm opening sieve size.

1 mm opening sieve

<p style="margin-left: 36pt; text-align: center; text-indent: 36pt;">       P = 76.45 – 0.01T – 0.001T2           (R2 = 0.949)                                 (21)

0.5 mm opening sieve

<p style="text-align: center;">                                        P = 77.36 + 0.003T – 0.001T2            (R2 = 0.941)                            (22)

<p style="text-align: center;">

<p style="text-align: justify;">Specific Energy Requirement (Es)

<p style="text-align: justify;">Fig. 3 shows variations in specific energy requirement with variations in grinding temperature.

<p style="text-align: justify;">

<p style="text-align: center;">

<p style="text-align: center;">Fig. 3 Variation in specific energy requirement with varying temperature

<p style="margin-left: 72pt;"> 0.5 mm opening sieve                         1 mm opening sieve

<p style="text-align: center;">Eq. (23) and (24) shows the variation in specific energy with different temperature while grinding with 1 and 0.5 mm opening sieve size.

<p style="text-align: center;">Murlidhar Meghwal, T K Goswami: Continental J. Food Science and Technology 4: 24 – 37, 2010

1 mm opening sieve

<p style="text-align: center;">                              Es = 118.3 + 0.012T – 0.002T2               (R2 = 0.954)                                  (23)

At 0.5 mm opening sieve

<p style="text-align: center;">                                Es = 123.2 + 0.060T - 0.002T2                (R2 = 0.944)                     (24)

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<p style="text-align: justify;">Energy Constant

It is another way to know the amount of power required to grind the material. Table 2 shows Rittenger’s, Kick’s and Bond’s law constants value.

Table 2 Rittenger’s, Kick’s and Bond’s law constants value

<p style="text-align: justify;">

<p style="text-align: justify;">The data are expressed as mean and the coefﬁcient of variation was <5%.

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<p style="text-align: justify;">Particle Size Analysis

<p style="text-align: justify;">Table 3 shows the different diameter calculated for the material ground under ambient and cryogenic grinding. It shows true density, initial specific surface and final specific surface area of the sample and its ground powder. It is evident from the table that cryogenic grinding produces fine particles. The mass mean diameter of the cryogenically ground powder was lies between 248 μm (0.25mm) to 502 μm (0.50mm) (Table 3). A low feed rate could produces with low average particle sizes (Indira, 2006).

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<p style="text-align: justify;">Table 3 Different types of diameter, specific area before and after grinding <p style="text-align: center;">

The data are expressed as mean and the coefﬁcient of variation was <5%.

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<p style="text-align: center;">Murlidhar Meghwal, T K Goswami: Continental J. Food Science and Technology 4: 24 – 37, 2010

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<p style="text-align: justify;">Volatile oil content

<p style="text-align: justify;">Interest in the production of high-quality spice products has encouraged by scientists to investigate the effects of cryogenic milling on spice quality. Results indicated that cryogenically milled spices retained more of the volatiles of the natural spice. The volatile oil content in cryogenically ground powder varied between 1.98 to 2.15 ml/100 g of powder at -60 to -110 oC (Table 2) respectively. Whereas, in ambient grinding oil was obtained 0.87 to 0.96 ml/100g at 60 to 65 oC product temperature (Fig. 4). Thus, cryogenic grinding helps to avoid the loss of volatile oil of the spice that also helps in retaining aroma and medicinal value of product.

<p style="text-align: center; text-indent: 36pt;">Fig. 4. Volatile oil content obtained from ambient and cryogenically ground sample

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<p style="text-align: justify;">Colour

<p style="text-align: justify;">Colour value and colour indexes results of the ambient and cryogenically ground black pepper powder. It can be observed that cryogenic grinding has improved the whiteness and yellowness indexes (Fig. 5), whereas ambient grinding produces ash coloured powder with high whiteness and low yellowness indices thus cryogenic grinding produces improved colour of spice.

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<p style="text-align: center; text-indent: 36pt;">Fig. 5. Colour indices for ambient and cryogenically ground sample

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">                               Whiteness index                       Yellowness index

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<p style="text-align: center;">Murlidhar Meghwal, T K Goswami: Continental J. Food Science and Technology 4: 24 – 37, 2010

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<p style="text-align: justify;">Liquid nitrogen requirement for cryogenic grinding

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">LN2 obtaining and storage is need very careful attention. We can’t totally sealed the LN2 because it starts at boiling -192oC and continuously evaporates from the vessel. So has as soon brought the LN2 to from central plant used it to avoid losses. The requirement of LN2 for cryogenic grinding at different very low temperature is shown in Fig. 6.

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">

<p style="text-align: center;">

<p style="margin: 0cm 0cm 0.0001pt; text-align: center; line-height: normal;">Fig. 6. Liquid nitrogen requirement at different cryogenic grinding temperature

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">It was observed that as we carried out grinding towards more and more negative temperature LN2 requirement increased and it is represented in eq. (25)

<p style="margin: 0cm 0cm 0.0001pt 72pt; text-align: center; line-height: normal;">LN2 = 0.828 + 0.007T +0.000T2      (R2 = 0.992)                               (25)

<p style="text-align: justify;">Comparison between cryogenic and ambient grinding

<p style="text-align: justify;">

<p style="text-align: justify;">Power requirement

<p style="text-align: justify;">Ambient grinding needs more power to grind the commodity compare to cryogenic grinding (Fig. 2) it is due to that in ambient for high oil bearing material, oil comes out from oil bearing material during grinding at high temperature (40 to 90 oC) which makes ground product gummy and sticky; gummy powder sticks on grinding surfaces that results in high power consumption and also chocking of sieves through which the product passes.

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<p style="text-align: justify;">Specific energy requirement

<p style="text-align: justify;">In case of ambient grinding specific energy consumption was high compare to cryogenic grinding. In ambient grinding at high temperature (40 to 90 oC) oil comes out of cells and makes the product viscous and sticky in nature. This leads to high consumption of power in ambient grinding while in case of cryogenic grinding due to low temperature (-60 to -110 oC) material becomes brittle and frugal in nature and requires less amount of energy for grinding (Fig. 3) (Ghorbani et al., 2010).

<p style="text-align: justify;">

<p style="text-align: justify;">Energy constant

<p style="text-align: justify;">It is evident from Table 3 that cryogenic grinding need less power for grinding of commodity compare to ambient grinding.

<p style="text-align: justify;">

<p style="text-align: justify;">Particle size of powder

<p style="text-align: justify;">The mass mean diameter in the case of ambient and cryogenic grinding were 326 μm (0.326) to 352 μm (0.352 mm) and 276 μm (0.276 mm) to 202 μm (0.202 mm) which shows that cryogenic grinding produces fine particles compare to ambient grinding (Santos et al., 2002). It was observed that size of product particle is function of feed rate, product temperature and moisture content of the sample.

<p style="text-align: justify;">

<p style="text-align: center;">Murlidhar Meghwal, T K Goswami: Continental J. Food Science and Technology 4: 24 – 37, 2010

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<p style="text-align: justify;">

<p style="text-align: justify;">Colour indexes

<p style="text-align: justify;">In case of ambient grinding due to high production temperature powder turns into dull in colour, became ash like in colour and lost its brightness. On the hand, in cryogenic grinding, high colour indexes were obtained due to preservation of brightness and natural lust of powder.

<p style="text-align: justify;">Yield of volatile oil

<p style="text-align: justify;">

<p style="text-align: justify;">A significant difference was observed in yield of volatile from ambient and cryogenic grinding. The yield of volatile oil in cryogenic grinding was obtained in the range of 1.98 to 2.15 ml/100g whereas in ambient grinding it was obtained in the range of 0.87 to 0.96 ml/100g of pepper powder.

<p style="text-align: justify;">

<p style="text-align: justify;">Sieve clogging

<p style="text-align: justify;">Sieve clogging is shown in Fig. 7. It is clear from the figure that as the temperature of grinding decreases the clogging of the sieve decreases for the same sieve opening.

<p style="margin: 0cm 0cm 0.0001pt 72pt; vertical-align: baseline;">1 mm sieve                                                                 0.5 mm sieve

<p style="margin: 0cm 0cm 0.0001pt 36pt; text-indent: 36pt; vertical-align: baseline;">1 mm sieve                                                            0.5 mm sieve

<p style="margin: 0cm 0cm 0.0001pt; text-indent: 36pt; vertical-align: baseline;">Ambient Grinding, 60oC                   Ambient Grinding , 60oC

<p style="text-align: center;">

<p style="margin: 0cm 0cm 0.0001pt 36pt; text-indent: 36pt; vertical-align: baseline;">1 mm sieve                                                                          0.5 mm sieve

<p style="margin: 0cm 0cm 0.0001pt; text-indent: 36pt; vertical-align: baseline;">Cryogenic Grinding -110 oC                              Cryogenic Grinding  -110 oC

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<p style="text-align: center;">Murlidhar Meghwal, T K Goswami: Continental J. Food Science and Technology 4: 24 – 37, 2010

<p style="margin: 0cm 0cm 0.0001pt; text-indent: 36pt; vertical-align: baseline;">

<p style="margin: 0cm 0cm 0.0001pt; text-indent: 36pt; vertical-align: baseline;">

<p style="text-align: center;">Fig. 7 Sieve chocking characteristics in ambient and cryogenic grinding

<p style="text-align: justify;">

<p style="text-align: justify;">Health and hygienic condition

<p style="text-align: justify;">A significant loss of volatile oil in ambient grinding may be because of high-grinding temperature that leads to vapourisation of volatile compounds. These fine vaporised oils molecules and spreading of very fine spice powder in surrounding causes eye, nose and throat irritations if inhaled, thereby leading to fatigue of workers. The dust and volatile oil produced during ambient grinding in working atmosphere of worker or mill operator can create respiratory problems (Murthy et al., 1999). Some time, there may fire accident in ambient grinding due to high operating temperature. Eye burning, sneezing and nose watering are common problem arise due to ambient grinding. During cryogenic grinding, the vapourisation of oils was found minimum as most of the oil compounds are retained within the powder itself because of low temperatures, and the oils are present mostly as solid. The dust formation is also very insignificant and the problem of eye burning, sneezing and nose watering are not observed in cryogenic grinding. Such positive aspects of cryogenic grinding show the practical usefulness of this novel technology.

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<p style="text-align: justify;">CONCLUSION

<p style="text-align: justify;">It is concluded from study that less specific energy and power were required for cryogenic grinding; improved colour powder was obtained from cryogenic grinding; higher volatile oil content was obtained from cryogenic grinding; clogging of sieve was found to be serious in ambient grinding. LN2 requirement varied between 1 to 1.4 kg kg-1 for grinding temperature of -60 to -110oC. Cryogenic grinding is free from eye burning, sneezing and nose watering and it is a hygienically novel technique.

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<p style="text-align: justify; vertical-align: baseline;"> REFERENCES

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<p style="margin-left: 21.3pt; text-align: justify; text-indent: -21.3pt;">technology for traditional Chinese herbal medicine. Cryogenics, Vol. 31, China.

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<p style="margin-left: 21.3pt; text-align: justify; text-indent: -21.3pt;">New Delhi, 150-320pp.

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<p style="margin-left: 21.3pt; text-align: justify; text-indent: -21.3pt;">Santos, D., Barbosa, F., Tomazelli, A.C., Krug, F.J., Nóbrega, J.A., Arruda, M.A.Z. (2002). Determination

<p style="margin-left: 21.3pt; text-align: justify; text-indent: -21.3pt;">of Cd and Pb in food slurries by GFAAS using cryogenic grinding for sample preparation. Anal Bioanal

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<p style="margin-left: 21.3pt; text-align: justify; text-indent: -21.3pt;">Singh, K.K. & Goswami, T.K. (1997). Studies on cryogenic grinding of spices. IIT Kharagpur(India).

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<p style="text-align: justify;">Received for Publication: 03/06/2010

<p style="text-align: justify;">Accepted for Publication: 20/07/2010

Corresponding Author:

T K Goswami

Agricultural and Food Engineering Department (AgFE), Indian Institute of Technology (IIT) Kharagpur,

E-mail: [mailto:tkg@agfe.iitkgp.ernet.in tkg@agfe.iitkgp.ernet.in]

<p style="text-align: justify;">Continental J. Food Science and Technology 4: 38 – 45, 2010                                ISSN: 2141 – 422X

<p style="text-align: justify;">©Wilolud Journals, 2010                                                                                  http://www.wiloludjournal.com

NUTRIENTS AND ANTI-NUTRIENTS CONTENT OF DEHULLED SOYBEAN

<p style="text-align: center;">

<p style="text-align: center;">

<p style="text-align: center;">Odumodu C.U.

<p style="text-align: center;">Department of Pediatrics, University of Jos, P.M.B. 2084 Jos, Plateau State

e-mail: odumoduc@unijos edu.ng.

= = =ABSTRACT= =The nutrient and anti-nutrient contents of dehulled soybean were investigated, using proximate composition, amino acid profile, mineral, micronutrient and anti-nutrient values. The soybean was boiled for 10 minutes, dried, dehulled and milled into a fine flour. The dehulled soybean (DSB) had high value for protein and fat (49.38 and 25.97g/100g respectively). The carbohydrate content of the DSB was very low (12.89g/100g). Leucine and glutamic acid were the most abundant essential and non-essential amino acids (6.62 and 16.25g/16g N respectively). The sulphur containing amino acids methionine and cystine were 1.22 and 2.09g/16g N respectively. The DSB is a good source of phosphorous (1193.3mg/100g). It had almost equal amount of calcium and magnesium (17.5 and  17.3 mg/100g respectively). It was a good source of thiamin (408.0mg/100g), riboflavin (394.1mg/100g) and niacin (603.0mg/100g).The DSB had very high level of trypsin inhibitor (2786.00 TIU/100g). The tannin and phytate content were 34.92 and 42.00mg/00g respectively. The dehulled soybean was found to have high nutritional values but was underscored by its high anti-nutrients; trypsin inhibitor (2786.00 TIU/100g). tannin (34.92mg/100g), phytate (42.00mg/100g), oxalate (27.62mg/100g) and saponin (34.60mg/100g).= <p style="margin: 0cm 43.2pt 0.0001pt;">

<p style="margin: 0cm 43.2pt 0.0001pt;">KEYWORDS:  Soybean, nutrient, anti-nutrient, amino acid, micronutrients, proximate compositions = = =INTRODUCTION= ==Soybean (Glycine max (L), is a legume that grows in tropical, subtropical and temperate climate. Approximately half of the world’s soybean is produced in the developing world and the other half in the developed world (IITA 2002). Soybean is an important source of inexpensive and high quality protein and oil (Khalid et al ; 2008). Extensive works have been reported on the  nutrients and anti-nutrients content of soybean but none on the quantity  of the anti-nutrients content of dehulled soybean.. == <p style="text-align: justify;">

<p style="text-align: justify;">Soybean nutritionally emerged as one single food crop with nutrients content almost equal to that of animal product. It contains all the essential amino acids and is an excellent source of dietary fiber. Messina (2004) reported that soybean was relatively high in iron, phosphorous, copper, magnesium and manganese. Soybean contains very little starch and about half of the total polysaccharides is composed of oligosaccharides

<p style="text-align: justify;">

<p style="text-align: justify;">It also contains several phytochemicals such as  lectins, (hemagglutinins), phytosterols, phytic acid, saponin, protease inhibitors,  a variety of phenolic acids and importantly isoflavones in relatively high amount (Messina, 2004). These anti-nutrients are compounds with adverse nutritional and physiologic effects. In animal model, protease inhibitors have been shown to cause growth retardation and enhanced chemically induced  pancreatic cancer (Messina, 2004). Most of the interest in these phytochemicals focused on their possible anti- nutrient effects especially the protease inhibitors and phytic acid.

<p style="text-align: justify;">

<p style="text-align: justify;">This work was undertaken to establish quantitatively the nutrients and anti-nutrients content of the dehulled soybean.

<p style="text-align: justify;">

<p style="text-align: justify;">

<p style="text-align: center;">Odumodu C.U: Continental J. Food Science and Technology 4: 38 – 45, 2010

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<p style="text-align: justify;">

MATERIALS AND METHODS
<p style="text-align: justify;">Soybean (Glycine max (L)) was purchased from Jos Main Market, manually cleaned to remove foreign materials. Four hundred grains of raw soybean was placed in 10 litres of boiling water containing 5.0g sodium bicarbonate. The soybean was boiled for 10 minutes and the water drained off. It was dried in the oven at 80oC for 24h and dehulled mechanically using laboratory  hammer mill. It was milled in a laboratory hammer mill into a fine flour (300μm mesh screen) and packaged in a labeled low density polyethylene  bag. The bag was placed in a plastic bucket with cover and stored in a deep freezer at - 18oC for analysis.

<p style="text-align: justify;">

Proximate Composition
Proximate analysis was carried out on the dehulled soybean by the methods of Association  of  Official  Analytical   Chemists (AOAC,  1980).

Determination of Amino Acid Profile
<p style="text-align: justify;">The amino acid profile of  the dehulled soybean, was determined using the method by (Spackman  et  al ; 1958). The sample was defatted by weighing 5g of the sample into extraction thimble. The fat was extracted with chloroform and methanol mixture (2:1) using the soxhlet extraction apparatus as described by AOAC, (1980). Two hundred milligram of the fat free sample was weighed into a glass ampoule. Seven millilitre of hydrochloric acid was added and oxygen was expelled by passing nitrogen gas through the solution. This was to avoid possibility of oxidation of cystine and methionine (Benitez, 1989). The glass ampoule was then sealed with Bunsen burner flame. The ampoule and its contents was hydrolysed in an oven at 105oC for 22 hours. Immediately after cooling, it was filtered through non-absorbent cotton wool. The filterate was dried at 40oC using Heidolyph evaporator (Model W1). The amino acids in the flask was diluted with 5ml of sodium citrate buffer (PH 2.0). About 5 to 10 microlitre was loaded into the cartridge of Technicon Sequential Multi-sample amino-acid analyzer (TSM). Precise amounts of ninhydrin and hydrazine sulphate reagents were combined on the manifold of the TSM to form hydrin dantin. A segmented reagent stream flowed under each analyzer column to join with the eluting buffers. The stream carrying the amino acid reagent mixture went through a heating bath where development of the colour reaction product occurs. The absorbance was proportional to the concentration of each amino acid and was measured by colorimeter. The determination was done in triplicate.

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<p style="margin-bottom: 0.0001pt; line-height: normal;">Elemental Determination

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">The levels of selenium, zinc, iron, copper, molybdenum, calcium, magnesium, manganese, and lead in the dehulled soybean, were determined by the method of Atomic Absorption spectrophotometric method  of  Association  of  Official  Analytical  Chemists  (AOAC, 1980) .One gram of  the sample was accurately  weighed  and  ashed for  2h  at 500oCand  allowed  to cool. The ash was made wet with 10drops of distilled water  and  3-4ml HNO3 (1+1). Excess HNO3 was evaporated. The content was ashed   for additional  1h at  500oC. It was dissolved in 10ml NHCl (1+1) and made up to 50ml. The mixture was forced into the flame with sufficiently high temperature to cause the atomization of the analyte element.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Total phosphate content was determined by the modified method of  Fiske and Subbarow, (1935). One gram of the sample was digested with 5ml of a mixture of  nitric acid, perchloric  acid and sulphuric acid in the ratio 4:1:1 v/v in a micro  Kjeldahi  flask. The absorbance of the blue colour produced was read at 720nm.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">The iodine determination was by the method of Muir and Lambert,  (1973). Sodium thiosulphate (thiosulphate vi) reacted with iodine to produce sodium iodide and sodium tetrathionate. The free iodine produced a deep blue colouration which disappeared  as  soon as  sufficient  thiosulphate was added to react with all the iodine.

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<p style="text-align: center;">Odumodu C.U: Continental J. Food Science and Technology 4: 38 – 45, 2010

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Vitamins Determination

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Determination of  various  vitamins  in the sample were done in accordance with  the  methods of  the  following  authors : vitamin  A  (Pearson, 1976) ,thiamin and  riboflavin ( Stroebecker  and Henning, 1965),  niacin  (AOAC,1980)  and  vitamin  C  (Roe and Kuether, 1943).

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Determination of Anti-nutrients

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">The levels of phytates, trypsin inhibitor, oxalate, tannin and saponin were  determined in the dehulled soybean.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Phytic acid (inositol hexaphosphate) content of the sample was determined by the modified method  of McCance and Widdowson ,(1935).Ten grams of the sampel was extracted in 200ml of 1NHCl  for 12h at room  temperature. Fifty milliliters of 1%  ferric chloride in 1NHCl  was added and boiled. This procedure precipitated phytic  acid  as  ferric phytate. The concentration of phytic phosphate  was calculated  from  the standard  inorganic phosphate  curve prepared.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Trypsin inhibitor activity (TIA) was estimated essentially according to the method Kakade et al, (1974), except that 2.0g of the finely ground sample was extracted with 10ml 0.01 NaOH for 1h. Five milliliter of BAPNA ( benzoyl-DL arginine-p-nitro anilide hydrochloride) solution was hydrolyzed with 2ml of 0.2mg/ml trypsin (Sigma  Type 11) in 0.0001MHCl. P-nitro anilide  was released as  a  coloured  product  and  absorbance  read  at 410 nm.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">The total oxalic acid content of the  sample was determined by the modified  method  of  Abeza et al, (1968). Two grams of the sample was digested for 1h  with 10ml  of 6M  of hydrochloric acid. The digest was boiled  and filtered. The precipitate was dissolved  in  5ml  of 1:4 sulphuric acid. The resultant solution was titrated against  0.05KMNO4  untill  the pink  colour persisted  for one minute. A blank was run  along with  the test sample.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">The tannic acid content of the sample was determined colorimetrically by the method of Price and Bulter,( 1977).Two grams of the sample was mixed  with 10ml of 2MHCl. The content was shaken and made up to 50ml and filtered. Five milliliters of the filterate was mixed with 3ml of 0.1MFeCl3  in 0.1NHCl and 3ml of 0.008M potassium ferrocynide   (K3Fe(CN)6.  Absorbance was read at 720 nm.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Saponin was determined according to the method of Birk et al (1963), modified by Hudson and El-Difrawi, ( 1979). Ten grams of the sample was mixed in 20ml of 20%  aqueous ethanol and filtered. The residue was re-extracted with  20ml  of 20% aqueous ethanol. The aqueous layer was recovered while the ether layer was discarded. The process was continued until a colourless aqueous extract was obtained. The extract was washed and evaporated to dryness to obtain crude saponin.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">The mean and standard error of the mean were calculated using the procedure  of  Steel and Torrie, (1960).

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<p style="margin-bottom: 0.0001pt; line-height: normal;">RESULTS

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">The proximate composition of the dehulled soybean is presented in Table 1. The dehulled soybean had 3.7g/100g moisture and 3.99g/100g ash. Soybean is an oil seed as such it had high fat value (25.97g/100g). It had fibre and protein values for 4.06 and 49.38g/100g  respectively. The carbohydrate content of soybean was very low (12.89g/00g).

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Table 2 shows the amino acid profile of the dehulled soybean. Seventeen amino acids were detected. The DSB was a source of nine essential amino acids. Leucine and glutamic acid were the most abundant essential and non-essential amino acids with values 6.62 and 16.25g/16g N respectively. The sulphur containing amino acids, methionine and cystine were 1.22 and 2.09g/16gN respectively.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">The minerals and micronutrients content of dehulled soybean are presented in Table 3. The dehulled soybean is a good source of phosphorous (1193.3mg/100g) and calcium (17.5mg/100g). It had almost equal

<p style="text-align: center;">Odumodu C.U: Continental J. Food Science and Technology 4: 38 – 45, 2010

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">amount of calcium and magnesium (17.5 and 17.3mg/100g respectively). The zinc and iron concentrations were 8.4 and 9.3mg/100g respectively. Molybdenum was found in traces in the dehulled  soybean. Selenium and iodine concentrations were 0.08 and 0.27mg/100g respectively. Manganese content in the dehulled  soybean was low (0.057mg/100g) while lead was not detected. The DSB was a good source of thiamin, riboflavin and  niacin (408.0, 394.1 and 603.0mg/100g respectively). The vitamin A content was 5.9 RE.

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Table 4 shows  the anti-nutrients content of dehulled soybean. The DSB had a very high level of trypsin inhibitor (2786.00 TIU/100g). The tannin and phytate content were 34.92 and 42.00mg/100g respectively. The oxalate and saponin content were 27.62 and 34.60mg/100g respectively.

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<p style="margin-bottom: 0.0001pt; line-height: normal;">DISCUSSION

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">The proximate composition results of the DSB in Table 1 differed from the USAID (1986) results; moisture 8.54, ash 4.87, fat 19.94, fibre 4.96, protein 36.49 and carbohydrate 30.16g/100g. The results (Table I) revealed higher values for moisture, ash, fibre and carbohydrate but lower values for fat and protein. This could be attributed to the effects of absence of  seed coat of the  soybean. The lower carbohydrate but higher fat values confirmed the report of Mayer  et al  (1960) that  during the maturation of many oil seeds an increase in oil content occurs concurrently  with a decrease in the quantity of carbohydrate. This indicated that its carbohydrate could  have  been  converted to fat. The lower moisture for the dehulled soybean supported the higher fat value because high fat products have low moisture and high protein.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">The results presented in table 2 differed from the Lathia and Kruchten (1996) results; histidine 28, isoleucine 50, leucine 85, lysine 70, methionine +  cystine 28, phenylalanine + tyrosine 88, threonine 42,  tryptophan 14 and valine 53mg/g protein but similar to the FAO  (1973) patterns. This is a confirmation of the statement by Messina  2004,  that soybean  nutritionally  emerged as one single food crop with nutrients content almost equal to animal product.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Khalid et al ; (2008) reports  on the minerals and micronutrients content of soybean, Ca 225, Cu 0.75, Fe 6.90, Mg 200, Mn 2.20, K 1190, Na 9.42 and Zn 1.65mg/100g were higher than the results presented in Table 3. In the present study the dehulled soybean had higher phosphorous value. This agreed with the reports of Linder (1997), which stated that phosphorous is generally abundant in foods with high protein content. Berk (1992) reported that the major mineral constituents of soybean  were mainly  potassium, calcium and magnesium and the trace elements (iron, zinc and copper) were the minor constituents. The traces of molybdenum in the dehulled soybean could be attributed to adverse of anti-nutrients and food toxicants (phytate, tannin, oxalate, cyanide and heamagglutinin).

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">A lot of works have been published on anti-nutrients content of soybean but none on  the  quantity  of  each   anti-nutrient e.g. Rackis et  al (2004), Hunter (2001) and Berk (1992)  reported  generally on the anti-nutrients content of soybean but this study quantified each anti-nutrient  as shown in Table 4. The phytate value of the DSB was 42mg/100g. This organic acid is present in the bran or hull of all seeds and legumes but no seed has higher  level of phytate which soybean has. Phytate blocks the body’s uptake of essential minerals like magnesium, calcium, iron and especially zinc (Ice, 2000). The trypsin inhibitor of the dehulled soybean was 2786. 00 TIU. This is a potent enzyme – inhibitor which blocks uptake of trypsin and other enzymes  which the body needs for protein digestion.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">In conclusion the DSB was found to have excellent nutritional values like that of animal product but was underscored by its high anti-nutrient values; trypsin inhibitor (2786.00 TIU/100g), tannin (34.92mg/100g), phytate (42.00mg/100g) oxalate (27.62mg/100g) and saponin (34.60g/100g). Based on the above results  soybean is  a good,  cheap source  of nutrients  but under scored by its  high anti-nutrients content.

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<p style="margin-bottom: 0.0001pt; line-height: normal;">REFERENCES

<p style="margin: 0cm 0cm 0.0001pt 36pt; text-align: justify; text-indent: -36pt; line-height: normal;">Abeza, R.H. Blake, J.J. and Fisher, E.J. (1968), Oxalate determination: Analytical problems encountered

<p style="margin: 0cm 0cm 0.0001pt 36pt; text-align: justify; text-indent: -36pt; line-height: normal;">with certain plants species. J. Assoc Agric. Chem. 51; 963 – 965.

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<p style="text-align: center;">Odumodu C.U: Continental J. Food Science and Technology 4: 38 – 45, 2010

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<p style="margin: 0cm 0cm 0.0001pt 36pt; text-align: justify; text-indent: -36pt; line-height: normal;">AOAC (1980). Official Methods of Analysis 13th Edition. Association of Official Analytical Chemists,

<p style="margin: 0cm 0cm 0.0001pt 36pt; text-align: justify; text-indent: -36pt; line-height: normal;">Washington, D.C.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Benitez, L.V. (1989). Amino acid and fatty acid profiles in aquaculture nutrition studies. In S.S De Salva (ed). Fish Nutrition Research in Asia. Proceedings of the Third Asian Fish. Nutrition Net Work Meeting. Manila Asian Fishery Society 23 – 35.

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Berk, Z. (1992) Technology of Production of edible flours and protein products from soybeans. Publisher FAO. Viale delle Terme di caracolla. 00100 Rome, Italy. 12 pp.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Birk, Y. Bondi, A.Gestetner, B. and Ishaya, I.A. (1963). Thermostable hemolytic  factor in  sobeans. Nature, 197:1089-1090.

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">FAO (1973). Energy and protein requirement. Report of joint FAO/WHO. Ad HOC Expert Committee. WHO Tech. Rep. Series No. 522 Geneva.

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Fiske, D and Subbarow, M. (1935). Colorimetric determination of phosphorus. Journal of Biological chemistry, 26: 375 – 382.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Hudson, B.J.F. and El-Difrawa, W.A. (1979) The sapogenine of the seeds of four  lupin species. J. Plant Food 3: 181 – 186.

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Hunter, B. T. (2004). The downside of soybean consumption. In consumers Research Magazine 1 – 7.

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Ice, B. (2000) Modern Soy Anti-nutrient and Toxins block vitamins and minerals. Extracted from Nexus Magazine, vol. 7. No. 3 pp 88 – 110.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">IITA (2002) International Institute of Tropical Agriculture Project includes work on soybean crops and Farming system. Nigeria. Pp 1 – 3.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Kakade, M.L. Rackis, J.J. McGhee, J.E. and Pushi, G. (1974). Determination of trypsin inhibitor activity of soy products. A collaborative analysis of an improved procedure. Cereal chem. 51: 376 – 379.

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Khalid A.I. Elsiddig A.E. and Elfadil E.B. (2008). Minerals composition of soybean (Glycine max L.) seed as influenced by Bradyrhizobium Inoculation and Chicken Manure or Sulphur Fertilization.

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Lathia, D. and Kruchten, S. (1996). Potential nutritional and health benefits of newly developed fermented soy milk desserts. Second International Symposium on the Role of Soy in Preventing and Treating  Chronic Disease. Brussels Belgium

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Linder, M.C. (1997). Nutritional Biochemistry and Metabolism with Clinic Application. 2nd Edition. Publisher Appleton and Lange Norwalk, Connecticut 142 – 276.

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">McCance, R. A. and Widdowson E.M. (1935). Estimation of phytic acid. Bioch, J. 29: 2694 – 2700.

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Messina, M. (2004) Nutritional value of soyfood. Ph.D. Thesis. Department of Nutrition Michigan State University, United State of America 6 pp.

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Meyer, B.S. Anderson, D.B. and Bohing, R.H. (1960). Introduction to Plant Physiology. Published by D.Van Nostrand Com,pany (Canada) LTD. 230 – 520.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Muir, Y.A. and Lamberts, J. 1973). Practical chemistry (3rd ed) Heinem ann Educational Books Ltd. Bulter and Tanner Ltd Fone and London. 339 – 340.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Pearson, D. (1976) The Chemical Analysis of Food. Other vitamins A. 7th Edition Church//Livingstone, New York Manual of Analysis, CBA – 301 series.

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Price, M.I. and Bulter, LG. (1977) Rapid usual estimation and Spectrophoto metric determination of tannin content of sorghum grain. J. Agric Food chem. 25:1269 – 1273.

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Rackis et al (2004). Evaluation of the Health Aspects of Soy protein Isolates as food ingredients, prepared for FDA by life sciences Research Office, Federation of America Societies for Experimental Biology, 9650 Rockville Pike, Bethseda, MD 20014, U.S.A.

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Roe, J.H. and Kuether, C.A. (1943) Photometric assay of Vitamin C. Journal of Biological chemistry, 147:399 – 400.

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Spackman, D.H. Stein, E.H. and Moore, S. (1958) Automatic Recording Apparatus for use in chromatography of amino acids.

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Steel, R.G.D. and Torrie, J.H. (1960) Principles and procedures of statistics with special reference                                           to the biological sciences. MacGraw-Hill Book Co. Inc, New York, Tronto and London.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Stroebecker, R. and Henning, M.H. (1965) vitamin Assay – Tested Methods Verlag Chemic GMBH Welnheim/Bergster 120 – 140.

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">USAID (1986) Composition of foods legume and legume products. USDA Human Nutrition Information Services Agriculture Handbook No. 8 – 16. United States Department of Agriculture, Washington D.C.

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<p style="margin: 0cm 0cm 0.0001pt 72pt; text-align: justify; line-height: normal;">Table 1: Proximate composition of dehulled soybean (g/100g, DRY WT)

<p style="margin: 0cm 0cm 0.0001pt 72pt; text-align: justify; line-height: normal;"> <p style="margin-bottom: 0.0001pt; text-align: justify; text-indent: 36pt; line-height: normal;">Values are means + standard deviation of triplicate determinations.

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<p style="text-align: center;">Odumodu C.U: Continental J. Food Science and Technology 4: 38 – 45, 2010

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<p style="margin-bottom: 0.0001pt; text-align: justify; text-indent: 36pt; line-height: normal;">Table 2: Amino Acid Profile of dehulled Soybean (g/16gN <p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">                Values are + standard deviation of triplicate determinations.

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<p style="margin: 0cm 0cm 0.0001pt 36pt; text-align: justify; line-height: normal;">Table 3:    Mineral and Micronutrients content of dehulled soybean (mg/100g, DRY WT.)

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;"> <p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Values are means + standard deviation of triplicate determinations,

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;"> N.D =  Not detected

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<p style="text-align: center;">Odumodu C.U: Continental J. Food Science and Technology 4: 38 – 45, 2010

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<p style="margin: 0cm 0cm 0.0001pt 36pt; text-align: justify; line-height: normal;">Table 4: Anti-nutrients content of dehulled soybean (TIU and mg/100g DRY WT)

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<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;"> <p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">

<p style="margin-bottom: 0.0001pt; text-align: justify; line-height: normal;">Values are means + standard deviations of triplicate determination

<p style="text-align: justify;">Received for Publication: 03/06/2010

<p style="text-align: justify;">Accepted for Publication: 20/07/2010

<p style="text-align: justify;">Continental J. Food Science and Technology 4: 46 – 52, 2010                                ISSN: 2141 – 422X

<p style="text-align: justify;">©Wilolud Journals, 2010                                                                                  http://www.wiloludjournal.com

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<p style="text-align: center;">EFFECT OF ILLUMINATION AND SCLEROTIA SIZE ON THE GROWTH AND DEVELOPMENT OF Pleurotus tuberregium (Fr.)Singer

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<p style="text-align: center;">A.O Oghenekaro, E.O Akpaja and J.O Samuel

<p style="text-align: center;">Department of Plant Biology and Biotechnology, University of Benin, P.M.B 1154, Benin City, Nigeria.

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<p style="margin: 0cm 43.2pt 0.0001pt; text-align: justify;">ABSTRACT

<p style="margin: 0cm 43.2pt 0.0001pt; text-align: justify;">Effect of illumination and sclerotia size on the growth and development of Pleurotus tubberregium was investigated. The cultivation was done using the sclerotia on garden soil. Mean fresh weight production by the mushroom was highest, 28.751.24g in the set up under shade and from the 25g sclerotia. While the least was 8.60± 0.44g in the dark from 10g sclerotia. Mean stipe length was greatest 7.30±0.71cm in the dark from the 20g sclerotia, while the least stipe length, 2.06±0.45cm from 25g sclerotia. 9.95±0.39cm was the highest cap diameter observed in 25g sclerotia with the least cap diameter of 2.90±0.59cm in the dark and from 25g sclerotia. This results shows that extremely high temperature and darkness has a retarding effect on the stipe length and cap development respectively. Therefore shade is the ideal condition for the growth and development of Pleurotus tuberregium regardless of sclerotia size.

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<p style="margin: 0cm 43.2pt 0.0001pt; text-align: justify;">KEYWORDS: Illumination, Darkness, Pleurotus tuberregium, Sclerotia, Garden soil,

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<p style="text-align: justify;">INTRODUCTION

<p style="text-align: justify;"> A Mushroom is a macrofungus with a distinctive fruiting body which can be either epigeous (above ground) or hypogeous (underground) and large enough to be seen with the naked eye and to be picked by hand. (Chang and Miles, 2004).The edible ones are highly nutritive when compared with meat, egg and milk. They are known to contain a lot of water, protein, lipids, sugars, amino-acids, glycogen, vitamins (B, C and D) and mineral elements (Ghosh and Chakravarty, 1990). Mushrooms represent one of the world’s greatest untapped resources of nutritious food. The cultivation of saprophytic edible mushrooms may be the only current economical biotechnology for Lignocellulose organic waste recycling that combines production of protein-rich food with the reduction of environmental pollution (Obodai et al, 2004). Cultivation of these edible fungi in the continent is very limited, it is only common in Zimbabwe, Kenya and South Africa (Declaire, 1978). It is still very skeletal in Nigeria. Although many eat mushrooms, they collect them from the wild. This practice is fraught with danger of collecting poisonous species along with the edible ones.

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<p style="text-align: justify;">Pleurotus is a genus of gilled mushrooms which includes one of the most widely eaten mushroom, the Oyster mushroom. Like all other basidiomycetes, it is distinguished by having clamp connections along its hyphal length. Pleurotus means “side ear”. The genus Pleurotus has many edible species that have been extensively studied. They are found in both temperate and tropical countries of the world.Pleurotus tuberregium is a very popular mushroom in Nigeria that has the potential of being commercialized. It produces tuberous sclerotia in dead wood (Oso, 1977). The fruit body and sclerotia are eaten in Nigeria where they are used for soup and tradomedical preparations, (Oso, 1977).

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<p style="text-align: justify;">P. tuberregium is indigenous to tropical Africa and the Australasian pacific regions of the wood ( Isikhuemhen et al, 2000) including sub-Saharan Africa, Madagascar, Malaysia, Papua New Guinea, Northern Australia, New Caledonia, Indonesia, Myanmar and the Yunnan province of China. Demand for sclerotia all over the world remains high while harvest from the wild is reduced. Deforestation and the conversion of forest into agricultural fields have led to the decline of natural habitat of this mushroom. The resultant scarcity of this mushroom has made it expensive, especially during the dry season (October to April), when it is not easy to collect them in the wild.

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<p style="text-align: center;">A.O Oghenekaro et al.,: Continental J. Food Science and Technology 4: 46 – 52, 2010

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<p style="text-align: justify;">During the rainy season, it is possible to see a decaying log filled with fruiting mushrooms during early years of its colonization. Once the substrate is fully colonized and has reached an advanced stage of decay, sclerotia will form at the end of the rainy season. The sclerotia survives the dry and hot season until the rain returns, at which point the sclerotia will either continue to enlarge or form sporophores (mushroom).

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<p style="text-align: justify;">Sclerotia are compact masses of mycelia tissue that serves to store food during unfavourable conditions and able to fruit when favourable conditions return. Sclerotia are dark brown on the surface, white inside, spherical to ovoid in shape and as large as 30cm in dameter.

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<p style="text-align: justify;">The sclerotia stage is not common among white-rot fungi. Among cultivated mushrooms, only Morchella spp. is known to form sclerotia as part of their life cycle. However, only the sclerotia of P. tuberregium are valued as food and medicine independent of its ability to produce mushroom. Although it is a sclerotia forming mushroom, some strains from Australia and Indonesia have been observed to fruit directly without ever forming sclerotia (Isikhuemhen 2000a). The sclerotia stage provides a unique advantage to the cultivation of this species. The reason is that it stores for years without loss of viability and eliminates the need for costly fruiting conditions.  

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<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">Indigenous people of Nigeria incorporate collected mushrooms, including P.tuberregium into their diet and medicine (Oso, 1977; Ogundana and Fagade, 1982; Akpaja et al; 2003). Similarly, people in Ghana, Cameroun, and the republic of Benin (Obodai et al, 2004; Kuyper et al; 2002) collect wild mushrooms and use them as food and medicine. Studies have shown that the Igbo people of Nigeria are the major consumers of P.tuberregium throughout the country using the fungus in place of meat as well as for its medicinal properties (Akpaja et al, 2003). The whitish inner tissue is milled into paste which is used to substitute in part or whole for melon seed (Citrutus lanatus) in the preparation, of ‘Egusi’ soup or mixed with corn flour and fried (Isikhuemhen and Okhuoya, 1995; Nwokolo, 1987). Analysis of both sclerotia and sporophores shows that they are rich in carbohydrate, protein, vitamins and minerals while low in fats (Nwokolo, 1987; Okhuoya and Ajerio, 1988)

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<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;"> Pleurotus tuberregium is cultivated either by inoculation of substrate with the pure spawn or by the use of sclerotia to induce sporophore production. Okhuoya and Okogbo (1990) reported that the use of casing material increases yield on a variety of substrates. Mushroom yield is approximately 25% of sclerotia weight and 90% moisture.

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<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">Okhuoya and Okogbo (1990) reported studies on sclerotium production in the wild and the induction of sclerotium production in the laboratory.

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<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">This study was undertaken to determine the effect of illumination and sclerotia size on the growth and development of P.tuberregium.

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<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">MATERIALS AND METHODS

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">Source of sclerotia and preparation

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">Sclerotia of Pleurotus tuberregium were bought from a woman who collected them from decaying logs in the Okomu forest in the Benin region and brought them to the Ekiuwa market for sale.45 plastic rubber plates of dimension 6cm by 16cm each were obtained from Oba market in Benin City. Knife was also purchased from Oba market. The knife was used to, not only, peel off the brown outer covering of the sclerotia but also to cut the sclerotia into various masses. The weights were then obtained with the aid of the electric weighing balance. The various weight of the sclerotia that were taken are 5g, 10g, 15g, 20g, and 25g. Three replicates were taken for each weight. The weighed sclerotia were then soaked in water for 5 hours.

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<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">Preparation of substratum

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">Loamy soil obtained from the Junior staff Quarter (JSQ) of the University of Benin, Benin-city. Electric soldering iron was purchased from the market at ring road, Benin City. Electric weighing balance obtained

<p style="text-align: center;">A.O Oghenekaro et al.,: Continental J. Food Science and Technology 4: 46 – 52, 2010

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<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">from the Departmental laboratory, Department of Plant Biology and Biotechnology, University of Benin, Benin City. The sieve was used to filter the soil for fine particles. The electric soldering iron was connected to a power source and was used to bore holes at the bottom of the plastic plates to provide for proper aeration and prevent water-logging of the plastic container. The perforated plates were filled with the sieved soil to the brim and then watered a little.

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<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">Sowing of sclerotia pieces and planting

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">After soaking, the various sclerotia were cultivated in the soil inside the perforated plastic rubber plates at a depth of 4cm from the soil surface.

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<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">Post planting operation

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">Distilled water was used for watering throughout the period of experiment. The experimental set-up was three (3) replicates for each mass for three (3) different lightening conditions which are given thus:

<p style="margin: 0cm 0cm 0.0001pt 54pt; text-align: justify; text-indent: -18pt; line-height: normal;">1. Total darkness condition, which was provided for by covering with black cellophane.

<p style="margin: 0cm 0cm 0.0001pt 54pt; text-align: justify; text-indent: -18pt; line-height: normal;">2. Shade which was obtained by placing in the mushroom house in the Department.

<p style="margin: 0cm 0cm 0.0001pt 54pt; text-align: justify; text-indent: -18pt; line-height: normal;">3. Direct sun light which was obtained by placing in the open field.

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<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">Distilled water was used for watering at a rate of 40ml/sclerotia during the experiment. The following data were subsequently obtained; time of primordial emergence, cap diameter, stipe length, fresh weight and dry weight.

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<p style="text-align: justify;">Data Analysis

<p style="margin: 0cm 0cm 0.0001pt; text-align: justify; line-height: normal;">The results obtained were analyzed using simple  descriptive statistical parameters like mean and standard error. Analysis of variance (ANOVA) was also calculated.

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<p style="text-align: justify;">RESULTS

<p style="text-align: justify;">The sclerotia of Pleurotus tuberregium grew successfully and directly into the sporophores regardless of the different illumination systems and the variation in sclerotia size. There was variation in the time of primordial emergence, morphometric measurements and sporophore yield for the different illumination system and sclerotia sizes.

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<p style="text-align: justify;">Primordial emergence was first observed simultaneously in the open field and shaded experiments. In the open field, sclerotia of size 20g emerged first with a value of 12.00±1.20 days. In the shaded experiment, it was the sclerotia of size 10g that emerged first with the value 12.00±1.50 days (Table 1). The primordial of the sclerotia size 10g in the dark had the longest time of emergence of 35.00±0.75 days (Tables 1). The differences in the illumination treatments of primordial emergence was insignificant at (P=0.05) while the difference in the masses of the time of primordial emergence was found to be highly significant at the same probability level.

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<p style="text-align: justify;">Mean stipe length was greatest, 7.30±0.71cm in the dark and was from sclerotia size, 20g. The mean stipe length was lowest in the experimental set up in the open field, with a value of 2.06 ± 0.45cm (Table 2). There was no significant difference in the illumination treatment on the stipe length at 5 percent probability level. The reverse is the case for the difference in sclerotia size on the stipe length at the same probability level.

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<p style="text-align: justify;">The mean highest cap diameter was observed in the shaded experiment of sclerotia size 25g with a value of 9.95±0.39cm (Table 2). The least value of 2.90±0.59cm was observed in the 25g set up in the dark. The difference in these values were found to be significance and insignificant for sclerotia size and illumination treatments respectively at P=0.05.

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<p style="text-align: justify;">Mean fresh weight production by the mushroom was highest, 28.75±1.24g in the 25g sclerotia in the shaded experiment while the least value of 8.6±0.44g was observed from the 10g sclerotia in the dark

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<p style="text-align: center;">A.O Oghenekaro et al.,: Continental J. Food Science and Technology 4: 46 – 52, 2010

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<p style="text-align: justify;">(Table 3). The difference in weight for all the treatment was insignificant and significant for the various masses of sclerotia (P=0.05)

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<p style="text-align: justify;">Mean dry weight production by the mushroom was highest, 3.76±0.71g and 3.76±0.30 in 25g of both the dark and shaded experiment respectively while the least value of 1.26±0.01 was observed in the 5g sclerotia in the open field. (Table 3). The difference in the illumination treatment was found to be significant as per dry weight at 5% probability level.

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<p style="text-align: justify;">Table 1: Effect of illumination and sclerotia size on the time of Primordial emergence of Pleurotus tuberregium <p style="text-align: justify;">* Time of primordial emergence (days) = Mean of 5 replicates), ± Standard error

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<p style="text-align: center;">A.O Oghenekaro et al.,: Continental J. Food Science and Technology 4: 46 – 52, 2010

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<p style="text-align: justify;">Table 2: Effect of illumination and sclerotia size on the  Morphometric measurement of Pleurotus tuberregium

<p style="text-align: justify;"> <p style="margin-left: 36pt; text-align: justify; text-indent: 36pt;">* Mean of 5 replicates,± Standard error

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<p style="text-align: justify;">Table 3: Effect of illumination and sclerotia size on the sporophore yield of ''Pleurotus tuberregium. ''

<p style="text-align: justify;"> <p style="margin-left: 36pt; text-align: justify; text-indent: 36pt;">* Mean of 5 replicates,± Standard error

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<p style="text-align: center;">A.O Oghenekaro et al.,: Continental J. Food Science and Technology 4: 46 – 52, 2010

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<p style="text-align: justify;">DISCUSSION

<p style="text-align: justify;">In general, there was significant difference between the illumination treatments and various sclerotia sizes for ''Pleurotus tuberregium. ''

<p style="text-align: justify;">'' ''

<p style="text-align: justify;">Pleurotus tuberregium grew successfully with sporophore production under the different illumination condition. The different treatments and sclerotia sizes gave different results. This may be due to differences in light intensity, oxygen available, carbon dioxide presence and relative humidity (Isikhuemhen and Lebauer, 2004.)

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<p style="text-align: justify;">The highest sporophore yield was at 7.80 0.49 days in the shaded experiment and from sclerotia size 20g. This is due to the optimum room temperature of 25⁰C that as provided in the mushroom house. This observation could also be attributed to the ability of P. tuberregium to utilize the nutrient in soil for growth and development (Laborde, 1992).

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<p style="text-align: justify;">The quick emergence of the 20g sclerotia in the open field was probably due to high oxygen concentration and optimum light intensity which are quite essential for sclerotia development. The long time of primordial emergence in the dark could be said to result from the lack of oxygen and extremely high humidity within the enclosed system. The fluctuation in the time of primordial emergence with increasing mass of the sclerotia could be attributed to the short period within which the sclerotia were soaked. But in general, the time in days of primordial emergence decreased with increasing mass of sclerotia.

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<p style="text-align: justify;">The stipe with the greatest length was observed in the dark. This is due to the fact that high concentration of carbon dioxide does not favour cap development but rather stimulates elongation of the stipe. This same reason could also be responsible for the observed mould infection along the stipe in the dark. The high oxygen concentration could be said to be responsible for the quick development of cap and short stipe length that were observed.

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<p style="text-align: justify;">Fluctuation in the stipe length and cap diameter was also observed which could be due to the effect of environmental factors mainly humidity and temperature on the time of primordial emergence (Laborde 1992)

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<p style="text-align: justify;">Mean fresh weight was highest, 28.75±1.24g under shade and least, 8.62±0.53 in the dark. The highest mean weight occurred from the sclerotia mass 25g while the least weight was from the sclerotia mass of 15g as shown in Table 3. For the illumination system, shade provides not only the optimum temperature for sclerotia development but also the high relative humidity for sporophore and mycelia production. The largest mass of 25g sclerotia gave the highest fresh weight which is by convention expected.

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<p style="text-align: justify;">CONCLUSION

<p style="text-align: justify;">This experiment showed that various levels of illumination produced sporophores (fruit bodies) at different times from different masses of sclerotia. The study however showed that shade with moderate environmental conditions favours the growth, development and sporophore yield of Pleurotus tubberegium. More Research should therefore be further encouraged into the cultivation of mushrooms under moderate environmental conditions as this may help make available the mushroom as well as its inherent nutrients for the growing population.

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<p style="text-align: justify;">REFERENCES

<p style="text-align: justify;">Akpaja,E.O.;Isikhuemhen,O.S. and Okhuoya,J.A.(2003). Ethnomycology and uses of edible and medicinal Mushrooms among the Igbo people of Nigeria. International Journal of Medicinal Mushrooms.5:313-319

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<p style="text-align: justify;">Chang, S.T and Miles, P.G (2004). Mushrooms: Cultivation, Nutritional Value, Medicinal Effect, and Environment Impact. CRC Press. Boca Raton. 415p.

<p style="text-align: justify;">

<p style="text-align: center;">A.O Oghenekaro et al.,: Continental J. Food Science and Technology 4: 46 – 52, 2010

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<p style="margin-left: 85.5pt; text-align: justify; text-indent: -85.5pt;"> Declaire, J. R. (1978).Economics of cultivated mushrooms.In: The biology and cultivation of edible

<p style="margin-left: 85.5pt; text-align: justify; text-indent: -85.5pt;">mushrooms. Chang, S.T. and Hayes,W.A.(Eds).Academic Press, New York 819 p

<p style="margin-left: 85.5pt; text-align: justify; text-indent: -85.5pt;">

<p style="text-align: justify;">Ghosh,N.and Chakravarty,D.K.(1990). Predictive analysis of the Protein quality of ''Pleurotus citrinopeleatus. Journal of Food Science Technology. ''27:236-238.

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<p style="text-align: justify;">Isikhuemhen,O.S.;Nerud,F.and Vilgalys,R.(2000).Cultivation Studies on wild and hybrid strains of Pleurotus tuberregium (Fr.)Singer on wheat straw substrate''. World Journal of Microbiology and Biotechnology''. 16:431-435

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<p style="text-align: justify;">Isikhuemhen,O.S.and Okhuoya,J.A.(1995).A low-cost technique for the cultivation of Pleurotus tuberregium (Fr.)Singer in developing tropical countries. Mushroom Growers Newsletter.4:2-4.

<p style="text-align: justify;">

<p style="text-align: justify;">Isikhuemhen,O.S. and Lebauer, D.S.(2004).Mushroom for the tropics:Growing Pleurotus tuberregium. In Mushroom Growers Handbook (Oyster mushroom cultivation). Mushworld. pp 6-24.

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<p style="text-align: justify;">Kuyper, T. W.; van Dijk, J.F.W.and   Onguene, N. A. (2002). Knowledge and utilization of edible mushrooms by local populations of South Cameroun,Oslo, Norway 115p.

<p style="text-align: justify;">

<p style="text-align: justify;">Nwokolo,E.(1987).Composition of nutrients in the sclerotium of the mushroom Pleurotus tuberregium. Plant Foods for Human Nutrition.37:133-139.

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<p style="margin-left: 85.5pt; text-align: justify; text-indent: -85.5pt;">Obodai, M.; Vowotor, K.A. and Marfo, K.(2004).Performance of various strains of Pleurotus species under

<p style="margin-left: 85.5pt; text-align: justify; text-indent: -85.5pt;">Ghanaian conditions. Available at http://www.mushworld.com

<p style="margin-left: 85.5pt; text-align: justify; text-indent: -85.5pt;">

<p style="text-align: justify;">Ogundana, S.K. and Fagade, O.I. (1982).Nutritive value of some Nigeria mushrooms. Food Chemistry.8:263-268.

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<p style="text-align: justify;">Okhuoya,J.A. and Ajerio,C.(1988).Analysis of sclerotia and sporophores of Pleurotus tuberregium (Fr.)Singer an edible mushroom in Nigeria''. Korean Journal of Mycology. ''16:204-206.

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<p style="text-align: justify;">Okhuoya,J.A.and Okogbo,F.O.(1990).Induction of edible sclerotia of Pleurotus tuberregium(Fr.)Singer, in the laboratory. Analysis of Applied Biology.117:295-298

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<p style="text-align: justify;">Oso, B. A. (1977). Pleurotus tuberregium from Nigeria. Mycologia. 69 (2): 271-279

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<p style="text-align: justify;">Received for Publication: 03/06/2010

<p style="text-align: justify;">Accepted for Publication: 20/07/2010

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<p style="text-align: justify;">Corresponding Author:

A.O Oghenekaro,

Department of Plant Biology and Biotechnology, University of Benin, P.M.B 1154, Benin City, Nigeria.

<p style="text-align: justify;">Email- [mailto:abbotkaro@yahoo.com abbotkaro@yahoo.com]

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<p style="text-align: justify;">Continental J. Food Science and Technology 4: 53 – 59, 2010                                ISSN: 2141 – 422X

<p style="text-align: justify;">©Wilolud Journals, 2010                                                                                  http://www.wiloludjournal.com

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<p style="text-align: center;">EFFECT OF HEAT TREATMENT ON ANTIOXIDANT ACTIVITY OF SOME SPICES

<p style="margin: 0cm 0cm 0.0001pt 54pt; line-height: normal;">Ademoyegun Olufemi Temitope1 Adewuyi Gregory Olufemi2 Fariyike Timothy Alaba3

1Crop Utilization Unit, National Horticultural Research Institute, Ibadan, 2Department of Chemistry, University of Ibadan, Ibadan, 3Spice Programme, National Horticultural Research Institute, Ibadan.

<p style="margin: 0cm 43.2pt 0.0001pt;">ABSTRACT:

<p style="margin: 0cm 43.2pt 0.0001pt; text-align: justify;">Spices show potential health benefits as they possess antioxidant activity. The study was to determine the effect of cooking on the antioxidant activity of some selected spices. The total phenol content of five spices (Onion, Garlic, Ginger, Turmeric, and Basil) was determined at different heating periods (1h and 2 h) at 1000c. Although these dietary spice are resistant to thermal denaturation, interestingly, in the case of onion shows reduction in all the tested activities and others shows different variation in all the activity. Total antioxidant activity was measured, based on the reduction of Mo (VI) to Mo (v) by the extract and subsequent formation of green phosphate / Mo (v) complex at acid pH. The extracts were found to have different level of antioxidant properties at different heating time. Considering all the activities, turmeric has good activity amongst the five spice materials screened for their antioxidant properties. Lowest activity was found in onion. In addition to the antioxidant activity of these spices, the total phenolic compounds, flavonoids were measured in the extracts. A correlation between the antioxidant activity and total phenolic content was observed.

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<p style="margin: 0cm 43.2pt 0.0001pt; text-align: justify;">KEYWORDS: Free radical scavanger, cooking, bioactivity and total phenol, total flavonoid

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<p style="text-align: justify;">INTRODUCTION

<p style="text-align: justify;">Free radicals, such as superoxide radical (02-.), hydroxyl radical (OH.) and non-free radical, such as H202 and singlet oxygen (102), e.t.c.), are produced in the body, primarily as a result of metabolism. These radicals cause chronic diseases such as cancer, cardiovascular diseases, diabetes, cataract, etc Halliwell et al. (1992); Aruoma (1994).  Free radicals create chain reactions, which cause cell membrane damage, DNA mutation, lipid and protein damages, and immune cell damage and cell death.  Natural antioxidants are known to scavenge free radicals, enhance the immune system prevent diseases and improve general health and life quality.  Epidemiological studies show an inverse correlation between cardiovascular disease risk and dietary antioxidant consumption Waring (2001).  Well known antioxidants include a number of enzymes (superoxide, catalase, gluthathione peroxidase, etc) vitamin C, vitamin E, carotenoids, phenolic compounds, etc. Phenolic compounds are the major bioactive compounds found in spices. Phenolic compounds act as antioxidant to scavenge reactive oxygen species and to chelate metals.

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<p style="text-align: justify;">Bioactive components of spices such as Curcumin (turmeric), Zingerone (ginger), Allicin (garlic) inhibit lipid peroxidation Lokesh (1992) and Noguchi et al. (1994). Curcumin acts as an anticarcinogen and anti-mutagenic agent Nagabhushau and Bhide (1997). A great number of aromatic, spicy, medicinal and other plants contain chemicals compounds exhibiting antioxidant properties. Numerous studies were carried out on some of these plants, e.g. rosemary, sage and oregano, which resulted in a development of natural antioxidant formulations for food, cosmetic and other application. Therefore, the assessment of such properties remains an interesting and useful task, particularly for finding new sources for natural antioxidant, functional foods and neutraceuticals.

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<p style="text-align: justify;">Spices are usually consumed after thermal cooking. Therefore antioxidant activity of spices may be affected by thermal cooking. As far as our literature survey could ascertain, scarce information was available on the effect of heat treatment on the in vitro antioxidative activities of spices. Therefore, the aim of this study was to investigate the in vitro antioxidant capacities of the methanol extracts of spices before heating and the changes of antioxidant activity after heating for 1 and 2 hours. The antioxidant activity was examined for all the fives spices (Garlic, Onion, Turmeric, Ginger and Basil) using different antioxidant

<p style="text-align: center;">Ademoyegun Olufemi Temitope et al.,: Continental J. Food Science and Technology 4: 53 – 59, 2010

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<p style="text-align: justify;">assays such as total antioxidant activity and free radical scavenging. Furthermore, the total phenolic content, and total flavoniods contents were also measured from the spices extracts and the correlation with the antioxidant activities were ascertained.

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<p style="text-align: justify;">MATERIALS AND METHODS:

<p style="text-align: justify;">Five spices were collected from Spice Programme of National Horticultural Research Institute, Ibadan. The spices were Ginger (Zingiber officinale L), Turmeric (Curcuma longa L), Onion (Allium Cepa),   Garlic (Allium Sativum),  Basil (Ocimum gratissmum)

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<p style="text-align: justify;">Heat Treatment:

<p style="text-align: justify;">The skin of ginger and turmeric were peeled with knife, the outer layer of onion and garlic were removed and the basil was washed and chopped. Each spice was pounded with mortal and pestle to have a thoroughly mined and fine powder spices. Each spice (1 g) was put in a light – capped test tube which was placed in a boiling water bath and healed at 1000c for 1 and 2 hour to prevent oxidation and loss of active components by evaporation. One treatment was also done without heating as a control experiment.

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<p style="text-align: justify;">Extraction:

<p style="text-align: justify;">After heat treatment, the tube was allowed to cool, and then the bioactive compounds were extracted with 20 ml of methanol by shaking for 20 min and centrifuging at 2,000 g for 20 min. The supernatant was used to determine the antioxidant activity, total phenol and total flavonoid content were measured using this extract solution. Three measurements were performed for each spice sample, and the results were expressed as the mean value± SD.

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<p style="text-align: justify;">ANALYSIS PROCEDURE

<p style="text-align: justify;">Determination of total phenolic component

<p style="text-align: justify;">Total soluble phenolic with methanol extract was determined with Folin Ciocalteu reagent, according to the method of Spanos and Wrolstad (1990).

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<p style="text-align: justify;">Determination of the total flavonoid content

<p style="text-align: justify;">Total flavonoid content was determined by using a method described by Sakanaka and Okada (2005).

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<p style="text-align: justify;">Determination of total antioxidant capacity

<p style="text-align: justify;">The antioxidant activity of the extracts was evaluated by the phosphomolybdenum method according to the procedure of Prieto et al. (1999).

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<p style="text-align: justify;">DPPH radical scavenging activity

<p style="text-align: justify;">The free radical scavenging activity of the extracts, based on the scavenging activity of the stable 1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radical, was determined by the method described by Braca et al.( 2001).

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<p style="text-align: justify;">Statistical Analysis

<p style="text-align: justify;">All data were recorded as means ± SD and analyzed by SAS (2003). One – way analysis of variance (ANOVA) and Duncan comparisons were carried out to test any significant differences between raw and cooked vegetables.

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<p style="text-align: justify;">RESULTS AND DISCUSSION

<p style="text-align: justify;">The Effect of heat treatment on total phenolic and total flavonoid content of spices

<p style="text-align: justify;"> Spice phenolics constitute one of the major groups of compounds acting as primary antioxidant or free radical scavengers. Therefore, it is worthwhile to determine their total amount in the spices chosen for the study. Flavonoids as one of the most diverse and widespread group of natural compounds, are likely to be the most important natural phenolics Agrawal (1989). These compounds posses a wide spectrum of chemical and biological activities including radical scavenging properties. Hence, flavonoids are phenolic compounds which are very effective antioxidants. The total phenolic content differed among the different

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<p style="text-align: justify;">spices and each plant contained lower total flavonoid content than the phenolic content (Table 1 and Table 2).Since other compounds besides flavonoids are phenolic substances in plants. Ademoyegun & Fariyike (2008).

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<p style="text-align: justify;">The content of total phenolics in the extracts of spice is determined using the folin – ciocalteu assay, calculated from regression equation of calibration curve (y = 0.0IIIx + 0.0008, r2 = 0.9958) and is expressed as garlic acid equivalents (GAE). It can be observed that the content of phenolics in the extracts correlates with the total antioxidant activity, (rundefined 0.9667). The fresh spices contained 5.96 – 41.10 mg GAE/g Fw total phenolics and the rankling were Turmeric > Ginger > Basil > Garlic > Onion. The total flavonoids ranged from 0.81 – 27.96 mg catechin equivalent / g Fw and in this decreasing order Turmeric > Ginger > Basil > Onion > Garlic.

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<p style="text-align: justify;">In (Table 1), after cooking procedures for 1 hour, the total phenolics content of basil was significantly (P < 0.05) reduced and reduction was the same for the 2 hours cooking. Conversely, of Ginger and Turmeric total phenolic content was significantly (P<0.05) increased to various extents for 1 hour. Heat treatment and increment was the same for ginger 2 hours cooking but turmeric reduce its total phenolic content at 2 hours cooking.

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<p style="text-align: justify;">For onion and garlic there was not significant difference (P < 0.05) for the unheated and heated treatment at 1h and 2 hrs. Data on total phenolic in heat treatment spices are very limited. Khatun et al, (2006) reported that ginger and turmeric contain 20.00 mg GAE/g and 14.5 mg GAE/g uncooked and retained 75% and 172.41% for 1 hour respectively. However, in the present study it was found that raw ginger and turmeric contained 18.01 mg GAE/g Fw and 41.10 mg GAE/g Fw and retaintion for 1 hour at 184.45 % and 141.87 % respectively. The difference may have been due to the differences in the extraction method, solvent used and cooking method.

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<p style="text-align: justify;">To our knowledge, this is the first time that the total flavonoids content of the cooked spices has been reported. The effect of heated spices are significant (p<0.05) for turmeric, onion and garlic. The turmeric show an increase in the total flavonoid 1h at 160.94% and further heated for 2h was observed for 2h. It was reported that heat treatment increased the level of free flavonoid Stewart et al. (2000). For Basil and ginger the effect of heat on the total flavonoid content has no significant (p<0.05), which show that the flavonoid content is relatively stable under thermal heat.

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<p style="text-align: justify;">Table 1: Effect of different cooking period on total phenolic and retention factors of spices <p style="text-align: justify;">

<p style="margin-left: 36pt; text-align: justify; text-indent: -36pt;">a)            Data are expressed as means ± SD of triplicate experiments (on Fresh basis); mean values in a row with different letters are significantly different at P < 0.05

<p style="text-align: justify;">b)           Fresh = 100, GAE = Gallic acid equivalent, FW = Fresh weight

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<p style="text-align: justify;">Table 2: Effect of different cooking period on total flavonoid and retention factors of spices <p style="margin-left: 36pt; text-align: justify; text-indent: -36pt;">

<p style="margin-left: 36pt; text-align: justify; text-indent: -36pt;">a)            Data are expressed as means ± SD of triplicate experiments (on Fresh basis), mean values in a row with different letters are significantly different at P < 0.05

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<p style="text-align: justify;">b)           Fresh = 100, eqv = equivalent, FW = Fresh weight

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<p style="text-align: justify;">The effect of heating times on antioxidant activity of spices

<p style="text-align: justify;">Total antioxidant activity:

<p style="text-align: justify;">The study reveals that the antioxidant activities of the five spices are in the order: Turmeric> Ginger> Basil > Garlic > Onion (Fig. 1).

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<p style="text-align: justify;">The total antioxidant capacity ranged from 79.49 to 29.35 equivalent to ascorbic acid mg/g extract for the unheated spice and for the heated spices range from 101.91 – 20.30 eqv AA mg/g spice extract. There is a strong relationship between the total correlation equation between total antioxidant capacity and total phenolic is 0.9667 in (fig.2). Many reported that high total phenol content increase the antioxidant activity velioglu et al. (1998); Holasova et al. (2002) and there is a linear correlation between phenolic content and antioxidant activity Gheldof & Enjeseth (2002).

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<p style="text-align: justify;">Total antioxidant activity of turmeric, ginger and garlic significantly (P< 0.05) increased during the 2h heating compared to the values for the fresh ones however, total antioxidant activity of onion and basil decreased during the 1h and 2h heating time.

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Fig. 1: Total Antioxidant Activity:                                  Fig 2: Correlation between Total Antioxidant

Phosphomolybdenum Method                                                        and Total Phenolic

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<p style="text-align: justify;">EFFECT OF HEATING TIME ON THE DPPH RADICAL SCAVENGING ACTIVITY

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<p style="text-align: justify;">The DPPH radical scavenging method decreased in the order Turmeric > Ginger > Basil > Garlic > Onion in (Fig. 3).

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<p style="text-align: justify;">Among the five spices, Turmeric showed highest scavenging activity with a inhibition of 89.58% whereas onion had lowest activity with 9.39%. Shobana and Naldn (2000) reported the relative antioxidant activities of some spices: the order of the activities was clove, ginger, pepper and onion, which is related to this study.

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<p style="text-align: justify;">Changes in the DPPH radical scavenging activity of spices for different heating times are shown in (fig. 3). After heating, a significant change in the activity occurring in many spices. An increase in radical – scavenging activity was found in Turmeric and Ginger. The main active component of spices such as Curcumin (were found in ginger and turmeric is usually fat – soluble. But the present study was carried out in absolute methanol. So, the active components of these spices might not – dissolve completely in this solution before heating. After heating, the solubilities of the components probably increased because of decomposition of the cell wall and by radical scavenging activity total antioxidant activating and total phenolic of two spices might be observed after heating. Sohobana and Naidu (2000) reported that the bound antioxidants might be released due to heat treatment, resulting in the higher antioxidant activity compared that in that of fresh spices extract. Dewanto et al. (2002) reported that thermal processing disrupt the cell membranes and cell walls to release lycopene from the insoluble portion of tomato, which might cause the antioxidant activity of tomato to increase. Takamuru et al., (2002) found a decrease of radical – scavenging activity of curry paste and cooked curry, possibly due to decomposition or evaporation of the active compounds. Since the spices were heated with butter at high temperature.

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<p style="text-align: justify;">In this study, a decrease in the radical – scavenging activity was observed in garlic and basil. After heating, coagulation of these spices was observed therefore, the extraction ability might be decreased by coagulation after heating, resulting in a reduction in the radical – scavenging activities of these spices. Onion showed no significant change due to heating.

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<p style="text-align: justify;">The DPPH radical – scavenging activity of all the spices was highly correlated (R2 = 0.8) with total phenol content fig 4). Form these results the active components of spices are considered to be mostly polyphenol compounds. (Several polyphenols did not show any DPPH radical – scavenging activity) is reported by Parejo et al, (2001); Oktay et al.( 2003) found a high correlation between phenolic context and DPPH radical – scavenging activity.

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Fig. 3 Antioxidant activity: DPPH Scavenging             Fig. 4: Correlation between Total Phenolic

<p style="text-align: justify;">Assay                                                                                                    and Free Radical Scavenging Activity

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<p style="text-align: justify;">From the results of this study, it is clear that that spices have strong antioxidant activities in a methanol extract solution. All the tested activities of spices remained after heating, suggesting that the bioactive components are relatively stable during thermal heating at boiling point of water. But we have results which show apparent change in antioxidant properties of these spices. In conclusion, the results illustrated that the health benefits from plant sources remained in the products after thermal process that it, heat do not denatured the antioxidant activities in all the selected spices studied. Spices are expected to be a valuable food constituent for promoting good health in our daily lives.

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<p style="text-align: justify;">REFERENCES:

<p style="text-align: justify;">Ademoyegun OT, Fariyike TA (2008) Antioxidant activity of phenolic components presents in basil (Ocimum basilicum L.) cultivars. Proceeding of Nigerian Institute of Food Science and Technology, pp 165-166.

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<p style="text-align: justify;">Agrawal PK (1989) Carbon – 13 NMR of flavonoids. New York: Elesvier.

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<p style="text-align: justify;">Aruoma OI (1994) Nutrition and health aspects of free radicals and antioxidants''. Food Chemistry and Toxicology ''32, 671 -683.

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<p style="text-align: justify;">Braca A, Nunziatina, De Tommasi, Lorenzo, Di Bari, Cosimo, Pizza, Mateo, Politi, Ivano, Morelli (2001) Antioxidant principles from Bauhinia terapotensis. Journal of Natural Products 64, 892–895.

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<p style="text-align: justify;">Dewanto V, Wu X, Adom KK, Liu RH (2002) Thermal processing enchances the nutritional valve of tomatoes by increasing total antioxidant activity. Journal of Agricultural and Food Chemistry 50, 3010 – 3014.

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<p style="text-align: justify;">Gheldof N,  Engeseth NJ (2002) Antioxidant capacity of honeys from various floral sources based on the determination of oxygen radical absorbance capacity and inhibition of in vitro lipoprotein oxidation in human serum samples. Journal of Agricultural and Food Chemistry 50, 3050 – 3055.

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<p style="text-align: justify;">Gordon MH (1990) The mechanism of antioxidant action in vitro. In B.J.F. Hudson (Ed), Food  antioxidants, pp. 1 - 18. London: Elsevier Applied Science.

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<p style="text-align: justify;">Halliwell B, Gutteridge JMC, Cross CE (1992) Free radicals, Antioxidants and human disease:Where are we now? Journal of Laboratory Clinical Medicine 119, 598 – 620.

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<p style="text-align: justify;">Holasovia M, Fiedlerova V, Smrcinova  H, Orsak  M, Lachman  J,  Vavreinova  S (2002) Buckwheat – the source of antioxidant activity in functional foods. Food Research International 35, 207 – 211.

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<p style="text-align: justify;">Khatun M, Eguchi S, Yamaguchi T, Takamura H, Matoba T(2006) Effect of Thermal Treatment on radical – Scavenging activity of some spices. Food science Technology Research 12 (3), 178 – 185.

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<p style="text-align: justify;">Noguchi N, Komuro E, Niki  E, Willson  RL (1994) Action of curcumin as an antioxidant against lipid peroxidation. Journal of Japan Oil Chemist Society 43, 1045 – 1051.

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<p style="text-align: justify;">Nagabhushan M, Bhide  SV (1997) Antimutagenicity and anticarcinogenicity of turmeric (Curcuma longa). Journal of Nutrition and Growth Cancer 4, 83 – 89.

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<p style="text-align: justify;">Oktay M, Gulçin I, Kufrevioglu O (2003) Determination of in vitro antioxidant activity of fennel (Foeniculum vulgare) seed extracts. ''Lebensm.-Wiss. u.-Technol ''36, 263 – 271.

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<p style="text-align: justify;">Parejo I, Viladomat F, Bastida  J, Schmeda-Hirschamann  G, Burillo J ,Codina C (2004) Bioguided isolation and identification Of the nonvolatile antioxidant compounds from fennel (Foeniculum vulgare Mill.) Waste. Journal of Agricultural and Food Chemistry 52, 1890 - 1897.

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<p style="text-align: justify;">Prieto P, Pineda M, Aguilar M (1999) Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: specific application to the determination of vitamin E. Analytical Biochemistry 269, 337–341.

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<p style="text-align: justify;">Sakanaka SA, Okada Y (2005) Preparation and antioxidant properties of leaf tea. Food Chemistry 89, 569 – 75.

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<p style="text-align: justify;">Shobana S,  Naidu  KA (2000) Antioxidant activity of selected Indian spices. Prostaglandins Leukot Essential Fatty Acids 62, 107 – 110.

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<p style="text-align: justify;">Spanos GA, Wrolstand RF (1990) Influence of processing and storage on phenolic composition of grape juice. Journal of Agricultural and Food Chemistry 38, 1565 – 1571.

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<p style="text-align: justify;">Stewart A J, Bozonnet S, Mullen W, Jenkins G I, Michael E J, Crozier A (2000) Occurrence of flavonols in tomatoes and tomato-based products. Journal of Agricultural and Food Chemistry 48, 2663–2669.

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<p style="text-align: justify;">Takamura H, Yamaguchi T, Terao J, Matoba T (2002) Change in radical-scavenging activity of spices and vegetables during cooking. In “Bioactive compounds in foods: Effect of processing and storage.” ed. by Lee, T.C. and Ho, C.T., American Chemical Society, Washington, D.C., USA, pp 34 – 43.

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<p style="text-align: justify;">Velioglu Y S, Mazza G, Gao L, Oomah  B D (1998) Antioxidant activity and total phenolics in selected fruits, vegetables, and grain products. Journal of Agricultural and Food Chemistry, 46(10), 4113–4117.

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<p style="text-align: justify;">Waring WS (2001) Antioxidants in prevention and treatment of cardiovascular disease. Proceeding of Royal College Physical Edinburge 31, 288 - 292.

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<p style="text-align: justify;">Received for Publication: 03/07/2010

<p style="text-align: justify;">Accepted for Publication: 20/09/2010

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<p style="text-align: justify;">Corresponding Author:

<p style="margin: 0cm 0cm 0.0001pt; line-height: normal;">Ademoyegun Olufemi Temitopeundefined

Crop Utilization Unit, National Horticultural Research Institute, Ibadan

''E- mail: [mailto:femtopyankee@yahoo.com femtopyankee@yahoo.com] ''

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<p style="text-align: justify;">Continental J. Food Science and Technology 4: 60 – 64, 2010                                ISSN: 2141 – 422X

<p style="text-align: justify;">©Wilolud Journals, 2010                                                                                  http://www.wiloludjournal.com

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<p style="text-align: center;">COMPARATIVE STUDIES OF THE NUTRITIONAL QUALITIES OF THREE FERMENTED AFRICAN LEGUME SEEDS USING Bacillus subtilis and Bacillus pumilus AS STARTERS

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<p style="text-align: center;">1Gberikon, G.M., 2Ameh,J.B., 2Ado, S.A., 2Umoh,V. J.

<p style="text-align: center;">1Biology Department, Federal College of Education, Zaria, 2Microbiology Department, Ahmadu Bello University, Zaria

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<p style="margin: 0cm 17pt 0.0001pt;"> ABSTRACT

<p style="margin: 0cm 17pt 0.0001pt; text-align: justify;">Analysis into three African legume seeds; Prosopis africana, Parkia biglobosa and Glycine max was carried out. Unprocessed seeds of Prosopis africana were purchased from Otukpo market in Benue state of Nigeria and unprocessed seeds of Parkia biglobosa and Glycine max were purchased from Sabon-Gari market, Zaria, Kaduna state. These seeds were processed and fermented using 5% mixed Bacillus subtilis and Bacillus pumilis as inoculum into 300g of unfermented seeds. Seeds were allowed to ferment naturally along side with seeds inoculated with inoculum in the Department of Microbiology Ahmadu Bello University Zaria, for 48h. For P. biglobosa and G.max, P.africana seeds were left to ferment for 118h. Fermented seeds from the analysis showed that seeds with inoculum fermented faster giving higher protein and lipids values than seeds allowed to ferment naturally. Fermented seeds of P. africana were richer in protein with a crude protein value of 40.07% followed by G.max with a crude protein value of 38.29% and P. biglobosa with a crude protein value of 36.13%. Fermented seeds of G.max were richer in lipids with a crude lipid value of 18.42%, followed by P.biglobosa with a crude lipid value of 13.01% and P.africana with 12.01%. These seeds were fermented into “daddawa” and “okpehe” condiments as flavour enhancers. The analysis of variance showed that the moisture content, Ash, Protein and Carbohydrate content of the seeds vary significantly (F < 0.0001). On the other hand, the lipid and fibre contents did not vary significantly, (F > 0.0001).

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<p style="margin: 0cm 17pt 0.0001pt; text-align: justify;">KEYWORDS: P.africana, P.biglobosa, G.max, inoculum, protein, lipids, “daddawa”, “okpehe”, condiments.

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<p style="text-align: justify;">INTRODUCTION

<p style="text-align: justify;">Seeds of some legumes such as Glycine max, (soyabean seeds) Prosopis africana,(African mesquite) and Parkia biglobosa (locust bean seeds) may account for up to 80% of dietary protein. This may be the only source of protein for some group of people. Their cooked forms may be eaten as meals and are commonly used in fermented forms as condiments to enhance flavour of foods (Oniofiok, 1996). Fermented condiments remain key constituents of diet throughout many parts of Asia and Africa. Fermentation processes evolved for the development of taste and aroma, often result in enhanced nutrition and detoxification of antinutrient factors (Ogunshe et al., 2006). Traditionally, fermented condiments (“daddawa” and “okpehe”) are based on vegetable proteins and are consumed by different ethnic groups in Nigeria. They serve not only as nutritious non-meat protein substitutes, but also as flavour enhancers in soups and other dishes (Achi, 2005). Traditional diet in West Africa consists of large quantities of staple foods (cassava, yam and maize). The staple foods provide the calories, but are poor in other nutrients. Soups are the main source of protein and minerals and one of the ways to improve the diet is to improve the nutrients in the soups (Achi, 2005).

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<p style="text-align: justify;">Recent microbiological information on Nigerian plant protein fermentations includes, those reported by Omafuvbe et al., 2002; Ouoba et al., 2003; Dakwa et al., 2005. In all their reports the involvement of genus Bacillus has been established in fermentation of locust bean seeds and other legumes. Fermentation in developing countries does not use inoculum, these processes could be improved by using starter cultures (Holpzapfel, 2002). A pure starter culture is essential for controlled fermentation. Controlled fermentation of soya beans was achieved by using pure single culture of B. subtilis, B.licheniformis or in combination (Sarkar et al., 1993). The use of a mixture of microorganisms with complimentary physiological properties seems to be the best approach for obtaining a product with the nutritional and sensory properties desired

<p style="text-align: center;">Gberikon, G.M et al.,: Continental J. Food Science and Technology 4: 60 – 64, 2010

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<p style="text-align: justify;">(Ouoba et al., 2003). Therefore the objective of this study is to carry out fermentation of leguminous seeds using standard and test strains of B.pumilus and B.subtilis to compare the nutritional properties/proximate analysis.

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<p style="text-align: justify;">MATERIALS AND METHODS:

<p style="text-align: justify;">Unprocessed samples of P.africana were purchased from Otukpo market, Benue state of Nigeria and samples of G.max and P. biglobosa were purchased from Sabon-Gari market, Zaria, Kaduna state of Nigeria. These seeds were transported to the laboratory, Department of Microbiology, Ahmadu Bello University, Zaria in polyethylene bags for processing.

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<p style="text-align: justify;">Isolation and characterization of Bacillus species

<p style="text-align: justify;">Test strains of B. pumilus and B. subtilis isolated from fermented legume seeds were obtained from Department of Microbiology, Ahmadu Bello University, Zaria. Standard strains of B. pumilus and B. subtilis were obtained from Federal Institute of Industrial Research, Oshodi (FIIRO). These organisms were identified biochemically using various tests. These include the following biochemical tests as described by Gordon et al., 1973. Indole test, coagulase test, motility test, catalase test, hydrogen sulphide production, starch hydrolysis, sugar fermentations and utilization, growth at different sodium chloride (NaCL) concentrations (5%,10%,15% 20% and 25%).

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<p style="text-align: justify;">Preparation of Bacillus inoculum

<p style="text-align: justify;">The inoculum used for the fermentation contains 2.1×107 cells/ml, the cell population was calibrated using McFarland standard (No 7), and the amount used formed 5.0% for fermentation comprising of 15ml of 24h old cultures of organism consortium A (Standard strains of B.subtilis and B.pumilus combined) and consortium B (Test strains of B.subtilis and B.pumilus combined)

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<p style="text-align: justify;">Controlled Fermentation of locust bean, soyabean and African mesquite seeds

<p style="text-align: justify;">The fermentation process was set up using both organism consortiums respectively. These organisms were inoculated into 300g of processed unfermented seeds. The seeds were wrapped with sterile aluminum foil paper and placed in an earth pot with cover. The fermentation was allowed to progress at room temperature (28 ± 20C) for 48h for P.biglobosa and G.max, 118h for P.africana in the Department of Microbiology, Ahmadu Bello University Zaria. After fermentation, proximate analyses for crude protein, carbohydrates, crude lipids, fiber, ash and moisture content were carried out adopting the AOAC (1980) methods.

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<p style="text-align: justify;">Analysis of results

<p style="text-align: justify;">The data were analyzed for significance using Analysis of Variance, by the Statistical Analysis System version 12.

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<p style="text-align: justify;">RESULTS

<p style="text-align: justify;">The moisture and ash content were higher in unfermented seeds of P.biglobosa with values of 67.50% than in other seeds. Crude lipids were higher in unfermented seed of G.max with a value of 10.00% than other seeds. Crude protein in unfermented seeds of P.africana was higher with a value of 17.13% than other seeds. Crude fiber value in unfermented P.africana seeds was much lower with a value of 0.01% while P.biglobosa and G.max had fiber values of 0.38%. Soluble carbohydrate was higher in unfermented seeds of P.biglobosa than the rest of the seeds with a value of 10.63%. The analysis of variance showed that the moisture content, Ash, Protein and Carbohydrate content of the seeds vary significantly (F < 0.0001). On the other hand, the lipid and fibre contents did not vary significantly, (F > 0.0001).

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<p style="text-align: justify;">The results of the fermented seeds with the organism (consortium A and B) are also presented in Table 1. The moisture content of seeds of P.africana and P.biglobosa fermented with consortium A and consortium B was 65.00% respectively, while moisture content was lower in fermented seeds of G.max subjected to the same condition, with a value of 62.00% respectively. Ash content was higher in consortium B fermented seeds of P.africana with values of 2.30%, and lower in consortium B fermented seeds of G.max with values of 0.01%. During fermentation, crude lipid and crude protein increased significantly as shown in Table 1.

<p style="text-align: center;">Gberikon, G.M et al.,: Continental J. Food Science and Technology 4: 60 – 64, 2010

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<p style="margin-left: 54pt; text-align: justify; text-indent: -54pt;">'Table 1: Proximate Analysis of Unfermented, Naturally Fermented and starter assisted Fermented Seeds of P.africana, P. biglobosa and G. max'' (%) '''

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<p style="margin-left: -9pt;">*All values are means of triplet readings. The analysis of variance showed that the moisture content, Ash, Protein and Carbohydrate content of the seeds vary significantly (F < 0.0001). On the other hand, the lipid and fiber contents did not vary significantly, (F > 0.0001).

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<p style="margin-left: -9pt;"> Keys

<p style="margin-left: -9pt;">NFS4 - Naturally fermented seeds of P. africana

<p style="margin-left: -9pt;">NFS5 - Naturally fermented seeds of P.biglobosa

<p style="margin-left: -9pt;">NFS6 - Naturally fermented seeds of G.max.

<p style="margin-left: -9pt;">FSSS1 - Fermented seeds of P.africana with consortium A.

<p style="margin-left: -9pt;">FSTS1 - Fermented seeds of P.africana with consortium B.

<p style="margin-left: -9pt;">FSSS2 - Fermented seeds of  P.biglobosa with consortium A

<p style="margin-left: -9pt;">FSTS2 - Fermented seeds of P.biglobosa with consortium B.

<p style="margin-left: -9pt;">FSSS3 - Fermented seeds of G.max with consortium A.

<p style="margin-left: -9pt;">FSTS3 - Fermented seeds of G.max with consortium B.

<p style="text-align: center;">Gberikon, G.M et al.,: Continental J. Food Science and Technology 4: 60 – 64, 2010

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<p style="text-align: justify;">Values of crude lipids were higher in both consortiums fermented seeds of G.max with a value of 18.43% and lower in consortium A and B fermented seeds of P.africana with values of 12.03% and 12.01% respectively. Protein value was higher in consortium B and A, fermented seeds of P. africana with values of 40.06% and 40.05% followed by consortium B fermented seeds of G.max with values of 38.29% and consortium A fermented seeds of G.max with values of 38.27%. Percentage fiber was higher in consortium B fermented seeds of P.biglobosa with values of 1.93% and lower in consortium B fermented seeds of G.max with values of 0.02%. Soluble carbohydrate was higher in consortium A fermented seeds of P.biglobosa with values of 7.99% and consortium B with values of 7.95% and lower in consortium A and B fermented seeds of G.max with a value of 1.81%. Nutritional information of P.africana seeds fermented naturally had a higher moisture content of 65.00%, while those of P.biglobosa and G.max had a moisture content of 62.20%. Values of ash content were higher in naturally fermented seeds of P.biglobosa which had 2.50% and lower in G.max with a percentage value of 0.22%. Crude lipids value was higher in naturally fermented seeds of G.max with a value of 17.43% and lower in naturally fermented seeds of P.africana with a value of 11.96%. Crude proteins were higher in naturally fermented seeds of P.africana with a value of 38.70% and G.max with a value of 38.03% and lower in naturally fermented seeds of P.biglobosa with a value of 36.00%. Crude fiber was higher in naturally fermented seeds P.biglobosa seeds with a value of 1.50% and lower in G.max with a value of 0.21%. Values of soluble carbohydrate were higher in naturally fermented seeds of P.biglobosa with a value of 12.09% and lower in naturally fermented seeds of P.africana with a value of 0.15%

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<p style="text-align: justify;">DISCUSSION

<p style="text-align: justify;">Fermentation remains an effective, inexpensive method for extending the shelf life of foods and increasing their nutritional content (Tamang, 2009). Lipids content and protein increased significantly as shown in Table 1. This is due to the fact that fermentation results in increased nutritional values which are in agreement with the results of Odunfa (1984). In Table 1, the three legume seeds fermented with both consortiums gave higher values of lipids and proteins than those fermented naturally. This is due to the fact that when organisms responsible for legume seeds fermentation are used as inoculum or starters in the right percentage, it will guarantee consistency and product quality (Holpzapfel, 2002). Seeds fermented without starters had higher moisture content, higher fiber and carbohydrates values. Protein values were low compared to seeds fermented with starters. Inconsistencies in naturally fermented seeds aroused because there was no appropriate starters to initiate fermentation. The use of consortium A and B for fermentation resulted in products of high nutritional properties desired which is in agreement with Holpzapfel 2002.

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<p style="text-align: justify;">CONCLUSION

<p style="text-align: justify;">It was concluded from this research that fermented legume seeds have higher nutritional values than unfermented legume seeds. Seeds of P.africana, P.biglobosa and G.max fermented with consortium A and B as starters gave higher nutritional values than seeds allowed fermenting naturally. Therefore, legume seeds fermented with both consortiums can be use as condiments and flavour enhancers in soups and other dishes.

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REFERENCES

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<p style="text-align: justify;">AOAC (1980). Official methods of analysis of the Association of Official Analytical     Chemist 11th Edn. Washington D.C

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<p style="text-align: justify;">Dakwa,.S.;Sakyi-Dawson .E.; Diako. C.; Annan.N.T.and Amoa-Awua. W.K. (2005). Effect on the fermentation of soyabeans into daddawa (soydaddawa). Int .J. Food Microbiol. 104:69-82.

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<p style="text-align: justify;">Gordon, .R.E.; Hayes, .W.C. and Pang, .C.H.N. (1973).The Genus Bacillus. Agricultural Hand Book.U.S Department of Agriculture Washington D.C.

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<p style="text-align: center;">Gberikon, G.M et al.,: Continental J. Food Science and Technology 4: 60 – 64, 2010

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<p style="text-align: justify;">HolpzapfelW.H. (2002).Approriate starter technologies for small scale fermentation in developing countries. ''Int. J. food Microbiol''. 78:119-131.

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<p style="text-align: justify;">Ogunshe,.A.A.S.; Ayodele,.A.E. and Okwonko,I.O.(2006). Microbial studies on Asia: a potential Indigenious Laboratory fermented Food Condiments from Albizia saman (Jacq).F. Mull. Parkistan Journal of Nutrition 5(1):51-58.

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<p style="text-align: justify;">Omafuvebe, B.O.; Abiose, S.H. and Shonukan,O.O.(2002). Fermentation of soybean (Glycine max) for soydaddawa production by starter cultures of Bacillus. Food Microbiol. 19:561-566.

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<p style="text-align: justify;">Ouoba,.L.I.I.;Rechinger,.K.B.;Barkholt,.V.;Diawara,.B.;Traore,.A.S.and Jakobsen, M. (2003). Degradation of proteins in the fermentation of African locust bean (Parkia biglobosa) using strains of Bacillus subtilis and Bacillus pumilus for production of soumbola''. J. of Appl.Microbiol''. 94:396-402.

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<p style="text-align: justify;">Sarka,.P.K.; Cooks,.P.K. and Owens,.J.C.(1993). Bacillus fermentation of soybeans. World .''J. Microbiol. Biotecnol''.9:232-259.

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<p style="text-align: justify;">SAS® System Version 12 for Microsoft® Window® (SAS Institute Inc., Carry, NC,USA).

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<p style="text-align: justify;">Tamang,.J.P.(2009). Himalayan fermented foods. Microbiology, Nutrition and Ethnic Values. Glyde Corporations. PP.90-112.

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<p style="text-align: justify;">Received for Publication: 03/07/2010

<p style="text-align: justify;">Accepted for Publication: 20/09/2010

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<p style="text-align: justify;">Corresponding Author:

Gberikon, G.M..,

Biology Department, Federal College of Education, Zaria,

<p style="text-align: justify;">Email: gberikon.grace@yahoo.com.

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