Pakistan Journal of Nutrition 3 (3): 140-145, 2004

© Asian Network far Scientific lnfarmation,2004

Chemical and BiochemicaCl hanges in AfricanLocustBean (Parkia biglobosa)

and Melon (Citrullusvulgaris)Seeds During Fermentation to Condiments
Bridget O. Omafuvbe1, Olumuyiwa S. Falade2, Bolanle A. Osuntogun3 and Steve R.A. Adewusi2*
1Department 2Department
of Microbiology, of Chemistry, Obafemi Awolowo University, lle-lfe, Nigeria
3Department
of Chemistry, University of Lagos, Akoka, Lagos, Nigeria
E-mail: sadewusi@oauife.edu. ng

Abstract:The chemical (proximate composition, pH, total tocopherol, acid and iodine values, total and reducing
sugars) and biochemical (a-amylase, sucrase and protease) changes occurring during the fermentation of locust
bean and melon to the condiments - iru and ogiri respectively were monitored. Processinglocustbeanto the
condiment involved boiling far 12 hours, soaking the seeds in water, dehulling, boiling far another 6 hours and
fermentation far 3-4 days while melon is boiled far 6 hours, cooled and fermented. Boiling, soaking (in water) and
dehulling of locust bean resulted in reduction of ash, crude fibre (CF) and total tocopherol but increased ether
extract. Fermentation increased the pH value, crude protein (CP), reducing sugar, a-amylase, sucrase, protease and
free amino acid, decreasedthe total sugar in both samples and also increased the CF of locust bean sample
but had no effect on the CF in melon. Fermentation had no effect on total tocopherol content in both samples.
Heat treatment and fermentation employed in processing both samples increased the acid value with a
corresponding decrease in iodine value. High negative correlation was observed between acid and iodine values
while a high positive correlationwas observed betweentocopherol and iodine value.

Key words: Proximate composition, tocopherol, sugar, free amino acids, sucrase, amylase, protease
lntroduction meat substitute by low-income families in parts of
Fermentation is one of the oldest methods of faod Nigeria (Odunfa, 1985).
preservation known to man. 1 n Africa, the art of Sorne research has been carried out on the production of
fermentation is widespread including the processing of fermented condiments-iru-from African locust bean (Eka,
fruits and other carbohydrate sources to yield alcoholic and 1980; Odunfa, 1986), melon seed fermented ogiri (Od unfa,
non-alcoholic beverages, the production of sour- tasting 1981; Barber and Achinewhu, 1992) and soybean
ogi - the fermented cereal products, which provide produced daddawa (Omafuvbe et al., 2000;
instant energy in breakfast and convalescent diets 2002). Oil seeds are generally processed to yield
(Adewusi et al., 1991; 1992). Oil seeds such as African condiments but so far, no investigation has been carried out
locust bean, melon seed, castor oil seed, mesquite bean on the changes in the oil component of these seeds as a
and soybean are also fermented to give condiments. result of fermentation. Furthermore, comparative chemical
The production of condiments is largely on a traditional and biochemical changes occurring during fermentation
small-scale, household basis under highly variable have been neglected especially in the fermentation of the
conditions (Odunfa, 1985). 1 n addition, the fermentation is two most popular condiments iru and ogiri. The main
usually carried out in a moist salid state, involving contact objective of the present study is to compare the
with appropriate inocula of assorted proximate composition of the raw African locust bean and
microorganisms and is accomplished by the natural melon seeds fermented far the production of iru and
temperatures of the tropics. The desired state of ogiri and the chemical and biochemical changes occurring
fermentation of the condiments is indicated by the during the fermentation process. The nutritive value of iru
farmation of mucilage and overtones of ammonia and ogiri is also compared with that of monosodium
produced as a result of the breakdown of amino acids glutamate based salts.
during the fermentation (Omafuvbe, 1998). The
characteristic ammoniacal odour and flavour of Materials and Methods
condiments enhance the taste of traditional soups and Materials: African locust bean and melon seeds were
sauces especially the various soups used as bought at the local market in lle-lfe. Seasoning salts were
accompaniment to the starchy root and tuber diets. bought off the shelf in supermarkets or from the local
Condiments are also known to contribute to the calorie and market also in lle-lfe, Nigeria. The processing of both
protein intake (Simmons, 1976; Umoh and Oke, African locust bean and melon seeds to condiments
197 4) and are generously added to soups as low-cost were carried out in the local factories of the

140
140
 Omafuvbe et et.: African Locust Bean and Melon Seeds

two major producers of these condiments in lle-lfe, Nigeria glycine as standard. The total soluble sugar was determined
as outlined below: by the anthrone reagent method of Morris (1948) while
reducing sugar was estimated by the calorimetric method
Productionof iru: Raw African locust bean was boiled for (Somogyi, 1945) using glucose as a standard.
12 hours and further soaked in the boiling water for
another 12 hours (preferably overnight). Excess water was Extraction of extracellular enzymes: Extracellular
drained off and the seeds were dehulled by marching enzymes (a-amylase, sucrase and protease) in the
the seeds by foot in a large wooden mortar and further fermenting material (5.0 g) were extracted with appropriate
removal of the seed coat was achieved by rubbing the buffer (50 mi) as previously described (Omafuvbe et al.,
cotyledons between the palms of the hand and washing 2000).
with water. The cotyledons were again cooked for a-Amylase activity was determined by the blue value
another 6 hours, the hot boil water was drained off and method of Fuwa (1954) on 0.1 M potassium phosphate
the cotyledons were then spread into calabash trays, buffer (pH 6.0) extract of the samples. One unit of a-
covered with wooden trays, wrapped with jute sacks and amylase activity was defined as the amount that produced
fermented for 3-4 days to produce iru. a 10% reduction in the intensity of blue colour of starch -
iodine complex under the experimental conditions.
Productionof ogiri:Shelled melon seeds were sorted out Protease activity was measured on 0.1 M sodium phosphate
to remove grit, dirt and decomposing seeds; rinsed with buffer (pH 6.5) extract of the samples. One unit of
warm water twice and boiled for one hour in 1 O times proteolytic activity was defined as the amount that produced
its volume of water. The boil water was then drained 100 µg of tyrosine in 1.0 mi of trichloroacetic acid soluble
and replaced by cold water and boiled again for peptides under the assay conditions (Omafuvbe et al., 2000).
6 hours to softness. The melon seeds were thereafter Sucrase activity was measured on 0.05M sodium citrate (pH
transferred into a clay pot and covered with Thaumaloccus 6.5) extract of the samples. One unit of enzyme activity
demoel/i leaves and wrapped in sack clothe for 3 days. The was defined as the amount of enzyme that produced 100
fermented product is then ground with stone mortar and µg of glucose under the assay conditions (Omafuvbe et al.,
pestle to almost a smooth paste sold as ogiri. 2000).

Sam pling: The raw seeds, samples collected at the Results and Discussion
different processing stages and the products thus produced Proximate composition and pH value of raw and fermented
were kept in a freezer at -17°C until required for analysis. African locust bean (Parkia biglobosa) and melon seeds
(Citrullus vulgaris) are presented in Table
Analytical technique: Proximate composition was 1. The moisture content of raw African locust bean and
determined by the Association of Official Analytical melon was 8.6 and 5.3% respectively. There was an increase
Chemists (AOAC, 1990) method. in the moisture content of processed African locust bean
Tocopherol content was estimated by the method of and processed melon, which ranged between 51.9 and
Contreras-Guzman and Strong (1982a, b) using cuproine (2, 2- 56.7% and 43.0 and 44.1% for both samples respectively as
biquinoline) as the complexing agent for the colour formation. a result of boiling in water followed by further soaking in
All rac-a-tocopherol (Merck) was used as a standard. Solvents water in the case of African locust beans.
used for the extraction and subsequent assay were The ash content of raw African locust bean and melon seeds
purified and redistilled as per Contreras-Guzman et al. (1982). was 5.4 and 3.3% respectively. The value reported for
lodine and Acid values were determined by the AOAC African locust bean agreed favourably with
(1990) method. 5.1 o/o reported earlier by Eka (1980). Boiling, soaking in
pH of the fermented products was determined using a water and dehulling of African locust beans led to a loss of
PYE Unicam pH meter (Model 290 MK2). 41% ash. This means that about 41% of the total mineral
content of African locust bean may reside in the hu 11 of the
Determination of sugars and free amino acids: The raw seed and or leached during processing. When the African
seeds and samples collected at the different processing locust bean seed was boiled for additional 6 h during
stages were dried in hot air oven at 70ºC, ground and processing, 13% of the mineral was further leached into
defatted. The soluble sugars and free amino acids in the the broth. Fermentation seemed to have increased the
defatted samples were extracted with 80% ethanol (v/v) ash content of African locust bean seed by 29% in direct
following the method of Odibo et al. (1990). The free amino agreement with the observation of Eka (1980) of about 30%
acid in the ethanolic extract was estimated by the ninhydrin increase in ash
calorimetric method (Rosen, 1957) using
Table 1: Proximate Composition and pH Value of Raw and Fermented African locust bean (Parkia biglobosa) and
Melon Seeds (Citrul/us vulgaris). o/o dry sample
Moisture* Ash Crude Et her Crude CHO pH
(%) (%) fiber (%) Extrae!(%) Protein (%) (%)
African 1 ocust bean

Raw 8.6 ± 0.6 5.4 ± 0.3 11.7±0.2 17.7 ± 0.2 31.0 ± 0.7 35.0 ± 0.5 6.2
Boiled 12 h and Dehulled 51.9 ± 0.7 3.2 ± 0.3 4.6 ± 0.1 20.9± 0.3 31.4±0.2 42.0 ± 0.4 6.3
Boiled 6 h 56.7 ± 0.7 2.8 ±O.O 4.4 ± 0.9 28.4 ± 0.1 31.1±0.9 33.3 ± 0.5 6.4
Fermentation Period (h)
24 55.7 ± 0.8 3.6 ± 0.1 5.7 ± 0.1 32.6 ± 0.7 31.7 ± 0.4 25.1±1.1 8.3
48 55.5 ± 0.4 3.5 ± 0.1 5.4 ± 0.1 35.2 ± 0.1 31.3±0.2 21.0 ± 0.4 8.4
72 52.0 ± 5.0 3.6 ± 0.1 4.0 ± 0.1 37.2 ± 0.2 32.9 ± 0.1 16.3 ± 0.8 8.4
Melon seed
Raw 5.3 ± 0.2 3.3 ± 0.2 15.8 ± 0.6 37.5 ± 0.6 19.3 ± 0.4 24.1 ± 0.2 7.0
Fermentation** period (h)
24 43.1±0.8 3.2 ± 0.2 20.7 ± 0.5 34.1 ±2.7 20.6 ± 0.4 21.4±0.7 7.2
48 43.0 ± 0.7 3.2 ± 0.3 20.0 ± 0.7 35.3 ± 0.4 19.5 ± 0.7 22.0 ± 0.9 7.5
72 44.1±0.8 3.0 ±O.O 15.6 ± 0.4 36.3 ± 0.5 19.9 ± 0.8 25.2 ± 1.2 7.9
*Moisture content was determined on the fresh materials.
**Fermentation of melon seed was carried out after boiling for 6 h (See Experimental for details).

Table 2: The Tocopherol, Acid and lodine Values of African locust bean and Melon Seeds during Different Stages of
Processing to Condiments in Comparison to the Seasoning Salts
African locust bean Total tocopherol (mg / 100 g) Raw Acid Value lodine Value
21.6 ±O.O 1.06 148.4
Boiled (12 h) & Dehulled 18.3±0.1 1.23 142.8
Scaled & Boiled (6 h) 18.2±0.1 1.40 137.3
Fermentation period (h)
24 17.2 ±O.O 1.46 134.5
48 17.1 ±O.O 1.57 131.7
72 17.2±0.1 1.63 125.2
Melon seed
Raw 25.0 ± 0.1 1.68 127.8
Fermentation period (h)
24 20.7±0.1 1.85 124.3
48 20.8 ± 0.1 1.96 114.1
72 20.9 ± 0.1 2.19 111.3
Seasoning salts
Knoorcube 1.7±0.0 ND ND
Royco cube 1.3 ±O.O ND ND
Doyin cube 0.6 ±O.O ND ND

after fermentation. In contrast, boiling and fermentation of observations of Gle (1992). At the end of the boiling
melon did not seem to have any significant effect on the ash process, the boil water of the African locust bean was more
content. viscous than it was at the beginning of the process. This
Crude fibre (CF) content of 11.7 and 15.8% are now is an indication of the presence of mucilaginous materials
reported for raw African locust bean and melon seeds in the boil water, which would explain in part the reduction
respectively (Table 1); these values were higher than the of the crude fibre content of the African locust bean seed
3.6% value reported for cowpea (Ojimelukwe et al., on boiling. On the other hand, the crude fibre content of
1999); 0.2% crude fibre (but 10.9% total dietary fibre) for
melon seed increased by 31 o/o after the boiling process
soybean (Suarez et al., 1999) but close to the 8.8% value probably dueto soluble compounds in the seed being leached
reported for African locust bean seed by Oyenuga (1968). out. The crude fibre content of melon seed like that of the
Boiling and dehulling the African locust bean seed reduced African locust bean seed decreased during the last 24
its CF by 61 o/o but fermentation increased the CF by 30% hours of fermentation (by 22%) probably due to the
during the first 48 hours and then reduced the amount production of extracellular enzymes.
by about the same level (30%) during the last 24 hours. The ether extract of raw African locust bean seed
This pattern agrees with the
Table 3: Sorne Biochemical Properties of Fermenting African locust bean and Melon Seeds for lru and Ogiri
Production respectively
Sample Total soluble sugar Reducing sugar Free amino acids

mg glucose s:' dry wt mg glycine s' dry wt

African MeIon*** African Melon*** African MeIon***
locust locust locust
bean bean bean
Raw undehulled seeds 57.0±0.6 33.4±0.5 11.8±0.2 4.9±0.1 47.8±0.4 15.8 ± 0.2
Cooked dehulled seeds* 10.9 ± 0.3 NA 6.0 ± 0.3 NA 22.9 ± 0.3 NA
Cooked dehulled seeds** 9.3 ± 0.2 16.2 ± 0.3 3.3 ± 0.2 2.1 ± 0.1 12.9 ± 0.2 9.0 ± 0.224
h fermentation 7.5 ± 0.3 11.4 ± 0.2 4.5 ± 0.2 2.0 ± 0.3 23.5 ± 0.2 10.4 ± 0.248
h fermentation 8.1 ± 0.2 8.5 ± 0.2 7.6 ± 0.3 3.0 ± 0.1 49.2 ± 0.4 12.3 ± 0.172
h fermentation 9.0 ± 0.4 7.5 ± 0.1 8.2 ± 0.5 1.5 ± 0.1 55.6 ± 4.2 30.0 ± 0.396
h fermentation 9.9 ± 0.3 NA 8.8 ± 0.4 NA 61.6 ± 0.3 NA
NA - Samples not available. *Raw African locust bean was cooked for 12 h and left in the boil water for another 12 h.
**African locust bean seed was dehulled and cooked again for 6 h before fermentation. ***Melon was shelled to give the
raw sample, cooked for 6 h to give the second set of samples and then fermented for 72 h.

Table 4: The Activity of Extracellular Enzymes in Fermenting African locust bean and Melon Seeds for lru and Ogiri
Production Respectively
Processing Steps a-Amylase Sucrase Protease

Enzyme activities expressed as unit m1-1

African MeIon African MeIon African Me Ion

locust seed locust seed locust seed
bean bean bean
Raw undehulled seeds ND*** ND ND ND ND ND
Cooked dehulled seeds* ND NA ND NA ND NA
Cooked dehulled seeds** 0.28± 0.03 0.34 ± 0.04 O.DO O.DO O.DO 0.0024
h fermentation 0.30± 0.02 0.54±0.03 0.47±0.03 0.25±0.02 0.23±0.01 0.14±0.02
48 h fermentation 0.66±0.05 0.72±0.06 0.81±0.09 0.31±0.03 0.31±0.02 0.19 ±0.03
72 h fermentation ND 3.4 ± 0.1 ND 0.16 ± 0.02 ND 0.28±0.10
96 h fermentation 3.0±0.1 NA**** 0.45±0.04 NA 0.51±0.04 NA
*Raw African locust bean was cooked for 12 h and left in the boil water for another 12 h. **African locust bean seed
was dehulled and cooked again for 6 h before fermentation. ***Melon was shelled to give the raw sample, cooked for 6 h
to give the second set of samples and then fermented for 72 h. ***ND - Not determined. ****NA - Samples not available.
increased by boiling and fermentation in agreement with property which makes it indispensable for the
earlier findings (Eka, 1980) while that of melon seed maintenance of the cell integrity and prevent oxidation of
decreased after boiling and thereafter increased by 3- lipid in vivo (Stephen et e!., 1996; Losonczy et al., 1996). The
5% every 24 hours during fermentation. total tocopherol content of the raw and processed
CP content was 30.2 and 19.3% for raw African locust bean products presented in Table 2 indicated that melon seed had
and melon seed respectively. Fermentation also led to the highest value of 25 mg/100 g tocopherol closely followed
increase in CP for the African locust bean but not for melon by African locust bean seed with 21.6 mg/100 g. These
(Table 1). The CP reported for African locust bean agreed values are higher than 2 and 8 mg/100 g reported for lima
favourably with 30.6% reported earlier by Eka (1979) while bean and banana respectively (Cataldo et al., 1999). Boiling
that of melon was close to the 18.4% value reported for and dehulling reduced total tocopherol of African locust
another sample of melon by Adewusi et al. (1992). The bean by 15%. Fermentation for 24 hours reduced
increase in ether extract and CP with increase in fermentation tocopherol by 5% while subsequent fermentation period
period of locust bean seed was probably due to the reduction had no effect on this vitamin. Boiling and fermentation
in the content of ash, CF and carbohydrate. of melon seed for the first 24 hours reduced tocopherol by
Fermentation of both African locust bean and melon seed 17% after which there was no effect with fermentation.
samples led to increase in pH (Table 1) as earlier observed by The total tocopherol of seasoning salts used in recent times
Odunfa (1981). as a replacement for these local condiments ranged between
Tocopherol is a fat soluble vitamin with antioxidant 0.6 and 1.7mg/100 g (Table 2). Rural communities and
 low-income families use iru and ogiri liberally often as a the level of sugar with fermentation may be related to its
substitute for meat and thereby consume a fair amount of utilization by the fermenting microorganisms for their
these condiments on a daily basis. 50 g of both condiments metabolic activities.
level will provide more than the Recommended Dietary The level of total free amino acids increased in both
Allowance (ROA) of 1 O and 8 mg a-tocopherol for adult men seeds with fermentation (Table 3). Similar increases in the
and women respectively (FNB, 1979) while the seasoning level of free amino acids with fermentation have been
salts cannot be regarded as a source of this essential reported in other seeds (Omafuvbe et al., 1999,
vitamin. 2000). This rapid increase in the total free amino acids
Acid and iodine values are presented in Table 2. Heat is a reflection of the increased protease activity observed in
treatment during processing of African locust bean the fermenting seeds (Table 4). Protease activity has been
increased acid value. Acid value in both African locust bean reported to be abundant in the fermentation of similar
and melon seeds also increased with protein rich foods (Sarkar et al., 1993; Omafuvbe et al.,
prolongation of the fermentation period. Heat treatment 2002). Of significant note is the high level of amino acids and
employed in the manufacture of the condiments reduced the protease activity in African locust bean though the protein
iodine value in both African locust bean and melon seed. content of both seeds are the same.
There was a high negative correlation between acid and
Conclusion:Fermentation of African locust bean and
iodine values (r = - 0.98 and - 0.94; p < 0.05 for African locust
melon seeds, to their respective condiments iru and
bean and melon seed respectively). A high positive
ogiri, is desirable nutritionally since the process
correlation was also observed between the total tocopherol
increases the crude protein and ether extract content of the
level and iodine value (r = 0.85 and 0.68; p <
products. The liberal use of these condiments is expected
0.05 for African locust bean and melon seed respectively). to increasethe intake of these essential dietary components
The pattern of change in the levels of soluble sugars and appreciably. 1 n addition, the presence of a high level of
free amino acid content of African locust bean and melon vitamin E in addition to vitamins A and D (Oyenuga, 1968)
seeds were quite different with processing and in these condiments is an added advantage over seasoning
fermentation into iru and ogiri respectively (Table 3). salts, which now tend to replace the local condiments in our
Fermentation of both seeds resulted in substantial kitchens.
decrease in the total sugar and free amino acid levels. This
was probably due to the removal of the hulls that contain Acknowled g ment
higher level of sugar and amino acids. The cooking stage The authors are grateful to Muyiwa Oladele and
of the seeds that have been reported as the most Bamidele Olawumi for their involvement in the
important step in the preparation of raw materia Is for preliminary stages of this work.
fermentation (Wang et al., 1979) may have also resulted in
References
partial loss of soluble solids in the cooking water. For
Adewusi, S.R.A., B.O. Orisadare and O.L. Oke, 1991.
example, Kan no et al. (1982) reported a partial loss of
Studies on Weaning Diets in Nigeria. 1. Carbohydrate
oligosaccharides in cooking and soak water during soaking
Sources. Cereal Chem., 68: 165-169.
and steaming of soybean for natto production. Further
Adewusi, S.R.A., B.O. Orisadare and O.L. Oke, 1992.
decrease in total soluble sugar level was observed with
Studies on weaning diets in Nigeria: 2 protein sources.
fermentation in melon seeds while the level decreased in
Plant Foods for Human Nutr., 42: 183-192.
the first 24 h of fermentation and increased thereafter in
Association of Official Analytical Chemists (AOAC), 1990.
African locust bean. These patterns of change in soluble
Official Methods of Analysis 151h edition, AOAC, Arlington,
sugar level have been reported in similar fermented
USA.
condiments (Omafuvbe and Oyedapo, 2000; Omafuvbe et al.,
Barber, L.I. and S.C. Achinewhu, 1992. Microbiology of
2000).
og iri production from meIon seeds (Citrul/us
The reducing sugar level decreased during the
vulgaris). Nig. Food J., 10: 129-135.
processing stages of both seeds but increased with Cataldo, C. B., L. K. DeBruyne and E.N. Wh itney, 1999.
fermentation in African locust bean and fluctuated in melon
Nutrition and Diet Therapy Principies and Practice
seeds (Table 3). The increased level of reducing sugar is a
5th edition. Wadsworth Publishing Company and
reflection of the activities of a-amylase and sucrase in the
lnternational Thomason Publishing Company U S A. pp:
fermenting seeds (Table 4). Alpha amylase increased with
fermentation in both seeds while sucrase increased, 35-204.
reached its peak at the 481h hour of fermentation and then Contreras-Guzman, E.S. and F.C. Strong 111, 1982a.
dropped. This is however not unusual since similar pattern Determination of Tocopherols (Vitamin E) by reduction
have been reported (Odunfa, 1983; Omafuvbe et al., 2002). lt of Cupric ion. J. Assoc. off. Anal. Chem.,
is worthy of note that sucrase activity was higher in African 65: 1215-1221.
locust bean than melon seeds. This may have been Contreras-Guzman, E.S. and F.C. Strong 111, 1982b.
responsible for the high reducing sugar level in the Determination of Total Tocopherols in grains, grain
fermented African locust bean seeds. The fluctuation in products, and commercial oils, with only slight
saponification, and by a new reaction with Cupric ion.
J. Agri. Food Chem., 30: 1109-1112.
Contreras-Guzman, E.S., F.C. Strong 111 and J.d-S. 173-189.
Williams, 1982. Fatty acid and Vitamin E content of
Nutrimaiz,a Sugary/Opaque-2Corn Cultivar. J. Agri. Food
Chem., 30: 1113-1117.
Eka, O.U., 1979. Effect of fermentation on the nutrient
status of locust beans. Food Chem., 5: 305-308.
Eka, O.U., 1980. Effect of fermentation on the nutritional
status of African locust beans. Food Chem., 5: 303-
308.
Food and Nutrition Board, National Research Council,
1979. Recommended dietary allowances, 9th ed.
Washington, D. C. National Academy of Sciences.
Fuwa, H., 1954. A simple new method for micro
determination of amylase by the use of amylose as
substrate. J. Biochem., 41: 583-603.
Gle, 1992. Please replace this reference with Enujiugha,
V. N. (2003). Nutrient Changes During the
Fermentation of African Oil Beans (Pentaclethra
macrophylla Benth) Seeds. Pak. J. Nut., 2: 320-323.
Heinonen, O.P., D. Albanes and J. Virtano, 1998.
Prostate cancer and supplementation with a-
tocopherol and a-carotene: incidence and mortality
in a controlled trial. J. National Cancer 1 nst., 90: 440-
446.
Kanno, A., H. Takamatsu and T. Akimoto, 1982. Studies on
natto.1. Changes in soybeans during
manufacturing of nafta. J. Japanese Soc. Food Sci., Tec.,
29: 105-110.
Losonczy, K.G., T.B. Harris and R.J. Havlik, 1996.
Vitamin E and vitamin C supplements use and risk of ali
- cause and coronary heart disease mortality in older
persons: The Established Populations (or Epidemiologic
studies of the Elderly). Am. J. Clin. Nutr., 29: 569-57 4.
Morris, D.L., 1948. Quantitative determination of carbohydrate
with Dreywoods anthrone reagent. Sci.
107: 245-255.
Odibo, F.J.C., F. Nwabunnia and D.I. Osuigwe, 1990.
Biochemical changes during fermentation of
Te/faria seeds for ogiri production. World J. Microbio!.
Biotech., 6: 425-427.
Odunfa, S.A., 1981. Microbiology and amino acid
composition of ogiri, a food condiment from fermented
melon seeds. Die Nahrung, 25: 811-816.
Odunfa, S.A., 1983. Biochemical changes during
production of ogiri, a fermented melon (Citrul/us
vulgaris Schrad) product. Qualities Planetarium: Plant
Food for Human Nutr., 32: 11-18.
Odunfa, S.A., 1985. African fermented foods, In: Wood, B.J.B.
(ed.) Microbiology of Fermented Foods, vol.
11, Amsterdam, Elsevier Applied Science
Publishers. p: 155-191.
Odunfa, S.A., 1986. Daddawa, In: Reddy, N.R., Pierson, M.O. and
Salunkhe, D.K. (eds.) Legume-based Fermented Foods.
Boca Raton, CRC Press lnc., p:
Ojimelukwe, P.C., J.C. Onweluzo and Okechukwu, 1999. 1510-1513.
Effects of infestation on the nutrient content and
physiochemical properties of two cowpea (Vigna
unguiculata) varieties. Plant foods for Human Nutr.,
53: 321-332.
Omafuvbe, B.O., 1998. Evaluation of the microbiological and
biochemical changes during the fermentation of
soybean (G/ycine max (L) Merrill) for soy- daddawa.
Ph.D Thesis, Obafemi Awolowo
University, l le-lfe, Nigeria.
Omafuvbe, B.O. and O.O. Oyedapo, 2000. Observed
biochemical changes during natural fermentation of
African oil bean (Pentaclethra macrophyl/a) seeds. Nig.
J. Nutr. Sci., 21: 19-23.
Omafuvbe, B.O., S.H. Abiose and O.O. Adaraloye, 1999.
The production of kpaye - a fermented condiment
from Prosopis africana Taub. seeds. lnt. J. Food
Microbio!., 51: 183-186.
Omafuvbe, B.O., O.O. Shonukan and S.H. Abiose, 2000.
Microbiological and biochemical changes in the
fermentation of soybean for soy-daddawa - Nigerian
food condiment. Food Microbio!., 17: 469-474.
Omafuvbe, B.O., S.H. Abiose and O.O. Shounkan, 2002.
Fermentation of soybean (G/ycine max) for soy-
daddawa production by starter cultures of Bacillus.
Food Microbio!., 19: 561-566.
Oyenuga, V.A., 1968. Nigeria's Food and Feeding-stuffs;
Their Chemistry and Nutritive Value. 1 badan
University Press, 1 badan.
Rosen, H., 1957. A modified ninhydrin calorimetric analysis
for amino acids. Archives of Biochemistry and
Biophysics, 67: 10-15.
Sarkar, P. K., P. E. Cook and J.D. Owens, 1993. Bacillus
fermentation of soybean. World J. Microbio!. and
Biotec., 9: 295-299.
Simmons, E. B., 1976. Calorie and protein intake in three
villages of Zaria Province, May 1970-July
1971.Samaru Miscellaneous Paper, lnstitute of
Agricultura! Research, Ahmadu Bello University, Zaria, No
55, 151 p.
Somogyi, M., 1945. A new reagent for the determination
of sugars. J. Bio. Chem., 160: 61-68.
Stephen, N.G., A. Parsons and P.M. Schofield, 1996.
Random controlled trial of Vitamin E in patients with
coronary disease: Cambridge Heart Antioxidant Study
(CHAOS). Lancet, 347: 781-786.
Suarez, F.L., J. Springfield, J.K. Furne, T.T. Lohrmann, P.S.
Kerr and M.D. Levitt, 1999. Gas production in hu mans
ingesting soybean flour derived from beans
naturally low in oligosaccliarides. Am. J. Clin. Nutr., 69:
135-140.
Umoh and Oke, 197 4. Nutritive value of lesser known oil
seeds in rats. Nutrition Report lnternational 9: 453-
456.
Wang, H.L., E.W. Swain, C.W. Hesseltine and H.D.
Heath, 1979. Hydration of whole soybeans affects solid losses
and cooking quality. J. Food Sci., 44: