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THE EFFECT OF DIFFERENT PARTICLE SIZES OF AFRICAN YAMBEAN (SPHENOSTYLIS STENOCARPA) SEED FLOUR ON ITS FUNCTIONAL PROPERTIES AZEKE, M. A., EKPO1, K. E. AND IHIMIRE I. G.1 Department of Biochemistry, Ambrose Alli University, P. M. B. 14, Ekpoma, Edo State
1

Corresponding author; e-mail: m_azeke@yahoo.com

ABSTRACT This study was conducted to determine the effect of particle sizes on some important functional properties of seed flour of the underutilized African yambean. The seeds were milled after which flour was separated into different particle sizes using meshes of different pore sizes (10, 20, 40, 60, 80 and 100 mesh respectively). The functional properties determined were ptotein solubility, bulk density, water and oil absorption capacities, emulsion properties (emulsion activity and stability) and foaming properties (foam capacity and stability). From the results obtained, it was found that the smaller particle size flour had the lower protein solubility, while the flour with larger particle size had the higher protein solubility (p<0.05). Particle size also significantly affected water absorption capacity, foam capacity and emulsion activity (p<0.05). The results implied that when high water absorption is desired, yambean flour of larger particle size would be appropriate while the flour of smaller particle size would be appropriate in food formulations where higher foam capacity and emulsion activity are desired. Key Words: Particle size, functional properties, African yambean, protein solubility in soybean (Evans and Boutler, 1974; Edem et al., 1990; Ene-Obong and Carnovale, 1992; Agunbiade and Longe, 1999). Yambean seeds are used to make products such as akara, a fried paste, and moin-moin, a steamed paste made from seed flour. The steamed paste is made by soaking overnight after which the coats are removed. The seeds are then ground into a paste to which palm oil, pepper and other condiments may be added to taste. When this praparation is boiled, usually wrapped in banana-leaves, it turns out as a yellowish food which is reputed to be very filling and lasting. After eating it people generally drink a lot of water, probably as an aid to the digestion. The desirability of these products depends largely on their texture, which is a function of the particle size of seed flour. A lot of work has been done on similar products from cowpea seed flour. For example, akara made from commercial cowpea flour exhibited poor water absorption, was too dense, and lacked crispness (Dovlo et al., 1976). McWatters (1983) related the poor performance of commercial Nigerian cowpea flour to the mean

INTRODUCTION The African yambean (Sphenostylis stenocarpa) is a relatively inexpensive but underutilized source of protein that is widely recognized for its high nutritional and excellent functional properties (Azeke et al., 2005; Obatolu et al., 2006). The chemical composition of African Yambean (AYB) has been evaluated by several authors (Edem et al., 1990; Apata and Ologhobo, 1990; Azeke et al., 2007). Its seeds are potential sources of nutritious snack foods, particularly because they are rich in protein. African yambean seeds contain approximately 11% moisture, 23% protein, 1.3% fat, 57% carbohydrate, 1.5% dietary fibre and 3.0% ash (Azeke et al., 2007). A crude protein content of between 20 and 29% has been reported (Edem et al., 1990; Apata and Ologhobo, 1990; Azeke et al., 2007), which is lower than that of soybean (38%). The amino acid composition of African Yambean protein has been reported in several literatures and the lysine proportion in the protein of AYB is reported to be equal to or higher than

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particle size of the flour. Akara made from cowpea flour in which 48% of the particle distribution was in the 400 mesh range was dry and dense, and exhibited a tough outer surface. Flour in which 42% of the particles were from the 100 mesh screen produced a more acceptable product. Ngoddy et al. (1986) studied the effects of successive milling of cowpea flour through a 1 mm screen, and observed that if fine flours were used to prepare akara or moin-moin, the foods were less hydrated, denser, and exhibited an undesirable spongy texture as well as an unacceptable appearance. This work was conducted to evaluate the effect of particle sizes on the functional properties of African Yambean seed flour. MATERIALS AND METHODS African yambean seeds (Sphenostylis stenocarpa), purchased from the Ekpoma main market, were decorticated as described by Hung et al. (1991) and ground in a Thomas-Wiley laboratory mill (Model 4, Arthur H. Thomas Co., Philadelphia, PA). The yambean seed flour was then passed through a series of sieves (20, 30, 40, 60, 80, 100 mesh screens) to separate it into fractions of various particle size. Functional properties Protein solubility: The pH-dependent protein solubility was determined by the method Were et al. 1997. 25 mg of seed flour was blended with 25 ml distilled water and the pH of solution was adjusted to 6.0, 7.0 or 9.0 using 0.5 M NaOH or HCl. The solutions were mixed thoroughly and centrifuged. The supernatant was filtered and

nitrogen was estimated by Kjeldahls method (Humphries, 1956). The soluble protein (%) profile was thus determined (nitrogen solubility X 6.25) Bulk Density The bulk density was determined according to the method of Okaka and Potter (1977). A 50-g sample was put into a 100 mL graduated cylinder. The cylinder was tapped about 30 times until there was no further change in volume. The bulk density was calculated as weight per unit volume of the sample. Water and Oil Absorption Capacities Water and oil absorption capacities (WAC and OAC) were determined in triplicate according to the method Beuchat (1977). One gram of flour was mixed by shaking with 10 ml distilled water or refined vegetable oil for 30 s in centrifuge tubes. The solution was allowed to stand at room temperature for 30 min, centrifuged (5000g, 30 min) and volume of supernatant was determined. Water and oil absorption capacities were expressed as ml/g flour. Emulsion Properties Emulsion Activity (EA) and Stability (ES) were determined in triplicate following the methods of Neto et al. (2001). Five millilitre of flour dispersion in distilled water (10 mg/ml) was homogenized (1 min) with 5 ml refined vegetable oil. The emulsions were centrifuged (1100g, 5 min) and the height of the emulsified layer and the total contents in the tube was determined. The emulsifying activity was calculated:

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Emulsifying Activity (%) =

Height of the emulsified layer Height of the total content

X 100

Emulsion stability was determined by heating the emulsion (80 oC, 30 min) before centrifuging (1100g, 5 min)

Height of the emulsified layer after heating Emulsifying Stability (%) = Height of emulsified layer before heating X 100

Foaming properties Foaming Capacity and Stability of flour was determined in triplicate according the method outlined by Seena and Sridhar (2005). Two grams of the flour was dispersed in 100 ml

distilled water and whipped vigorously for 2 min in a kitchen blender at speed 1. The volumes of foam were recorded before and after whipping and the percentage volume increase was calculated:

Volume after whipping Volume before whipping Foam Capacity (%) = Volume before whipping X 100

Foam stability was determined as the volume of foam that remained after 8 h at room temperature

(27 2o C) and expressed as the percentage of initial foam volume

Foam Stability (%) = Volume after 120 min Initial foam volume
RESULTS AND DISCUSSION The protein solubility profile of yambean seed flour is as shown in table 1. Particle size had a significant effect on protein solubility. It was found that the smaller the particle size, the lower the protein solubility. This is unexpected and difficult to explain considering the fact that the smaller particle size flour has larger surface area

X 100

of contact with the extracting solvent. This result is contrary to the report of Kerr et al. (2000) who worked on the effect of particle size on protein solubility of cowpea seed flour and found that there was an increase in solubility as particle size decreased. One possible reason for this contrast could be that more of the protein is present in the flour with larger particle size

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Table 1: Protein Solubility Index (%) of yambean bean flour as affected by pH and particle size. Mesh Sizes 10 20 40 60 80 100 > 100 Particle size (mm) 2.0 0.84 0.42 0.297 0.177 0.149 < 0.149 Water (pH 7.0) 10.41 3.40a 10.27 3.35a 9.73 3.17ab 9.51 3.10b 9.29 3.03bc 8.82 2.88c pH 6.0 pH 7 pH 9

6.02 0.01a 6.63 0.01a 8.12 0.01b 8.73 0.01b 4.82 0.09cd 4.51 0.09c 5.13 0.01d

5.42 0.01a 4.22 0.01b 2.12 0.01c 1.82 0.01cd 2.12 0.01c 2.43 0.01c 1.52 0.09d

5.82 0.01a 5.71 0.09a 6.72 0.01b 6.91 0.01b 6.93 0.01b 9.02 0.09c 9.92 0.01c

Results are means of three independent determinations Standard Deviation; Values in a column with different alphabets are significantly different The effect of particle size on protein solubility was found to be pH dependent. While there was a gradual reduction in PSI as particle size decreased at pH 7.0, the reverse was the case at pH 9.0. At both acidic and alkaline pH, the flour recorded a higher PSI than at pH 7.0 regardless of particle size. Similar trend was reported for Canavalia species (Seena and Sridhar, 2005) bambarra ground nut and Mucuna beans (Adebowale and Lawal, 2004). At acidic and alkaline pH the protein gets net positive and negative charge, respectively, which favours the repulsion of molecules and thereby increasing the solubility of proteins (Adebowale and Lawal, 2004). The protein concentrates of seeds that are soluble at pH 4 7 are normally suitable for use as additives in beverages such as vegetable milk. The effects of particle size on some functional properties of yambean seed flour are shown in table 2. With the exception of water absorption capacity, foam capacity and emulsion activity, most functional properties did not show any pattern of effect with particle size. The bulk densities of yambean seed flour of different . particle sizes ranged from 0.57 0.66 g/ml. These results are similar to that reported for cowpea (0.6 g/ml) by Okaka and Porter (1979) and that reported for bambara groundnut (0.6 0.75 g/ml) by Onimawo et al. (1998) but higher than that reported for pigeon pea (0.27 g/ml) by Onimawo and Akpojovwo (2006). Bulk density is a reflection of the load the flour samples can carry if allowed to rest directly on one another (Onimawo and Akpojovwo, 2006). Water absorption capacity (WAC) of yambean flour decreased with decrease in particle size from 6.47 5.84 ml/g (table 2). The oil absorption capacity (OAC) of the flour was found to be 6.02 6.22 ml/g. These values are high compared to earlier reports from other legumes. For example Onimawo and Akpojovwo (2006) reported WAC and OAC of 4.47 and 2.50 respectively for pigeon pea. Seena and Sridhar (2005) reported WAC and OAC of 2.3 and 1.4 respectively for Canavalia species. Khattab and Arntfield (2009) working with different legumes reported a range of 3.2 7.9 and 2.1 2.6 g/ml for WAC and OAC respectively

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Table 2: Some functional properties of yambean seed flour as affected by particle size Mesh Size Particle Size (mm) Bulk density (g/ml) Water absorption (ml/g) 6.47 0.06a 6.57 0.02a 6.17 0.10ab 5.97 0.04b 5.95 0.11b 5.86 0.12b 5.84 0.04b Oil absorption (ml/g) Foam capacity (%) Foam stability (%) Emulsion activity (%) Emulsion stability (%)

10 20 40 60 80 100 > 100

2.0 0.84 0.42 0.297 0.177 0.149 -

0.57 0.004a 0.61 0.005a 0.66 0.006a 0.57 0.004a 0.61 0.018a 0.60 0.005a 0.58 0.016a

6.22 0.13a 6.13 0.06a 6.11 0.14a 6.25 0.11a 6.02 0.03a 6.20 0.03a 6.18 0.13a

14.00 1.00a 15.67 0.88b 30.67 0.88c 33.67 1.76c 37.67 0.88d 40.00 1.16e 40.00 0.58e

5.67 1.20a 12.67 3.71b 11.67 1.20b 13.67 1.76c 9.67 0.88d 10.33 1.45d 14.00 3.06e

52.00 4.58a 57.00 5.13b 65.67 4.06c 70.33 4.09d 77.33 3.76e 81.67 3.84f 86.67 4.06g

8.63 0.11a 8.22 0.15a 8.62 0.40a 8.85 0.06a 8.70 0.06a 8.77 0.05a 8.92 0.02a

Results are means of three independent determinations Standard Deviation; Values in a column with different alphabets are significantly different

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The differences in the WAC and OAC may be due to the variation in the presence of non-polar side chains, which bind the hydrocarbon side chain of oil. The enhanced ability of the meal to absorb and retain water and oil may help to improve binding of the structure, enhance flavor retention, improve mouth feel and reduce moisture and fat losses of food products (Sreerama et al., 2008). WAC enabled bakers to add more water to doughs so as to improve handling characteristics and maintain freshness in bread. Oil absorption capacity is desired in ground meat formulations, flavour retention, improvement of palatability, extension of shelf life of bakery or meat products, meat replaces and extenders, doughnuts, pancake, baked goods and soups. The foaming capacities of flour were found to increase from 14 40 % with decrease in particle size. Owing to a large increase in the surface area in the liquid/air interphase, proteins denature and aggregate during whipping. This is important for flour used in many leavening food products such as baked goods, cakes and biscuits (Sreerama et al., 2008). There are wide variations in the foaming capacities reported for different flours. While Onimawo and Akpojovwo (2006) reported 600 % foam capacity for pigeon pea flour, Khattab and Arntfield (2009) reported 17 56 % for different legume varieties. The foam from the yambean flour of smaller particles was found to be more stable than that from flour of larger particle. Emulsifying activity (EA) increased from 52 to 86 % with decrease in particle size while emulsion stability (ES) was not dependent on particle size (table 2). These values agree with previous reports for other legumes (Seena and Sridhar, 2005; Onimawo and Akpojovwo, 2006; Khattab and Arntfield, 2009). EA and ES are important functional properties of food proteins. They vary with the type of protein, its concentration, pH, ionic strength, and viscosity of the system. Many chemical and physical factors are involved in the formation, stability, and textural properties of proteinfatwater emulsions (Khattab and Arntfield, 2009). Superior emulsifying properties are desired to make milk like beverages and meat analogues.

CONCLUSION Milling and subsequent sieving of African yambean seeds into flours of different particle sizes had various effects on the functional properties of yambean seed flour. When high water absorption is desired, yambean flour of larger particle size would be appropriate while the flour of smaller particle size would be appropriate in food formulations where higher foam capacity and emulsion activity are desired. Therefore, the present study shows the potential of yambean flour for incorporation into new value-added products. The detailed toxicological properties of this meal still remain to be evaluated and further research is needed on the reaction of protein of yambean seeds with other constituents of food products such as starch and lipids

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