International Journal of Scientific Research in Knowledge, 2(2), pp. 92-104, 2014 Available online at http://www.ijsrpub.

com/ijsrk ISSN: 2322-4541; 2014 IJSRPUB http://dx.doi.org/10.12983/ijsrk-2014-p0092-0104

Full Length Research Paper Nutritional and Anti-Nutritional Composition of Bridelia Ferruginea Benth (Euphorbiaceae) Stem Bark Sample
Adesina Adeolu Jonathan1*, Akomolafe Seun Funmilola2
2

Department of Chemistry, Ekiti State University, PMB 5363, Ado Ekiti, Nigeria Department of Biochemistry, Ekiti State University, PMB 5363, Ado Ekiti, Nigeria *Corresponding Author: Email: unclejoshua2012@gmail.com
Received 06 December 2013; Accepted 11 January 2014

Abstract. Nutritional composition of Bridelia ferruginea Benth (Euphorbiaceae) stem bark sample was evaluated. The percentage protein, fat, fibre and carbohydrate contents were: 15.7 0.30, 5.45 0.05, 4.35 0.06 and 60.7 0.71 respectively while the total gross energy was 1501kJ/100g. The mineral composition ranged from 0.25 to 74.2 mg/100g, with phosphorus being the most concentrated. The K/Na ratio (1.80) was higher than 1.0 recommended. The mineral safety index computation showed only Zn to be in excess based on the recommended daily allowance (RDA). The amino acid contents ranged between 0.597 13.1 g/100g. All the essential amino acids were present in varying amount with Isoleucine being the most concentrated. The % TEAA and % TNEAA were: 47.8 and 52.2 respectively. The P-PER (predicted protein efficiency ratio), pI (Isoelectric point) and EAAI (essential amino acid index) values were: 1.69, 5.14, 1.18 respectively. Based on the whole hens egg amino acid scoring pattern, methionine was limiting while with respect to FAO/WHO provisional amino a cid scoring pattern and essential amino acid scoring pattern based on the requirements of pre-school child Met + Cys was limiting. The anti-nutritional factors analyzed; phytates, tannins, oxalates, phenolic content, saponins, phytin phosphorus and alkaloids in the sample were lower than the range of values reported for most vegetables. This study revealed that the Bridelia ferruginea stem bark consumed in Ekiti State and other states in the South-western part of Nigeria can contribute useful amount of nutrients to human diet. Keywords: Nutritional, anti-nutritional composition, Bridelia ferruginea stem bark

1. INTRODUCTION Forage trees and shrubs play an essential and multiple roles in the balance of the Sahelian and Sudanian ecosystems exploited by man and his animals. This role becomes more important as the dry season grows longer, and decreases as the mean annual rainfall increases. It therefore grows less important from north to south according to the rainfall gradient, which is about 1 mm per km, or 110 mm per each degree of latitude. Throughout West Africa, especially in areas prone to drought, previous studies demonstrate the importance of edible wild plants as food sources (Grivetti et al., 1987; Sena et al., 1998). Commonly, drought is associated with inadequate food intake and disease, where food scarcity and inadequate dietary intakes clearly have led to increased incidence of malnutrition and famine (Franke and Chasin, 1980). Use of edible wild species in combination with domesticated foods has remained a hallmark of many African agro-pastoral societies (Grivetti, 1978, 1979; Grivetti et al., 1987). Traditional medicine practiced

in rural Nigeria, reveals well-documented uses of plant barks and bark extracts, fruits, leaves, nuts and seeds, and tubers, but with few exceptions, dietarymedical uses of edible wild plants as components of West African traditional medicine have not been widely documented (Ogugbuaja et al., 1997). Bridelia ferruginea Benth (Euphorbiaceae) is a medicinal plant that is widely used in African folkloric medicine. In Nigeria, it is commonly called Kirni, Kizni (Hausa); Marehi (Fulani); Iralodan (Yoruba), Ola (Igbo), Kensange abia (Boki). Its habitat is the savannah especially in the moister regions extending from Guinea to Zaire and Angola. The tree is 6-15m high, up to1.5m in girth and bole crooked branching low down. The bark is dark grey, rough and often markedly scaly (Kolawole and Olayemi, 2003). The stem bark decoction is used in African traditional medicine to treat diarrhea, dysentery and gynaecological disorders (including sterility). A decoction of the leaves is used to treat diabetes. It has even been evaluated for antimalarial (Kolawole and Adesoye, 2010), antimicrobial

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(Kareem et al., 2010), analgesic (Akuodor, et al., 2011) and antidiabetic (Bakoma et al., 2011) activities. B. ferruginea bark is used for treatment of bacterial infections on wounds (Irobi et al., 1994). A decoction of the leaves is used as a purgative and also in the treatment of diabetes (Cimanga et al., 1999). Roots and leaves extracts are used to cure piles, diarrhea and dysentery (McNeely, 1990) and also confirmed for anti-inflammation activities (Olajide et al., 1999). Galocatechin has been isolated from the bark (De Bruyne et al., 1997). In western Nigeria, B. ferruginea is used as a mouthwash and remedy for candidal oral thrush; whereas in Northern Nigeria, the bark is used for treatment of infections caused by poisoned arrow wounds (Irobi et al., 1994). A decoction of the bark extract has also been proven to have antibacterial effect (Kolawole et al., 2006). It is also used as purgative and a vermifuge. A macerated extract of the bark is used in Northern Nigeria to harden beaten laterite and mud floors (Kolawole and Olayemi, 2003). The chemical constituents of Bridelia ferruginea have not been thoroughly examined. The present study is therefore aimed at exploiting the proximate, minerals, anti-nutritional and amino acid

profile of the back of Bridelia ferruginea as this would promote its dietary-medical uses. 2. MATERIAL AND METHODS 2.1. Collection and preparation of samples The sample was collected from local farms around Iworoko- Ekiti, Irepodun-Ifelodun local Government area of Ekiti State. It was properly sorted, washed, dried, milled into fine powdered form and kept in an air-tight plastic bottle prior to analysis. 2.2. Proximate analysis Moisture, total ash, fiber and ether extract of the samples were determined by the methods of the AOAC 2005. Nitrogen was determined by a microKjeldahl method and the crude protein content was calculated as N x 6.25 (Pearson, 1976). Carbohydrate was determined by difference. All the proximate results were reported in g/100 g dry weight. The energy values obtained for carbohydrates (x 17 kJ), crude protein (x 17 kJ) and crude fat (x 37 kJ) for each of the samples. Determinations were in duplicate.

Table1: Proximate and some calculated parameters in the sample of Bridelia ferruginea stem bark

PEP = Proportion of total energy due to protein PEF = Proportion of total energy due to fat PEC = Proportion of total energy due to protein UEDP = Utilizable energy due to protein

2.3. Mineral analysis The mineral elements were determined in the solutions obtained above-Na and K by flame photometry, Model 405 (Corning, Halstead Essex, UK) using NaCl and KCl to prepare standards. Minerals were analysed using the solutions obtained by dry ashing the samples at 550 oC and dissolving it

in 10 % HCl (25 ml) and 5 % lanthanum chloride (2 ml), boiling, filtering and making up to standard volume with deionized water. Phosphorus was determined colorimetrically using a Spectronic 20 (Gallenkamp, London, UK) instrument, with KH2PO4 as a standard. All other elements (Ca, Mg, Zn, Fe, Mn, Cu and Cr) were determined by atomic absorption spectrophotometry, Model 403 (Perkin-Elmer,

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Norwalk, Connecticut, USA). All determinations were made in duplicate. All chemicals used were of analytical grade, and were obtained from British Drug House (BDH, London, UK). The detection limits for the metals in aqueous solution had been determined just before the mineral analyses using the methods of Varian Techtron, giving the following values in g/ml: Fe (0.01), Cu (0.002), Na (0.002), K(0.005), Ca(0.04), Mg(0.002), Zn (0.005), Mn (0.01) and Cr (0.02) (Varian Techtron, 1975). The optimal analytical range was 0.1 to 0.5 absorbance units with coefficients of variation from 0.9-2.2 %. The coefficients of variation per cent were calculated (Steel and Torrie, 1960). The percentage contribution to energy due to protein (PEP), due to total fat (PEF) and due to carbohydrate (PEC) as PEP %, PEF % and PEC % respectively were calculated. The percentage utilizable energy due to protein (UEDP %) was also calculated. Ca/P, Na/K, Ca/Mg and the millequivalent ratio of [K/(Ca +Mg)]; the mineral safety index (MSI) of Na, Mg, P, Ca, Fe and Zn were also calculated (Hathcock, 1985). To calculate MSI, we have: RAI is recommended adult intake; CV in the Table will represent calculated value

(CV) of calculated MSI from research results. The differences between the standard MSI and the MSI of the samples were also calculated. 2.4. Determination of Anti-nutritional factors 2.4.1. Determination of tannin 200mg of the sample was weighed into a 50ml sample bottle. 10ml of 70% aqueous acetone was added and properly covered. The bottles were put in an orbital shaker and shaken for 2 hours at 300C. Each solution was then centrifuged and the supernatant stored in ice. 0.2ml of each solution was pipetted into test tubes and 0.8ml of distilled water was added. Standard tannic acid solutions were prepared from a 0.5mg/ml stock and the solution made up to 1ml with distilled water. 0.5ml folin reagent was added to both sample and standard followed by 2.5ml of 20% Na2CO3. The solutions were then vortexed and allowed to incubate for 40 minutes at room temperature after which absorbance was red against a reagent blank concentration of the sample from a standard tannic acid curve (Makkar and Goodchild, 1996).

Table 2: Composition and some calculated mineral ratios in the Bridelia ferruginea bark sample

*milliequivalent ratio

2.4.2. Determination of oxalate 1g of the sample was weighed into 100ml conical flask. 75ml of 1.5N H2SO4 was added and the solution was carefully stirred intermittently with a magnetic stirrer for about 1 hour and then filtered using Whatman filter paper. 25ml of sample filtrate was collected and titrated hot (80-900C) against 0.1N

KMnO4 solution to the point when a faint pink colour appeared that persisted for at least 30 seconds (Day and Underwood, 1986). 2.4.3. Determination of alkaloid Alkaloid determination was carried out following the procedure of Harborne(Harborne, 1973). 5.0g of the

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sample was weighed into a 250ml beaker and 200ml of 10% acetic acid in ethanol was added and covered and allowed to stand for 4h. This was filtered and the extract was concentrated on a water bath to one quarter the original volume. Concentrated ammonium hydroxide was added drop wise to the extract until the precipitation was complete. The whole solution was allowed to settle and the precipitate was collected and washed with dilute ammonium hydroxide and then filtered. The residue is the alkaloid which was dried and weighed. 2.4.4. Determination of saponin The method used was that of Obadoni and Ochuko, 2001). 5g of the sample was put into a conical flask and 100cm3 of 20% aqueous ethanol were added. The sample was heated over a hot water bath for 4h with continuous stirring at about 550C. The mixture was filtered and the residue re-extracted with another 200ml 20% ethanol. The combined extracts were reduced to 40ml over water bath at about 900C. The concentrate was transferred into a 250ml separating funnel and 20ml of diethyl ether was added and shaken vigorously. The aqueous layer was recovered while the ether layer was discarded. The purification process was repeated. 60ml n-butanol was added. The combined nbutanol extracts were washed twice with 10ml of 5% aqueous sodium chloride. The remaining solution was heated in a water bath after evaporation; the sample was dried in the oven to a constant weight. The saponin content was calculated as percentage. 2.4.5. Determination of flavonoid The method of Boham and Kocipai-Abyazan (Boham and Kocipai-Abyazan, 1974) was followed in the determination of flavonoid. 5g of the sample was extracted repeatedly with 100ml of 80% aqueous methanol at room temperature. The whole solution was filtered through whatman filter paper (125ml). The filtrate was later transferred into a crucible and evaporated into dryness and weighed to a constant weight. 2.4.6. Oxalate determination The titration method as described by Day and Underwood (1986) was followed. 1g of sample was weighed into 100 ml conical flask. 75 ml 3M H2SO4 was added and stirred for 1 h with a magnetic stirrer. This was filtered using a Whatman No 1 filter paper.

25 ml of the filtrate was then taken and titrated while hot against 0.05 M KMnO4 solution until a faint pink colour persisted for at least 30 s. The oxalate content was then calculated by taking 1ml of 0.05 M KMnO4 as equivalent to 2.2 mg oxalate (Chinma and Igyor, 2007; Ihekoronye and Ngoddy, 1985).

2.4.7. Phytate content determination This was determined by the method of Wheeler and Ferrel (1971).100 ml of the sample was extracted with 3% trichloroacetic acid. The extract was treated with FeCl3 solution and the iron content of the precipitate was determined using Atomic Absorption spectrophotometer (Pye Unicam 2900). A 4:6 Fe/P atomic ratio was used to calculate the phytic acid content (Okon and Akpanyung, 2005). Phytin phosphorus (Pp) was determined and the phytic acid content was calculated by multiplying the value of Pp by 3.55 (Young and Greaves, 1940). Each milligram of iron is equivalent to 1.19 mg of Pp. Phytin phosphorus as percentage of phosphorus (Pp % P) = Pp/P 10 2.5. AMINO ACID ANALYSIS The amino acid profile was determined using the method described by Sparkman et al. (1958). Each sample was dried to constant weight, defatted, hydrolyzed, evaporated and loaded into the techno sequential multi-sample amino acid analyzer (TSM). Following the steps described below: 2 g of the dried sample was weighed into extraction thimble and the fat extracted with chloroform: methanol (2:1) mixture using soxhlet extraction apparatus (AOAC, 2005). Then, 1 g of the defatted sample was weighed into glass ampoule. 7 ml of 6 N HCl were added and oxygen expelled by passing nitrogen into the ampoule. The glass ampoule was sealed and placed in an oven preset at 1050C for 22 h. The ampoule was allowed to cool before breaking open at the tip and the content filtered. The filterate was then evaporated to dryness and the residue dissolved with 5 ml acetate buffer (pH 2.0) and stored in plastic specimem bottles. 10 l was dispensed into the cartridge of the analyser which is designed to seperate andanalyse free, acidic, neutral and basic amino acids of the hydrolysate. The amount of each amino acid present in the sample was calculated in g/100 g protein from the chromatogram produced.

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Table 3: Mineral safety index of Na, Mg, P, Ca, Fe, and Zn for the Bridelia ferruginea stem bark sample

TV = table value, CV = calculated value D = difference (TV - CV)

Table 4: Anti-nutritional content of the Bridelia ferruginea stem bark samples

2.5.1. Determination of quality parameters 2.5.1.1. Determination of amino acid scores Determination of the amino acid scores was fir

st, based on whole hens egg (Paul et al., 1976). In this method, both essential and nonessential amino acids were scored. Secondly, amino acid score was calculated using the following formula (FAO/WHO, 1973):

Amino acid score = (amount of amino acid per test protein (mg/g)) / (amount of amino acid per protein in reference pattern (mg/g)). In this method, Met + Cys and Phe + Tyr were each taken as a unit. Also, only essential amino acids determined were scored. Amino acid score was also calculated based on the composition of the amino acids obtained in the samples compared with the suggested pattern of requirements for pre-school children (2-5 years). Here, Met + Cys and Phe + Tyr were each taken as a unit. Also, only essential amino acids with (His) were scored. 2.5.1.2. Determination of the essential amino acid index The essential amino acid index (EAAI) was calculated by using the ratio of test protein to the reference protein for each of the eight essential amino acid plus histidine in the equation that follows (Steinke et al., 1980):

Methionine and cystine are counted as a single amino acid value in the equation, as well as Phe + Tyr. 2.5.1.3. Determination of the predicted protein efficiency ratio The predicted protein efficiency ratio (P-PER) was determined using one of the equations derived by Alsmeyer et al. (1974), i.e. P-PER = 0.468 + 0.454 (Leu) 0.105 (Tyr). 2.5.1.4. Other determinations Determination of the total essential amino acid (TEAA) to the total amino acid (TAA), i.e. (TEAA/TAA); total sulphur amino acid (TSAA); percentage cystine in TSAA (% Cys/TSAA); total aromatic amino acid (TArAA), etc; the Leu/Ile ratios were calculated while the isoelectric point (pI) was calculated using the equation of the form (Olaofe and Akintayo, 2000):

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pIm IPiXi Where pIm is the isoelectric point of the mixture of amino acids, IPi is the isoelectric point of the ith amino acid in the mixture and Xi is the mass or mole fraction of the ith amino acid in the mixture (Finar, 1975).
Table 5: Amino acids profile of the Bridelia ferruginea bark samples (g/100g cp)

physiological state while that of infants is 740 kCal (3094.68 kJ) (Bingham, 1978). This implies that while an adult man would require between 590-709 g (taking the calculated energy of 1501 kJ/100 g) of his energy requirement, infants would require 174.9 g (taking the calculated energy of 1501 kJ/100 g). On the whole this meant that samples with higher energy value would require lower quantity of sample to satisfy the energy needs of man and infants. The utilizable energy due to protein (UEDP %) for the sample (assuming 60 % utilization) 10.7 %. The UEDP % compared favourably with the recommended safe level of 8 % for an adult man who requires about 55 g protein per day with 60 % utilization. The PEF % values were generally low in the sample (13.4 %) and far below the recommended level of 30 % (NACNE, 1983) and 35 % (COMA, 1984) for total fat intake; this is useful for people wishing to adopt the guidelines for a healthy diet.
Table 6: Concentrations of essential, non-essential, acidic, neutral, sulphur, aromatic, basic, etc. (g/100g crude protein) of Bridelia ferruginea stem bark samples (dry matter of sample)

3. RESULTS AND DISCUSSION The results of proximate composition of the Bridelia ferruginea back sample are shown in Table 1. The crude protein content (15.7 0.30 %) was comparably higher than the values reported for some vegetables consumed in Nigeria: 5.91 % (Cnidoscolus chayamansa), 3.31 % (Solanium nodiflorum), 3.03 % (Senecio biafrae) (Adeleke and Abiodun, 2010); 4.60% (A. hydridus), 4.30 % (T. occidentallis) (Fafunso and Basir, 1977) but favourably compared with the values reported for C. maxima (18.6 %), A. viridis (19.2 %) and B. alba (18.0%) (Adesina, 2013) and S. indicum (18.59%), B.aegypiaca (15.86 %) (Kubmarawa et al., 2008). The ash content (6.54 %) of Bridelia ferruginea back sample is an indication of the levels of minerals or inorganic component of the sample. These minerals act as inorganic co-factors in metabolic processes which mean in the absence of these inorganic cofactors there could be impaired metabolism (Iheanacho and Udebuani, 2009). Table 1 still contains other parameters calculated from the proximate values. It shows the various energy values as contributed by protein, fat and carbohydrate. The daily energy requirement for an adult is between 2500-3000 kCal (10455-12548 kJ) depending on his

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Table 2 gives the list of the nutritionally important minerals as well as the computed mineral ratios in Bridelia ferruginea back sample. Minerals are important in human nutrition. It is well known that enzymatic activities as well as electrolytic balance of the blood fluid are related to the adequacy of Na, K, Mg and Zn. Potassium is very important in maintaining the blood fluid volume and osmotic equilibrium. Metal deficiency syndrome like rickets and calcification of bone is caused deficiency. Appreciable levels of all the essential minerals were present in the the sample. The sample was apparently high in phosphorus (74.2 0.105 mg/100g), a value which was comparably higher than what was reported for F. asperifolia and F. sycomorus (Nkafamiya et al., 2010). The levels of K, Na, Ca and Mg were comparably higher than Mn and Cu levels. The Ca/P (0.326) was comparably lower than 0.5 which is the minimum ratio required for favourable calcium absorption in the intestine for bone formation (Nieman et al., 1992) although the level of Ca/P has been reported to have some effects on calcium in the blood of many animals (Adeyeye et al., 2012). The value of ratio (0.556) was lower than 0.6, the value that favours non-enhancement of high blood pressure disease in man. Although for normal retention of protein during growth and for balancing fluid a K/Na ratio of 1.0 is recommended (Helsper et al., 1993), the high value of K/Na ratio (1.80) obtained in the present report suggests that bringing the ratio down would require the consumption of food sources rich in Na. the Ca/Mg value obtained for the present sample (1.14) was fairly higher than the 1.0 recommended. It means both that both Ca and Mg would need adjustment for normal for normal health. The milliequivalent ratio of [K/(Ca+Mg)] (1.31) was comparably lower than 2.2 recommended, which means the sample would not promote hypomagesaemia in man (NRC, 1989; Adeyeye and Adesina, 2012). Iron and Zinc are among the required elements for humans and their daily requirements for adults are 10 and 15 respectively. Levels obtained in the present report (4.55 0.02 mg/100g) (Fe) and 25.7 mg/100g (Zn)) compared favourably with the values reported for F. asperifolia and F. sycomorus (Nkafamiya et al., 2010). However zinc requirements can easily be met by consuming this sample (25.7 mg/100g). Generally from the recommendation set out by NRC/NAS, the daily requirement of Zn, Mn and Cu can easily be met while the diets may need be supplemented with foods high in K, Na, Ca and P. The mineral safety index (MSI) values of the sample are shown in Table 3. The standard MSI for the elements are Na (4.8), Mg (15), P (10), Ca (10), Fe (6.7) and Zn (33). For Ca, P, Mg, Fe, Cu and Na, the MSI values ranged from 0.16 2.75, with all the

differences between the standard and calculated MSI values being positive. Let us take Na for an example, the calculate MSI value is 0.16 and the difference is +4.64, this meant that no amount of the sample might be overloading the body with sodium that can lead to secondary hypertension. For Ca, Mg and P all the calculated MSI were lower than standard MSI and hence within the USRDA (Hathcock, 1985). For Zn, the odd sample out, was 56.5 and the difference was -23.5. The implication of the above is that abnormally high level Zn was present in the sample. The sample could cause the reduction of Zn absorption in the small intestine in children. The Zn MSI greater than 33 are above the recommended adult intake. The minimum toxic dose is 500 mg, or 33 times the RDA (Hathcock, 1985). High doses of Zn can be harmful. Zinc supplements can decrease the amount of high density lipoprotein (HDL) circulating in the blood, increasing risk of heart disease. Excess Zn interacts with other minerals, such as Cu and Fe, decreasing their absorption. In animals, Zn supplements decrease the absorption of Fe so much that anaemia is produced (Adeyeye et al., 2012).When patients are given 150 mg of Zn per day, Cu deficiency results. Intakes of Zn only 3.5 mg/day above the RDA decrease Cu absorption (Nieman et al., 1992). In animals, Cu deficiency causes scarring of the heart muscle tissue and low levels of Ca in the bone (Adeyeye et al., 2012). The antinutrient content of the sample are listed in Table 4. These are compounds that limit the wide use of many plants due to their ubiquitous occurrence of them as natural compounds capable of eliciting deleterious effect in man and animals (Kubmarawa et al., 2008). The antinutrient factors; oxalate, tannin, saponin, phytate, alkaloids, phenolic content, phytin phosphorus and flavonoids were present in varying amounts in the sample. These anti nutritional factors tend to bind to mineral elements there by forming indigestible complex (Nkafamiya and Manji, 2006). Oxalate for instance tends to render calcium unavailable by binding to the calcium ion to form complexes (calcium oxalate crystals). These oxalate crystal formed prevent the absorption and utilization of calcium. The calcium crystals may also precipitate around the renal tubules thereby causing renal stones (Ladeji et al., 2004; Nkafamiya and Manji, 2006). In general the levels of antinutrients in Bridelia ferruginea bark sample are low to significantly interfere with nutrients utilization. They are below the established toxic level (Nkafamiya and Manji, 2006). The amino acid profile of the Bridelia ferruginea back sample is shown in Table 5. The levels of the

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amino acid ranged between 0.597 0.001 13.1 0.450 g/100g. All the essential amino acids were present in the sample, with Isoleucine having the highest concentration (6.20 0.014 g/100g). Table 6 shows the concentrations of total AA (TAA), total essential AA (TEAA), total acidic AA (TAAA), total neutral AA (TNAA), total sulphur AA (TSAA), total aromatic AA (TArAA) and their percentage values. The Leu/Ile ratios, their differences and percentage differences are contained in Table 6. Non- essential amino acids have the highest % concentration (52.2) while TEAA total essential amino acids had a % concentration of 47.8. The content of TEAA of 43.1 g/100g crude protein was close to the value for egg reference protein (56.6 g/100g cp) (Paul et al., 1976); comparably close to the values reported for soya bean (44.4 g/100g cp) (Altschul, 1958). The TAA in the current report was 90.1 g/100g cp, this value was comparably close to the values of reported for the dehulled African yam bean (AYB) (70.3 91.8 g/100g cp) (Adeyeye, 1997) but comparably higher than the values reported for raw and processed groundnut seeds (Adeyeye, 2010). The content of TSAA (1.42 g/100g) was lower than the 5.8 g/100g cp recommended for infants (FAO/WHO/UNU, 1985) The ArAA range suggested for ideal infants protein (6.8 11.8 g/100g cp ) (FAO/WHO/UNU, 1985), the present report has its value better than the minimum, i.e 8.96 g/100g cp. The ArAA are the precursors of epinephrine and thyroxin (Robinson, 1987). The % ratios of TEAA/ TAA in the sample was 47.8 % which was well above 39 % considered to be adequate for ideal protein food for infants ; 26 % for children and 11 % for adults (FAO/WHO/UNU, 1985). The TEAA/ TAA was strongly comparable to that of egg (50 %) (FAO/WHO, 1990), and 43.6 % reported for pigeon pea flour (Oshodi et al., 1993). Most animal protein are low in Cys and hence in Cys in TSAA (Adeyeye and Adamu, 2005). In contrast many vegetable proteins contain substantially more Cys than Met. The reverse is the case in the present in which the % Cys in TSAA was 42.0 %. Information on the agronomic advantages of increasing the concentration of sulphur-containing amino acid in staple foods shows that Cys had

positive effects on mineral absorption, particularly zinc (Mendoza, 2002). The P-PER value (1.69) was higher than 1.21 (cowpea), close to (Salunkhe and Kadam, 1989); 1.62 (millet ogi) and 0.27 (sorghum ogi) (Oyarekua and Eleyinmi, 2004). A common feature of sorghum and maize is that the proteins of these grains contain a relatively high proportion of leucine. It was therefore suggested that an amino acid imbalance from excess leucine might be a factor in the development of pellagra (FAO, 1995). Clinical, biochemical and pathological observations in experiments conducted in humans and laboratory animals showed that high leucine in the diet impairs the metabolism of tryptophan and niacin and is responsible for niacin deficiency in sorghum eaters (Ghafoorunissa and Narasinga Rao, 1973). High leucine is also a factor contributing to the pellagragenic properties of maize (Belavady and Gopalan, 1969). These studies suggested that the leucine/isoleucine balance is more important than dietary excess of leucine alone in regulating the metabolism of tryptophan and niacin and hence the disease process. The present Leu/Ile ratios were low in value. The pI value was 5.14. The pI of any organic matter is important when the protein isolate is to be prepared. The EAAI can be useful as a rapid tool to evaluate food formulations for protein quality. The EAAI for soy flour is 1.26 (Nielsen, 2002) which is better than the current result of 1.18. Table 7 showed that Met had the lowest score with a value of 0.26. to correct for the AA needs from the sample, it becomes 100/26 or 3.85 times as much raw sample protein to be taken (eaten) when they are the sole source of protein in the diet (Bingham,1977). In Table 8 two different scoring patterns were presented: Scorea = Essential amino acid scores based on FAO/WHO (1973) scoring pattern, Scoreb = Essential amino acid scores based on requirements of preschool child (2-5 years)(FAO/WHO/UNU, 1985). In both patterns Met + Cys had the lowest score (limiting amino acid), with values 0.41 and 0.57. and would need a correction of 100/41 or 2.44 according to the essential amino acid scoring pattern 100/57 or 1.75 times as much raw sample protein to be taken (eaten) when they are the sole protein in the diet (Bingham, 1977).

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Table 7: Amino acid score of the Bridelia ferruginea stem bark samples based on whole hen's egg amino acid

4. CONCLUSION This study has revealed that the Bridelia ferruginea back consumed in Ekiti State and other states in the South-western part of Nigeria can contribute useful amount of nutrients including amino acids to human diet. Interestingly, the anti-nutritional content of the sample was low, much lower than is obtainable in most Nigerian vegetables. This implies that, its overall nutritional value will not be affected. Indeed, this part of the plant consumed largely by the rural populace in Ekiti State is not inferior to the conventional popular Nigerian vegetables. There is need, however, to determine the vitamins and fatty acids present in the sample. Understandably, nutrient loss is of great concern during blanching and cooking of vegetables, therefore there is need to study the effects of cooking and processing procedures on nutrient availability of the Bridelia ferruginea back sample. This will help to adequately establish their importance in human nutrition and provide basis for maximum utilization of the plants part.

Table 8: Essential amino acid scores of the Bridelia ferruginea stem bark samples

Scorea = Essential amino acid scores based on FAO/WHO (1973) scoring pattern, Scoreb = Essential amino acid scores based on requirements of pre-school child (2-5 years)(FAO/WHO/UNU, 1985)

REFERENCES Adeleke RO, Abiodun OA (2010) Chemical Composition of Three Traditional Vegetables in Nigeria. Pakistan Journal of Nutrition, 9 (9): 858-860. Adesina AJ (2013). Proximate, Minerals and Antinutritional Compositions of Three Vegetables Commonly Consumed in Ekiti State,Nigeria. International Journal of Pharmaceutical and Chemical Sciences, 2(3): 1631-1638. Adeyeye EI, Adamu AS (2005). Chemical composition and food properties of

Gymnarchus niloticus (Trunk fish). Biosci. Biotechn. Res. Asia, 3(2): 266-272. Adeyeye EI (1997). Amino acid composition of six varieties of dehulled African yam bean (Sphenostylis stenocarpa) flour. Int. J. Food Sci. Nutr., 48: 345-351. Adeyeye EI (2010). Effect of cooking and roasting on the amino acid composition of raw groundnut (Arachis Hypogaea) seeds. Acta Sci. Pol., Technol. Aliment., 9(2): 201-216. Adeyeye EI, Adesina AJ (2012). Nutritional composition of African breadfruit (Treculia africana) seed testa. Journal of Agric. Res. & Dev., 11(1): 159- 178.

100

Jonathan and Funmilola Nutritional and Anti-Nutritional Composition of Bridelia Ferruginea Benth (Euphorbiaceae) Stem Back Sample

Adeyeye EI, Orisakeye OT, Oyarekua MA (2012). Composition, mineral safety index, calcium, zinc and phytate interrelationships in four fastfoods consumed in Nigeria. Bulletin of Chemical Society of Ethiopia, 26(1): 43-54. Akuodor GC, Mbah C C, Anyalewechi NA, IdrisUsman M, Iwuanyanwu TC, Osunkwo, UA (2011). Pharmacological profile of aqueous extract of Bridelia ferruginea stem bark in the relief of pain and fever. Journal of Medicinal Plants Research, 5(22): 5366-5369. Alsmeyer RH, Cunningham AE, Happich ML (1974). Equations to predict PER from amino acid analysis. Food Technol., 28, 34-38. Altschul AM (1958). Processed plant protein foodstuff. Academic Press, New York. AOAC (2005). International Official Methods of Analysis (18th edition). Association of Analytical Chemists, Washington DC. Bakoma B, Eklu-Gadegkeku K, Agbonon A, Aklikoku K. Bassene E, Gbeassor M. (2011). Preventive effect of Bridelia ferruginea against highfructose diet induced tolerance, oxidative stress and hyperlipidamia in male wistar rats. Journal of pharmacology and toxicology, 6(3): 249 - 257. Belavady B, Gopalan C (1969). The role of leucine in the pathogenesis of canine blacktongue and pellagra. Lancet 2, 956-957. Bingham S (1977). Dictionary of nutrition. Barrie and Jenkins London. Bingham S (1978). Nutrition: A consumers guide to good eating. Transworld Publishers.London. Boham BA, Kocipai-Abyazan R (1974). Flavonoids and condensed tannins from leaves of Hawairan vaccinium valiculatum andV. calycinium. Pacific Sci., 48: 458-463. Chinma CE, Igyor MA (2007). Micronutrients and anti-nutritional contents of selected tropical vegetables grown in Southeast, Nigeria. Niger. Food J., 25(1): 111- 116. Cimanga K, DeBruyne T, Apers S, Pieter L, Totte J, Kambu K, Tona L, Gill LS Akinwumi C (1999). Nigeria medicinal plants practice and belief of Ondo people.J. Ethnopharmacol, 18: 257-266. Committee on Medical Aspects (COMA) (1984). Food Policy Diet and cardiovasculardisease. HMSO. London. Day (Jnr) RA, Underwood AL (1986). Quantitative analysis 5th ed., Prentice HallPublication,London.. De Bruyne T, Cimanga K, Pieters L, Claeys M, Dominisse R, Vlietinck A (1997). Galocatechin; Epigallocatechin. A New

Biflavonoid Isolated from Bridelia ferruginea. Nat. Prod. Let., 11: 47- 52. Fafunso M, Bassir O (1977). Variations in the Loss of Vitamins in Leafy vegetables with various methods of food preparation, Food Chem, 21: 51-55. Finar IL (1975). Organic chemistry. ELBS and Longman London. FAO/WHO (1990). Protein quality evaluation Report of Joint FAO/WHO Expert Consultation.FAO Food and Nutrition Paper 51. FAO Rome. FAO/WHO/UNU (1985). Energy and protein requirement. WHO Technical Report Series 724, WHO Geneva. FAO/WHO (1973). Energy and protein requirements. Technical Report Series 522. WHO, Geneva Switzerland. FAO (1995). Sorghum and millets in human nutrition. FAO Food Nutrition Series 27. Food and agriculture Organization of the United Nations. Rome Italy. Franke RW, Chasin BH (1980). Seeds of Famine: Ecological Destruction and the Development Dilemma in the West African Sahel. Montclair, NJ: Allanheld and Osmun. Ghafoorunissa S, Narasinga-Rao BS (1973). Effect of leucine on enzymes of the tryptophan niacin metabolic pathway in rat liver and kidney. Biochem. J., 134: 425-430. Grivetti LE, Frentzel CJ, Ginsberg KE, Howell KL, Ogle BM (1987). Bush foods and edible weeds of agriculture: perspectives on dietary use of wild plants in Africa, their role in maintaining human nutritional status and implications for agricultural development.Health and disease in Tropical Africa. Geographical and Medical Viewpoints, ed. R Akhtar, pp. 5181. London: Harwood. Grivetti LE (1979): Kalahari agro-pastoral huntergatherers. The Tswana example. Ecol. Food Nutr., 7: 235 256. Grivetti LE (1978). Nutritional success in a semi-arid land. Examination of Tswana agro-pastoralists of the Eastern Kalahari, Botswana. Am. J. Clin. Nutr., 31: 12041220. Harborne JB (1973). Phytochemical methods. Capman and Hall, Ltd., London, 49-188. Hathcock JN (1985). Quantitative evaluation of vitamin safety. Pharmacy Times. 104-113. Helsper JPFG, Hoogendijk M, Van Norel A and Burger-Meyer K (1993). Antinutritional factors in faba beans (Vicia faba L.) as affected by breeding toward the absence of condensed tannin. J Agric Food Chem.41:1058-1061. Iheanacho ME, Udebuani AC (2009).Nutritional Composition of Some Leafy Vegetables

101

International Journal of Scientific Research in Knowledge, 2(2), pp. 92-104, 2014

Consumed in Imo State, Nigeria. J. Appl. Sci. Environ. Manage., 13(3): 3538. Ihekoronye AI, Ngoddy PO (1985). Integrated Food Science and technology or tropics. Macmillian publisher. London, pp.257-264. Irobi ON, Moo-Young M, Anderson WA, Daramola SO (1994). Antimicrobial activity of bark extract of Bridelia ferruginea (Euphorbiaceae). Journal of Ethnopharmacology, 43 (3): 185190. Kareem KT, Kareem SO, Adeyemo OJ, Egberongbe RK (2010). In vitro antimicrobial properties of Bridelia ferruginea on some clinicalisolates. Agriculture and Biology Journal of North America, 1(3): 416-420. Kolawole OM, Olayemi AB (2003). Studies on the efficacy of Bridelia ferruginea Benth bark in water purification. Nigerian Journal of Pure & Applied Science, 18: 1387-1394. Kolawole OM, Oguntoye SO, Agbede OO, Olayemi AB (2006). Studies on the efficacy of Bridelia ferruginea Benth bark extract on reducing the coliform load and BOD5 of domestic wastewater. Ethnobotanical Leaflet, 10: 228 238. Kolawole OM, Adesoye AA (2010). Evaluation of the antimalarial activity of bridelia ferruginea benth bark. SENRA Academic Publishers, Burnaby, British Columbia, 4(1): 1039-1044. Kubmarawa D, Andeyang IF, Magomya H (2008). Amino Acid Profile of Two Non- conventional Leafy Vegetable, Sesamum and Balanites aegyptiaca. Afr. J. Biotechnol., 7(19): 35023504. Ladeji O, Ahin CU, Umaru HA (2004). Level of antinutritional factors in Vegetables commonly eaten in Nigeria. Afr.J. Nat. Sci.7:71-73. Makkar AOS, Goodchild AV (1996). Qualification of tannins. A laboratory manual, ICARDA, Aleppo, Syria. Mendoza C (2002). Effect of genetically modified low phytic acid plants on mineral absorption. Int. J. Food Sci. Nutr., 37: 759-767. McNeely JA (1990). Conserving the Worlds Biological diversity. Transaction Books. New Brunsick. 208. National Advisory Committee on Nutrition Education (NACNE) (1983). Proposal for nutritional guidelines for healthy education in Britain. Health Education Council.London. National Research Council (NRC) (1989). Food and Nutrition Board Recommended Dietary Allowances (10th edition). National Academy Press. Washington DC.

Nelson SS (1994). Introduction to the Chemical Analysis of Foods. Jones and Bartletes Publishers, Londoon pp. 93-201. Nieman DC, Butterworth DE, Nieman CN (1992). Nutrition. Wm. C. Brown Publishers.Dubuque. Nielsen SS (2002). Introduction to the chemical analysis of foods. CBS Publ. Distrib. NewDelhi. Nkafamiya II, Osemeahon SA, Modibbo UU, Aminu A (2010). Nutritional status of non conventional leafy vegetables,Ficus asperifolia and Ficus sycomorus. African Journal of Food Science, 4(3): 104-108. Nkafamiya II, Manji AJ (2006). A Study of Cyanogenetic Glucoside Contents of some Edible Nuts and Seeds. J. Chem. Soc. Niger. 31(1 and 2): 12-14. Ogugbuaja VO, Akinniyi JA, Ogarawu VC, Abdulrahman F (1997). Elemental Contents of Medicinal Plants. Faculty of Science Monograph Series, No. 1, pp. 142. Faculty of Science, University of Maiduguri, Nigeria, pp. 142. Obadoni BO, Ochuko PO (2001). Phytochemical studies and comparative efficacy of the crude extracts of some Homostate plants in Edo and Delta States of Nigeria. Global J Pure Appl Sci.:8b:203-208. Okon EU, Akpanyung EO (2005). Nutrients and Antinutrients in selected Brands of Malt- drinks Produced in Nigeria. Pak. J.Nutr., 4(5):352-355. Olaofe O, Akintayo ET (2000). Prediction of isoelectric points of legume and oilseed proteins from their amino acid compounds. J. Techno-Sci., 4: 49-53. Olajide OA, Makinde JM, Awe SO (1999). Effect of aqueous extract of Bridelia ferruginea stem bark corrageenan-induced oedema and grand coma tissue formation rats and mice. J. Ethnopharmacol., 66 (1): 113-117. Osagie AU, Offiong AA (1998). Nutritional Quality of Plant Foods. Ambik Press, Benin City, Edo State Nigeria pp. 131-221. Oshodi AA, Olaofe O, Hall GM (1993). Amino acid, fatty acid and mineral composition of pigeon pea (Cajanus cajan). Int. J. Food Sci. Nutr., 43: 187-191. Oyarekua MA, Eleyinmi AF (2004). Comparative evaluation of the nutritional quality of corn,sorghum and millet ogi prepared by modified traditional technique. Food Agric.Environ. 2 (2), 94-99. Paul AD, Southgate AT, Russel J (1976). First supplement to McCance and Widdowsons. The composition of foods. HMSO,London.

102

Jonathan and Funmilola Nutritional and Anti-Nutritional Composition of Bridelia Ferruginea Benth (Euphorbiaceae) Stem Back Sample

Pearson D (1976). Chemical Analysis of Foods (7th edition). Churchill. London. Varian Techtron (1975). Basic AtomicAbsorption Spectroscopy-A Modern Introduction. Dominica Press. Victoria. Robinson DE (1987). Food biochemistry and nutritional value. Longman Sci. Techn. London. Salunkhe DK, Kadam SS (1989). Handbook of world food legumes, nutritional chemistry, processing technology and utilisation. Boca Raton, CRC Press Florida. Sena LP, VanderJagt DJ, Rivera C, Tsin AT, Muhamadu I, Mahamadu O, Millson M, Pastuszyn A, Glew RH (1998). Analysis of nutritional components of eight famine foods of the Republic of Niger. Plant Foods Hum. Nutr. 52: 17 30. Sparkman DH, Stein EH, Moore S (1958). Automatic Recoding Apparatus for Use in Chromotography of Amino acids. Anal.Chem., 30: 119.

Sravan PM, Venkateshwarlu GK, Vijaya BP, Suvarna D, Dhanalakshmi CH (2011). Effects of anti inflammatory activity of Amaranthus viridis Linn. Annals Biological Research, 2(4): 435438. Steel RGD, Torrie JH (1960). Principles and Procedures of Statistics. McGraw-Hill. London. Steinke FH, Prescher EE, Hopkins DT. (1980). Nutritional evaluation (PER) of isolated soybean protein and combinations of food proteins. J. Food Sci., 45: 323-327. Wheeler H, Ferrel J (1971). In:Okon, E.U and Akpanyung EO (2005).Nutrients and Antinutrients in selected Brands of Malt Drinks Produced in Nigeria. Paki. J. Nutr., 4(5): 352355. Young SM, Greaves JS (1940). Influence of variety & treatment onphytin content of wheat, Food Res., 5: 103-105.

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Adesina, Adeolu, J. is a Ph.D candidate in Food/ Analytical Chemistry of the Department of Chemistry, Ekiti State University, Ado Ekiti, Nigeria. He received his first degree from University of Ilorin, Kwara State, Nigeria 2001 awarded with Bachelor of Science in Pure Chemistry. He obtained degree in Master of Science in Analytical Chemistry from University of Ibadan, Nigeria in 2006. His current research focuses on Food chemistry and quality. To date, he has published several scientific journal articles related to Food chemistry and quality evaluation field.

Akomolafe Seun Funmilola is a Ph.D candidate in Food biochemistry and toxicology of the Department of Biochemistry, Federal University of Technology, Akure, Nigeria. She received her first degree from Ekiti State University, Ado Ekiti Nigeria. 2005 awarded with Bachelor of Science in Biochemistry. She obtained degree in Master of Science in Biochemistry from University of Ibadan, Nigeria in 2009. Her current research focuses on Food Biochemistry and quality. To date, she has published several scientific articles related to food Biochemistry and toxicology field.

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