Food Sci. Biotechnol. 20(1): 143-149 (2011) DOI 10.

1007/s10068-011-0020-y

RESEARCH ARTICLE

Nutritional Analysis and Antioxidant Activity of Palmyrah (Borassus flabellifer L.) Seed Embryo for Potential Use as Food Source
Karuppusamy Arunachalam, Shanmugam Saravanan, and Thangaraj Parimelazhagan

Received: 18 August 2010 / Revised: 15 October 2010 / Accepted: 9 November 2010 / Published Online: 28 February 2011 KoSFoST and Springer 2011

Abstract The present study was carried out with ungerminated seed embryos of palmryah to evaluate nutritional quality with respect to minerals and fiber components, total phenols, and antioxidant properties. It is found to be good source of carbohydrate, fiber, fat, amino acids, and protein. Analysis of macro and micronutrient composition showed potent source of sodium, potassium, calcium, magnesium, zinc, and iron. In vitro antioxidant activity was evaluated by different assays, including 2,2-diphenylpicryl-1-picrylhydrazyl (DPPH) radical scavenging, 2,2'azinobis (3-ethylbenzothiozoline-6-sulfonic acid) disodium salt (ABTS+) assay, ferric reducing antioxidant power (FRAP) assay, phospomolybdenum reduction assay, metal chelating activity, and hydroxyl radical scavenging activity. The results indicate that this plant seed embryo possesses micro, macro nutrients and antioxidant properties and has nutraceuticals potential for the treatment of malnutrition. Keywords: nutritional value, mineral content, antioxidant, food supplement, palmyrah (Borassus flabellifer) seed embryo

Introduction
Plants provide the basic diet for over 5 billion people in our world today (1). Even today, isolated populations of the world such as the Mbuti hunter-gatherers of the Angolan tropical rainforest consume a large variety of plants (of which 41 are fruits, 25 seeds, 13 tubers, and 21 leaves) (2).
Karuppusamy Arunachalam, Shanmugam Saravanan, Thangaraj Parimelazhagan ( ) Bioprospecting Laboratory, Department of Botany, Bharathiar University, Coimbatore-641 046, Tamil Nadu, India Tel: +91-422-2428305; Fax: +91-422-2425706 E-mail: drparimel@gmail.com

In contrast, western civilizations rely on about 6 major staples (wheat, potato, rice, oats, barley, and corn) as their source of dietary energy. This reduction in plant use has both nutritional and agricultural implications. A reduction in plant biodiversity and use means a significant loss of valuable food sources for mankind. Moreover, the use of a smaller range of plants as staples may reduce the amount and quantity of micronutrients required by man. Insufficient intake of micronutrients such as zinc, iron, calcium, or vitamin A may lead to deficiency diseases in man. Societies of the world that use a large range of plant species and a variety of staples are therefore to be encouraged in order to meet optimum nutritional requirements. The palmyrah, or toddy palm (Borassus flabellifer L.) belonging to Arecaceae, grows wild from the Persian Gulf to the CambodianVietnamese border. It is commonly cultivated in India, Southeast Asia, Malaysia, and occasionally in other warm regions including Hawaii and southern Florida. It has innumerable medicinal uses for all parts of the Palmyra palm. The plant is said to relieve biliousness, dysentery, and gonorrhea. Young roots are diuretic and anthelmintic, and a decoction is given for certain respiratory diseases. The ash of the spadix is taken to relieve heart burn and enlarged spleen and liver. The bark decoction with salt is used as mouth wash, and charcoal made of the bark serves as a dentifrice. Sap from the flower stalk is prized as tonic, diuretic, stimulants, laxative and anti phlegmatic, and amebicide. Sugar made from this sap is used to counteract poisoning and it is remedy for coughs and various pulmonary compliant. Fresh toddy heated to promote fermentation, is bandaged on to all kinds of ulcers. The pulp of the mature fruit relieves dermatitis. Its fruit is a kind of drupe, large, and fibrous with usually 3 to 5 nutlike portions, each of which encloses a seed when it is tender. The seeds contain a soft sweet glutinous pulp with a little liquid in them. The tender pulp gradually hardens

144

Arunachalam et al.

and develops a fibrous kernel. The presence of galactomannan from the soft kernel of palm was first reported by Subrahmanyan et al. (3) and show that it consisted of mannose (78.32%) and galactose (27.64-28.06%). Recently Pathberiya and Jansz (4) studied the in vitro antioxidant capacity of carotenoids of palmyrah fruit pulp from mannar and also demonstrated the nature of free sugars and polysaccharides present in the kernel. Palmyrah is a good source of carbohydrate, calcium, magnesium, iron, and fiber but, limited in fat and protein. Further research is needed to exploit its full potential that may influence its extensive consumption (5). No information is available regarding nutritional and antioxidant activity of the seed embryo. Therefore, the present study was carried out to evaluate nutritional quality, particularly with respect to minerals and fiber components, of selected new adapted cereals for the food industry.

collected during the month of January, 2009, from Coimbatore, Tamil Nadu, India. The freshly collected seed embryo materials were washed thoroughly in tap water, shade dried at room temperature (25oC), powdered, and used for solvent extraction. The powder sample was successively extracted with petroleum ether (for disposing lipid and pigments), chloroform, acetone, and methanol using Soxhlet apparatus and the air dried. Each time before extracting with the next solvent, the material was dried in hot air oven at 40C. The solvents were evaporated using a rotary vacuum-evaporator (RE300; Yamato, Tokyo, Japan) at 50C and the remaining water was removed by lyophilization (4KBTXL-75; VirTis Benchtop K, NY, USA). The extract recovery in different solvents was expressed as percent of the sample dry matter. The freezedried extracts thus obtained were dissolved in the respective solvents at the concentration of 1 mg/mL and used for assessment of antioxidant capacity through various chemical assays. Determination of total phenolic and tannin contents The total phenolic content of palmyrah seed embryo extracts was determined by Folin-Ciocalteu method. Using the same extract the tannins were estimated after treatment with polyvinyl polypyrrolidone (PVPP). The amount of total phenolics and tannins were calculated as the tannic acid equivalents (TAE) as described by Siddhuraju and Becker. (11). DPPH radical scavenging property The 2,2-diphenylpicryl-1-picryl-hydrazyl radical (DPPH) scavenging activity of palmyrah seed embryo extracts was measured according to the method of Blios (12). IC50 values of the extract i.e., concentration of extract necessary to decrease the initial concentration of DPPH by 50% was calculated. Ferric reducing antioxidant power (FRAP) assay The antioxidant capacity of solvent extracts of samples was estimated as described as Pulido et al. (13), FRAP reagent (900 L), prepared freshly and incubated at 37oC, was mixed with 90 L of distilled water and 30 L of test sample or methanol (for the reagent blank). The test samples and reagent blank were incubated at 37oC for 30 min in a water bath. The final dilution of the test sample in the reaction mixture was 1/34. The FRAP reagent contained 2.5 mL of 20 mol/L 2,4,6-tripyridyl-2-triazine (TPTZ) solution in 40 mol/L HCl plus 2.5 mL of 20 mol/L FeCl36H2O and 25 mL of 0.3 mol/L acetate buffer (pH 3.6) as described by Siddhuraju and Becker (11) at the end of incubation, the absorbance readings were taken immediately at 593 nm, using a spectrophotometer. Methanolic solutions of known Fe (II) concentration, ranging from 100 to 2,000 mol/L, (FeSO47H2O) were

Materials and Methods

Proximate composition Moisture, crude fat, protein, dietary fiber, and ash were determined according to standard AOAC method (6). The protein content of the samples was calculated using the factor N6.25. Carbohydrate was estimated by subtracting the sum of moisture, protein, fat, fiber, and ash content from 100 g. Calcium, potassium, and sodium were estimated using a flame photometer. Iron and magnesium were determined by use of an absorption spectrophotometer (Baird Alpha 2AA; Baird Corporation, Bedford, UK). Gross energy was determined using bomb calorimetry (7). Samples were analyzed in triplicate and the results reported are the mean valuesstandard deviation (SD). Amino acid analysis Samples (300 mg), in triplicate from each cultivar, were hydrolyzed with 6 N HCl in an evacuated test tube for 24 hr at 105oC. The dried residue was dissolved in citrate buffer (pH 2.2) after flash evaporation. Aliquots were analyzed in an automatic amino acid analyzer (KLA 3B; Hitachi Perkin-Elmer, Stuttgart, Germany), using the buffer system described earlier (8). Methionine and cystine were analysed separately after performic acid treatment and subsequent hydrolysis with HCl (9). Tryptophan was determined after alkali (NaOH) hydrolysis by the colorimetric method (10). Essential amino acids score was calculated with reference to the FAO/WHO reference amino acid pattern (19), Amino acid score= Test amino acid 100 Reference amino acid

Preparation of extracts Palmyrah seed embryo, is

Nutritional Analysis of Palmyrah Seed Embryo

145

used for the preparation of the calibration curve. The FRAP value is expressed as mmol Fe (II) equivalent/mg extract. Antioxidant activity by the ABTS+ assay 2,2'Azinobis (3-ethyl-benzothiozoline-6-sulfonic acid) disodium salt (ABTS) was dissolved in water to a 7 mM concentration. ABTS radical cation (ABTS+) was produced by reacting ABTS stock solution with 2.45 mM potassium persulfate (final concentration) and allowing the mixture to stand in the dark at room temperature for 12-16 hr before use. Prior to assay, the solution was diluted in ethanol (about 1:89 v/ v) and equilibrated 30C to give an absorbance at 734 nm of 0.700.02 in a 1-cm cuvette (14). The concentration of the extracts that produced between 20-80% inhibitions of the blank absorbance was determined and adapted. After the addition of 1 mL of diluted ABTS+ solution to 10 L of palmyrah seed embryo extracts or Trolox standards (final concentration 0-15 M) in ethanol, optical density (OD) was taken at 30C exactly 30 min after the initial mixing. The unit of total antioxidant activity (TAA) is defined as the concentration of Trolox having equivalent antioxidant activity expressed as mol/g sample extracts on dry matter. Metal chelating activity The chelating of ferrous ions by various extracts of palmyrah seed embryo was estimated by the method of Dinis et al. (15). Briefly the extract samples (250 L) were added to a solution of 2 mmol/L FeCl2 (0.05 mL). The reaction was initiated by the addition of 5 mmol/L ferrozine (0.2 mL) and the mixture was shaken vigorously and left standing at room temperature for 10 min. Absorbance of the solution was then measured spectrophotometrically at 562 nm. The chelating activity of the extracts was evaluated using ethylenediamine tetraacetic acid (EDTA) as standard. The results were expressed as mg EDTA equivalent/g extract. Phosphomolybdenum assay The antioxidant activity of samples was evaluated by the green phosphomolybdenum complex formation according to the method of Prieto et al. (16). An aliquot of 100 L of sample solution (in 1 mM dimethyl sulfoxide, DMSO) was combined with 1 mL of reagent solution (0.6 M sulphuric acid, 28 mM sodium phosphate, and 4 mM ammonium molybdate) in a 4-mL vial. The vials were capped and incubated in a water bath at 95oC for 90 min. After the samples had cooled to room temperature, the absorbance of the mixture was measured at 695 nm against a blank. The results reported (ascorbic acid equivalent antioxidant activity) are mean values expressed as g of ascorbic acid equivalents/100 g extract. Hydroxyl radical scavenging activity The scavenging

activity of petroleum ether, chloroform, acetone, and methanol extracts of palmyrah seed embryo on hydroxyl radical was measured according to the method of Klein et al. (17). Various concentrations (50, 100, 150, and 200 g) of extracts were added with 1.0 mL of iron-EDTA solution (0.13% ferrous ammonium sulfate and 0.26% EDTA), 0.5 mL of EDTA solution (0.018%), and 1.0 mL of DMSO (0.85%, v/v in 0.1 M phosphate buffer, pH 7.4). The reaction was initiated by adding 0.5 mL of ascorbic acid (0.22%) and incubated at 80-90oC for 15 min in a water bath. After incubation, the reaction was terminated by the addition of 1.0 mL of icecold trichloroacetic acid (TCA) (17.5%, w/v). Three mL of Nash reagent (75.0 g of ammonium acetate, 3.0 mL of glacial acetic acid, and 2 mL of acetyl acetone were mixed and raised to 1 L with distilled water) was added and left at room temperature for 15 min. The reaction mixture without sample was used as control. The intensity of the color formed was measured spectroscopically at 412 nm against reagent blank. The % hydroxyl radical scavenging activity (HRSA) is calculated by the following formula: %HRSA=[(control ODsample OD)/control OD]100 Statistical analysis All analyses were carried out in triplicate and the data were reported as meansSD. The data were subjected to one-way analysis of variance (ANOVA) and the significance of the difference between means was determined by Duncans multiple range test (p<0.05) using statistical (Stat Soft Inc., Tulsa, OK, USA).

Results and Discussion

The proximate composition of seed embryo (g/100 g) dry matter examined is shown in Table 1. Palmyrah seed embryo yielded an energy level of 1,398 kJ, with a high content of carbohydrate (71.5 g), protein (12.5 g), and fiber (4.3 g). In contrast, its fat content was found to be low (1.9 g). The low moisture content recorded (65.5%), makes it a stable product for prolonged period of time. The results obtained shows seed embryo to possess better nutritional quality and comparable with that of palmyrah odiyal (18). This indicates that the sample is endowed with essential nutrients required for human consumption. The analysis of mineral contents is of nutritional importance as they offer a potential benefit in the normal embryo metabolism of human body. The mineral level of seed embryo estimated is presented in Table 1. The sample was found to contain highest amount of potassium (68 mg), sodium (52 mg), and calcium (48 mg). Magnesium recorded a moderate level of 23 mg whereas the iron content was low containing 0.5 mg. The amino acid composition of the seed embryo sample

146
Table 1. Proximate composition and mineral profiles of palmyrah seed embryo (g/100 g) Sample Seed embryo
1)

Arunachalam et al.

Energy (kJ/100 g) 1,398

1)

Moisture Protein 65.5 12.5

Fat 1.9

Carbohydrate 71.5

Fiber 4.3

Sodium Potassium Calcium (mg/100 g) 52 68 48

Magnesium 23

Iron 0.5

(g/100 g)

Mean of 3 replicates, expressed on dry weight basis

Table 2. Amino acid profile of palmyrah seed embryo Amino acid (g/100 g protein) Histidine Lysine Leucine Isoleucine Methionine+cysteine Phenylalanine+tyrosine Threonine Tryptophan Valine
1) 2)

Seed embryo
2)

Reference pattern FAO/WHO (1985)1) 1.9 5.8 6.6 2.8 2.5 6.3 3.4 1.1 3.5

Table 3. Solvent extract recovery, total phenolics, and tannins content of palmyrah seed embryo Sample1) BFEP BFEC BFEA BFEM
1)

5.8 1.2 8.7 5.8 1.2 8.6 6.2 1.8 8.2

2)

Extract yield (%) 0.8 1.0 6.5 7.8

Total phenolics (g/100 g extract) 10.5560.727 13.7570.684 37.4341.5090 18.3942.083

Tannins (g/100 g extract) 0.690.12 1.800.89 16.723.350 6.993.19

BFEP, palmyrah petroleum ether extract of seed embryo; BFEC, palmyrah chloroform extract of seed embryo; BFEA, palmyrah acetone extract of seed embryo; BFEM, palmyrah methanol extract of seed embryo. Values are meanSD (n=3).

Amino acid reference pattern of protein for children (2-5 years old) diet Values are % of protein. Each amino acid in the reference pattern was presumed to score a value=100; Values for each cultivar is expressed relatively to the reference pattern.

along with the essential amino acid requirements pattern suggested by FAO/WHO is shown Table 2. The amino acid profile of the sample revealed that the proteins of the seed embryo contained adequate levels of amino acid as compared with FAO/WHO (19). The amount of amino acids, histidine, leucine, iso leucine, phenylalanine+tyrocine, threonine, tryptophan, and valine, were found to be higher, whereas the amino acid lysine, methonine+cysteine, recorded lower level but appreciable content than the reference pattern of FAO/WHO (19). Amino acids in the form of proteins make up the greatest portion of our body weight. Apart from this, cysteine was found to have the ability to quench DPPH radical (20). Arginine plays vital roles in nutrition and metabolism. It is classified as an essential amino acid in infants and young children and as a conditionally essential amino acid in adults at times of trauma or disease. Arginine is metabolized to nitric oxide, creatine, ornithine, proline, polyamines, and protein (21). Amino acid composition generally indicates the nutritive value of a protein source (22). The chemical score and amino acid index are widely used for screening potential protein foods. Amino acid deficiency can be met by consuming large amounts of legumes, or by taking a mixture of legumes, or by employing the complimentarily that exists between high sulphur amino acid cereals and legumes. Recovery percent, total phenolics, and tannin contents Polyphenols, widely distributed in plants, have gained

much attention, due to their antimutagenic, antitumor, antioxidant, and free radical scavenging abilities, which potentially have beneficial implications for human health (23). The maximum extractable total phenolics and tannins were recorded in acetone extracts followed by methanol extracts. Petroleum ether gave the lowest levels of yield, total phenolics, and tannins for the plant samples used Table 3. DPPH radical scavenging property The results on DPPH radical scavenging activity of the different solvent extracts along with the reference standards quercetin, butylated hydroxyl anisole (BHA), butylated hydroxyl toluene (BHT), and -tocopherol are shown in Fig. 1. The model of stable DPPH free radicals can be used to evaluate the antioxidative activities in a relatively short time. The absorbance decreases as a result of a color change from purple to yellow as radical is scavenged by antioxidants through donation of hydrogen to form the stable DPPH molecule. Concentration of the sample necessary to decrease initial concentration of DPPH by 50% (IC50) under the experimental condition was determined. Therefore, lower value of IC50 indicates a higher antioxidant activity. All solvent extracts showed excellent DPPH radical scavenging activity except petroleum ether extract. Acetone extract of (171.23 g/mL) seed embryo showed higher levels of free radical scavenging activity among the solvent extract tested. The DPPH radical scavenging activity was found to be the least in petroleum ether and chloroform extracts of seed embryo (IC50 793.65 and 684.93 g/mL, respectively). The scavenging effect of various solvent extracts and standards with the DPPH radical is in the following order: quercetin>BHA>BHT>-T>BFEA> BFEM>BFEC>BFEP. Several studies have evaluated the

Nutritional Analysis of Palmyrah Seed Embryo

147

Fig. 1. DPPH radical scavenging activity of various solvent extracts from palmyrah seed embryo. Values are meanSD of 3 determinations; Values are amount of sample necessary to decrease initial concentration of DPPH by 50% (IC50).

relationship between antioxidant activity of plant products and their phenolics content. The obtained data revealed that all the extracts of palmyrah seed embryo contain powerful inhibitor compounds, which may act as primary antioxidants that react with DPPH radicals. FRAP Antioxidants, explained as reductants and inactivation of oxidants by reductants, are involved in redox reactions in which one reaction species is reduced at the expense of the oxidation of another antioxidant. The antioxidant potential of various extracts of seed embryo were estimated from their ability to reduce TPTZ-Fe(III) complex to TPTZ-Fe(II) complex and the results are expressed as concentration of substance having ferric-TPTZ reducing ability equivalent that of 1 mmol concentration of Fe(II). The FRAP values for different solvent extracts of seed embryo is depicted in Table 4. Among the various solvent extracts, methanol extract of seed embryo (863.8 mmol Fe(II)/mg extract) registered higher antioxidant activity. All extracts showed reducing power but not at the same level. The order of FRAP activity of various sample extracts is as follows: BFEM>BFEC>BFEP>BFEA. Similarly, Tachakittirungrod et al. (24) have also reported that

methanol extract of guava leaves exhibited a higher FRAP reducing property. Hence they should be able to donate electrons to free radicals stable in the actual biological and food system. ABTS+ cation radical scavenging activity ABTS+, a protonated radical, has characteristic absorbance maximam at 734 nm which decreases with the scavenging of the proton radicals. ABTS+ was generated by incubating ABTS with potassium persulfate. The presence of chemical compounds in the tested extracts that inhibit the potassium persulfate activity may reduce the production of ABTS+. The efficacy of ABTS cation radical scavenging activity of various solvent extracts of seed embryo is shown in Table 4. In the present investigation, the acetone and methanol extracts of seed embryo registered the highest TAA (5,206.9 and 3,438.8 mol/g). The petroleum ether extracts of the sample showed lower level of activity while the chloroform extracts registered moderate activity. Metal chelating activity Iron is essential for life because it is required for oxygen transport, respiration, and activity of many enzymes. In complex systems, such as food and

Table 4. FRAP assay, ABTS+ scavenging activity, iron chelating activity, and phosphomolybdenum assay of petroleum ether, chloroform, acetone, and methanol extracts of palmyrah seed embryo Sample BFEP BFEC BFEA BFEM
1) 2)

FRAP1) (mol Fe (II)/mg extract) 0614.051.64) 637.341.1 239.614.7 863.825.5

TAA2) (mol/g extract) 589.1750.0 0,1,4061,819.0 5,206.96,815.3 3,438.84,498.6

Metal chelating property (mg EDTA/g extract) 46.733.14 53.798.93 30.011.58 20.066.62

AEAC3) (g AA/100 g extract) 10.60.5 16.31.7 40.75.6 28.25.6

Concentration of substance having ferric-TPTZ reducing ability expressed as mmol Fe(II) equivalents Total antioxidant activity (mol equivalent Trolox performed by using ABTS+ radical cation) 3) Ascorbic acid equivalent antioxidant capacity (g equivalent of ascorbic acid/100 g extract) through the formation of phosphomolybdenum complex 4) Values are meanSD (n=3); Mean values followed by different superscripts in a column are significantly different (p<0.05).

148

Arunachalam et al.

food preparations, various different mechanisms may contribute to oxidative processes, such as Fenton reaction, where transition metal ions play a vital role. Different reactive oxygen species might be generated and various target structures such as lipids, proteins, and carbohydrates, can be affected. Therefore, it is important to characterize the extracts by a variety of antioxidant assays (25). The chelating effect on the ferrous ions by the various solvent extracts of seed embryo is presented in Table 4. All the sample extracts exhibited the ability to chelate metal ions. Among the different sample extracts, the chloroform extract of seed embryo showed higher activity (53.79 mg EDTA/g extracts). Further, the activity decreased in methanol extract of seed embryo part (20.06 mg EDTA/g). Metal chelating capacity was significant as they reduced the concentration of the catalyzing transition metal in lipid peroxidation (26). It was already reported that chelating agents which form -bonds with a metal, are effective as secondary antioxidants because they reduce the redox potential, thereby stabilizing the oxidized form of the metal ion (27). Antioxidants inhibit interaction between metal and lipid through formation of insoluble metal complexes with ferrous ion. Hence the data obtained for seed embryo reveals that some of the extracts demonstrate an effective capacity for iron binding, suggesting that its action as antioxidant may be related to its iron binding capacity. Phosphomolybdenum assay The phosphomolybdenum method is based on the reduction of Mo(VI) to Mo(V) by the antioxidant compounds and the formation of green phosphate/Mo(V) complex with the maximal absorption at 695 nm. Among the various extracts evaluated, the acetone palmyrah seed embryo had the strongest phosphomolybdenum reduction (56.7 g AA/100 g) and the values were significantly higher (p<0.05). All the other sample extracts registered moderate phosphomolybdenum reduction (10.6 to 28.2 g AA/100 g), (Table 4). Hydrogen/electron transfer from antioxidants to DPPH radical and Mo(VI) complex occur in the DPPH radical and phosphomolybdenum assays, respectively (28). Hydroxyl radical scavenging activity Scavenging of hydroxyl radical is an important antioxidant activity because of very high reactivity of the OH radical, enabling it to react with a wide range of molecules found in living cells, such as sugars, amino acids, lipids, and nucleotides (29). Thus, removing OH is very important for the protection of living systems. The hydroxyl radical scavenging potential of various solvent extracts of palmyrah seed embryo is shown in Fig. 2. Each extract showing hydroxyl radical scavenging activity was increased with increasing concentration of sample extracts. In the present investigation, the hydroxyl radical scavenging activity observed were in the range of

Fig. 2. Hydroxyl radical scavenging activity of petroleum ether, chloroform, acetone, and methanol extracts of palmyrah seed embryo. Values are meanSD (n=3).

(28.2-42.8%) in seed embryo part at the concentration of 200 g/mL. While scavenging hydroxyl radical, the ability of acetone extract of seed embryo (50.5%) was found to be higher than other sample extracts. In the present study, the nutritional composition of palmryah seed embryo exhibited a good source of carbohydrate, protein, and fiber but limited in fat. In the micronutrient composition, the calcium, magnesium, and iron are valuable sources. In addition to the nutritional composition, the seed embryo exhibited higher antioxidant potential. The antioxidant foods are valuable research contribution, expanding the research evidence base for plant-based nutritional research and may be utilized in epidemiological studies where reported food intakes can be assigned antioxidant values. It can also be used to test antioxidant effects and synergy in experimental animal and cell studies or in human clinical trials. The ultimate goal of this research is to combine these strategies in order to understand the role of dietary phytochemical antioxidants in the prevention of cancer, cardiovascular diseases, diabetes, and other chronic diseases related to oxidative stress.

References
1. Harris DR. An evolutionary continuum of people-plant interaction. pp. 11-26. In: Foraging and Farming, the Evolution of Plant Exploitation. Harris DR, Hillman GC (eds). Unwin Hyman, London, UK (1989) 2. Ichikawa M. Diversity and selectivity in food of the hunter-gatherer in Zaire. Vol. XII, pp. 487-496. In: Tropicd Forest, People, and Foods. Hladik CM, Hladik C, Linarcs OF, Pagezy H, Semple A, Hadlcy M (eds). Lanes. The Parthenon Publishing Group Ltd., London, UK (1993) 3. Subrahmanyan GS, Bains CP, Natarajan DS, Bhatia V. The carbohydrates of tender kernel of palmyra palm (Borassus

Nutritional Analysis of Palmyrah Seed Embryo

flabellifer, L.). Arch. Biochem. Biophys. 60: 27-34 (1956) 4. Pathberiya LG, Jansz ER. Studies on the carotenoids and in vitro antioxidant capacity of palmyrah fruit pulp from mannar. J. Nat. Sci. F. Sri Lanka 33: 269-272 (2005) 5. Mason D, Henry CJK. Chemical composition of palmyrah (Borassus flabellifer) seed shoots-odiyal. Int. J. Food Nutr. 45: 287290 (1994) 6. AOAC. Official Methods of Analysis. 16th ed. Method 1094. Association of Official Analytical Chemists, Arlington, VA, USA (1995) 7. Miller DS, Payne DR. A ballistic bomb calorimeter. Brit. Nutr. 13: 501-508 (1959) 8. Zarkadas CG, Yu Z, Voldeng HD, Minero-Amador. Assessment of the protein quality of new high-protein soya bean cultivars by amino acid analysis. J. Agr. Food Chem. 41: 616-623 (1993) 9. Khalil LA, Durani FR. Haulm and hull of peas as a protein source in animal feed. Sarhad J. Agr. 6: 219-225 (1990) 10. Freidman M, Finely JW. Methods of tryptophan analysis. J. Agr. Food Chem 19: 626-631 (1971) 11. Siddhuraju P, Becker K. Studies on antioxidant activities of mucuna seed (Mucuna pruriens var. utilis) extracts and certain non-protein amino/imino acids through in vitro models. J. Sci. Food Agr. 83: 1517-1524 (2003) 12. Blios MS. Antioxidants determination by the use of a stable free radical. Nature 26: 1199-1200 (1958) 13. Pulido R, Bravo L, Sauro-Calixto F. Antioxidant activity of dietary polyphenols as determined by a modified ferric reducing/antioxidant power assay. J. Agr. Food Chem. 48: 3396-3402 (2000) 14. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice- Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Bio. Med. 26: 1231-1237 (1999) 15. Dinis TCP, Madeira VMC, Almeida LM, Action of phenolic derivatives (acetoaminophen, salycilate, and 5 aminosalycilate) as inhibitors of membrane lipid peroxidation and as peroxyl radical scavengers. Arch. Biochem. Biophys. 315: 161-169 (1994) 16. Prieto P, Pineda M, Aguilar M. Spectophotometric quantitative of antioxidant capacity through the formation of a phosphomolybdenum complex, specific application to the determination of vitamin E. Anal. Biochem. 269: 337-341 (1999) 17. Klein SM, Cohen G, Cederbaum AI. Production of formaldehyde during metabolism of dimethyl sulphoxide by hydroxyl radical

149
generating system. Biochemistry 20: 6006-6012 (1991) 18. Jansz ER, Nadi T, Wickremasekara, Sumuduni KAV. A review of chemistry and biochemistry of seed shoot flour and fruit of the palmyrah palm (Borassus flabellifer L). J. Nat. Sci. F. Sri Lanka 30: 61-87 (2002) 19. FAO/WHO/UNU. Energy and protein requirements. WHO Technical Report Series No. 724. World Health Organization, Geneva, Switzerland (1985) 20. Yokozawa T, Chen CP, Dong E, Tanaka T, Nonaka GI, Nishioka I. Study on the inhibitory effect of tannins and flavonoids against DPPH radical. J. Food Agr. Chem. 56: 213-222 (1998) 21. Wu G, Meininger CJ. Arginine nutrition and cardiovascular function. J. Nutr. 130: 2626-2629 (2000) 22. Bodwell CE, Satterlee LD, Hackler LR. Protein digestibility of the same protein preparations by human and rat assays and by in vitro enzymatic digestion methods. Am. J. Clin. Nutr. 33: 677-686 (1980) 23. Govindarajan R, Singh DP, Rawat AKS. High-performance liquid chromatographic method for the quantification of phenolics in Chyavanprash a potent Ayurvedic drug. J. Pharm. Biomed. Anal. 43: 527-532 (2007) 24. Tachakittirungrod S, Okonogi S, Chowwanapoonpohn S. Study on antioxidant activity of certain plants in Thailand: Mechanism of antioxidant action of guava leaf extract. Food Chem. 103: 381-388 (2007) 25. Halliwell B. Antioxidants the basics-what they are and how to evaluate them. Adv. Pharmcol. 38: 3-20 (1997) 26. Duh PD, Tu YY, Yen GC. Antioxidant activity of water extract of Harng Jyur Chrysanthemum morifolium Ramat. Lebensm.-Wiss. Ttechnol. 32: 269-277 (1999) 27. Keowmaneechai E, McClements DJ. Influence of EDTA and citrate on thermal stability of whey protein stabilized oil-in-water emulsions containing calcium chloride. Food Res. Int. 39: 230-239 (2006) 28. Hyder RC, Lio ZD, Khodr HH. Metal chelation of polyphenols. Method Enzymol. 335: 192-203 (2001) 29. Loo AY, Jain K, Darah I. Antioxidant activity of compounds isolated from the pyroligneous acid, Rhizophora apiculata. Food Chem. 107: 1151-1160 (2008) 30. Wang H, Gao XD, Zhou GC, Cai L, Yao W B. In vitro and in vivo antioxidant activity of aqueous extract from Choerospondias axillaries fruit. Food Chem. 106: 888-895 (2008)