Comparative analysis of the phytochemical and nutrient

Chemistry and Materials Research www.iiste.org
ISSN 2224- 3224 (Print) ISSN 2225- 0956 (Online)
Vol.3 No.9, 2013
91
Comparative Analysis of the Phytochemical and Nutrient
Evaluation of the Seeds and Leaves of Plukenetia
conophora plant
P. B. Ayoola1
, O. O. P. Faboya2
and O. O. E. Onawumi2
1
Department of Science Laboratory Technology, Ladoke Akintola University of Technology, P.M.B.4000,
Ogbomoso, Oyo State, Nigeria.
E-mail: pbayoola@yahoo.com
2
Department of Chemistry, Ladoke Akintola University of Technology, P.M.B.4000, Ogbomoso, Oyo State,
Nigeria.
2
Department of Chemistry, Ladoke Akintola University of Technology, P.M.B.4000, Ogbomoso, Oyo State,
Nigeria.
E-mail: estherdr@rocketmail.com
ABSTRACT
Samples of Plukenetia conophora fresh seeds and fresh leaves were analysed for Proximate composition,
secondary metabolites, Vitamins and Mineral constituents. The result of proximate analysis shows that the fresh
seeds and leaves respectively contained 46.03±0.03% and 29±0.71% moisture, 4.28±0.01% and 5.63±0.08%
fat, 7.34±0.71% and 14.92±0.04% fibre, 21.65±0.5% and 16.62±0.30% protein, 5.27±1.35% and 12.89±0.02%
Ash, 18.10±0.01% and 20.94±0.01% carbohydrate. The phytochemical screening and subsequent quantification
of the seed and leaves revealed the presence of tannin, 0.089±0.01mg/kg and 0.560±0.01mg/kg, alkaloid,
2.380±0.2mg/kg and 2.670±0.02, gallic acid, 0.024±0.01mg/kg and 0.180±0.02mg/kg. There was no traces of
saponin and anthraquinones in the fresh seed, while flavonoids, cardiac glycosides, anthraquinones, ellagic acid
and caffeinic acid were absent in both part of the plant samples. The mineral analysis revealed the constituents to
be K, 6250±0.2mg/kg and 15937±0.02, Na, 4830±0.1mg/kg and 7980±0.01mg/kg, Ca, 4337.5±0.08mg/kg and
18700±0.02mg/kg, Mg, 1711.25±0.8mg/kg and 1766.25±0.1mg/kg, Fe, 110±0.04mg/kg and 4610±0.10mg/kg,
Zn, 40.10±0.10mg/kg and 61.15±0.08mg/kg, Mn, 22.00±0.01mg/kg and79.50±0.03mg/kg, Cu, 16.45±0.03mg/kg
and 8.60±0.10mg/kg for the fresh seed and leaves respectively.
Vitamin composition results showed that the fresh seed and leaves respectively contained Thiamine (B1)
0.06±0.008μg/100g and 0.29±0.01µg/100g, Riboflavin (B2) 0.02±0.008μg/100g and 0.34±0.01µg/100g, Niacin,
0.05±0.02μg/100g and 0.12±0.3µg/100g, Cyanocobalamin (B12),0.12±0.1μg/100g and 0.23±0.03µg/100g.
Ascorbic acid (C) 4.15±0.02mg/100g and 16.28±0.04mg/100g, tocopherol (E), 122.57±0.20µg/100g and
2.67±0.008µg/100g. The results proved that Plukenetia conophora fresh seeds and leaves can serve as foods and
could also be potential sources of useful drugs formulation.
Key words: Plukenetia, conophora, phytochemical, nutrient,

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92
INTRODUCTION
Plukenetia conophora , formerly called Tetracarpidium conophorum (family Euphorbiaceae) is found
in Nigeria and Cameroon. It is a climbing shrub 10-20ft long, it is known in the Southern Nigeria as ukpa (Igbo),
Western Nigeria as awusa or asala (Yoruba). It is found in Uyo, Akamkpa, Akpabuyo, Lagos, Kogi , Ajaawa-
Ogbomoso and Ibadan. This plant is cultivated principally for the nuts which are cooked and consumed as
snacks [20]. A bitter taste is usually observed upon drinking water immediately after eating the nuts. This could
be attributed to the presence of chemical substances such as alkaloid. [3] reported the presence of oxalate,
phylates and tannin in the raw Tetracarpidium conophorum nuts. [9] also reported the proximate composition,
ascorbic acid and heavy metal contents of the nut. The work done by [25] revealed the presence of amino acid
and fatty acid components of the nut and the use of its leaf juice for the treatment of prolonged and constant
hiccups.
The impact of traditional processing on the nutrient and sensory qualities of the nut. the methods of
processing the Tetracarpidium conophorum nuts [21], and in processing waste in livestock feed formulation
have been reported. [10] compared the level of toxicant in the seeds of Terminalia catappa (Indian almond) and
Coula edulis (African walnut). They also determined the lipid content of the fresh seed. [4] reported the
phytochemical and nutrient evaluation of the T.conophorum root.
Though the nuts are generally eaten in Nigeria as snack not minding is medicinal properties. Some data are
available on the phytochemical constituents of the seeds but the leaves are of little or not available. The objective
of this work therefore is to compare and evaluate the secondary metabolites, proximate, mineral contents and
vitamin compositions of Plukenetia conophora seeds and leaves in order to ascertain their possible usefulness as
food and in formulation of drugs.
MATERIALS AND METHODS
Fresh seeds and leaves samples of study plant were collected at Oshu village in Oko area, Alagbayen
farm, Surulere Local Government Area of Oyo State, Nigeria. The materials were washed, cut into small pieces
to facilitate dryness, and air-dried for fourteen days. The dried samples were ground into fine powder and stored
in an air tight bottle and stored in the desiccator prior to analysis.
Proximate analysis:
Moisture content was determined by drying to constant weight at 100-105o
C in an oven, ash content by
ignition at 550o
C in a muffle furnace for 4hr, oil content by soxhlet extraction with hexane as solvent, protein by
the kjeldahl method, and crude fibre by the acid and alkaline digestive methods all described by [15]. The
carbohydrate content was estimated by difference, subtracting the sum of water, protein, fat, crude fibre and ash
percentages from one hundred.
Phytochemical analysis:
The phytochemical screening was done on the sample using methods as described by [2, 27] Alkaloids were
extracted using a slightly modified method of [17]. Here, the dried sample was homogenized and the alkaloid
extracted from 10g of the sample for 4h using 20% v/v acetic acid in ethanol. The extract was filtered to remove
cellulose debris and then concentrated to about one quarter of the original volume. One percent NH4 OH was
added drop wise until a precipitate occurred. The crude alkaloid was dried to constant weight in an oven and the
percentage alkaloid calculated.

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93
Mineral analysis:
The [2] method was used for the determination of minerals in the test sample. Calcium, sodium,
potassium, magnesium were determined by flame photometric method while iron, zinc, manganese, copper and
chromium were determined by atomic absorption spectrophotometric method.
Vitamin analysis:
The composition of the water-soluble vitamins such as thiamine (B1), riboflavin (B2), niacin (B3),
cyanocobalamin (B12) were determined by the method of scalar analyzer (2000) While ascorbic acid (vitamin c)
content was determined by the method of [2]. [5] was used for the determination of tannins, while saponin was
analyzed using that of [26].
RESULTS AND DISCUSSION
Based on the results of the proximate composition of the seeds and leaves as shown in Table 1, the moisture
content was much in the seed than the leaves which is an indication that the leaves could have longer shelf life
than the seeds. The fat content in the leaves was higher (5.63 ±0.08%) than that of the seed and can be compared
to the reported value by [9]. The higher ash content of the leaves is an indication of the presence of high
inorganic materials than the seed. Leaves contained more nutrients than the seed except the protein content
where the seeds have higher value but lower than the reported value [9].
The result for the mineral analysis (Table 2) shows that leaves of Plukenetia conophora plant contained higher
values of minerals except copper that was higher in the seed. Both parts are good sources of important minerals
that are useful to the human body. This result become so important when the usefulness of such minerals like Ca,
Mg, Na, K in the body are considered and their usefulness in bone management. The presence of copper is
responsible for the absorption of iron, it is therefore often seen with iron naturally. Copper is important for
cellular defence and protection of the mucous membrane, antianaemic and essential for the formation of
haemoglobin from iron [6]. Manganese is necessary for the functioning of the pituitary gland, the pineal gland
and the brain, it promotes hepatorenal function, combat anaemia and also essential for growth.
The presence of zinc is an indication that the seed and leaves may have some effect on the nerve function
and male fertility. It is important for normal sexual development, especially for the development of testes and
ovaries, it is also essential for reproduction. Zinc stimulates the activity of vitamins, formation of red and white
corpuscles [6], healthy functioning of the heart and normal growth [11].
The phytonutrients present in the seed and leaves as shown in Table 3 and 4, shows that leaves were rich
in alkaloids, saponin, tannin and anthraquinones while seeds contained less alkaloids and traces of tannins,
saponin and anthraquinones were absent in the seed. Alkaloids are the most efficient plant substances used
therapeutically. Pure isolated alkaloids and the synthetic derivatives are used as the basic medicinal agent
because of their analgesic, antispasmodic and bacterial properties. The presence of tannins in both seed and leaf
support their strong use for healing of haemorrhoids, frost bite and varicose ulcers in herbal medicine [13].

Vol.3 No.9, 2013
94
PLATE 1.0: P.CONOPHORA LEAVES AND A POD OF THE NUT
PLATE 2: WALNUT SEEDS IN THE POD: SINGLE AND DOUBLE PODS

Vol.3 No.9, 2013
95
PLATE 2: WALNUT SEEDS RELEASED FROM THE POD
PLATE 3: WALNUT SEED RELEASED FROM THE SHELL
Table 5 shows the vitamin composition of both the seed and leaves. The seed contained higher amount of
Vitamin E than the leaf, while the leaf contained higher amount of vitamin C than the seed (Both vitamins are
antioxidants). Vitamin B groups are present in the leaf in trace amount yet more than that of the seed.
Considering the importance of all these mentioned vitamins, vitamin C can be used for the treatment of common
cold and other diseases like prostrate cancer [23, 24]. Higher amount of vitamin E in the seed supports the use of
the seed in southern Nigeria ethnomedicine as a male fertility agent [1].

Vol.3 No.9, 2013
96
Other vitamins though in trace amount are essential for body metabolism. There is also an interesting
ability of ascorbic acid as an antioxidant, to prevent or at least minimize the formation of carcinogenic
substances from dietary material. Deficiency of ascorbic acid is associated with pains in the joint and defect in
skeletal calcification, anaemia, manifestation of scurvy haemorrhage from mucous membrane of the mouth and
gastrointestinal track [12].
CONCLUSION
The present study compared the phytochemicals, proximate, vitamins and minerals composition of
Plukenetia conophora (African walnut) seeds and leaves. This partly shows that the leaf is also useful and
important as the seed and can equally be used as food and drug in herbal medicine or drug formulation.
REFERENCE
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[3] Ayodele , O. B., Nutrition in Ibadan. Catoon Publishers, Nigeria. 2003.
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[8] D’ Amelio F.S, Botanical; A phytocosmetic Desk Reference, Boca Raton, FL,CRC Press. p: 209, 1999.
[9] Edem, Christopher A. Dosunmu, Miranda,I. and Bassey Francesca, I. Determination of Proximate
Composition, Ascorbic Acid and Heavy Metal Content of African Walnut (Tetracarpidium conophorum).Pak .J.
Nutrit., 8: 225-226, 2009.
[10] Ekpo, A. S. and Eddy, N. O. Comparative Studies of The Level of Toxicant in the Seeds of Terminalia
catappa (india almond) and Coula edulis (African walnut). Chem Class Journal , 2, p: 74, 2005.
[11] Elizabeth, K., Immense help from Natures workshop. 1st
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[12] Hunt, S., Goff ,J.L. and Holbrook, J., Nutrition Principles and Chemical Practices. John Wiley and Sons.
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[13] Igboko, D.O., Phytochemical studies on Garcinia kola Heckel. M.sc. Thesis. University of Nigeria
Nsukka., pp: 202, 1983.

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[14] James, N. Roitman. Volatile Components of Green Walnut Husks. J. Agric. FoodChem. 48(7) pp: 2858-
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[16] Maduiyi, I., Biochemical and pharmacological studies of active principles of the seeds of Garcinia kola
Heckel. M.sc. Thesis University of Nigeria, Nsukka. , pp: 108, 1983.
[17] Maxwell, A., Seepers, M.P., and Mootoo, D.R., Aminospirosola NE steroidal alkaloids from solanum
trists. J .Nat. Product, 56, pp: 821-825, 1995.
[18] Nwaoguikpe, RN, Ujowundu, CO and Wesley, B. Phytochemical and Biochemical Compositions of
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[19] Okafor, B.B., Chemical Studies on some Nigerian Foodstuffs. Nigeria Cone Press Ltd., London, 1988.
[20] Oke, O.L., Leaf Protein Research in Nigeria Ibadan. University of Ibadan Press, Nigeria. 1995
[21] Okpero, A.O., The Nutritive Value of Conophor Seed. University of Ibadan Press, Nigeria. 2001.
[22] Okwu, D.E., Evaluation of Chemical Composition of Indigenous Spices and Flavouring Agents. Global J.
Pure Appl. Sci., pp: 458-459, 2001.
[23] Okwu, D.E.,and Okeke, O.,(2003). Phytochemical Screening and Mineral Composition of Chewing Sticks
in South Eastern Nigeria. Global J. Pure Appl. Sci., 9: 235-238. 2003.
[24] Okwu, D.E., Okwu, M.E., Chemical Composition of Spondia mombin plants .J. Sustain. Agri. Environ., 6:
140-147, 2004.
[25] Oyenuga, V.A., Nigeria Food and Feeding Stuffs Ibadan. University Press, Ibadan. 1997
[26] Peng, J.P. and Kobayasli, H. Novel Furastonol Glycosides from Allium Anacros Tenon Plant Media., 6:
58-61, 1995
[27] Sofowara, A., (1993). Medicinal Plants and Traditional Medicine in African. Spectrum book. 2nd
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[28] Stray, F., The Natural guide to medicinal herbs and plants. Tiger Book International, London, pp: 12-16,
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[29] Tapsell, L. C, Gillen, I. J., Patch, C. S. Including Walnuts in a low-fat/Modified-fat diet improves HDL
cholesterol-to-total cholesterol ratios in patients with type 2 diabetes care. 27 (12). Pp: 2777-2783, 2004
[30] Tidjani Amza, Issoufou Amadou, Mohammed, T. Kamara, Kexue Zhu and Huiming Zhou. Chemical and
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Technology, 2(4):191-195, 2010.

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RESULTS AND DISCUSSION
Table 1: Proximate composition of the fresh seeds and leaves of Plukenetia conophora
(Tetracarpidium conophorum) plant
Component Fresh seeds Leaves Literature
Edem et al, 2009
Moisture content (%) 46.03±0.30 29±0.71 48.70
Crude Fat content (%) 4.28±0.01 5.63±0.08 6.21
Crude Protein content (%) 21.65±0.5 16.62±0.30 35.22
Crude Fibre content (%) 7.34±0.71 14.92±0.04 3.34
Ash content (%) 5.27±1.35 12.89±0.02 2.03
Carbohydrate content (%) 18.10±0.01 20.94±0.01 53.20
Total energy value (J) 197.52±2.13 200.91±1.96 409.57±0.5
Results are means (±SD) of three determinations
Table 2: Mineral compositions of the raw seeds and leaves of the Plukenetia conophora plant on a dry
weight basis.
Mineral
Mg/Kg
Fresh seeds
Mg/kg
Leaves
Mg/kg
Literature
Tidjain et al, 2010
Potassium 6250.00±0.2 15937.00±0.02 12500.59±0.53
Sodium 4830.00±0.1 7980.00±0.01 1360.03±0.30
Calcium 4337.5±0.08 18700±0.02 6669.99±0.10
Magnesium 1711.25±0.8 1766.25±0.1 20999.65±0.65
Iron 110.00±0.04 4610.00±0.10 166.06±0.06
Zinc 40.10±0.10 61.15±0.08 110.84±0.79
Manganese 22.00±0.01 79.50±0.03 22.66±0.11
Copper 16.45±0.03 8.60±0.10 45.08±0.10
Table 3: Phytochemical screening of the fresh seeds and leaves of Plukenetia conophora plant
Constituent Fresh seeds Leaves Literature
(Ajaiyeoba and
Fadare,2006)
Alkaloids +++ +++ ++
Flavonoids -ve -ve -ve
Cardiac glycosides -ve -ve -ve
Saponins -ve +++ ++
Tannins +++ ++ ++
Anthraquinones -ve + +
Gallic acid + ++ -ve
Ellagic acid -ve -ve -ve
Caffeic acid -ve -ve -ve
KEY: -ve= absent
+ = present in a minute amount
++ = present in a moderate amount
+++ = present in an appreciable amount

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99
Table 4: Quantitative estimates of secondary metabolites present in the fresh seeds and leaves of the
Plukenetia conophora plant
Constituent Fresh seed Leaves Literature
Nwaoguikpe et al, 2012.
Alkaloids 2.380±0.2 2.670±0.02 0.41±0.01
Flavonoids 0.000 0.000 2.70±0.1
Cardiac glycosides 0.000 0.000 0.000
Saponins 0.000 1.080±0.01 5.03±0.01
Tannins 0.089±0.01 0.560±0.01 0.51±0.2
Anthraquinones 0.000 0.130±0.01 -
Gallic acid 0.024±0.01 0.180±0.02 -
Ellagic acid 0.000 0.000 -
Caffeinic acid 0.000 0.000 -
Table 5: Vitamin compositions of the fresh seeds and leaves of Plukenetia conophora plant on a dry
weight basis
Vitamin Fresh seeds Leaves Literature
Nwaoguikpe et al,2012
Ascorbic acid (C)
mg/100g
4.15±0.02 16.28±0.04 17.57±0.02
Tocopherol (E)
μg/100g
122.57±0.20 2.67±0.008 0.27±0.02
Thiamine (B1)
μg/100g
0.06±0.008 0.29±0.01 0.12±0.01
Riboflavin(B2)
μg/100g
0.02±0.008 0.34±0.01 0.13±0.01
Niacin (B3)
μg/100g
0.05±0.02 0.12±0.3 2.91±0.10
Pantothenic acid (B5)
μg/100g
0.00 0.00 -
Pyridoxine(B6)
μg/100g
0.00 0.00 -
Folic acid (B9)
μg/100g
0.00 0.00 -
Cyanocobalamin (B12)
μg/100g
0.12±0.1 0.23±0.03 -

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