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IJPSR (2011), Vol. 2, Issue 7 (Review Article)

Received on 21 February, 2011; received in revised form 02 June, 2011; accepted 28 June, 2011

IN-VITRO & IN-VIVO EFFICACY OF MORINGA OLEIFERA PLANT CONSTITUENTS IN UROLITHIASIS AS ANTILITHIATIC DRUG

Dipti Sachan*, S. K. Jain and Nandlal Singh

Institute of Pharmacy, Bundelkhand University, Jhansi, Uttar Pradesh, India

ABSTRACT

Ethanolic extract of Moringa oleifera was tested for its in-vitro antilithiatic/anticalcification activity by the homogenous precipitation method. The extract was compared with an aqueous extract of cystone (a marketed preparation) for their activities. The stones normally consist of calcium oxalate and calcium phosphate and the activity of the extract was evaluated by measuring the ability of the extract to inhibit the formation of calcium and phosphate precipitate. Ethylene glycol induced hyperoxaluria model was used to assess in-vivo antilithiatic activity.

INTRODUCTION: Moringa oleifera Lam. belonging to the single genus family Moringaceae is a small fast-growing ornamental tree originally belongs to India. Root, bark, pods and leaves of this tree are used in traditional medicine for the treatment of human diseases, whereby the leaves enriched in vitamin A and C. In the traditional medicinal system, almost all parts of this plant have been used in the treatment of various ailments. Fried leaves of this plant are useful in cold and fever 1. The leaves are nutritive and cooling, and are useful in removing all kinds of excessive pain. The aqueous extract of the leaves has been found to possess antifertility activity 2.

The juice of the leaves is used in cases of headache and it is also applied in the eyes in fainting fits due to nervous debility. Moreover, the local people of the Gangetic plain of West Bangal, India used this leaf juice as a cooling agent and refrigerant and also in anxiety. Adequate intake of vitamins and minerals is essential for preventing common micronutrient disorders such as vitamin A, iron and iodine deficiencies.

Several diseases associated with micronutrient deficiency could be prevented if amongst other interventions, adequate information on the existence and uses of indigenous plants that provide these nutrients were made available to rural households through better designed and implemented nutrition education and agricultural interventions. The problem of vitamin A deficiency and the widespread availability of Moringa in Malawi, one of the indigenous plants rich in vitamin A as well as other nutrients, is a typical case 3.

Synonyms: Assamese : Saijna, Sohjna Bengali : Sajina, Sajna English : Horse Radish Tree Gujrati : Saragavo Hindi : Sahajan Kannada : Neegge Malayalam : Muringa Marathi : Sevaga, Segat Sala Oriya : Sajina Punjabi : Sohanjana

Keywords:

Moringa oleifera,

Antilithiatic activity,

Calcium,

Phosphate

Correspondence to Author:

Dipti Sachan

Department of Pharmacognosy, Institute of Pharmacy, Bundelkhand University, Jhansi, Uttar Pradesh, India

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Tamil : Murungai Telugu : Munaga, Mulaga Urdu : Sohanjana, Sahajan

Scientific Classification: Kingdom : Plantae Subkingdom : Tracheobionta Super division : Spermatophyta Division : Magnoliophyta Class : Eudicots Subclass : Rosidsae Order : Brassicales Family : Moringaceae Genus : Moringa Species : oleifera

Description:

a) Macroscopic: Drug occurs in pieces of variable sizes, external surface, light greyish-brown, rough, reticulated, marked with transverse row of lenticels; outer bark, thin, peeling off, internal surface white.

b) Microscopic: Mature bark shows a very wide zone of cork, consisting of 25 or more rows of rectangular cells, arranged radially, a few inner layers, larger and cubicular in shape; secondary cortex composed of rectangular, thin-walled cells, a few containing starch grains and rosette crystals of calcium oxalate and a few others containing oil globules and coloured resinous matter; starch grains mostly simple and rarely compound, composed of 2 or 3 components, round to oval in shape, measuring 6 to 28μ in diameter, groups of stone cells, round to rectangular, of various sizes, present in secondary cortex; mucilaginous cavities found scattered towards inner secondary cortical region; secondary phloem appreciably wide, consisting mainly of phloem fibres and phloem parenchyma; phloem fibres in large patches, alternating with phloem parenchyma; numerous starch grains and cell contents as described above also present in phloem cells; phloem rays numerous, long, 2 to 4 seriate, consisting of radially elongated, thin-walled cells containing numerous starch grains, similar to those present in secondary cortex.

Powder - Pinkish-brown; shows stone cells, phloem fibres, starch grains, measuring 6 to 28 μ in dia., rosette crystals of calcium oxalate and oil globules.4

Identity, Purity and Strength:

Foreign matter not more than 2 per cent,

Total Ash not more than 18 per cent

Acid-insoluble ash not more than 10 per cent,

Alcohol-soluble extractive not less than 3 per cent,

Water-soluble extractive not less than 11 per cent.

T.L.C.: T.L.C. of the alcoholic extract on Silica gel 'G' using Toluene: Ethylacetate (9:1) shows under U.V. (366 nm) two fluorescent zones at Rf. 0.06 and 0.52 (both green). On exposure to Iodine vapour seven spots appear at Rf. 0.06, 0.33, 0.43, 0.54, 0.70, 0.78 and 0.87 (all yellow). On spraying with Vanillin- Sulphuric acid reagent and heating the plate at 105°C for ten minutes six spots appear at Rf. 0.33, 0.43, 0.54, 0.70, 0.78 and 0.87 (all violet).

Origin & Geographical Distribution: It is grown mainly in semi-arid, tropical, and subtropical areas, corresponding in the United States to USDA hardiness zone. It grows best in dry sandy soil, tolerates poor soil, including coastal areas. It is a fast-growing, drought.

Cultivation: The plant is propagated by planting limb cuttings 1–2 m long, from June to August. The plant starts bearing pods 6–8 months after planting & bears for several years. It does not tolerate freeze or frost. It can also be propagated by seed. Optimum cultivation depends on producing the right environment for the plant to thrive 5.

Chemical Constituents: Leaves contain all essential amino acids, protein, Vitamin A, Vitamin B, Vitamin C, and minerals, 2 nitrile glycosides (niazirin & niazirinin), 3 mustard oil glycosides .The seeds contain 30-50% oil. The oil contains 65-75% oleic acids. Also contain β-sitosterol, glycerol-1-(9-octadecanoate), kaempherol, magnesium, tocopherol, L-glutamic acid.

Plant parts used: Leaves : Ascorbic acid, tocopherol Root : kaempferol Stem : β- sitosterol Shoot : Glutamic acid

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Flowers : Kaempferol Fruits : Magnesium Seeds : Oleic acid

Species of Moringa:

Moringa arborea Verdc. (Kenya) Moringa borziana Mattei Moringa concanensis Nimm Moringa drouhardii Jum.– Bottle Tree Moringa hildebrandtii Engl. Moringa (south western Madagascar) Moringa longituba Engl.-Moringa oleifera Moringa ovalifolia Dinter & Berger Moringa peregrina (Forssk.) Fiori Moringa pygmaea Verdc. Moringa ruspoliana Engl. Moringa stenopetala (Baker f.) Cufod.

Antioxidant Activity: The crude extract of phenolic compounds was obtained from Moringa oleifera Lam. The content of total phenolics in the extract was determined by using the Folin- ciocalteu’s phenol reagent. Phenolic compounds present in the crude extract show antioxidant activity as revealed by the Total Antioxidant Activity (TPA), DPPH radical scavenging activity and reducing power. The content of total phenolics in the Moringa oleifera extract is 118mg/ g. The extract of Moringa oleifera also has strong antiradical activity against DPPH radical. Phenolic acids (derivatives of caffeic, p-coumaric and ferulic acids) are the dominant phenolic constituents of Moringa oleifera leaves extract 6.

Analgesic Activity: Methanolic extract (ME) of the root of Moringa oleifera was tested for possible pharmacological effects on experimental animals. ME potentiated significantly the sleeping time induced by pentobarbitone sodium, diazepam and meprobamate, showed analgesic properties and also potentiated analgesia induced by morphine and pethidine. Analgesic activity was tested in mice using various doses orally. Acetic acid-induced writhing episodes were significantly and dose-dependently reduced.

Anti-Inflammatory Activity: Moringa contains Vitamin A, Vitamin B1 (Thiamine), Vitamin C, Vitamin E, Arginine, Beta-sitosterol, Caffeoylquinic Acid, Calcium, Chlorophyll, Copper, Cystine, Omega 3, Omega 6,

Omega 9, Fiber, Glutathione, Histidine, Indole Acetic Acid, Indoleacetonitrile, Isoleucine, Kaempferal, Leucine, Magnesium, Oleic-Acid, Phenylalanine, Potassium, Quercetin, Rutin, Selenium, Stigmasterol, Sulfur, Tryptophan, Tyrosine, Zeatin, Zinc. The seeds of malunggay, Moringa oleifera, were extracted with distilled ethanol and concentrated under reduced pressure at 40oC.

The resulting extract was partitioned between hexane, ethylacetate, butanol and water. The solvent fractions were likewise concentrated under reduced pressure. The crude ethanol extract of dried seeds inhibited the carrageenan-induced inflammation in the hind paw of mice by 85% at a dosage of 3 mg/g body weight while the mature green seeds by 77%The hexane fraction of the crude ethanol extract of the dried seeds also inhibited inflammation by 77% at the same dosage while both butanol and water fractions inhibited inflammation by only 34% 7.

Anti-helmintic Activity: Preliminary phytochemical screening has shown the presence of saponins, carbohydrates, alkaloids, tannins, proteins, flavonoids in methanolic extracts of plants. The methanolic extracts of seeds of the plant Moringa oleifera showed significant anthelmintic activity at 100 mg/ ml concentrations whereas Chloroform, showed moderate activity and Petroleum ether extract is having least anthelmintic activity.

Cancer Protective Activity: Moringa species have long been recognized by folk medicine practitioners as having value in tumour therapy. We examined compounds for their cancer preventive potential. The related compounds were shown to be potent inhibitors of phorbol ester (TPA)-induced Epstein-Barr virus early antigen activation in lymphoblastoid (Burkitt’s lymphoma) cells (57,104). In one of the study, it also inhibited tumour promotion in a mouse two-stage DMBA-TPA tumour model.

In an even more recent study, Bharali and colleagues have examined skin tumour prevention following ingestion of drumstick (Moringa seedpod) extracts. In the mouse model, which included appropriate positive and negative controls, a dramatic reduction in skin papillomas was demonstrated.

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Antipyretic Activity: The antipyretic activity of ethanolic, petroleum ether and ethyl acetate extracts of seeds was screened using yeast induced hyperpyrexia method. Paracetamol was used as a standard for comparison. The ethanolic and ethyl acetate extracts of seeds showed significant antipyretic activity.

Wound Healing Activity: When assessed for healing activity in excision, incision and dead space wounds the ethanolic and ethyl acetate extracts of leaves showed significant wound healing activity that is comparable with the standard vicco turmeric cream. Phytosterol & Phenolic compounds present in these extracts promote the wound healing activity.

Hepatoprotective Activity: The methanolic and chloroform extracts of leaves of Moringa have shown very significant hepatoprotection against CCL induced hepatotoxicity. Whole plant play important role in hepatoprotective activity. Aqueous and ethanolic extract of roots and flower &Ethanolic extract of leaves used in hepatoprotective activity 8.

Antiulcer activity: Methanolic extract of leaves and flower buds of Moringa oleifera is used as antiulcer activity.

Antispasmodic Activity: The fresh leaves ethanol extract of Moringa was tested in guinea-pig ileum and mouse duodenum. There were significant dose and time dependent reductions of the acetylcholine response with initial stimulatory effects in both the guinea-pig ileum and the mouse duodenum preparations. Spontaneous rhythmic contractions were greatly reduced, suggesting an antispasmodic property of the crude LE extract. The LE extract showed some oxytocic activity on uterus strips of guinea- pigs and mice. The results are indicative of the traditional use of the leaves of Moringa stenopetala for stomach pain and to expel retained placentae by women.

Antidiabetic Activity: The present study deals with the scientific validation of glycemic potential of aqueous extract of M. oleifera leaves in normal, sub, mild and severely diabetic rats since M. oleifera have been claimed to possess hypoglycemic effect in Indian traditional system of medicine.

The leaves of other species of Moringa stenopetala are used traditionally in Ethiopia for treating diabetes mellitus and had already been explored for their hypoglycemic action. It was considered therefore worthwhile to investigate the aqueous extract of M. oleifera leaves for its glycemic potential by evaluating its effect on blood glucose level of normal, sub, mild and severely diabetic rats. Its effect on haemoglobin, total protein, urine sugar, urine protein and body weight was also examined in the severely diabetic models. Thus, this is the first reporting of hypoglycemic and antidiabetic effect of aqueous extract of Moringa oleifera leaves.

Anti-mycobacterial Activity: From aqueous extract of seeds of Moringa oleifera Lam. (Moringaceae) 4-(l-Rhamnosyloxy) benzyl isothiocynate, isolated. The minimum bactericidal concentration (MBC) was found to be 17.5_g/ml (56_mol/l) in tube dilution test. It is used to treat tuberculous glands, leprosy, asthma, expectorant, bronchitis.

Water Purifying Properties: Moringa oleifera is a tropical plant belonging to the family Moringaceae. The seeds from this plant contain active coagulating agents characterized as dimeric cationic proteins having a molecular weight of 13 kDa and an isoelectric point between 10 and 11. During the last 5 years there has been a growing interest in using the water extract of M. oleifera seeds (WEMOS) as a natural coagulant to treat drinking water 9.

Chemomodulatory Activity: The modulatory effects of a hydro-alchoholic extract of drumsticks of Moringa oliefera Lam at doses of 125 mg/ kg bodyweight and 250 mg/ kg body weight for 7 and 14 days, respectively, were investigated with reference to drug metabolising Phase I (Cytochrome b5 and Cytochrome P450) and Phase II (Glutathione-S- transferase) enzymes, anti-oxidant enzymes, glutathione content and lipid peroxidation in the liver of 6-8 week old female Swiss albino mice. Further, the chemopreventive efficacy of the extract was evaluated in a two stage model of 7, 12- dimethylbenz(a) anthracene induced skin papillomagenesis.

Antifungal Activity: The fungi live in the dead or top layer of skin cells in moist areas of the body, such as between the toes, the groin, and under the breasts.

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These fungal infections cause only a minor irritation. Other types of fungal infections could be more serious. They can penetrate into the cells and cause itching, swelling, blistering and scaling. In some cases, fungal infections can cause reactions elsewhere in the body. The seeds and leaves of Moringa oleifera Lam as herbal medicines. Ethanol extracts showed anti-fungal activities in vitro against dermatophytes such as Trichophyton rubrum, Trichophyton mentagrophytes, Epidermophyton Xoccosum, and Microsporum canis.

Hypocholestraemic Activity: The leaves of M. oleifera were used by the Indians in their herbal medicine as a hypocholesterolaemic agent in obese patients. They tested the crude extract of leaves of M. oleifera and showed that it possessed hypocholesterolaemic activity. This led them to conclude that there is a valid pharmacological basis for employing the leaves for this purpose. As the fruit of M. oleifera is widely consumed for its hypocholesterolaemic activity. Thus, the aim of the study was to evaluate the fruit of M. oleifera for its effect on the lipid profile in normal and hypercholesterolaemic rabbits.

Antihypertensive, Diuretic and Cholesterol Lowering Activities: The widespread combination of diuretic along with lipid and blood pressure lowering constituents make this plant highly useful in cardiovascular disorders. Moringa leaf juice is known to have a stabilizing effect on blood pressure. Nitrile, mustard oil glycosides and thiocarbamate glycosides have been isolated from Moringa leaves, which were found to be responsible for the blood pressure lowering effect. Most of these compounds, bearing thiocarbamate, carbamate or nitrile groups, are fully acetylated glycosides, which are very rare in nature.

As Biosorbent: Moringa seeds could be used as a less expensive biosorbent for the removal of cadmium (Cd) from aqueous media 10. The aqueous solution of Moringa seed is a heterogeneous complex mixture having various functional groups, mainly low molecular weight organic acids (amino acids). These amino acids have been found to constitute a physiologically active group of binding agents, working even at a low concentration, which because of the ability to interact with metal ions is likely to increase the sorption of metal ions 11.

Medicinal Uses:

Root: Antilithiatic, rubefacient, vesicant, carminative, antifertility, anti-inflammatory, stimulant in paralytic afflictions; act as a cardiac/circulatory tonic, used as a laxative, abortifacient, treating rheumatism, inflammations, articular pains, lower back or kidney pain and constipation.

Leaves: Purgative, applied as poultice to sores, rubbed on the temples for headaches, used for piles, fevers, sore throat, bronchitis, eye and ear infections, scurvy and catarrh; leaf juice is believed to control glucose levels, applied to reduce glandular swelling.

Stem Bark: Rubefacient, vesicant and used to cure eye diseases and for the treatment of delirious patients, prevent enlargement of the spleen and formation of tuberculous glands of the neck, to destroy tumors and to heal ulcers. The juice from the stem bark is put into ears to relieve earaches and also placed in a tooth cavity as a pain killer, and has anti-tubercular activity.

Gum: Used for dental caries, and is astringent and rubefacient; Gum, mixed with sesame oil, is used to relieve headaches, fevers, intestinal complaints, dysentery, asthma and sometimes used as an abortifacient, and to treat syphilis and rheumatism.

Flower: High medicinal value as a stimulant, aphrodisiac, abortifacient, cholagogue; used to cure inflammations, muscle diseases, hysteria, tumors, and enlargement of the spleen; lower the serum cholesterol, phospholipid, triglyceride, VLDL, LDL cholesterol to phospholipid ratio and atherogenic index; decrease lipid profile of liver, heart and aorta in hypercholesterolaemic rabbits and increased the excretion of faecal cholesterol.

Seed: Seed extract exerts its protective effect by decreasing liver lipid peroxides, antihypertensive compounds thiocarbamate and isothiocyanate glycosides have been isolated from the acetate phase of the ethanolic extract of Moringa pods.

Industrial uses:

1. The oil content of de-hulled seed (kernel) is approximately 42%. The oil is brilliant yellow. It is used as a lubricant for fine machinery such as

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timepieces because it has little tendency to deteriorate and become rancid and sticky 12, 13. It is also useful as vegetable cooking oil. The oil is known for its capacity to absorb and retain volatile substances and is therefore valuable in the perfume industry for stabilising scents. The free fatty acid content varies from 0.5 to 3%. The seed oil of Moringa contains approximately 13% saturated fatty acids and 82 % unsaturated fatty acids. It has a particularly high level of oleic acid (70%). Other vegetable oils normally contain only about 40% oleic acid.

2. Water purification- Moringa seeds contain between 30-42% oil and the press cake obtained as a by-product of the oil extraction process contains a very high level of protein. Some of these proteins (approximately 1%) are active cationic polyelectrolytes having molecular weights between 7-17 K Dalton. The cationic polyelectrolytes neutralize the colloids in muddy or dirty water since the majority of these colloids have a negative electrical charge. This protein can therefore be used as a non-toxic natural polypeptide for sedimenting mineral particles and organics in the purification of drinking water, for cleaning vegetable oil, or for sedimenting fibers in the juice and beer industries.

It thus, works as a primary coagulant as natural bridges are continuously formed between the colloid particles. In contrast, industrial coagulants such as alumina can be toxic. Their proper use requires qualified personnel and the majority of underdeveloped countries don’t have the means of producing them. In addition, these industrial coagulants are expensive and represent a considerable drain on the hard currency reserves of developing countries.

3. Forage plant -The nutritional characteristics of the Moringa tree are excellent so it can easily be used as a fresh forage material for cattle. The leaves are rich in protein, carotene, iron and ascorbic acid and the pod is rich in the amino acid lysine 14, 15, 16. Another important advantageous characteristic of Moringa is its

high productivity of fresh material per unit area compared with other forage crops 17, 18. Moringa is especially useful as forage for cattle both economically and productively given the problems facing typical cattle breeders (70% of the national herd in Nicaragua is in the hands of these small cattle producers) 19, 20.

In-vitro study on antilithiatic activity of moringa oleifera plant: Ethanolic extract of Moringa oleifera were tested for its in-vitro antilithiatic/anticalcification activity by the homogenous precipitation method. The extract was compared with an aqueous extract of cystone (a marketed preparation) for its activity. The aqueous extract of cystone was prepared by grinding a tablet to powder. This powder was mixed with 5mL water and kept for 2-3 h and then centrifuged at 1000 rpm. The clear supernatant was used for the study. The extracts of the plants were compared with the aqueous extract of cystone (a marketed herbal formulation for urolithiasis) for their antilithiatic activity.

In-vivo study on antilithiatic activity of moringa oleifera plant: The effect of oral administration of aqueous and alcoholic extract of Moringa oleifera root-wood on calcium oxalate urolithiasis has been studied in male Wistar albino rats. Ethylene glycol induced hyperoxaluria model was used to assess the antilithiatic activity. Most calculi in the urinary system arises from a common component of urine, e.g. calcium oxalate (CaOx), representing up to 80% of analysed stones. Stone formation in the kidney is one of the oldest and most widespread diseases known to men.

CONCLUSION: M. oleifera is a tropical tree whose numerous economic applications and facility of propagation are growing international interest. The Moringa tree is cultivated to use as a vegetable (leaves, green pods, flowers, roasted seeds), for spice (mainly roots), for cooking and cosmetic oil (seeds) and as a medicinal plant (all plant organs). Moringa oleifera, Medicinal and Socio-Economic uses.

Furthermore, the raw seeds are valuable because extracts have a flocculating protein that works as a coagulant of surface muddy and turbid water to tap-

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water in many African and Asian countries and Central America. Now that research and pilot scale tests have been carried out, consideration is being given to the production and use of the Moringa’s coagulants at national and international levels. In Tanzania, a Franco-Swiss company has started large-scale production of Moringa seeds to that end. The oil extracted from the seeds is not only comparable in terms of quality to olive oil as oil for human consumption, but is also a raw material used by the cosmetics industry.

M. oleifera is also a food-producing plant: In India, it is cultivated for the production of its fruit, while in the Sahelian zones of Africa its leaves are eaten as a vegetable. The planting of Moringa trees by smallholder farmers should be encouraged because it will improve both their health and income.

REFERENCES:

1. Bhatnagar SS, Santapau H, Desai JDH, Yellore S, Rao TNS.: Biological activity of Indian medicinal plants. Antibacterial, antitubercular and antifungal action. Indian J Med Res 1961; 49: 799–805.

2. Bhattacharya SB, Das AK, Banerji N.: Chemical investigations on the gum exudates from Moringa oleifera. Carbohydr Res 1982; 102: 253–262.

3. Faizi S, Siddiqui B, Saleem R, Saddiqui S, Aftab K. : Isolation and structure elucidation of new nitrile and mustard oil glycosides from Moringa oleifera and their effect on blood pressure. J Nat Prod 1994 a; 57: 1256–1261.

4. Fahey JW, Zalcmann AT, Talalay P.: The chemical diversity and distribution of glucosinolates and isothiocyanates among plants. Phytochemistry 2001; 56: 5–51.

5. Ruckmani K, Kavimani S, Anandan R, Jaykar B.: Effect of Moringa oleifera Lam on paracetamol-induced hepatoxicity. Indian J Pharm Science 1998; 60: 33–35.

6. Atawodi SE et.al.: Evaluation of the polyphenol content and Antioxidant properties of methanol extracts of the leaves,

stem, and root barks of Moringa oleifera Lam. J Med Food 2010; 13(3): 710-716.

7. Cheenpracha S et.al. Potential anti-inflammatory phenolic glycosides from the medicinal plant Moringa oleifera fruits. Bioorg Med Chem. 2010: 379-381.

8. Hamza AA: Ameliorative effects of Moringa oleifera Lam seed extract on liver fibrosis in rats. Food Chem Toxicol 2010; 48(1): 345-355.

9. Firth J et.al. Point-of-use interventions to decrease contamination of drinking water: a randomized, controlled pilot study on efficacy, effectiveness, and acceptability of closed containers, Moringa oleifera, and in-home chlorination in rural South India. Am J Trop Med Hyg 2010; 82(5): 759-765.

10. Sharma P, Kumari P, Srivastava MM, Srivastava S.: Removal of cadmium from aqueous system by shelled Moringa oleifera Lam. seed powder. Bioresour Technol 2006; 97: 299–305.

11. Brostlap AC, Schuurmans J.: Kinetics of valine uptake in tobacco leaf disc. Comparison of wild types the digenic mutant and its monogenic derivatives. Planta 1988; 176: 42–50.

12. Ferrao, A.M.B.C. & Mendez Ferrao, J.E.: Acidos gordos em oleo de Moringueiro (Moringa oleifera Lam.). Agronomia Angolana 1970; 8: 3-16.

13. Ramachandran, C., Peter, K.V. & Gopalakrishnan, P.K.: Drumstick (Moringa oleifera): a multipurpose Indian vegetable. Economic Botany 1980; 34: 276-283.

14. Council of Scientific & Industrial Research: The wealth of India. A dictionary of Indian raw materials and industrial products. Raw materials 1962; 6: L-M, New Delhi, CSIR, India.

15. Chawla S., Saxena A. & Seshadri S.: In-vitro availability of iron in various green leafy vegetables. Journal of the Science of Food and Agriculture 1988; 46: 125-127.

16. Dogra P.D., Singh B.P. & Tandon S.: Vitamin content in Moringa pod vegetable. Current Science 1975; 44: 31.

17. Dahot MU. : Vitamin contents of flowers and seeds of Moringa oleifera. Pak J Biochem 1988; 21: 1–24.

18. Faizi S, Siddiqui B, Saleem R, Siddiqui S, Aftab K, Gilani A. : Novel variety of Malavi. J Am Oil Chem Soc 1994b; 79: 677–683.

19. Garimella T.S, Jolly CI, Narayanan S: In-vitro studies on Antilitiatic Activity of seeds of Dolichos biflorus Linn. and Rhizomes of Begenia ligulata Wall. Phytotherapy Research 2001; 15: 351-355.

20. Lalas S, Tsaknis J.: Extraction and identification of natural antioxidant from the seeds of Moringa oleifera tree variety of Malavi. J Am Oil Chem Soc 2002; 79: 677–683.

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