INTRODUCTION:

Medicines derived from plants have played a pivotal role in the health care of many cultures, both ancient and modern. Scientific evaluation of plants has often shown that active principles in these are responsible for therapeutic success. Medicinal plants may serve as a vital source of potentially useful new compounds for the development of effective therapy to combat a variety of ailments. Plant derived natural products such as flavonoids, terpenoids, steroids, etc. have diverse pharmacological properties.¹

Mineral elements are essential for human life; they may be beneficial or harmful. The human body contains at least 60 detectable chemical elements. However, only about 25 of these are believed to participate in the healthy functioning of the human body.²

Analysis of mineral elements is most important to understand the pharmacological and nutritional values of medicinal plants. Mineral elements play important role in biochemical and metabolic reactions in the living organism, which are mainly responsible for the formation of active organic constituents.³Over the past two decades, medicinal plant has gained attention as natures cure for human cancer, is a major reason for the human mortality rate,⁴ especially with a growing number of population in the world.

Boerhavia diffusa belongs to the family Nyctaginaceae. Boerhavia diffusa is known to possess medicinal value in traditional system and represented a wide range of pharmacological properties. Reports are scanty regarding radioprotective property of B.diffusa. B.diffusa in radiation induced haemopoetic injury increase haemoglobin And total RBC count in albino mice.⁴ The hydroalcoholic extract of B.diffusa has a radioprotective effect against gamma Radiation induced damage in mice.⁵Pharmacological studies have demonstrated that B. diffusa known to possessdiuretic,⁶ antifertility,⁷ antifibrinolytic,⁸ immunomodulatory,⁹ antidiabetic,¹⁰ antiviral,¹¹ antistress,¹² antimicrobial,¹³ antiinflammatory, hepatoprotective,¹⁴ antioxidant,¹⁵ antiurethritis¹⁶ and antimetastatic.¹⁷

MATERIALS AND METHODS:

The plant material of Boerhavia diffusa L leaf used for the investigationwas collected fromin and around the village nearby Thanjavur. The plant was identified and authenticated by Dr. P. Jayaraman, Director, Plant Anatomy Research Centre (PARC), Chennai and a voucher specimen is kept in the herbarium. The leaves were extracted by using methanol as a solvent.

Phytochemical analysis of different extract:

Different chemical tests were carried out for both types of extracts to identify the presence of various chemical constituents like alkaloids, flavonoids, carbohydrates, phenols, glycosides, saponins, Steroids and phytosteroids, Tannins, terpenoids, Fixed oils and Fats, Resins, Gums and Mucilage, Coumarins and Chlorogenic acid etc.¹⁸

Total Alkaloid,¹⁹ total flavonoid,²⁰ total phenolic content,²¹ total tannin content,²² total saponins,²³total terpenoids,²⁴ minerals like manganese, copper, iron and zinc²⁵were analysed.

Statistical analysis:

All the assays were carried out in triplicate. Experimental results are expressed as mean±standard deviation. The results were analyzed using one-way analysis of variance and the group means were compared using Duncan’s multiple range tests using SPSS version.

RESULT AND DISCUSSION:

PHYTOCHEMICAL ANALYSIS:

The medicinal value of these plants lies in some chemical substances that produce a definite physiological action on the human body. The most important of these bioactive constituents of plants are alkaloids, tannins, flavonoids, and phenolic compounds. Many of these indigenous medicinal plants are used as spices As flavonoids having antioxidant property, it protects tissues against oxygen free radicals, thus have a role in prevention of atherosclerosis, cancer, chronic inflammation and may inhibit aging. The polyphenols possess anti parasitic activity, and monoterpenes have been reported to constitute anti-plasmodic, anti-neoplasmic and anti-viral activities.²⁶

Table 1 shows in our study, are observed in a phytochemical screening of the plant revealed the absence of saponins, Resin, Gum and mucilage, Coumarin, Chlorogenic acid, glycosides and tested positive for alkaloids, phenols, flavonoids, tannins, terpenoids, glycosides, carbohydrate, protein, fixed oil and fat, amino acids and phytosterol. The presence of flavonoids and tannins in the plant extract is likely to be responsible for the free radical scavenging effects observed. Flavonoids and tannins are phenolic compounds and plant phenolics are a major group of compounds that act as primary antioxidants or free radical scavengers.²⁷

Table 1: Preliminary phytochemical screening of methanolic extracts of B.diffusa leaves

S. No	Phytochemical constituents	Methanolic extract
1	Alkaloids	+
2	Flavonoids	+
3	Carbohydrates	+
4	Phenol	+
5	Glycosides	-
6	Saponin	-
7	Phytosterol	+
8	Tannins	+
9	Terpenoids	+
10	Protein andaminoacids	+
11	Steroids	+
12	Resin	-
13	Gum and mucilage	-
14	Coumarin	-
15	Fixed oil and fat	+
16	Chlorogenic acid	-

‘+’ indicates the presence ofPhytochemical constituents

‘-‘indicates the absence of Phytochemical constituents

These results expose that the plant has quite a number of chemical constituents, which may be responsible for many pharmacological actions and have protective or disease preventive properties. Alkaloids are beneficial chemicals to plants with predator and parasite repelling effects.²⁸ In the present study, moderate concentration of phenols was observed in B.diffusa L. Phenolics play a vital role against several health disorders. They exhibit anti ulcer, anti inflammatory, antioxidant and cytotoxic potential. Many biological activities of phenolic compounds attributed by their antioxidant activity.²⁹

QUANTITATIVE PHYTOCHEMICAL ANALYSIS:

Quantitative estimation of phytochemical constituents of B.diffusa are represented in Table 2. The results indicated the phytochemical constituents as flavonoids, saponins, alkaloids, phenols, carbohydrates and tannins.

Table: 2: Quantitative determination of methanolic leaf extract of Boerhavia diffusa L.

S. No	Chemical constituents	Total contents in mg/g equivalents
1.	Alkaloids	115.05±3.50
2.	Flavanoids	210.65±1.40
3.	Total Phenols	280.15±2.45
4.	Tannins	90.90±5.85
5.	Saponins	116.25±3.15
6.	Erpenoids	135.25±2.85

Note: n=3, Values are given in Mean ± SEM

The flavonoids have long been recognized to possess antiallergic, anti-inflammatory, antibacterial, antiviral, anti-proliferative and anti-carcinogenic activities as well as to affect some aspects of mammalian metabolism. It gives protection against free radicals, platelet aggregation, microbes, ulcers and hepatoxins. The phytochemical analysis revealed the presence of flavonoids in the herbal plants. Our plant also contains alkaloids, terpenoids and other phenolic compounds hence it might has exhibited pharmacological activity.⁸

Plant steroids are known to be important for their cardio tonic, insecticidal and anti-microbial properties. They are also used innutrition, herbal medicine; cosmetics and they are routinely used in medicine because of their profound biological activities.

Tannin is reported to exhibit antiviral, antibacterial, anti-tumor activities. It was also reported that certain tannin are able to inhibit HIV replication selectivity and is also used as diuretic. Plant tannin has been recognized for their pharmacological properties and is known to make trees and shrubs a difficult meal for many caterpillars.³⁰

THIN LAYER CHROMATOGRAPHY OF METHANOLIC EXTRACT OF B.DIFFUSA:

In our study, the most suitable TLC system for analysis was shown to be chloroform: methanol with the largest discriminating power. Fig 1: Thin Layer Chromatography of methanolic extract of B.diffusa. Three bands found in this method and its Rf values were 0.38, 0.40 and 0.45. This values indicate the presence of phenolic compounds.

Fig 1: Thin Layer Chromatography of methanolic extract of B.diffusa

TLC indicated the presence of polar metabolites. The polar and water-soluble extracts possessed potent antimicrobial and cytotoxic activity. Proton NMR indicates that the compounds may be related to phenolic lipids produced by Boerhavia diffusa L.³¹ The highly polar compounds would not appear as a distinct spot by TLC but would remain at the origin. However, even these polar metabolites, when present in moderate to high concentrations, should be detectable as a large, spot at the origin of the TLC. They were not detected in our TLC analyses.

Quantitative Mineral Composition of leaf extract of B.diffusa L.:

The minerals have been categorized in to major, minor and trace elements on the basis of their availability in the plants. Their deficiency causes diseases whereas the excess presence may cause toxicity.³² Medicinal plants contain vital elements which are widely used as a precursor for regulated pharmaceutical products. Medicinal values of the pharmaceutical products are due to the presence of secondary metabolites.

Studies on quantitative mineral analysis of B.diffusa L leaves conducted and presented in Table 5 revealed that the plant contains appreciable amount of the basic food minerals like The amount of Mg 145.9±0.5, Na 82.5. ±2.5, Ca 76.25±0.65, while Cu is 4.2±0.5, Zn 6.25±2.0, Mn 19.65±0.35and Fe 22.5±0.25 mg/100g.

Table 3 depicts that B.diffusa L leaves were found to have high quantities of Mg, Na and Ca were present but not in very high concentration. Calcium and Mg which are present in high concentrations are essential minerals for life important in the formation of bones, nerves and teeth as a cofactor for enzymes and a component of ATP, DNA, RNA and cell membranes respectively. The minerals present in low concentration (zinc, copper and manganese) perform various important functions in humans like the formation of hemoglobin, growth and sexual maturation, facilitating iron intake, as cofactor for enzymes and so many other functions.

Table 3: Mineral composition of leaf extract of Boerhavia diffusaL

Parameter	Concentration (mg/100g)
Sodium	82.5. ±2.5
Potassium	61.35±0.85
Calcium	76.25±0.65
Magnesium	145.9±0.5
Copper	4.2±0.5
Iron	22.5±0.25
Manganese	19.65±0.35
Zinc	6.25±2.0

Note: n=3; Values are given in Mean ±SEM

In our study, The amount of Mg 145.9±0.5, Na 82.5. ±2.5, Ca 76.25±0.65, while Cu is 4.2±0.5, Zn 6.25±2.0, Mn 19.65±0.35and Fe 22.5±0.25mg/100g. Trace element plays a crucial role in the medicinal value of a plant, in health and to cure disease. They play a nutritive, catalytic and balancing function in plants.³³ Plants take them from the ground and incorporate them into organic compounds that we consume them by eating either the plants or the animals that ate the plants. In the present study Mg, Ca, Na are in large quantity while Cu and Zn are relatively low compared to the other elements. The Mg is found in large percentage that is (145.9±0.5). In this context,³⁴reported the macro and micro nutrients of leaves of C. intybus from Brazil. The main difference which is observed between the two studies is that Mg, Na and Ca are present in large amount in leaves of Boerhavia diffusa L while Zn and Cn are present in lesser amount. Similarly,³⁵ detected similar elements and showed the same pattern except with slight differences in values. Hence, the plant can be recommended as raw or in addition form for food products.

Magnesium, assist in the assimilation of phosphorus. Lack of magnesium can be responsible for tetany, tuberculosis, diabetes, cancer and all nervous diseases.³⁶ Maybe this is the reason why pawpaw leaves especially the green ones are being employed in the treatment of the above listed diseases.³⁷ Potassium is necessary for muscular weakness which is associated with malaria, and also slows down sclerosis of the vascular system. It contributes to the fight against bacteria and cleanses the digestive system. Sodium takes part in the metabolism of water, promotes digestion, assimilation, osmosis, cleanses the digestive system, combats stomach acidity and alkalize the blood. This is the reason behind the brown pawpaw leaf being used as a cleanser in herbal remedy.³⁷The presence of Iron signifies that the leaves can be used against anaemia, tuberculosis and disorder of growth.³⁶ Iron is an energizer but excess can cause fatigue but we hardly have excess if taken from natural source.^38,39

Manganese, according to Claude and Paule,³⁶ is necessary for the functioning of the pituitary gland, the pineal gland and the brain. It promotes hepato-renal function, combat anaemia and it is also essential for growth. Since these leaves contain these elements in different concentrations, the quantity and the type of leaf to be taken will depend on the therapeutic need of the individual and they can also be combined for an effective result in such cases as diabetes, cancer, fibroid and tuberculosis.³⁷

CONCLUSION:

This study has shown the phytochemicals and minerals composition of B.diffusa L extract by using TLC. This partly shows the use of this plant in herbal medicine, as a rich source of phytochemicals, coupled with the presence of the essential minerals. B.diffusa L leaves can be seen as a potential source of useful food and drug items. The presence of alkaloids in them explains the reason why it is being effectively used as an pharmacological properties. Therefore pawpaw leaves can be manipulated in the herbal treatment of various diseases and as a potential source of useful elements for drugs formulation.

ACKNOWLEDGEMENTS:

We thank the Rabiammal Ahamed Maideen College for Women – 610001, Tamilnadu, India

for providing laboratory facility for the study.

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Received on 20.10.2019 Modified on 29.12.2019

Research J. Pharm. and Tech. 2020; 13(10):4856-4860.

DOI: 10.5958/0974-360X.2020.00854.9