EVALUATION OF SELECTED MEDICINAL PLANTS FOR ANTIUROLITHIATIC ACTIVITY

EVALUATION OF SELECTED MEDICINAL PLANTS FOR ANTIUROLITHIATIC ACTIVITY

1. Sai Sindu*, M. Aparna, B. Sravani, M. Kranthi and T. Ratna Maheswari

Siddharth Institute of Pharmacy, Kantepudi (v), Guntur (dist), Andhra Pradesh, India

ABSTRACT

The main aim of our study is to evaluate the antiurolithiatic activity of pomegranate (punica granatum, Family: Puniaceae) and garlic (Allium sativum, Family: Liliaceae). For this, we have subjected the seeds of pomegranate and garlic bulbs to maceration using 95% of ethanol, ethyl acetate, petroleum ether and water at a ratio of 1:4 for 24 hours, followed by reflux over a water bath for 3 hours and the obtained extract was concentrated under vacuum. The prepared extracts were then subjected to titrimetry and colorimetry for the estimation of calcium and phosphate respectively. By comparing the result of different extracts, we have found that ethanol extract of garlic has showed better result in inhibiting precipitation of calcium and phosphate. But individual isolation and further studies are to be carried out to confirm this activity.

Colorimetry

INTRODUCTION: Urolithiasis is a condition that is characterized by formation of calculi in urinary tract. Stone disease is a multifactorial disease,ingestion of excessive amounts of purines,oxalates,calcium phosphate,sodium and other elements often results in excessive excretion of these components in urine thus increase in the risk of calculi formation ¹.

Medical conditions that are associated with increased risk of kidney stone formation include hyperparathyroidism, hyperthyroidism, gout, systic fibrosis ². Symptoms for urolithiasis include abnormal urine colour, blood in urine, fever, nausea and vomiting ³.

Though some synthetic drugs are available in market for treating urolithiasis, herbal medicines have their own emphasis. In the present scenario, the demand for herbal products is growing exponentially throughout the world and major pharmaceutical companies are currently conducting extensive research on plant materials for their potential medicinal value.

By knowing the importance of herbal medicine, we have choosen pomegranate and garlic to evaluate antiurolithiatic activity.

Punica granatum (Family: Puniaceae) is a tree that grows best in well drained ordinary soil. The edible fruit is a berry and is between a lemon and a grapefruit in size, 5-12 cm in diameter with a rounded hexagonal shape, and has thick reddish skin. The exact number of seeds in a pomegranate can vary from 200 to about 1400 seeds, contrary to some beliefs that all pomegranates have exactly the same number of seeds.

Each seed has a surrounding water-laden pulp-the edible aril-ranging in color from white to deep red or purple. The seeds are embedded in a white, spongy, astringent pulp.juice of pomegranate contains 85.4% water, 10.6% total sugars, 1.4% pectins, 0.2-1.0% polyphenols ⁴.

Allium sativum (Family: Liliaceae) is cultivated in well drained moderately clay loamy soil.

Garlic bulbs contain 29% of carbohydrates, about 56% of proteins (albumin), 0.1% of fat, mucilage, and 0.06 to 0.1% of volatile oil. It also contains phosphorus, iron and copper. Volatile oil of the drug is the chief active constituent, and contains allyl propyl disulphide, dially disulphide alliiin and allicin.

Alliin by action of enzyme allinlyase is converted into allicin. Garlic oil is yellow in colour and has specific gravity of 1.046. It is optically inactive ⁵

MATERIALS AND EQUIPMENTS:

Plant material used:

• Pomegranate (Punica grantum)
• Garlic (Allium sativum)

Apparatus used:

• EI Colorimeter with matched glass cuvettes
• Type A glass Burette
• Conical flasks
• Beakers
• Digital pH meter

TABLE 1: CHEMICALS USED

Reagents preparation:

0.1 M TRIS buffer: 0.1 M TRIS buffer was prepared by using solution A and solution B.

Solution A: 0.4 M TRIS 48.4 g of tris (trihydroxymethyl) amino methane was accurately weighed and dissolved in few ml of water and finally made upto1000 ml with water).

Solution B: 0.4 M hydrochloric acid (33.6ml of concentrated hydrochloric acid was measured and added in few ml of water and finally made upto1000ml with water).

A working solution was made by adding 25ml of solution A and 20.7 ml of solution B, made up to 100 ml by using water & then the pH was adjusted to 7.4.⁶

Sodium chloride preparation: It was prepared by dissolving 5.85 g of sodium chloride in 10 ml of water.

Sodium oxalate preparation: It was prepared by dissolving 1.3 g of sodium oxalate in 10 ml of water.

Preparation of extracts:

1. Pomegranate extract: The seeds of pomegranate are subjected to maceration using 95% of ethanol, ethyl acetate, petroleum ether and water at a ratio of 1: 4 for 24 hours, followed by reflux over a water bath for 3 hours and the obtained extract was concentrated under vacuum.
2. Garlic extract: The garlic bulbs are subjected to maceration using 95% of ethanol, petroleum ether, ethyl acetate and water at a ratio of 1: 4 for 24 hours, followed by reflux over a water bath for 3 hours and the obtained extract was concentrated under vacuum.

Methodology:

Plant authentication: The plants were collected from Kantepudi, sattenapalli and authenticated by Mrs.Ch. Jhansi vani, Assistant Professor, A.C. College, Guntur.

Methods:

Estimation of calcium: Two sets of test tubes (A₁, A₂) were taken. To these test tubes 2ml of TRIS-HCl buffer, 1ml of sodium chloride, 1ml of calcium chloride di hydrate, and 1ml of sodium oxalate were added. Now 2ml of extract (Ethanol, Ethyl acetate, Petroleum ether, Water) and vehicle under investigation was added. Amount of calcium was estimated by titrimetry using disodium edetate as titrant and solochrome black T as an indicator.

  Formula:

Amount= (titre volume × equivalent factor × actual molarity) / (wt. taken × expected molarity)

Estimation of phosphate: Two sets of test tubes (B₁, B₂) were taken. To these test tubes 2ml of TRIS-HCl buffer, 1ml of sodium chloride, 1ml of Calcium chloride di hydrate, and 1ml of disodium hydrogen phosphate were added. Now 2ml of extract (Ethanol, Ethyl acetate, Petroleum ether, Water) and vehicle under investigation was added. Amount of phosphate present was estimated by colorimetric analysis at 530 nm against blank.

Formula:

Amount= (sample absorbance/standard absorbance) × standard concentration

% inhibition = (Control - test) ×100

Control

RESULTS: The results of the present study (Table 2 to Table 9) indicates that the ethanol extract of both pomegranate and garlic was found to have more inhibiting property than the ethyl acetate, petroleum ether and aqueous extracts of plants. Among garlic and pomegranate, garlic has showed better result in inhibiting precipitation of calcium and phosphate.

Results of pomegranate:      

TABLE 2: PERCENTAGE INHIBITION USING ETHYL ACETATE EXTRACT OF PUNICA GRANATUM

TABLE 3: PERCENTAGE INHIBITION USING PETROLEUM ETHER EXTRACT OF PUNICA GRANATUM

TABLE 4: PERCENTAGE INHIBITION USING ETHANOL EXTRACT OF PUNICA GRANATUM

TABLE 5: PERCENTAGE INHIBITION USING AQUEOUS EXTRACT OF PUNICA GRANATUM

Results of Garlic:

TABLE 6: PERCENTAGE INHIBITION USING ETHYL ACETATE EXTRACT OF    ALLIUM SATIVUM

TABLE 7: PERCENTAGE INHIBITION USING PETROLEUM ETHER EXTRACT OF  ALLIUM SATIVUM

TABLE 8: PERCENTAGE INHIBITION USING ETHANOL EXTRACT OF ALLIUM SATIVUM         

TABLE 9: PERCENTAGE INHIBITION USING AQUEOUS EXTRACT OF ALLIUM SATIVUM

DISCUSSION AND CONCLUSION: In the present study, pomegranate and garlic were used to evaluate the antiurolithiatic activity by in vitro methods. Garlic stimulates the production of nitric oxide and hydrogen sulphide which relax the blood vessels; this improves the circulation of the kidneys and prevents the formation of infection stones. Garlic is also believed to contain vitamin B-6 which helps to reduce the levels of urinary oxalate and the seeds of pomegranate can be considered as natural remedy for kidney stones. This may be related to their astringent property. To determine the antiurolithiatic activity we have selected titrimetry for the estimation of calcium and colorimetry for the estimation of phosphate. Based on the results obtained in table 4 and table 8 we found that garlic has showed better results than pomegranate. But individual isolation and further studies are to be carried out to confirm this activity.

REFERENCES:

1. Holmes RP et al, (1993), Relationship of protein intake to urinary oxalate and glycolate excretion. Kidney Int. (44), 366-372.
2. Kramer HJ et al, (2003), the association between gout and nephrolithiasis in men: The health professionals’ followup study. Kidney Int. (64), 1022-6.
3. PubMed Health; Diseases and Conditions; Kidney Stones; Louis S. Liou, MD, PhD, et al.; January 2009
4. E.wallis; Text book of pharmacognosy; 44th edition; page no; 83.
5. K.Kokate, A.P.purohit, S.B. Gokhale; Pharmacognosy; 41st edition; May 2008; page no; 11.51
6. Bhavik A. Patel et al., In vitro evaluation of antilithiatic activity on Tectona grandis Linn seeds. Journal of Pharmacy Research 2011, 4(6), 1699-1700
   

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   How to cite this article:

   Sai Sindu PS, Aparna M, Sravani B, Kranthi M and Maheswari RT: Evaluation of Selected Medicinal Plants for Antiurolithiatic Activity. Int J Pharm Sci Res. 3(12); 5029-5032.

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