Evaluation of Diuretic Activity of Achyranthes aspera leaves extracts

 

Prashant D Warke*¹, Dr. Ashutosh Upadhayay¹, M. K. Kale²

¹Mahatama Jyoti Rao Phoole Univeristy, Department of Pharmacy, Jaipur, Rajasthan.

²Konkan Gyanpeeth Rahul Dharkar College of Pharmacy and Research Institute, Karjat, Maharashtra.

 

ABSTRACT:

Plants used in traditional medicine have stood up to the test of time and contributed many novel compounds for preventive and curative medicine to modern science. India is sitting on a gold mine of well recorded and traditionally well practiced knowledge of herbal medicine. Pet. ether, ethyl acetate, alcoholic and aqueous extracts of Achyranthes aspera leaves were tested for diuretic activity in rats. The parameters studied on individual rat were total urine volume, urine concentration of Na⁺, K^- and Cl^-. In the present Pet. ether, ethyl acetate, alcoholic and aqueous extracts of Achyranthes aspera leaves showed increase in urine volume, Na⁺, K^- and Cl^-. Furosemide was used as reference diuretic.

 

 

 

INTRODUCTION:

Achyranthes aspera is an erect plant and about 1 m in height. It is common plant as weed and found roadside throughout India. The various vernacular names of the plant are as Sanskrit Aghata, Apamarga, in hindi Chirchira, in marathi Aghada. The various parts like Herb, leaves, seeds and root used in various diseases. Decoction of leaves is a good diuretic found in renal dropsies and general anasarea. The leaf juice is also useful in stomach ache and bowl complaints, piles, boils skin eruptions. In large doses it produces abortion or labour pains¹. It is widely used for asthmatic cough, snakebite, hydrophobia, urinary calculi, rabies, influenza, piles, bronchitis, diarrhea, renal dropsy, gonorrhea and abdominal pain. The seeds are employed as an emetic, purgative, and cathartic, in gonorrhoea, for insect bite and in hydrophobia cough including whooping cough as an anti-asthmatic².

 

 

 

 

MATERIAL AND METHODS:

Collection and Identification of plant material:

In the present study, the leaves of Achyranthes aspera, Linn. (Family-Amaranthaceae), was collected from the local areas of Jalgaon district. The plants were authenticated from Agharkar Research Institute (ARI) Pune. (An autonomous, grant-in-aid research institute of the Department of Science and Technology (DST), Government of India).

 

Soon after authentication, all the crude drugs were dried at room temperature until they free from moisture and subjected to physical evaluation with different parameters. 

 

Preparation of extracts:

In the present study, the crude drugs were carefully selected and shade dried.  The dried material was reduced to coarse powder in a mechanical grinder and passed through a Sieve No.40 to obtain powder of desired particle size.

 

About 200 gms of powdered material was subjected to exhaustive extraction successively with petroleum ether, ethyl acetate and ethanol at temperature of 45°–50°C to about 40 cycles per batch for 8 batches.  The extraction was continued until the solvent in the thimble became clear then few drops of solvent were collected in a test tube during the completion of the cycle (during syphoning) and chemical test of that solvent was performed. Extraction was completed only when chemical test shows negative results.  Finally the drug was be macerated with chloroform water. After each extraction the solvent was distilled off rotary evaporator and the extract was concentrated at low temperature.  These extracts were used for phytochemical investigation.

 

Experimental animals:

Wistar albino rats of either sex weighing 150-250 g procured from the animal house were used for the study. The animals were maintained in polypropylene cages of standard dimensions at a temperature of 25±1° C and standard 12 hour: 12 hour day/night rhythm. The animals were fed with standard rodent pellet and water ad libitum. Prior to the experiment the animals were acclimatized to the laboratory conditions.

 

Animals and treatment:

Wistar albino rats were kept in metabolic cages. These cages were specially designed to separate urine and faeces of the study animals. In the cages, the study animals were housed under the standard conditions of temperature, humidity and dark/light cycle (12 h/12 h). The animals were given pelleted food and drinking water ad libitum. The bedding of the animal cages was changed after every 48 hours^3,4.

 

Acute toxicity studies:

Acute toxicity studies for Pet. ether, ethyl acetate, alcoholic and aqueous extracts of Achyranthes aspera was carried out in rats at different doses (500–3000 mg/kg, orally), showed no gross evidence of any abnormalities in the rats up to the end of 72 hr of the observation period. This indicates the safety of extract. Acute toxicity study was done as per OECD Guidelines. Hence we selected 250mg/kg and 500mg/kg as low and high doses 

Phytochemical screening:

The preliminary phytochemical screening of extracts were carried out as per standard procedures^5-11. The results are reported in Table No. 1

 

Diuretic Activity:

Wistar albino rats weighing 150 to 180gm were maintained under standard condition of temperature and humidity. The method of Lipschitz et al^12,13 was employed for the assessment of diuretic activity. The experimental protocols have been approved by the Institutional Animal Ethical Committee. Ten groups of six rats in each and were fasted and deprived of water for eighteen hours prior to the experiment. The first group of animals serving as control, received normal saline(25ml/Kg,); the second group received furosemide (100mg/Kg) in saline; other groups received the Pet. ether, ethyl acetate, alcoholic and aqueous extract at the doses of 250 mg/Kg and 500mg/kg. Immediately after administration the animals were placed in metabolic cages (2 per cage), specially designed to separate urine and feaces , kept at room temperature of 25± 0.5ºC throughout the experiment. The urine was collected in measuring cylinders up to 3 hrs after dosing. During this period, no food or water was made available to animals. The parameters taken for individual rat were body weight before and after test period, total concentration of Na⁺ , K⁺ , and Cl^- in the urine. Na⁺ , K⁺ concentrations were measured by Flame photometry¹⁴ and Cl- concentration was estimated by titration¹⁵ with silver nitrate solution(N/50)using three drop of 5% potassium chromate solution as indicator. Furosemide sodium salt was given by stomach tube. Optimal dose activity relation was found to be 20mg/Kg of furosemide per kg body weight in series of supportive experiments. Results are reported as mean ± SD, the test of significance (p<0.01 and p<0.001).

 

Table No 2: Diuretic activity of Achyranthes aspera  in albino rats

Treatment	Dose (ml/kg b.w.)/ (mg/kg b.w.)	Urine Volume (ml)	Electrolyte Excretion (M ± S.E.M) Na+        (mEq/lit)	Electrolyte Excretion (M ± S.E.M) K+(mEq/lit)	Electrolyte Excretion       (M ± S.E.M) Cl -(mMol/lit)
Normal saline	25	8.10±0.12	81.2±0.73	39.4± 0.52	96.7±2.72
Furosemide	100	15.6±0.43***	124±2.6***	59.6±0.8***	139.2±1.9***
(AAPEE)	250	9.2±0.08	98.2±2.1	49.6±2.0	103.1±2.5
(AAPEE)	500	9.9±0.5	102±2.1	51.2±1.9	109.2±1.6
(AAETAE)	250	10.4±0.16	99±2.3	50.2±2.1	108.2±1.9
(AAETAE)	500	11.23±0.14**	105±2.9	52.7±2.1	112.8±2.8
(AAAE)	250	13.1±0.15***	109.2±2.6**	52.2±1.9**	124.8±3.2**
(AAAE)	500	14.1±0.46***	119.2±1.7***	53.9±1.8***	136.2±2.9***
(AAAQE)	250	11.2±0.19**	106.2±2.0	53.6±1.56	105±2.5
(AAAQE)	500	12.9± 0.5***	112.5±2.3**	54.3±1.48**	117.2±1.9**

   n=6,           ml. of urine expressed in Mean ±S.E.M.

**= P <0.01(Statistically significant), ***= p < 0.001 (Statistically more significant) when compared with Normal Saline group.  

 

 

 

 

 

RESULT AND DISCUSSION:

Table No.1: Preliminary phytochemical investigation of Achyranthes aspera leaves.

Chemical Constituents	PEE	ETAE	ALCE	AQE
Carbohydrates	+	-	+	+
Flavonoids	-	+	+	+
Saponins	-	-	+	-
Tannins	+	+	+	+
Alkoloids	+	+	-	+
Glycosides	+	+	+	+
Steroids	+	+	-	+

+ Present – absent   PEE-Petroleum ether extract, ETAE-Ethyl acetate extract, ALCE-Alcoholic extract, AQE-Aqueous extract.

 

In the present study, we can demonstrate that Pet. ether, ethyl acetate, alcoholic and aqueous extracts may produce diuretic effect by increasing the excretion of sodium, potassium and chloride. The control of plasma sodium is important in the regulation of blood volume and pressure. The control of plasma potassium is required to maintain proper function of cardiac and skeletal muscles. The regulation of sodium, potassium balance is also intimately related to renal control of acid base balance. The Potassium loss that occurs with many diuretics may leads to hypokalemia. For this reason, generally potassium-sparing diuretics are recommended¹⁶ Active principles such as flavanoids, saponins are known to be responsible for diuretic activity¹⁷. The results from the Table 2 clearly showed that the extract of  alcoholic and aqueous extracts of Achyranthes aspera act as diuretic in a dose-dependent manner.

 

CONCLUSION:

The extracts of Achyranthes aspera has diuretic effect supporting the ethnopharmacological use as diuretics. This effect may be explored in the use of the plant in the management of some cardiovascular diseases.

 

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Accepted on 27.09.2018        © RJPT All right reserved

Research J. Pharm. and Tech 2018; 11(12): 5394-5396.