Diuretic Activity of Pongamia pinnata Linn. Leaf Extracts in Rats

 

Nagarathna PKM*, Prabhakar T and Vikram BS

Department of Pharmacology, Karnataka College of Pharmacy, #33\2, Thirumenahalli, Hegde Nagar Main Road, Yelahanka Hobli, Jakkur Post, Bangalore-560064, India

 

ABSTRACT

Pongamia pinnata Linn were evaluated by determination of urine volume, electrolyte concentration and diuretic potency in male albino rats. Different concentrations of petroleum ether, and methanol extract (250 mg/kg, 500mg/kg) were orally administered to hydrated rats and their urine out put was immediately measured after 5 h of treatment. Furosamide (10 mg/kg) was used as reference drug while normal saline (0.9%) solution was used as control. The petroleum ether extract exhibited dose dependent diuretic property. The onset of diuretic action was extremely prompt (within 1h) and lasted through out the studied period (upto 5h). The ethanol and petroleum ether extracts caused marked increase in Na+, K+ and Cl- level. The result of this experiment suggests that Pongamia pinnata Linn leaves extracts possessed significant diuretic activity in rats.

 

 

KEYWORDS: Diuretic activity, Pongamia pinnata Linn, ethanol petroleum ether extracts.

 

 

INTRODUCTION

Pongamia pinnata Linn is a plant belonging to family fabeceae  Pongamia pinnata  synonymous to Pongamia  glabra is a  handsome tree which is normally found growing along   river  and  stream sides.  The leaves of the tree are placed alternatively and is impari – pinnately compound.  The 5 to 9 leaflets are ovate – oblong.  The white   or   purplish papilionacius flowers are found in axillary racemes.  The pod is obliquely – oblong, bilaterally compressed and woody.  The single seed inside the pod is kidney-shaped. Pharmacognostical and  phytochemical screening  have confirmed the various chemical constituents given in table

 

 

Phytochemicals  of   Pongamia pinnata linn

Sl. No.

Name of the Phytochemicals

1

Karanjin,

2

Pongapin,

3

3-methoxy-pongapin,

4

Pongal,

5

Lanceolatin,

6

Betasitosterol,

7

Pongalbal, Pongachalone, IandII,

8

Glycoside,

9

Quercetin-glycosides,

 

 

 

 

MATERIALS AND METHODS:

Preparation of extracts: Leaves of Pongamia pinnata were collected from the herbal garden of our college and were identified by botanist.  Herbarium was prepared and submitted to the museum of our college. The leaves were collected in the month of October.  The leaves were dried in shade at room temperature.  The dried leaves were subjected to size reduction using mixy,  to  coarse powder.  The powder was packed in to column – percolator and extracted with petroleum ether (60 – 800C) for 48 hours and same marc was successively extracted with ethyl alcohol for 48 hours.  The obtained extract was dried at room temperature till semisolid mass was obtained and was stored in air tight container in refrigerator below 100C.

 

The suspension of petroleum ether, ethyl alcohol prepared using 2 % gum acacia were used for all experiments95.

 

These two extracts (Petroleum ether, ethyl alcohol) of Pongamia pinnata were subjected to following investigations.

1)      Preliminary phytochemical Screening.

2)      Pharmacological activities.

 

a)      Anti diuretic activity:

1). Preliminary Phytochemical Screening:

The Preliminary Phytochemical Screening  were carried out on the ethyl alcohol, petroleum ether of  Pongamia pinnata linn, for qualitative identification. Test for common phytochemicals were carried out by standard methods described in practical pharmacognosy  by  Dr. C.K. Kotate95 and  K.R.Khandelwal96

 

2) Pharmacological  activities:

a)      Determination of acute toxicity(LD­50)

The acute toxicity of ethylalcohol, Petroleum ether and Pongamia pinnata  were determined in albino mice of either sex (20 – 30 gm )  by  the fixed dose (OECD) method ( guideline No 420) of CPCSEA.

 

The individual extracts were collected and concentrated by evaporation in vacuum. The dried extracts were formulated as suspension using normal saline as a vehicle. Various extracts of each plant material were evaluated for their diuretic activity.

 

Animals used:

Male albino rats weighing 125-to150 gm bred in our laboratory were used for the study. The animals were housed and acclimatized under standard laboratory conditions and were supplied with standard laboratory feed and water was supplied ad libitum.  The animals were divided into 4 groups consisting of 6 animals each, the entire experimental model 1 to 4 served as petroleum ether, ethanol, while 3 and 4 received +ve control and solvent control respectively. All the experiments were carried out under the guidance of ethical committee of S.C.S. College of pharmacy (Registration CPCSEA/ CH/ CRG/ 2001/Harapanahalli).

 

Toxicological studies:

Preliminary oral LD50 doses of petroleum ether, ethanol extracts of Pongamia pinnata Linn in mice were found to be  400 mg / kg respectively.

 

Evaluation of diuretic activity:

The method of lipschitz16 et al was employed for the assessment of diuretic activity. The male albino rats weighing between 125 to 150 gm were procured from central animal house of S.C.S. College of pharmacy, Harapanahalli. The animals were maintained under standard conditions of temperature and humidity. The animals were divided into 4 groups consisting of 6 animals each were hydrated with 5ml / kg of water orally prior to drug/extract administration Normal saline (0.9%) and furosamide (10mg / kg) served as control and standard drug respectively. 250 mg and 500mg / kg of petroleum ether, ethanol extracts of Pongamia pinnata Linn were administered orally to animals in each group. Immediately after dosing the rats were placed in metabolic cages specially designed to separate urine and faeces and kept at room temperature. The urine was collected in measuring cylinder upto 5 hours after dosing. During this period no food or water was made available to the animals. The urine volume was measured with graduated measuring cylinder. The parameters taken were total urine volume, urine concentration of Na+, K+ and Cl-.  Concentration of Na+ and K+ was determined with flame photometer while Cl- concentration was estimated titrimetrically. The mean urine volumes were determined and diuretic potency was assessed by comparison of urine excretion due to the extracts with respect to the standard drug Furesamide.

 

Statistical analysis:

All values are shown as mean SEM. The results were statically analysied using one-way ANOVA followed by Dunnett’s test. P ≤ 0.01 was considered significant.

 

RESULTS AND DISCUSSION:

Preliminary phyto chemical screening indicated the presence of Alkaloids, Phyto sterols, Flavonoids and Saponins. The results of diuretic activity of Pongamia pinnata Linn shows that petroleum ether extract are active and displayed dose dependent diuretic activity.

 

The present study indicates that the petroleum ether, methanol extracts of Pongamia pinnata Linn leaves at doses of 250 and 500 mg / kg caused dose dependent diuretic activity. At the concentration of 250mg and 500 mg / kg, the petroleum ether, methanol extracts gave a mean urine volume of 1.60 ± 0.35, and 1.77 ± 0.5 after 5 hours respectively. The petroleum ether ,methanol extracts (250mg and 500 mg / kg) produced urine with Na+, K+ and Cl-content of73.3 ± 1.38, 91.3 ± 0.65, 96.7 ± 1.35, 130.2 ±3.46, 170 ± 2.35, 170 ±4.2 Na+, K+ and Cl- respectively. The diuretic potency of Pongamia pinnata Linn extracts was dose dependent (Table No 1).

 

ACKNOWLEDGEMENTS:

The authors are thankful to the management of Karnataka College of pharmacy, Bangalore for providing the necessary facilities for research work.

 

On the basis of above results, it can be concluded that the Pongamia pinnata Linn leaves extracts produced significant diuretic effect with increase in electrolyte concentration in urine.

 

However, further studies are necessary to identify and isolate the active constituents responsible for its diuretic activity and also there is a need to elucidate its mechanism of its diuretic action.

 

 

Table 1: Diuretic activity of Pongamia pinnata Linn leaves extracts in rat.

Extract

Dose

Mg/kg

Mean

Urine volume

Diuretic potency

Ecectrolyte concentration Drug

Na+                        K+                          Cl-

Petroleum ether

250

1.60 ± 0.35

0.64

73.3 ± 1.38              91.3 ± 0.65              96.7 ± 1.35

Ethanol

500

1.87  ± 0.5

0.76

130.2 ±3.46             170 ± 2.35               170 ±4.2

Furosamide

10

2.45 ± 0.56

1.05

146.0 ± 1.60            115.6 ± 1.20            162.0 ± 0.80

Normal saline

5ml/kg

kg  0.84 ± 0.37

0.42

92.3 ± 1.46              64.3 ± 0.85              125.6 ± 0.31

Values (except diuretic potency) are mean ±SEM (n = 6). * p ≤ 0.01 (ANOVA followed by Dunnett’s test) compared with control. Diuretic potency is a ratio of urine volume due to tested drug to that of standard drug.

 

 

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Received on 20.07.2009

Accepted on 15.09.2009     

© A&V Publication all right reserved

Research Journal of Pharmacognosy  and Phytochemistry. 1(3): Nov. -Dec. 2009, 185-187