Two Wavelength Method for Estimation of Indapamide and Perindopril Erbumine in Combined Tablet Dosage Form

 

S.S. Kale*, R.L. Bakal, A.V. Chandewar and R.S. Sakhare

Department of Quality Assurance, P. Wadhwani College of Pharmacy, Yavatmal-445 001 (M.S.)

*Corresponding Author E-mail: samruddhi.kale@gmail.com

 

ABSTRACT:

A new, simple, accurate and sensitive UV-spectrophotometric two wavelength method has been developed for simultaneous determination of Indapamide and Perindopril Erbumine in combined pharmaceutical dosage form. Two wavelength i, e 220 nm and 240 nm were selected for estimation of Indapamide where as wavelength 216 nm was selected for estimation of Perindopril Erbumine using Methanol as solvent. Indapamide and Perindopril Erbumine shows linearity in the concentration range of 2-20 μg/ml and 4-40 μg/ml respectively. The method was validated statistically.

 

KEYWORDS: Indapamide, Perindopril Erbumine Two wavelength method.

 


 

INTRODUCTION:

The chemical formula of indapamide(IND) is 3-(aminosulfonyl)-4-chloro-N-(2,3-dihydro-2-methyl-1H-indol-1-yl)-benzamide Indapamide is an oral antihypertensive diuretic agent indicated for the treatment of hypertensive and edema1 Indapamide inhibits carbonic anhydrase enzyme2.

 

Perindopril erbumine(PER),(2S,3(infinity)S,7(infinity)S)-1-[(S)-N-[(S)-1-Carboxy- butyl]alanyl]hexahydro-2-indolinecarboxylic acid, 1-ethyl ester, compound with tert-butylamine (1:1), belongs to a group called angiotensin converting enzyme (ACE) inhibitors.Inhibition of ACE results in decreased plasma angiotensin II, leading to decreased vasoconstriction, increased plasma rennin activity and decreased aldosterone secretion. The overall effect of this is a drop in blood pressure and a decrease in the workload of the heart3.

 

Several methods have been reported for the determination of indapamide in biological fluids and in pharmaceutical preparations including spectrophotometry4,5, high performance liquid chromatography6-8, LC-MS/MS9, LC-ESI-MS 10,capillary electrophoresis11 and several methods for perindopril including spectrophotometric12-14, gas chromatography15, LCMS16, HPLC17 .Indapamide officially in BP18 and USP19 and Perindopril Erbumine officially in BP.

 

However no method is reported for simultaneous estimation of these two drugs in tablet. Hence the present work was attempted to develop accurate, simple and sensitive method for simultaneous estimation of IND and PER. For this purpose marketed tablet Perigard –D containing 2 mg of PER and 0.625 mg of IND was used. The developed method has been validated by evaluation of the system suitability, linearity, precision and accuracy. The validated method was applied to the commercially available pharmaceutical formulations containing IND and PER.

 

MATERIAL AND METHODS:

Chemicals:
Pharmaceutical grade IND and PER were kindly supplied as a gift sample by Micro Labs and Cipla, Bombay, (India) respectively. Methanol AR grade purchased from Samar Chemicals, Nagpur, India.

 

Instrument:
Double beam UV –visible spectrophotometer UV 2401 PC (Japan) Thermo, with 1cm UV matched quartz cells was used. Citizen Balance was used for experimental purpose.

 

Preparation of IND Standard Stock Solution:

An accurately weighed quantity of IND 10 mg was transferred to the 25 mL volumetric Flask, dissolved in sufficient quantity of methanol and sonicate for 5 min. The volume was made up to the mark with methanol. (400 mg/mL). Further, diluted 5.0 mL of the above solution to 10 mL with methanol. (200ug/ml)

 

Preparation of PER Standard Stock Solution:

An accurately weighed quantity of PER 10 mg was transferred to the 25 mL volumetric flask dissolved in sufficient quantity of methanol and sonicate for 5 min. The volume was made up to the mark with methanol. (400 mg/mL).

Study of spectra and selection of wavelength:

The aliquot portions of stock standard solutions of IND and PER were diluted appropriately with solvent to obtain concentration 40 mg/mL of each drug. The solutions were scanned in the range of 400 – 200 nm in 1 cm cell against blank. The overlain UV absorbance spectrum of IND and PER is shown in Fig. No.1.

 

Fig. No. 1: Overlain spectra of IND and PER

 

From the overlain spectrum shown in Fig.1, the wavelength selected for estimation of IND was 220 nm and 240 nm, where as for PER was 216 nm. At 216 nm, there was considerable absorbance of Indapamide. The IND and PER obey Beer’s law in the concentration range of 2 to 20 μg/ml and 4 to 40 μg/ml.

Quantitative estimation of these drugs was carried out by using following formulae’s.

A concentration of IND is calculated from slope and intercepts equation of calibration curve of A 220.0 nm - A 240.0 nm against concentration of IND.

Cy                      µ  A 220.0 nm - A 240.0 nm.                              (1)

Cx                      = CAx 216.0 nm / A (1%, 1cm) 216.0 nm of PER.   (2)

CAx 216.0 nm   = A 216.0 nm - Ay 216.0 nm                                     (3)

Ay 216.0 nm      = Cy X A (1%, 1cm) 216.0 nm of IND                     (4)

Where, Cx and Cy are concentration (g /100ml) of PER and IND, respectively, A 220 nm, A 240.0 nm and A 216.0 nm are absorbance of mixture at 220.0 nm, 240.0 nm and 216.0 nm, respectively, CAx 216.0 nm is corrected absorbance of PER and Ay 216.0 nm absorbance of IND at 216.0 nm.

 

Preparation of marketed formulation:

Twenty tablets were accurately weighed. Average weight of tablet was calculated. The tablets were reduced to fine powder and mixed thoroughly. A quantity of tablet powder equivalent to weight of one tablet was transferred to 100 mL volumetric flask and dissolved in 25 ml of solvent i.e. methanol and sonicate for 5 min. and volume was made to 100 mL with the same solvent to get final concentration of about 75 mg/mL IND and 240 mg/mL PER. The solution was filtered through Whatman filter paper no. 41.The absorbance of sample solution was measured at 216 nm, 220 nm and 240 nm in 1 cm cell against blank.

 

VALIDATION:

The proposed method was validated on the basis of parameters namely accuracy, precision, ruggedness and linearity and range. The accuracy of the proposed method was ascertained by carrying out recovery studies using standard addition method. The recovery study was performed to determine if there was any positive or negative interference from excipients present in the formulation. Precision of an analytical method is expressed as SD or RSD of a series of measurements. It was ascertained by replicate estimation of drug by the proposed method. Test for ruggedness was carried out by repeating the procedure under different conditions, i.e., on different days, at different time and by different analysts. Linearity and range study was done by preparing concentration in the range of 80 -120 % of test concentration and absorbance values were recorded at 216 nm, 220 nm and 240 nm. The plot of linearity and range is shown in Fig. 2.

 

Fig. No. 2: Plot of linearity and range for IND and PER

 

RESULTS AND DISCUSSION:

An attempt has been made to develop a fast, sensitive, precise, reproducible and economical analytical method for simultaneous estimation of IND and PER in their combined dosage form. In this method IND and PER obey Beer’s law in the concentration range of 2 to 20 μg/ml and 4 to 40 μg/ml. It was observed that both the drugs showed additivity of absorbance at selected wavelengths indicating that both the drugs do not interact with each other in the solvent system used. A (1%, 1cm) values were also calculated for both the drugs. For PER, A (1%, 1cm) was found to be 196.8 at wavelength 216 nm and for IND, it was 625.0 at wavelength 216 nm respectively. The result of percentage estimation of drug is shown in Table No.1. The method was validated as per the ICH and USP guidelines. The results of recovery study were found to be within the prescribed limit of 98.9 – 99.8 %, proving the accuracy and showing that the method is free from interference from excipients. The results are shown in Table No. 2.


Table No.1: Result of estimation of IND and PER in tablet formulation

Brand name. – PERIGARD -D                Average weight =91.87 mg

Sr. No.

Weight of tablet powder (gm)

Absorbance

% Lable claim

A1-A2

A3

IND

PER

1

91.80

0.1016

0.9419

100.2

100.2

2

91.85

0.1008

0.9381

99.5

99.8

3

91.82

0.1011

0.638

99.8

99.7

 

Mean

99.8

99.9

 

± S.D.

0.35

0.5

 

C.V.

0.35

0.5

* Results are mean of three replicate

 

Table No.2: Results of recovery studies of IND and PER

SR.

No

Weight of Tablet    Powder in  mg.

Amt.Added in µg.

Absorbance

Amt.Recoverd in µg.

% Recovery

IND

PER

A1-A2

A3

IND

PER

IND

PER

1

 

91.80

0.75

2.4

0.110

0.907

0.74

2.38

99.2

99.3

2

1.5

4.8

0.120

0.993

1.49

4.79

99.7

99.8

3

2.25

7.2

0.129

1.075

2.22

7.17

98.9

99.6

 

Mean

99.2

99.5

± S.D.

0.41

0.26

C.V.

0.41

0.26

 

 

 

Table No.2: Results of Ruggedness studies of IND and PER

Parameter

Statistical data

Two wavelength method

IND

PER

Interday

Mean

99.2

99.3

± S.D.

0.50

0.5

C.V.

0.51

0.5

Intraday

Mean

99.3

99.1

± S.D.

0.51

0.36

C.V.

0.52

0.36

 

Different analyst

 

Mean

99.1

99.2

± S.D.

0.61

0.45

C.V.

0.62

0.46

 

 


For precision, replicate estimation of both IND and PER in the same batch of tablets was done by proposed method, which yielded quite concurrent results, indicating reliability of the method. The values of SD or RSD are within the prescribed limit of 2 %, showing high precision of the method, as shown in Table No. 1. For ruggedness the proposed method was repeated under different conditions like different time, on different day and by different analyst. The results shown in Table No. 3, prove that the method is reproducible. During the linearity study it was observed that absorbance values of IND and PER in the marketed formulation were linear in the range of 80 % to 120 % of the test concentration with R2 close to one for this method of analysis. From the study of validation parameters namely accuracy, precision (SD and RSD), ruggedness (interday, intraday and different analyst), linearity and range, it was observed that the method is specific, accurate, precise, reproducible and rugged. Hence, this method can be employed for routine analysis of tablet dosage form.

 

 

 

ACKNOWLEDGEMENTS:

The authors are thankful to Prof. (Dr.) A.V.Chandewar, Principal, P.Wadhwani College of Pharmacy, Yavatmal, for his valuable guidance and advice. The authors are grateful to Micro Labs, Banglore and Cipla, Bombay for providing gift sample of Indapamide and Perindopril Erbumine respectively.

 

 

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Received on 11.07.2010          Modified on 23.07.2010

Accepted on 31.07.2010         © RJPT All right reserved

Research J. Pharm. and Tech. 4(4): April 2011; Page 545-548