Abstract

HPTLC METHOD DEVELOPMENT AND VALIDATION OF IRBESARTAN AND HYDROCHLOROTHIAZIDE IN TABLET DOSAGE FORM.

Tabassum I. Hangad*, Pranav Ghatbhale, Asmita More, Nikita Barse, Dr. Rani Potawale, Dr. Dilnawaz Pathan

ABSTRACT

High performance thin layer chromatographic method has been developed and validated for irbesartan and hydrochlorothiazide in tablet dosage form. Both drugs were found to have a significant absorbance at 271 nm. Lambda maximum is 271 nm. Because Hydrochlorothiazide has a lower concentration in the formulation than Irbesartan so it was chosen for measurement. Chloroform: Ethyl Acetate (7:3 v/v), was chosen as the mobile phase. HPTLC pre-coated plates of silica gel G 60 F254 10x10 cm in size were used for the spotting of standard solutions.Solution of 300 μg/ml of Irbesartan and 25 μg/ml of Hydrochlorothiazide was prepared. 4 μl (1200 ng/band) of Irbesartan and 4 μl (100 ng/band) of Hydrochlorothiazide solution was applied on the TLC plate with the help of a Hamilton syringe. The plate was dried and was scanned over an 80-mm distance at 271 nm. The spot showed a good response at 271 nm. The linearity (relationship between peak area and concentration) was determined over the concentration range 600-3600 ng/band for IRB and 50-300 ng/band for HTZ. Intra-day and inter-day studies were used to demonstrate the method's precision. In the intra-day precision 2, 4, 6 μl of solution of concentration 300 μg/ml of IRB (600, 1200, 1800 ng/band) and 2, 4, 6 μl of solution of concentration 25 μg/ml of HTZ (50, 100, 150 ng/band) were prepared and three replicates were analyzed, % RSD was calculated. For each medication, a sample solution was injected and the area was recorded. Concentration and % purity was determined from linear equations. To check accuracy of the method, recovery studies were carried out by adding standard drug to sample at three different levels 50, 100 and 150 %.The drug concentrations of IRB and HTZ were calculated by using linearity equations of IRB and HTZ. Peak purity profiling experiments were used to determine the method's specificity.

Keywords: drug concentrations, linearity, Hydrochlorothiazide, accuracy.