Abstract

DEVELOPMENT AND VALIDATION OF A METHOD FOR THE SIMULTANEOUS HPLC ESTIMATION OF AMBRISENTAN IN TABLET DOSAGE FORM

P. Balakrishanaiah*, D. Srilatha, Ravindar Bairam and Manjunath S. Y.

ABSTRACT

The RP-HPLC method development of Ambrisentan was tested in a Kromosil C18 column based on the results of the trials. Different ratios of the different mobile phase compositions, such as methanol: water, acetonitrile: water, and potassium dihydrogen orthophosphate buffer: acetonitrile, were employed. An analytical approach was developed by experimenting with different chromatographic settings. The optimal approach was chosen for the development and validation of Ambrisentan in tablet dosage form after all of the above strategies were compared. Using a Kromosil ODS C18 Column, 5μ (250×4.6mm) as a stationary phase, Ambrisentan was measured byreverse phase HPLC technique using mobile phase A:B (30:70). Mobile phase A is phosphate buffer (pH 3.0), and mobile phase B is acetonitrile:water (90:10). At 210 nm, a VWD detector was used for detection. The method was developed and then validated for system applicability, linearity, specificity, precision, accuracy, robustness, limit of detection and limit of quantification, and stability of the solution. Chromatograms demonstrate Ambrisentan's specificity. This approach had good separation between all peaks and no interference. It indicates that there was no interference from impurities and also shows that the suggested procedures did not conflict with regularly used excipients and additives included in the tablet dosage form. The medicine complies with Beer's rule, according to the linearity table.16. In the 50–150 μg/ml range, the calibration plot for Ambrisentan was determined to be linear, and the correlation coefficient was 0.999.The accuracy of the approach is demonstrated by the findings displayed in accuracy table17, which showed that the recovery values of pure medication from the solution ranged from 98.0% to 102%.

Keywords: Ambrisentan, Rp-Hplc, Method development, Validation.