Analytical Method Validation and Simultaneous Determination of Ketotifen Fumarate and Cyproheptadine Hydrochloride in Divided Powder by RP-HPLC-UV Method

Abstract

Ketotifen fumarate and cyproheptadine HCl on divided powder dosage form are the most popular prescription for children at “X” hospital in Yogyakarta. The interest for studying this divided powder was related to the quality test towards human health. If the drug doesn’t meet the requirement of good quality drug, it can cause damage to the quality of life. Hence, it was important to develop a validated analytical method to quantify content of active drug from samples. The samples were prepared by a pharmacist at “X” hospital in Wonosari, Yogyakarta. Tablets from each compound were blended and divided by pulverizer into powder bags then were sealed. In this study, a reversed phase HPLC Ultraviolet (RP-HPLC-UV) method was optimized and validated then applied for determining the content of ketotifen fumarate and cyproheptadine HCl in divided powder. The optimal condition for HPLC separation with mobile phase of methanol were 0.04M Na2HPO4 buffer (60:40) and flow rate 0.8 mL/min. According to AOAC International 2019, the developed RP-HPLC-UV method was good and met the requirement of selectivity and sensitivity. The calibration curve formulas of ketotifen fumarate and cyproheptadine hydrochloride were y = 27714x-79111 (r=0.994) and y = 26324x+72581 (r=0.993) in the range of 10.9-54.5 and 10.6-53.0 µg/mL, respectively. Accuracy and intermediate precision were assessed on both standard solution and samples spiked with standard solution of ketotifen fumarate and cyproheptadine hydrochloride. It was found that both standard solution and the spiked samples solution met the requirement of recovery of 80-110% and RSD of less than 7.3% for intraday and interday analysis. Content of ketotifen fumarate and cyproheptadine hydrochloride in sample (n=7) were 16.574±0.098 and 30.640±0.035 µg/mL, respectively. It can be concluded that the RP-HPLC-UV method was successfully validated then applied in the determination of ketotifen fumarate and cyproheptadine HCl in divided powder.