Estimation of Fluralaner from Bulk and Dosage Form By Novel RP-HPLC Technique

Fluralaner is a systemic insecticide and acaricide that is administered orally or topically. It is a veterinary drug and a member of the novel chiral isooxazoline class of antiparasitic drugs possessing promising insecticidal, acaricidal activity. The work aims to develop a simple, accurate, precise, and reproducible RP-HPLC method for the evaluation of fluralaner in bulk and liquid dosage form. The primary objective is to give an overview of the mechanism of RP-HPLC of fluralaner and explain the basis of the retention mechanism and the secondary is to achieve high-speed separation without any loss of reproducibility. The chromatographic analysis was performed isocratically by using Zorbax Eclipse XDB- C18 (4.6 mm X 150 mm, 5 µm) column. The mobile phase consisting of acetonitrile and 0.1% Formic acid, with a ratio of 70:30 v/v, was passed through the column maintained at 40°C with a flow rate of 1 ml/min. Approximately 20 µl of the solution was injected and the analyte was eluted at 265 nm. The method was validated as per ICH Q2 R1 guidelines. The retention time of the fluralaner was around 5.2 min. The Percentage RSD of each parameter was found within the limit. The recovery of the fluralaner was determined as 100.06 %. LOD and LOQ values of the fluralaner were found to be 2 µg/ml and 6 µg/ml respectively. The method was linear over the range of 10-90 µg/ml with a regression coefficient of 0.9997. All the verification parameters were within the range according to ICH guidelines.