Implementation of Total Error Perception in the Analytical Method Validation for Quantification of Related Substance in Pharmaceutical Formulation

Abstract

Objectives: This study is initiated to develop a simple and reproducible liquid chromatographic method with ultraviolet detection for the estimation of Sisomicin in Gentamicin pharmaceutical formulation by application of total error statistical tool. Methods: Gentamicin has a weak UV chromophore it is not possible to detect low levels of known related substance of gentamicin using a UV detector. Hence derivatization technique was applied to detect those substances. Chromatographic separation was accomplished by using Thermo scientific Hypersil Gold column (150 x 4.6 mm) and 5µm particle size as stationary phase by isocratic elution with Methanol: Water: Glacial acetic acid: Sodium hexane sulfonate in the ratio 70:25:5:3% v/v/ v/w as mobile phase. Sisomicin was detected at 330 nm within 25 min with a flow rate of 0.5 mL/min. Results: Concurrent results were obtained in the developed analytical method based on total error measurement. Proposed method showed good linearity response (r> 0.995) with limit of quantification at 0.007495 mg/mL, % relative standard deviation less than 1% in repeatability and % recovery was found within 96 to 98 % in accuracy. Accuracy profile result found within the range of ±10% and risk profile ±5% between the two set. Conclusion: This method can separate all the analogues of Gentamicin including known related substances. The finding demonstrated that method could be suitable for quantification of related substances in parental liquid dosage form.