Development and validation of Spectrophotometric method and TLC Densitometric Determination of Irinotecan HCl in pharmaceutical dosage forms

Abstract

The study is focused to develop and validate a UV-Spectrophotometric and Densitometry method for simultaneous estimation of Irinotecan from their dosage form. Based on spectrophotometric characteristic of method are described for the simultaneous determination of Irinotecan HCl, at 247nm for simple UV spectrum and at 268nm for derivative spectrum was found adequate for quantification. The method was validated for linearity, accuracy, precision, reproducibility, and specificity as per International Conference on Harmonization guidelines. The linearity signal and concentration of Irinotecan in the range of 2–10μg/ml in aqueous solution present a correlation coefficient (r2) of 0.9999 for simple UV and 0.9997 for first order derivative spectrum. Second method is the high performance thin layer chromatographic (HPTLC) separation followed by densitometric measurements on normal phase silica gel 60F254. The chromatographic separation was carried out on precoated silica gel 60F254 aluminium plates using a mixture of toluene/ethyl acetate/methanol/carbon tetrachloride, in the volume ratio of 9.2:5:0.9:0.8 (v/v/v/v) respectively as mobile phase. Densitometric analysis was carried out at 317nm. The linear regression analysis data showed good linear relationship in the concentration range of 200–1200 (ng/band) for Irinotecan. The limits of quantitation for Irinotecan were 34 (ng/band). The limit of detection (LOD) and limit of quantitation (LOQ) were 36 and 57ng/spot, respectively. The drug was satisfactorily resolved with Rf value 0.34±0.08. The accuracy and repeatability of the proposed method were ascertained by evaluating various validation parameters like linearity (200–1200ng/spot). The method was found to be rapid, specific, precise and accurate and can be successfully applied for the routine analysis of Irinotecan bulk and marketed dosage form.