Abstract

ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR THE ESTIMATION OF SOFOSBUVIR AND LEDIPASVIR IN RAW MATERIAL AND TABLET FORMULATION BY HIGH PERFORMANCE THIN LAYER CHROMATOGRAPHIC (HPTLC) METHOD

Ramesh Jayaprakash*, Anandakumar Karunakaran, Kailasam Periyana Gounder, Kalaiselvi Ponnusamy, Anjana Elampulakkadu

ABSTRACT

A simple, sensitive and rapid high performance thin layer chromatographic (HPTLC) method has been developed and validated for quantitative determination of Sofosbuvir (SFB) and Ledipasvir (LDP) in bulk and formulations. The chromatographic development was carried out on HPTLC aluminum plates precoated with silica gel 60 F254 using a mixture of ethyl acetate, methanol, toluene, acetone and acetic acid (2:1.5:4.5:2.0:0.2) (v/v/v/v) as mobile phase. Detection was carried out densitometrically at 269 nm. The Rf value of SFB and LDP were found to be 0.19 and 0.06 respectively. The method was validated as per ICH guidelines with respect to linearity, accuracy, precision, robustness etc. the calibration curve were found to be linear over a range of 200-1200 ng/spot and 45-270 ng/spot for SFB and LDP respectively, with a regression coefficient of 0.9979 and 0.9986 for SFB and LDP respectively. The accuracy was found to be very high 99.41-100.04 and 99.64-100.64 for SFB and LDP respectively. %RSD values for intra-day and inter-day variation were less than 2. The method has demonstrated high sensitivity and specificity. The method is new, simple and economic for routine estimation of SFB and LDP in bulk and pharmaceutical formulation to help in industries as well as researchers for their sensitive determination of SFB and LDP rapidly at low cost in routine analysis.

Keywords: HPTLC; Sofosbuvir; Ledipasvir; Validation; Estimation; Raw material.