| Abstrakt: |
A high-performance thin-layer chromatography (HPTLC) method has been developed for the simultaneous estimation of abacavir sulfate (ABC), lamivudine hydrochloride (LAM), and dolutegravir sodium (DTG) in an in-house physical mixture and it was validated as per the International Conference on Harmonization (ICH) guidelines. A 23 full factorial design was utilized to aid in method development and optimization. Effective chromatographic separation was achieved using pre-coated silica gel aluminum HPTLC plate 60 F254 as the stationary phase and ethyl acetate-ethanol- acetone-ammonia (4.478:0.740:0.50:0.15, v/v) as the mobile phase. The optimized chamber saturation time was kept at 30 min. Densitometric scanning was performed in the absorbance-reflectance mode at a detection wavelength of 267 nm. Quality by design approach was applied to evaluate the effect of all three factors, i.e., volume of ethyl acetate, volume of ethanol, and volume of acetone, on the chromatographic response retardation factor (RF) of each drug. The RF values of ABC, LAM, and DTG were found to be 0.65, 0.34, and 0.26, respectively. The calibration curves were found to be linear by taking independently weighed standard solutions with concentrations of 4.8, 7.2, 9.6, 12.0, and 14.4 μg per band for ABC, 2.4, 3.6, 4.8, 6.0, and 7.2 μg per band for LAM, and 0.4, 0.6, 0.8, 1.0, and 1.2 μg per band for DTG. The limit of detection and limit of quantification values were found to be 0.9972 μg per band and 3.0218 μg per band for ABC, 0.2544 μg per band and 0.7711 μg per band for LAM, and 0.1004 μg per band and 0.3043 μg per band for DTG, respectively. The % recoveries of the drugs by the developed method were found in the range of 98.0955-100.9813% for ABC, 98.2616-99.9900% for LAM, and 98.4666-101.3000% for DTG, respectively. The proposed method was found to be novel, simple, accurate, reproducible, and robust. The developed HPTLC method was successfully applied for the quantitative determination of ABC, LAM, and DTG in an in-house physical mixture. [ABSTRACT FROM AUTHOR] |
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