HPTLC (high performance thin layer chromatography) is playing an important role in today analytical world, not in competition to HPLC but as a complementary method. This study presents the first report of sensitive, selective, precise and robust HPTLC method, which has been developed and validated for quantification of the rubiadin using the solvent system of toluene: acetonitrile: ethyl acetate: formic acid (10: 1: 2.5: 2 v/v/v/v). The method employed HPTLC aluminium plates precoated with silica gel 60 F254 as a stationary phase as the stationary phase. Densitometric analysis of rubiadin was carried out in the absorbance mode at 280 nm by using camag TLC scanner-3. This system was found to give compact spot of rubiadin at R f value of 0.70 with good linear relationship i.e. r2 = 0.9984. The method was validated using ICH guidelines in terms of specificity, precision, robustness, stability and repeatability. The repeatability of the method was found to be 1.25 RSD and recovery values from 98.40 to 100.28% for rubiadin. Statistical analysis proves that the method is repeatable and selective for the estimation of the said drug. Since the proposed mobile phase effectively resolves the rubiadin, this HPTLC method can be applied for identification and quantitation of these phytochemical in herbal extracts and pharmaceutical dosage form.
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