Recent progress in simultaneous estimation of rutin, quercetin and liquiritin in Cocculus hirsutus by HPTLC

Background and objectives: Rutin, quercetin and liquiritin arepolyphenol flavonoids which have shown anti-inflammatory, antihepatotoxic, antiulcer, antiallergic, antidiabetic, antiviral and antioxidant activities. They are found in many herbal plants, one of them is Cocculus hirsutus.  The objective of this research was to develop and validate a new, accurate, precise and economic HPTLC method for simultaneous estimation of rutin, quercetin and liquiritin in ethanol extract of Cocculus hirsutus leaves. Methods: The simultaneous estimation of rutin, quercetin and liquiritin has been done by HPTLC on silica gel 60 F254 TLC plate using n- butanol: acetic acid: water: formic acid (7:1:1:0.25) as the mobile phase and was quantified by densitometric scanning at 254 nm. The method was validated as well. Results: Rutin, quercetin and liquiritin were satisfactorily resolved with Rf values of 0.47±0.03, 0.63 ±0.03 and 0.82±0.02, respectively. The linearity was found to be 1500-4000, 500-3000 and 100-700 ng per spot for rutin, quercetin and liquiritin, respectively. The inter-day RSD values were always less than 2, accuracy was 99.25% ±5% for rutin, 99.29% ±5% for quercetin, and 94.04 ±6% for liquiritin. The LOD was found to be 310.234, 346.8421 and 11.5571 ng per spot and LOQ was found to be 940.1032, 451.037 and 35.0213 ng per spot for rutin, quercetin and liquiritin, respectively. Conclusion: The statistically validated results indicated that the proposed new method has good accuracy and precision. Thus this new HPTLC method could be successfully applied for simultaneous determination of rutin, quercetin and liquiritin in herbal plants and their product.