Optimization of Ethanol Extraction Process of Herba Clematis by Orthogonal Test
Objective To optimize the ethanol extraction process of Herba Clematis.Methods The high-performance liquid chromatography(HPLC)method was used to determine the content of quercetin in the Herba Clematis Decoction Piece samples,and the ultraviolet absorption spectroscopy was used to determine the content of total flavonoids in the decoction piece samples.The ethanol extraction process of Herba Clematis was optimized by the L9(34)orthogonal test with the ratio of material to liquid,extraction frequency and extraction time as the investigation factors,and with the yield of dry extract,total flavonoid content and quercetin content as the investigation indicators.Then the verification test was conducted.Results The mass concentration of quercetin showed a good linear relationship with the peak area in the range of 1.99-198.99 µg/mL(r=0.999 9,n=6).The limit of quantification and the limit of detection were 0.060 µg/mL and 0.018 µg/mL respectively.The RSDs of precision,stability and repeatability tests were all lower than 2.0% .The average recovery rate of quercetin was 101.74% with an RSD of 1.25% (n=6).The mass concentration of rutin showed a good linear relationship with the absorbance in the range of 7.42-44.52 µg/mL.The optimal process was as follows:adding 10 times of 40% ethanol,extracting three times,each time for 0.5 h.The yield of dry extract,total flavonoid content and quercetin content in the the verification test were 25.10%,56.78 mg/g,1.35 mg/g,with the RSDs of 2.43%,4.10%,2.13% (n=3),respectively.Conclusion The optimal ethanol extraction process in this study is stable,reliable and repeatable,which can provide a reference for the quality evaluation of Herba Clematis and the industrial development of its preparation products.
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