Study on HPLC Fingerprint and Content Determination of Huangqi Guizhi Wuwu Decoction
Objective To establish a high-performance liquid chromatography(HPLC)fingerprint of Huangqi Guizhi Wuwu Decoction and a content determination method of multi-indicator components.Methods The HPLC method was adopted.The chromatographic column was the Waters SunFireTM C18 column(250 mm×4.6 mm,5 µm),the mobile phase was acetonitrile-0.1% formic acid aqueous solution(gradient elution),the flow rate was 1.0 mL/min,the detection wavelength was 254 nm,the column temperature was 40℃,and the injection volume was 10 µL.The HPLC fingerprint of 10 batches of samples was established with cinnamaldehyde peak as a reference.The Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(2012A Version)was used for similarity evaluation.The common peaks were identified,and the medicinal herb attribution analysis was conducted.The contents of albiflorin,paeoniflorin,calycosin-7-O-β-D-glucoside,cinnamic acid and cinnamaldehyde were determined by the above chromatographic conditions.Results A total of 17 common peaks in 10 batches of samples were identified,with the similarity greater than 0.990.The verification test showed that the HPLC fingerprint of 10 batches of samples was consistent with the reference fingerprint.The linear ranges of albiflorin,paeoniflorin,calycosin-7-O-β-D-glucoside,cinnamic acid and cinnamaldehyde were 19.0-171.0 µg/mL,61.6-555.0 µg/mL,8.3-75.0 µg/mL,18.0-162.0 µg/mL and 28.4-255.9 µg/mL,respectively.The limit of detection was in the range of 0.1-1.2 µg/mL,and the limit of quantitation was in the range of 0.3-4.9 µg/mL.The RSDs of precision,stability and repeatability tests were all lower than 3.0% .The average recovery rates of the above five components were 99.31%,98.93%,100.89%,98.83%,100.85% with the RSD of 0.25%,0.30%,0.60%,1.99%,0.74% respectively(n=6).Conclusion The method is sample,stable,feasible and repeatable,which can be used for quality control of Huangqi Guizhi Wuwu Decoction.
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