Study on the HPLC Fingerprint of Zuogui Drink and Content Determination of Five Components
Objective To provide a basis for the quality control of Zuogui Drink.Methods Fingerprint of 15 batches of Zuogui Drink samples was established by the high-performance liquid chromatography(HPLC)method.The chromatographic column was the Agilent 5 HC-C18 column(250 mm×4.6 mm,5 μm),the mobile phase was 0.2%formic acid aqueous solution-acetonitrile(gradient elution),the flow rate was 1.0 mL/min,the detection wavelength was 250 nm,the column temperature was 30℃,and the injection volume was 20 μL.The Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(Version 2012.1307023)was used for similarity evaluation,the common peaks were marked,the components were identified,and the contents were determined.Results The similarity between the fingerprint of 15 batches of samples and the reference fingerprint was in the range of 0.975-0.999.A total of 33 common peaks were marked,and five of them were identified,namely 5-hydroxymethylfurfural(5-HMF),morroniside,loganin,liquiritin and glycyrrhizic acid.The linear ranges of the above five components were 0.017-0.266 mg/mL,0.049-0.784 mg/mL,0.034-0.539 mg/mL,0.016-0.250 mg/mL,and 0.009-0.150 mg/mL(R2>0.9990,n=5)respectively.The RSDs of precision,stability and repeatability tests were all lower than 3.0%.The average recovery rates of the above five components were 101.40%,98.72%,98.00%,101.53%,99.13%,with RSDs of 2.06%,2.09%,1.65%,1.56%,0.40%(n=6)respectively.The contents of 5-HMF,morroniside,loganin,liquiritin and glycyrrhizic acid in samples were in the range of 49.838-140.933 μg/g,131.060-340.103 μg/g,68.081-178.313 μg/g,12.108-47.326 μg/g,and 1.190-55.483 μg/g,respectively.Conclusion The established method can be used for the establishment of HPLC fingerprint of Zuogui Drink and the contents determination of the five components,which can provide a basis for the comprehensive quality evaluation of Zuogui Drink.
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