Establishment of UPLC Fingerprint and Determination of Four Components in Yinxing Yangnao Formula
Objective To establish the ultra-high performance liquid chromatography(UPLC)fingerprint of Yinxing Yangnao Formula and determine the contents of four identified components.Methods The chromatographic column was the ACQUITY UPLC® BEH C18 column(100 mm×2.1 mm,1.7 μm),the mobile phase was 0.1%phosphoric acid aqueous solution-acetonitrile(gradient elution),the flow rate was 0.4 mL/min,the detection wavelength was 321 nm,the column temperature was 30℃,and the injection volume was 1 μL.The UPLC fingerprint of 12 batches of samples was established and the similarity evaluation was performed by the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(Version 2012 A),the common peaks were marked and identified.The samples were classified and the quality was evaluated by the cluster analysis and principal component analysis.The contents of identified components in the samples were determined by the UPLC method.Results There were 30 common peaks in the UPLC fingerprint of 12 batches of samples,which from Ginkgo Folium,honey-baked Polygalae Radix and Chuanxiong Rhizoma;four components were identified,namely ferulic acid,3,6'-disinapoyl sucrose,isoginkgetin and sciadopitysin.The similarity of the samples was greater than 0.990.The samples were classified into four groups.The cumulative variance contribution rate of the seven principal components was 94.100%.The linear ranges of ferulic acid,3,6'-disinapoyl sucrose,isoginkgetin and sciadopitysin were 3.36-67.20 μg/mL,5.10-102.00 μg/mL,5.00-100.00 μg/mL,2.50-50.00 μg/mL(R2≥0.9980,n=5)respectively.The RSDs of precision,stability and repeatability tests were all lower than 3.0%.The average recovery rate of the above four components was in the range of 99.01%-101.19%,with the RSDs in the range of 0.22%-1.01%(n=6).Conclusion This method is simple,feasible,specific and repeatable,which can be used for the quality evaluation of Yinxing Yangnao Formula,lay the foundation for the quality control and secondary development of Yinxing Yangnao Tablets.