经典名方圣愈汤的UPLC指纹图谱建立及多成分定量分析

目的:建立圣愈汤的超高效液相色谱法(UPLC)指纹图谱及其11个成分的定量分析方法.方法:采用UPLC-二极管阵列检测器(PDA)/蒸发光散射检测器(ELSD),根据化学成分极性不同使用不同的检测器,建立2种色谱条件,指纹图谱1选择ACQUITY UPLC HSS T3色谱柱(2.1 mm×100 mm,1.8μm),...

Establishment of UPLC Fingerprint and Quantitative Analysis of Multi-components of Shengyutang

Objective:To establish ultra performance liquid chromatography(UPLC)fingerprint of Shengyutang and quantitative analysis method of 11 index components in this famous classical formula.Method:UPLC-diode array detector/evaporative light scattering detector(UPLC-PDA/ELSD)was used,two chromatographic conditions were established by different detectors according to the polarity of chemical components.Conditions of fingerprint 1 were as follows:ACQUITY UPLC HSS T3 column(2.1 mm×100 mm,1.8μm)with the mobile phase of acetonitrile(A)-0.6%formic acid solution(C)for gradient elution(0-4 min,0-4%A;4-8 min,4%A;8-9 min,4%-8%A;9-14 min,8%-9%A;14-21 min,9%-15%A;21-26 min,15%-17%A;26-30 min,17%-20%A;30-35 min,20%-32%A;35-40 min,32%-40%A;40-50 min,40%-80%A;50-55 min,80%A),the flow rate of 0.3 mL·min^-1,PDA with detection wavelengths of 280 nm and 321 nm,the column temperature at 30℃.Conditions of fingerprint 2 were as follows:the CORTECS C18column(3.0 mm×100 mm,2.7μm)with the mobile phase of acetonitrile(A)-water(D)for gradient elution(0-11 min,19%A;11-16 min,19%-25%A;16-34 min,25%-28%A;34-47 min,28%-47%A;47-60 min,47%-80%A),the flow rate of 0.4 mL·min^-1,ELSD with drift tube temperature of 95℃,the carrier gas(air)flow rate of 2.0 L·min^-1,and the column temperature at 30℃.UPLC-PDA/ELSD fingerprints of 15 batches of Shengyutang were established,and the similarity was evaluated by similarity evaluation system of chromatographic fingerprint of traditional Chinese medicine(2012 edition)issued by the Chinese Pharmacopoeia Commission,and the contents of eleven index components in this famous classical formula were determined.Result:The similarities of UPLC-PDA/ELSD fingerprints of 15 batches of Shengyutang were>0.98 by comparing with the control fingerprint,27and 16 common peaks were identified in fingerprint 1,2,respectively.It was tested and verified that the precision,repeatability,stability,linear relationship and other results of this method all met the requirements of the 2020 edition of Chinese Pharmacopoeia.The contents of chlorogenic acid,ferulic acid,calycosin glucoside,verbascoside,senkyunolide I,senkyunolide H,senkyunolide A,ginsenoside Rg1,ginsenoside Re,ginsenoside Rb1 and astragaloside A in 15 batches of Shengyutang were 0.063-0.193,0.509-0.638,0.160-0.318,0.012-0.056,0.394-0.519,0.110-0.143,0.031-0.097,0.382-0.595,0.292-0.505,0.590-0.803,0.142-0.367 mg·g-1,respectively.Conclusion:The established detection method meets the requirements of the 2020 edition of Chinese Pharmacopoeia,which can characterize the overall characteristics of chemical components in Shengyutang,and provide experimental basis for the quality standard research of this famous classical formula.