固相萃取-超高效液相色谱-串联质谱法测定六神曲中4种烷基间苯二酚

Determination of 4 Alkylresorcinols in Massa Medicata Fermentata by Solid Phase Extraction-Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry

建立固相萃取-超高效液相色谱-串联质谱法(UPLC-MS/MS)同时测定六神曲中5-十五烷基间苯二酚(AR-15)、5-十七烷基间苯二酚(AR-17)、5-十九烷基间苯二酚(AR-19)和5-二十一烷基间苯二酚(AR-21)4种烷基间苯二酚含量.分析采用Brownless Spp C_(18)色谱柱(2.1 mm×100 mm,2.7μm),流动相:A相为5 mmol/L乙酸铵溶液,B相为甲酸∶乙腈溶液(体积比为1∶1000),采用梯度洗脱方式,体积流速为0.5 mL/min.柱温为35℃,使用电喷雾离子源,负离子扫描,多反应监测模式.结合主成分分析与正交偏最小二乘法判别分析,寻找影响六神曲炮制前后质量的主要化学成分.结果表明,4种成分在各自范围内线性关系良好(r>0.9926),平均加样回收率在99.36%~101.36%之间,相对标准偏差(RSD)为0.85%~2.10%.通过化学模式识别分析发现造成主要差异的质量差异物是AR-21.方法简便可行,稳定可靠,可用于炮制不同六神曲的质量控制.

A method of solid phase extraction-ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)has been established for the simultaneous determination of 4 alkylresorcinols in Massa Medicata Fermentata,including 5-pentadecyl resorcinol(AR-15),5-heptadecylresorcinol(AR-17),5-nonadecylresorcinol(AR-19)and 5-heneicosylresorcino(AR-21).The analysis was performed on a Brownless Spp C_(18)column(2.1 mm×100 mm,2.7μm)with the mobile phase of 5 mmol/L ammonium acetate solution in phase A and formic acid:acetonitrile solution(volume ratio of 1:1000)in phase B at a flow rate of 0.5 mL/min with gradient elution.The column temperature was 35℃,and the electron spray ionization source was adopted in negative ion scanning with multiple reaction monitoring mode.Combined with principal component analysis and orthogonal partial least square-discriminant analysis,the main chemical components affecting the quality of Massa Medicata Fermentata before and after processing were found.The results showed that 4 components had good linear relationships in their respective ranges(r>0.9926),the average spiked recoveries ranged from 99.36%to 101.36%,and the relative standard deviations(RSD)were 0.85%~2.10%.The mass difference substance was AR-21 that caused the main difference through chemical pattern recognition analysis.The method is simple,feasible,stable and reliable,and can be used for the quality control of different products of Massa Medicata Fermentata.