UPLC-MS/MS快速测定茯苓中去氢土莫酸和茯苓酸的含量

Rapid Determination of Dehydrotumulosic Acid and Pachymic Acid in Poria by UPLC-MS/MS

摘要:

目的：建立一种测定茯苓药材中茯苓酸和去氢土莫酸含量的UPLC-MS/MS方法。方法：采用ZORBAX Eclipse plus C₁₈色谱柱（2.1 mm×100 mm，1.8 μm），流动相乙腈-1.0 mmol·L^-1乙酸铵（80：20），流速0.3 mL·min^-1，柱温40℃，进样量5.0 μL。ESI离子源，负离子模式，多反应监测模式（MRM），检测离子对分别为茯苓酸m/z 527.40/465.30，去氢土莫酸m/z 483.20/421.20。结果：茯苓酸和去氢土莫酸分别在26.2~524 μg·L^-1（r=0.999 8）和24.2~488 μg·L^-1（r=0.999 9）线性良好，分析时间为6 min，平均回收率分别为101.4%（RSD 1.2%）和99.8%（RSD 1.2%）。结论：该方法准确、快速、可靠、灵敏，可作为茯苓酸和去氢土莫酸的一种快速定量分析方法。

Abstract:

Objective: To establish a UPLC-MS/MS method for the content determination of dehydrotumulosic acid and pachymic acid in Poria. Method: Chromatographic separation was carried out on ZORBAX Eclipse XDB-C₁₈ column (2.1 mm×100 mm, 1.8 μm) with acetonitrile-1.0 mmol·L^-1 ammonium acetate (80:20) as the mobile phase. The flow rate was 0.3 mL·min^-1, and the column temperature was 40℃. The injection volume was 5.0 μL. Electrospray ionization (ESI^-) under negative ion mode was used, and the quantization of dehydrotumulosic acid and pachymic acid was performed with multiple reaction monitoring (MRM) mode. The detected ion-pairs were dehydrotumulosic acid m/z 483.20/421.20 and pachymic acid m/z 527.40/465.30. Result: The linear range was 24.2-488 μg·L^-1(r=0.999 9) for dehydrotumulosic acid, and 26.2-524 μg·L^-1 (r=0.999 8) for polyporenic acid. The retention time was 6 min. The average recoveries of dehydrotumulosic acid and polyporenic acid were 99.8% with RSD of 1.2% (n=6) and 101.4% with RSD of 1.2% (n=6), respectively. Conclusion: The proposed UPLC method is rapid, accurate, selective, sensitive, and suitable for the content determination of dehydrotumulosic acid and polyporenic acid in Poria.