UPLC-MS/MS快速测定茯苓中去氢土莫酸和茯苓酸的含量
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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,1.8μm),流动相乙腈-1.0 mmol·L-1乙酸铵(80∶20),流速0.3 m L·min-1,柱温40℃,进样量5.0μL。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26.2~524μg·L-1(r=0.999 8)和24.2~488μg·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。
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-C18column( 2. 1 mm × 100 mm,1. 8 μm) with acetonitrile-1. 0 mmol·L- 1ammonium acetate( 80 ∶ 20) as the mobile phase. The flow rate was 0. 3 m L·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 of1. 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.
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-C18column( 2. 1 mm × 100 mm,1. 8 μm) with acetonitrile-1. 0 mmol·L- 1ammonium acetate( 80 ∶ 20) as the mobile phase. The flow rate was 0. 3 m L·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 of1. 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.
引文
[1]国家药典委员会,中华人民共和国药典.一部[M].北京:中国医药科技出版社,2015:240-141.
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[9]杨改红,程昊,黄群,等.HPLC-MS/MS快速测定远志中远志口山酮Ⅲ和3,6'-二芥子酰基蔗糖的含量[J].中国实验方剂学杂志,2014,20(24):39-42.
[10]刘校妃,张志强,邹志琴,等.白参及红参中五种人参皂苷含量的HPLC-MS/MS法快速测定[J].时珍国医国药,2015,26(8):1801-1804.
[11]WANG W,DONG H,YAN R,et al.Comparative study of lanostane-type triterpene acids in different parts of Poria cocos(Schw.)Wolf by UHPLC-Fourier transform MS and UHPLC-triple quadruple MS[J].J Pharm Biomed Anal,2015,102:203-214.
[2]昝俊峰.茯苓三萜类成分抗肿瘤活性研究与茯苓药材质量分析[D].武汉:湖北中医药大学,2012.
[3]WANG Y,ZHANG J,ZHAO Y,et al.Mycology,cultivation,traditional uses,phytochemistry and pharmacology of Wolfiporia cocos(Schwein.)Ryvarden et Gilb:A review[J].J Ethnopharmacol,2013,147(2):265-276.
[4]WU Z,REN H,LAI W,et al.Sclederma of Poria cocos exerts its diuretic effect via suppression of renal aquaporin-2 expression in rats with chronic heart failure[J].J Ethnopharmacol,2014,155(1):563-571.
[5]仲兆金,许先栋.茯苓三萜化学成分及其光谱特征研究进展[J].中国药物化学杂志,1997,7(1):74-81.
[6]徐榕,许津.紫外分光光度法测定茯苓中三萜成分[J].药物分析杂志,2005,25(4):449-451.
[7]徐斌,昝俊峰,何丽姗,等.27种菌种茯苓中茯苓酸分析比较研究[J].中草药,2010,41(4):647-649.
[8]段启,姚丽梅,庄义修.茯苓与茯苓皮中茯苓酸含量比较研究[J].中药新药与临床药理,2010,21(6):654-655.
[9]杨改红,程昊,黄群,等.HPLC-MS/MS快速测定远志中远志口山酮Ⅲ和3,6'-二芥子酰基蔗糖的含量[J].中国实验方剂学杂志,2014,20(24):39-42.
[10]刘校妃,张志强,邹志琴,等.白参及红参中五种人参皂苷含量的HPLC-MS/MS法快速测定[J].时珍国医国药,2015,26(8):1801-1804.
[11]WANG W,DONG H,YAN R,et al.Comparative study of lanostane-type triterpene acids in different parts of Poria cocos(Schw.)Wolf by UHPLC-Fourier transform MS and UHPLC-triple quadruple MS[J].J Pharm Biomed Anal,2015,102:203-214.