LC-MS/MS法测定人血浆中隐丹参酮及丹参酮ⅡA的含量
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摘要
目的用LC-MS/MS法测定人血浆中隐丹参酮及丹参酮ⅡA的浓度。方法血浆样品经甲醇沉淀蛋白处理,色谱柱:Symmetry C18(150. 0 mm×4. 6mm,5μm),流动相:乙腈-水(含0. 1%甲酸),梯度洗脱,流速:1. 0 m L·min~(-1),柱温:35℃,进样量:10μL,采用电喷雾离子源(ESI),正离子模式,多反应监测方式。考察该方法的专属性、标准曲线和定量下限、精密度与回收率,以及稳定性。结果人血浆中隐丹参酮及丹参酮ⅡA分别为0. 50～100. 10和0. 51～101. 20 ng·m L~(-1)内线性关系良好,定量限分别为0. 50和0. 51ng·m L~(-1),日内、日间精密度(RSD)均小于8. 3%,低、中、高3种浓度隐丹参酮及丹参酮ⅡA的方法平均回收率均大于88. 3%。结论本法特异性好、灵敏度高、精密度好、简便快速,可用于人血浆中隐丹参酮和丹参酮ⅡA的含量测定。
Objective To develop a LC-MS/MS method for the determination of cryptotanshinone and tanshinone Ⅱ A in human plasma.Methods The plasma samples were precipitated with methano1. After sample preparation,the samples were separated on a Symmetry C18( 150 mm × 4. 6 mm,5 μm) column using acetonitrile and water( with 0. 1%formic acid) as mobile phase in gradient elution mode. Velocity of flow was 0. 1 m L·min~(-1). Column temperature was 35 ℃. Sample size was 10μL. The mass spectrometer was operated under the positive ion mode with the electrospray ionization source. And,the detection was performed by multiple reaction monitoring. The specificity,standard curve and lower limit of quantification,precision and recovery,stability of the method were investigated. Results The linear relationships for cryptotanshinone and tanshinone Ⅱ A were good in the concentration range of0. 50-100. 10 ng·m L~(-1) and 0. 51-101. 20 ng·m L~(-1),limit of quantitation were 0. 50 and 0. 51 ng·m L~(-1) and the intra-day and inter-day precision( RSD) were less than 8. 3%. The average recoveries for cryptotanshinone and tanshinone Ⅱ A at low,medium and high levels were greater than 88. 3%. Conclusion The analysis method was sensitive,simple and suitable enough to be applied for the determination of cryptotanshinone and tanshinone ⅡA in human plasma.
Objective To develop a LC-MS/MS method for the determination of cryptotanshinone and tanshinone Ⅱ A in human plasma.Methods The plasma samples were precipitated with methano1. After sample preparation,the samples were separated on a Symmetry C18( 150 mm × 4. 6 mm,5 μm) column using acetonitrile and water( with 0. 1%formic acid) as mobile phase in gradient elution mode. Velocity of flow was 0. 1 m L·min~(-1). Column temperature was 35 ℃. Sample size was 10μL. The mass spectrometer was operated under the positive ion mode with the electrospray ionization source. And,the detection was performed by multiple reaction monitoring. The specificity,standard curve and lower limit of quantification,precision and recovery,stability of the method were investigated. Results The linear relationships for cryptotanshinone and tanshinone Ⅱ A were good in the concentration range of0. 50-100. 10 ng·m L~(-1) and 0. 51-101. 20 ng·m L~(-1),limit of quantitation were 0. 50 and 0. 51 ng·m L~(-1) and the intra-day and inter-day precision( RSD) were less than 8. 3%. The average recoveries for cryptotanshinone and tanshinone Ⅱ A at low,medium and high levels were greater than 88. 3%. Conclusion The analysis method was sensitive,simple and suitable enough to be applied for the determination of cryptotanshinone and tanshinone ⅡA in human plasma.
引文
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[2]ZHOU L,ZUO Z,CHOW M S.Danshen:an overview of its chemistry pharmacology,pharmacokinetics,and clinical use[J].J Clin Pharmacol,2005,45(12):1345-1359.
[3]蒋晓蕊,苗琳,吴晓燕,等.丹参酮ⅡA对心血管系统保护作用及机制的研究进展[J].中国当代医药,2014,21(14):183-185.
[4]丁建刚,高允生,刘江林,等.HPLC-MS/MS法测定比格犬血浆中丹参酮ⅡA的含量[J].药物分析杂志,2009,29(5):769-772.
[5]宋玉乔,廖杰,黄学亮,等.液相色谱-串联质谱法快速测定人血浆中丹参酮ⅡA的含量[J].现代仪器,2011,17(97):22-24.
[6]周琳,刘宪英,郑金凤,等.丹参多组分渗透泵片在Beagle犬体内的药动学研究[J].中国中药杂志,2012,37(12):176l-1764.