人血浆7α-羟基-4-胆固醇烯-3-酮及胆汁酸的超高效液相色谱-串联质谱定量方法研究

Translated title of the contribution: Quantification of human plasma 7α-hydroxy-4-cholesten-3-one and bile acids by UPLC-MS/MS

葛坤, 瞿纯, 贾伟, 赵爱华*

*Corresponding author for this work

Research output: Contribution to journalJournal articlepeer-review

Abstract

目的
建立人血浆中7α- 羟基 -4- 胆固醇烯 -3- 酮(7α-hydroxy-4-cholesten-3-one,C4)以及胆酸(cholic acid,CA)、鹅去氧胆酸(chenodeoxycholic acid,CDCA)的定量方法。

方法
采用乙腈提取人血浆中C4、CA、CDCA,利用超高效液相色谱 - 串联质谱(ultra-high performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)技术,通过内标校正的标准曲线法定量C4、CA、CDCA含量。

结果
经方法学验证,C4、CA、CDCA的检出限(limits of detection,LOD)分别为0.16、0.02、0.04 nmol/L,均具有良好的线性关系(均R2>0.998);重复性的相对标准偏差(relative standard deviation,RSD)均低于3.0%;日内及日间精密度的RSD均小于6%;4 ℃保存7 d结果稳定,RSD均小于10%;C4、CA和CDCA的平均加标回收率分别为97.7%、113.3%、105.0%。

结论该方法具有较高的检测灵敏度、良好的精密度和稳定性,可用于血浆生物样本的定量检测。

ObjectiveTo develop a quantitative method of 7α-hydroxy-4-cholesten-3-one (C4), cholic acid (CA) and chenodeoxycholic acid (CDCA) in human plasma.

MethodsAfter extraction of C4, CA and CDCA with acetonitrile plasma, they were quantified with standard curve correctedthe internal standards based on Ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).

ResultsThe limits of detection (LOD) of C4, CA and CDCA were 0.16, 0.02 and 0.04 nmol/L respectively; All three metabolites had good linear relationships (correlation coefficients R2 were over than 0.998). The relative standard deviations (RSDs) of repeatabilities were below 3.0%. The RSDs of inter-day and intra-day precision were less than 6%, and the RSDs of stabilities at 4 °C were below 10% within 7 days. The average added recoveries of C4, CA and CDCA were 97.7%, 113.3% and 105.0%, respectively.

Conclusion
This method is of high detective sensitivity, good precision and stability, which meets the quantitative requirements of plasma biological samples.
Translated title of the contributionQuantification of human plasma 7α-hydroxy-4-cholesten-3-one and bile acids by UPLC-MS/MS
Original languageChinese (Simplified)
Pages (from-to)443-449
Number of pages7
Journal上海交通大学学报 (医学版)
Volume40
Issue number4
DOIs
Publication statusPublished - 28 Apr 2020

Scopus Subject Areas

  • Biochemistry, Genetics and Molecular Biology(all)

User-Defined Keywords

  • 7α-hydroxy-4-cholesten-3-one (C4)
  • Chenodeoxycholic acid (CDCA)
  • Cholic acid (CA)
  • Quantification
  • UPLC-MS/MS

Fingerprint

Dive into the research topics of 'Quantification of human plasma 7α-hydroxy-4-cholesten-3-one and bile acids by UPLC-MS/MS'. Together they form a unique fingerprint.

Cite this