上海交通大学学报(医学版) ›› 2020, Vol. 40 ›› Issue (4): 443-.doi: 10.3969/j.issn.1674-8115.2020.04.005

• 论著·基础研究 • 上一篇    下一篇

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

葛 坤,瞿 纯,贾 伟,赵爱华   

  1. 上海交通大学附属第六人民医院转化医学中心,上海200233
  • 出版日期:2020-04-28 发布日期:2020-05-22
  • 通讯作者: 赵爱华,电子信箱:zhah@sjtu.edu.cn。
  • 作者简介:葛 坤(1992—),男,学士;电子信箱:1462968575@qq.com。
  • 基金资助:
    国家自然科学基金项目(81772530,81974073)。

Quantification of human plasma 7α-hydroxy-4-cholesten-3-one and bile acidsUPLC-MS/MS

GE Kun, QU Chun, JIA Wei, ZHAO Ai-hua   

  1. Center for Translational Medicine, Shanghai Sixth Peoples Hospital, Shanghai Jiao Tong University, Shanghai 200233, China
  • Online:2020-04-28 Published:2020-05-22
  • Supported by:
    National Natural Science Foundation of China (81772530, 81974073).

摘要: 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。

关键词: 7&, alpha, - 羟基 -4- 胆固醇烯 -3- 酮, 胆酸, 鹅去氧胆酸, 超高效液相色谱 - 串联质谱, 定量

Abstract: Objective · To develop a quantitative method of 7α-hydroxy-4-cholesten-3-one (C4), cholic acid (CA) and chenodeoxycholic acid (CDCA) in human plasma. Methods · After 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). Results · The 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.

Key words: 7α-hydroxy-4-cholesten-3-one (C4), cholic acid (CA), chenodeoxycholic acid (CDCA), UPLC-MS/MS, quantification