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同位素稀释超高效液相色谱串联质谱法检测血浆18-羟皮质酮方法的建立及初步临床应用

尹逸丛 禹松林 于佳磊 王丹晨 马超超 邹雨桐 谢少伟 程倩 邱玲

尹逸丛, 禹松林, 于佳磊, 王丹晨, 马超超, 邹雨桐, 谢少伟, 程倩, 邱玲. 同位素稀释超高效液相色谱串联质谱法检测血浆18-羟皮质酮方法的建立及初步临床应用[J]. 协和医学杂志, 2021, 12(4): 510-517. doi: 10.12290/xhyxzz.2021-0301
引用本文: 尹逸丛, 禹松林, 于佳磊, 王丹晨, 马超超, 邹雨桐, 谢少伟, 程倩, 邱玲. 同位素稀释超高效液相色谱串联质谱法检测血浆18-羟皮质酮方法的建立及初步临床应用[J]. 协和医学杂志, 2021, 12(4): 510-517. doi: 10.12290/xhyxzz.2021-0301
YIN Yicong, YU Songlin, YU Jialei, WANG Danchen, MA Chaochao, ZOU Yutong, XIE Shaowei, CHENG Qian, QIU Ling. Quantification of Human Plasma 18-Hydroxycorticosterone by Isotope Dilution Ultra Performance Liquid Chromatography Tandem Mass Spectrometry[J]. Medical Journal of Peking Union Medical College Hospital, 2021, 12(4): 510-517. doi: 10.12290/xhyxzz.2021-0301
Citation: YIN Yicong, YU Songlin, YU Jialei, WANG Danchen, MA Chaochao, ZOU Yutong, XIE Shaowei, CHENG Qian, QIU Ling. Quantification of Human Plasma 18-Hydroxycorticosterone by Isotope Dilution Ultra Performance Liquid Chromatography Tandem Mass Spectrometry[J]. Medical Journal of Peking Union Medical College Hospital, 2021, 12(4): 510-517. doi: 10.12290/xhyxzz.2021-0301

同位素稀释超高效液相色谱串联质谱法检测血浆18-羟皮质酮方法的建立及初步临床应用

doi: 10.12290/xhyxzz.2021-0301
基金项目: 

北京市临床重点专科医学检验科卓越项目 ZK201000

详细信息
    通讯作者:

    邱玲  电话:010-69159712,E-mail: lingqiubj@163.com

  • 中图分类号: R446.1

Quantification of Human Plasma 18-Hydroxycorticosterone by Isotope Dilution Ultra Performance Liquid Chromatography Tandem Mass Spectrometry

Funds: 

Beijing Key Clinical Specialty for Laboratory Medicine-Excellent Project ZK201000

More Information
  • 摘要:   目的  建立一种同位素稀释超高效液相色谱串联质谱法(isotope dilution ultra performance liquid chromatography tandem mass spectrometry, ID-UPLC-MS/MS)检测血浆18-羟皮质酮(18-Hydroxycorticosterone,18-OHB)的方法。  方法  取血浆标本或标准溶液200 μL置于离心管中,然后加入同位素氚标记的18-OHB为内标,用甲醇沉淀血浆蛋白,离心后取上清液,用Prime HLB Elution 96孔SPE板进行萃取,收集洗脱液,在正离子电喷雾离子化的多离子反应监测模式下检测18-OHB。评价该方法的精密度、加标回收率、定量检测下限、线性和基质效应。2019年11月起招募表观健康志愿者,采用ID-UPLC-MS/MS检测其血浆18-OHB水平,并验证梅奥医学实验中心提供的18-OHB参考区间。  结果  该方法检测血浆18-OHB的分析时间约为3.0 min,检测低、中、高3个水平18-OHB的重复性变异系数和实验室内不精密度分别为2.2%~3.5%和3.7%~5.0%,平均加标回收率为98.1%~101.7%,定量检测下限为0.01 μg/L,在0.1~10 μg/L范围内线性良好(r>0.990),血浆基质效应为86.83%~119.00%。基于本研究建立的ID-UPLC-MS/MS方法,73名表观健康人群血浆18-OHB第2.5、97.5百分位数分别为0.01、0.60 μg/L,其中66%(48/73)血浆18-OHB水平处于梅奥医学实验中心提供的参考范围外。  结论  本研究建立了一种ID-UPLC-MS/MS测定血浆18-OHB的方法,该方法检测快速、结果准确可靠,性能满足临床需求。
    作者贡献:尹逸丛负责实验设计、病例招募、数据采集与整理、统计分析、论文撰写;禹松林参与实验设计、数据整理、论文撰写;于佳磊参与实验设计、实验操作;王丹晨、马超超、邹雨桐负责病例招募、数据采集与整理;谢少伟、程倩负责样本采集、实验操作;邱玲参与实验设计、病例招募、数据采集与整理,负责论文修订与审核。
    利益冲突:
  • 图  1  18-OHB标准品及血浆样本同位素稀释超高效液相色谱串联质谱法分析图

    A1~A4、B1~B4分别为18-OHB标准品(5 μg/L)、血浆样本色谱分析图; A1、B1为总离子对色谱图,A2、B2为定量离子对色谱图,A3、B3为定性离子对色谱图,A4、B4为内标色谱图; 18-OHB: 18-羟皮质酮

    图  2  18-羟皮质酮线性评估图

    图  3  18-羟皮质酮的基质效应评估

    表  1  18-OHB精密度评价结果

    不同浓度18-OHB样本(μg/L) 测定的18-OHB浓度(x±s,μg/L) 重复性CV(%) 实验室内不精密度(%)
    0.2 0.20±0.01 2.2 5.0
    0.5 0.50±0.02 3.5 4.5
    2.0 2.02±0.08 3.4 3.7
    18-OHB:同图 1;CV:变异系数
    下载: 导出CSV
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出版历程
  • 收稿日期:  2021-03-31
  • 录用日期:  2021-06-15
  • 刊出日期:  2021-07-30

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