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血浆中6种初级胆汁酸HPLC–MS/MS测定

牟燕 吴瑛瑕 张易天 曾颖炫 周枝凤

牟燕, 吴瑛瑕, 张易天, 曾颖炫, 周枝凤. 血浆中6种初级胆汁酸HPLC–MS/MS测定[J]. 中国公共卫生, 2023, 39(1): 102-106. doi: 10.11847/zgggws1138567
引用本文: 牟燕, 吴瑛瑕, 张易天, 曾颖炫, 周枝凤. 血浆中6种初级胆汁酸HPLC–MS/MS测定[J]. 中国公共卫生, 2023, 39(1): 102-106. doi: 10.11847/zgggws1138567
MU Yan, WU Ying-xia, ZHANG Yi-tian, ZENG Ying-xuan, ZHOU Zhi-feng. Simultaneous and rapid determination of 6 primary bile acids in plasma by high-performance liquid chromatography-tandem mass spectrometry[J]. Chinese Journal of Public Health, 2023, 39(1): 102-106. doi: 10.11847/zgggws1138567
Citation: MU Yan, WU Ying-xia, ZHANG Yi-tian, ZENG Ying-xuan, ZHOU Zhi-feng. Simultaneous and rapid determination of 6 primary bile acids in plasma by high-performance liquid chromatography-tandem mass spectrometry[J]. Chinese Journal of Public Health, 2023, 39(1): 102-106. doi: 10.11847/zgggws1138567

血浆中6种初级胆汁酸HPLC–MS/MS测定

doi: 10.11847/zgggws1138567
基金项目: 国家自然科学基金(21607070)
详细信息
    作者简介:

    牟燕(1995 – ),女,四川泸州人,硕士在读,研究方向:环境污染与健康

    通讯作者:

    周枝凤,E-mail:zfzhou@smu.edu.cn

  • 中图分类号: Q 548+.5

Simultaneous and rapid determination of 6 primary bile acids in plasma by high-performance liquid chromatography-tandem mass spectrometry

  • 摘要:   目的  建立高效液相色谱 – 串联质谱法快速检测血浆中6种初级胆汁酸的含量。  方法  血浆经甲醇沉淀蛋白后,用Thermo Accucore C18色谱柱(100 mm × 4.6 mm,2.6 μm)进行色谱分离,流动相为甲醇 – 水(0.1 %甲酸 + 5 mmol/L乙酸铵),梯度洗脱。负离子模式下多反应监测模式检测。实测6名健康成年人和6只成年SD大鼠样本。  结果  6种初级胆汁酸在6 min内实现基线分离,在5~2 000 μg/L范围内线性良好,相关系数>0.995,检出限为0.25~0.45 μg/L,定量限为0.84~1.49 μg/L。日内、日间加标回收率分别为86.3 %~111.5 %和87.7 %~104.9 %;日内、日间相对标准偏差分别为5.2 %~11.9 %和6.6 %~14.3 %。健康成年人和成年SD大鼠血浆中初级胆汁酸的浓度显著不同。  结论  该方法简单、快速、灵敏、准确,可用于临床检验和动物试验中初级胆汁酸的快速检测。
  • 图  1  水相中不同乙酸铵含量的响应强度

    图  2  6种初级胆汁酸及内标的色谱图(10 μg/L)

    表  1  6种初级胆汁酸及内标的母离子、子离子、去簇电压和碰撞电压

    物质母离子(m/z)子离子(m/z)去簇电压/(V)碰撞电压/(V)
    CA407.9407.9 a – 141 – 17
    343.3 – 47
    CDCA391.3391.3 a – 151 – 21
    373.3 – 47
    TCA514.3514.3 a – 120 – 10
    80.0 – 120
    TCDCA498.2498.2 a – 90 – 13
    80.0 – 120
    GCA464.3464.3 a – 140 – 10
    402.4 – 50
    GCDCA448.3448.3 a – 110 – 10
    368.2 – 46
    CA-d4411.2411.2 a – 125 – 10
    347.4 – 48
    TCA-d4518.3518.3 a – 120 – 10
    80.0 – 120
    GCA-d4468.3468.3 a – 127 – 30
      404.6  – 48
      注:a 为定量离子。
    下载: 导出CSV

    表  2  6种初级胆汁酸的线性方程、相关系数、检出限和定量限

    物质内标线性方程相关系数检出限/(μg/L)定量限/(μg/L)
    CACA-d4y = 1.13 × 10 – 3x + 1.22 × 10 – 30.998 30.451.49
    CDCACA-d4y = 4.45 × 10 – 3x + 1.25 × 10 – 20.998 70.391.30
    TCATCA-d4y = 6.19 × 10 – 3x + 9.46 × 10 – 30.995 10.351.17
    TCDCATCA-d4y = 7.38 × 10 – 3x + 8.33 × 10 – 30.995 60.381.26
    GCAGCA-d4y = 8.09 × 10 – 3x + 3.02 × 10 – 20.997 00.381.28
    GCDCAGCA-d4y = 1.61 × 10 – 2x + 5.18 × 10 – 20.995 30.250.84
    下载: 导出CSV

    表  3  血浆中6种初级胆汁酸的加标回收率及相对标准偏差

    物质加标量(μg/L)日内 日间
    回收率(%)相对标准偏差(%) 回收率(%)相对标准偏差(%)
    CA694.18.798.614.1
    800108.09.4102.910.4
    160095.75.298.27.7
    CDCA6101.811.9101.06.6
    800101.011.0102.910.2
    160091.47.995.89.8
    TCA6100.17.899.914.0
    800101.86.596.510.9
    160095.411.291.58.2
    TCDCA699.810.1104.912.1
    800100.87.094.18.4
    160099.27.887.77.3
    GCA692.47.097.69.6
    800102.011.1101.611.1
    160097.09.799.214.3
    GCDCA686.38.594.111.7
    800111.510.7102.611.4
     1600102.311.4 96.212.8
    下载: 导出CSV

    表  4  6种初级胆汁酸在健康成年人和SD大鼠血浆中的含量(n = 6)

    物质检出量/(μg/L) P
    健康成年人SD大鼠 
    CA9.7~270.0248.0~467.00.031
    CDCA43.4~406.0106.0~405.00.893
    TCA20.0~222.065.1~600.00.139
    TCDCA129.0~697.059.0~572.00.139
    GCA184.0~1020.05.9~128.00.005
    GCDCA1150.0~3144.0ND~9.80.005
      注: ND 为未检出。
    下载: 导出CSV
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    [3] 李明, 成淑梅. 氯乙烯作业工人血清甘胆酸含量的测定[J]. 中国公共卫生, 1998(12): 40 – 40.
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    [11] Wang X, Wang F, Lu Z, et al. Semi-quantitative profiling of bile acids in serum and liver reveals the dosage-related effects of dexamethasone on bile acid metabolism in mice[J]. Journal of Chromatography B, 2018, 1095: 65 – 74. doi: 10.1016/j.jchromb.2018.07.021
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出版历程
  • 收稿日期:  2022-03-20
  • 网络出版日期:  2023-01-17
  • 刊出日期:  2023-01-31

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