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基于有机元素分析的多肽纯度测量技术

李明 张伟

李明,张伟. 基于有机元素分析的多肽纯度测量技术[J]. 计量科学与技术,2022, 66(7): 18-21 doi: 10.12338/j.issn.2096-9015.2021.0558
引用本文: 李明,张伟. 基于有机元素分析的多肽纯度测量技术[J]. 计量科学与技术,2022, 66(7): 18-21 doi: 10.12338/j.issn.2096-9015.2021.0558
LI Ming, ZHANG Wei. Peptide Purity Measurement Technique Based on Organic Elemental Analysis[J]. Metrology Science and Technology, 2022, 66(7): 18-21. doi: 10.12338/j.issn.2096-9015.2021.0558
Citation: LI Ming, ZHANG Wei. Peptide Purity Measurement Technique Based on Organic Elemental Analysis[J]. Metrology Science and Technology, 2022, 66(7): 18-21. doi: 10.12338/j.issn.2096-9015.2021.0558

基于有机元素分析的多肽纯度测量技术

doi: 10.12338/j.issn.2096-9015.2021.0558
基金项目: 国家重点研发计划(2017YFF0205403)。
详细信息
    作者简介:

    李明(1979-),中国计量科学研究院副研究员,研究方向:多肽计量,邮箱:liming@nim.ac.cn

Peptide Purity Measurement Technique Based on Organic Elemental Analysis

  • 摘要: 多肽的纯度测量是临床化学、药物化学、食品化学等领域中多肽量值溯源的关键。除质量平衡法、核磁法、水解法外,有机元素分析技术是化学计量中用于多肽纯度测量的一种新手段。然而,对该技术的详细介绍鲜有报道。本文建立了一种基于氮、硫等元素分析的多肽纯度测量技术,采用缩宫素作为典型样品,首先对样品中氮、硫元素进行分析,扣减相关结构杂质中氮、硫元素的含量得到主成分中氮、硫元素的含量,利用氮、硫元素在缩宫素分子中的原子个数及分子量信息,通过计算获得了缩宫素的纯度结果。与权威方法质量平衡法的测量结果相比,相对标准偏差小于1%。本文建立的杂质校正有机元素分析技术结果准确、可靠,可用于多肽纯度标准物质定值。
  • 图  1  质量平衡法与元素分析法的测量结果

    注:A:质量平衡法;B:氮元素分析法;C:硫元素分析法;D:氮、硫元素分析法的平均值。

    Figure  1.  Measurement results from mass balance method and elemental analysis method

    表  1  杂质中氮元素含量的计算结果

    Table  1.   Calculated nitrogen content in the impurities

    名称氮含量 mg/g不确定度 mg/g
    OT acid2.390.39
    [Asp5] OT1.040.18
    [Glu4] OT0.850.14
    [Cya1,Cya6] OT0.1610.022
    endo-Gly10a-OT0.1260.022
    ent-OT10.1150.020
    [Csa1, Cya6] OT0.0920.017
    OT+O0.0900.014
    [Cya1, Csa6] OT0.03380.0046
    endo-βAln9a-OT0.03430.0071
    des-Gly9-OT0.01490.0032
    endo-Asn5a-OT0.00870.0087
    des-Pro7-OT0.00850.0085
    endo-Gln4a-OT0.00860.0086
    TFA OT0.00760.0076
    ent-OT 20.00830.0083
    ent-OT 30.00830.0083
    Unk491a0.00860.0086
    Unk491b0.00860.0086
    α dimer0.00830.0083
    β dimer0.00830.0083
    下载: 导出CSV

    表  2  杂质中硫元素含量的计算结果

    Table  2.   Calculated sulfur content in the impurities

    名称硫含量 mg/g不确定度 mg/g
    OT acid1.000.16
    [Asp5] OT0.4340.076
    [Glu4] OT0.3520.058
    [Cya1,Cya6] OT0.06130.0085
    endo-Gly10a-OT0.04450.0077
    ent-OT10.04380.0078
    [Csa1, Cya6] OT0.03520.0066
    OT+O0.03450.0053
    [Cya1, Csa6] OT0.01290.0018
    endo-βAln9a-OT0.01210.0025
    des-Gly9-OT0.00680.0015
    endo-Asn5a-OT0.00290.0029
    des-Pro7-OT0.00350.0035
    endo-Gln4a-OT0.00280.0028
    TFA OT0.00290.0029
    ent-OT 20.00320.0032
    ent-OT 30.00320.0032
    Unk491a0.00330.0033
    Unk491b0.00330.0033
    α dimer0.00320.0032
    β dimer0.00320.0032
    下载: 导出CSV
  • [1] JOSEPHS R D, STOPPACHER N, WESTWOOD S, et al. Concept paper on SI value assignment of purity - Model for the classification of peptide/protein purity determinations[J]. Journal of Chemical Metrology, 2017, 11(1): 1-8. doi: 10.25135/jcm.1.17.02.030
    [2] JOSEPHS R D, LI M, SONG D, et al. Key comparison study on peptide purity-synthetic human C-peptide[J]. Metrologia, 2017, 54(1A): 08007. doi: 10.1088/0026-1394/54/1A/08007
    [3] JOSEPHS R D, LI M, SONG D, et al. Pilot study on peptide purity-synthetic human C-peptide[J]. Metrologia, 2017, 54(1A): 080011.
    [4] 李明, 李红梅, 李孟婉. C肽的国际比对及C肽纯度标准物质的研制[J]. 计量技术, 2020(9): 112-114.
    [5] JOSEPHS R D, LI M, DAIREAUX A, et al. Key comparison study on peptide purity - synthetic oxytocin[J]. Metrologia, 2020, 57(1A): 08014. doi: 10.1088/0026-1394/57/1A/08014
    [6] JOSEPHS R D, LI M, DAIREAUX A, et al. Pilot study on peptide purity - synthetic oxytocin[J]. Metrologia, 2020, 57(1A): 08016. doi: 10.1088/0026-1394/57/1A/08016
    [7] LI M, JOSEPHS R D, CHOTEAU T, et al. Identification and accurate quantification of structurally-related peptide impurities for synthetic oxytocin by liquid chromatography–high resolution mass spectrometry[J]. Analytical and Bioanalytical Chemistry, 2021, 413(7): 1861-1870. doi: 10.1007/s00216-021-03154-5
    [8] WANG S, WU P, LI M, et al. Mass balance method for SI-traceable purity assignment of synthetic oxytocin[J]. Journal of Pharmaceutical and Biomedical Analysis, 2022, 207: 114401. doi: 10.1016/j.jpba.2021.114401
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    [10] ZHANG W, HUANG T, LI H, et al. Purity Measurement of Human C-Peptide by High Performance Liquid Chromatography and Quantitative Nuclear Magnetic Resonance International[J]. Journal of Peptide Research and Therapeutics, 2018, 24: 391-396. doi: 10.1007/s10989-017-9620-6
    [11] HUANG T, ZHANG W, LI H, et al. High performance liquid chromatography-Quantitative nuclear magnetic resonance-High performance liquid chromatography for purity measurement of human insulin[J]. Journal of Liquid Chromatography & Related Technologies, 2018, 41(4): 170-179.
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    [14] WU P, LI M, KAN Y, et al. Impurities identification and quantification for calcitonin salmon by liquid chromatography-high resolution mass spectrometry[J]. Journal of Pharmaceutical and Biomedical Analysis, 2020, 186: 113271. doi: 10.1016/j.jpba.2020.113271
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出版历程
  • 网络出版日期:  2022-01-24
  • 刊出日期:  2022-08-04

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