基于有机元素分析的多肽纯度测量技术

李明; 张伟

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

李明,
张伟

中国计量科学研究院，北京 100029

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

计量
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出版历程
- 网络出版日期: 2022-01-24

Peptide Purity Measurement Technique Based on Organic Elemental Analysis

LI Ming,
ZHANG Wei

National Institute of Metrology, Beijing 100029, China

摘要

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。
- 多肽 /
- 纯度测量 /
- 元素分析 /
- 氮 /
- 硫
Abstract: Purity measurement of the peptide is the key to building metrological traceability for the measurement value of peptides in clinical chemistry, pharmaceutical chemistry, food chemistry, and other fields. In addition to the mass balance method, nuclear magnetic resonance method, and hydrolysis method, organic elemental analysis is a new technique for measuring the purity of peptides in chemical metrology. However, a detailed introduction to this technique is yet to be reported. In this paper, a technique for peptide purity measurement based on the analysis of nitrogen, sulfur, and other elements has been established. Using oxytocin as a typical sample, the nitrogen and sulfur elements in the sample were analyzed, the content of nitrogen and sulfur elements in the main component was obtained by deducting the content of nitrogen and sulfur elements in the relevant structural impurities, and the purity results of the peptide were obtained by calculation using the information of the number of atoms and molecular weight of nitrogen and sulfur elements in the molecule of the peptide. The relative standard deviation was less than 1% compared with the measurement results of the authoritative mass balance method. The results of the impurity-corrected organic elemental analysis technique established in this paper are accurate and reliable and can be used for peptide purity standard substance determination.
- peptide /
- purity measurement /
- elemental analysis /
- nitrogen /
- sulfur

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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

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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 acid	2.39	0.39
[Asp⁵] OT	1.04	0.18
[Glu⁴] OT	0.85	0.14
[Cya¹,Cya⁶] OT	0.161	0.022
endo-Gly^10a-OT	0.126	0.022
ent-OT1	0.115	0.020
[Csa¹, Cya⁶] OT	0.092	0.017
OT+O	0.090	0.014
[Cya¹, Csa⁶] OT	0.0338	0.0046
endo-βAln^9a-OT	0.0343	0.0071
des-Gly⁹-OT	0.0149	0.0032
endo-Asn^5a-OT	0.0087	0.0087
des-Pro⁷-OT	0.0085	0.0085
endo-Gln^4a-OT	0.0086	0.0086
TFA OT	0.0076	0.0076
ent-OT 2	0.0083	0.0083
ent-OT 3	0.0083	0.0083
Unk491a	0.0086	0.0086
Unk491b	0.0086	0.0086
α dimer	0.0083	0.0083
β dimer	0.0083	0.0083

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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 acid	1.00	0.16
[Asp⁵] OT	0.434	0.076
[Glu⁴] OT	0.352	0.058
[Cya¹,Cya⁶] OT	0.0613	0.0085
endo-Gly^10a-OT	0.0445	0.0077
ent-OT1	0.0438	0.0078
[Csa¹, Cya⁶] OT	0.0352	0.0066
OT+O	0.0345	0.0053
[Cya¹, Csa⁶] OT	0.0129	0.0018
endo-βAln^9a-OT	0.0121	0.0025
des-Gly⁹-OT	0.0068	0.0015
endo-Asn^5a-OT	0.0029	0.0029
des-Pro⁷-OT	0.0035	0.0035
endo-Gln^4a-OT	0.0028	0.0028
TFA OT	0.0029	0.0029
ent-OT 2	0.0032	0.0032
ent-OT 3	0.0032	0.0032
Unk491a	0.0033	0.0033
Unk491b	0.0033	0.0033
α dimer	0.0032	0.0032
β dimer	0.0032	0.0032

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参考文献(15)

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