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Development of a human insulin certified reference material with SI-traceable purity

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Abstract

A human insulin (hINS) certified reference material (CRM) was developed by the National Institute of Metrology (NIM). Three milligrams of purified solid hINS was packed into a brown sealed tube. The candidate material was identified by de novo sequence using mass spectrometry and Edman degradation methods. The content of insulin-related impurities, aggregation, moisture, volatile organic compounds (VOCs), anions, and ignition residues was also determined. Both mass balance (MB) and amino acid analysis-based isotope dilution mass spectrometry (AAA-IDMS) were used for the certified value assessment, which was determined to be (0.857 ± 0.024) g/g. The certified value was validated by liquid chromatography-circular dichroism spectroscopy (LC-CD) and quantitative nuclear magnetic resonance (qNMR) methods, which were in good agreement. No inhomogeneity was observed during a homogeneity examination. A stability examination showed that the CRM was stable for at least 12 months when stored at − 70 °C, and for 7 days when stored at 4, 25, or 40 °C. The CRM is expected to be used as a primary calibrator for matrix insulin CRM development and for quality control in biopharmaceutical production and clinical diagnostics.

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Notes

  1. All the numbers that follow “ ± ” are expanded uncertainties with a coverage factor of 2 except “110.0 ± 0.5 °C” that appears later, which means the maximum permitted bias.

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Funding

This work was financially supported by the projects AKYZD1907, AKY1959, and 2018B02020707. Victoria Muir, PhD, from Liwen Bianji, (Edanz) (www.liwenbianji.cn/), edited the English text of a draft of this manuscript.

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Authors and Affiliations

  1. Beijing University of Chemical Technology, No. 15 North Third Ring Road, Chaoyang District, Beijing, 100029, China

    Xianxia Wang, Ping Su & Yi Yang

  2. National Institute of Metrology, No. 18 North Third Ring Road, Chaoyang District, Beijing, 100029, China

    Liqing Wu, Bin Yang & Ning Zhang

  3. South China National Centre of Metrology/Guangdong Institute of Metrology, No. 30, Songbai East Street, Baiyun District, Guangzhou, 510405, Guangdong, China

    Yanjie Huang

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  1. Xianxia Wang
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Correspondence to Ping Su or Yi Yang.

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Wang, X., Wu, L., Huang, Y. et al. Development of a human insulin certified reference material with SI-traceable purity. Anal Bioanal Chem 414, 3443–3457 (2022). https://doi.org/10.1007/s00216-022-03965-0

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