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Purity Measurement of Human C-Peptide by High Performance Liquid Chromatography and Quantitative Nuclear Magnetic Resonance

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Abstract

Purity determination of human C-peptide (hCP) with metrological traceability is fundamental to accurate and comparable clinical analysis of hCP. Based on high performance liquid chromatography and quantitative nuclear magnetic resonance (qNMR) with a purified hCP, a method was reported to determine the purity of an hCP sample used in an intercomparison. The composition of deuterium solvent for qNMR was optimized to improve the separation of the β-proton of an aspartic acid residue in the sequence of hCP, which were used for NMR integration. The calculated mean and uncertainty (77.86 ± 3.85%) are consistent with the results (80.18 ± 0.62%) of different methods reported by many metrological laboratories. This method extended the application of qNMR to peptides with high molecular weight of about 3020.

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References

  • Albrecht T, Schilperoort M, Zhang S, Braun JD, Rodriguez A, Pastene DO, Krämer BK, Köppel H, Baelde H, de Heer E, Altomare AA, Regazzoni L, Denisi A, Aldini G, van den Born J, Yard BA, Hauske SJ (2017) Carnosine attenuates the development of both type 2 diabetes and diabetic nephropathy in BTBR ob/ob mice. Sci Rep 7:44492

    Article  PubMed  PubMed Central  Google Scholar 

  • Arsene CG, Ohlendorf R, Burkitt W, Pritchard C, Henrion A, O’Connor G, Bunk DM, Güttler B (2008) Protein quantification by isotope dilution mass spectrometry of proteolytic fragments: cleavage rate and accuracy. Anal Chem 80:4154–4160

    Article  PubMed  CAS  Google Scholar 

  • Fierens C, Stöckl D, Baetens D, De Leenheer AP, Thienpont LM (2003) Application of a C-peptide electrospray ionization-isotope dilution–liquid chromatography–tandem mass pectrometry measurement procedure for the evaluation of five C-peptide immunoassays for urine. J Chromatogr B 792:249–259

    Article  CAS  Google Scholar 

  • Ghanim H, Batra M, Abuaysheh S, Green K, Makdissi A, Kuhadiya N, Chaudhuri A, Dandona P (2017) Anti-inflammatory and ROS suppressive effects of the addition of fiber to a high fat high calorie meal. J Clin Endocrinol Metab 102:858–869

    PubMed  Google Scholar 

  • Giraudeau P (2017) Challenges and perspectives in quantitative NMR. Magn Reson Chem 55:61–69

    Article  PubMed  CAS  Google Scholar 

  • Holzgrabe U (2010) Quantitative NMR spectroscopy in pharmaceutical applications. Prog Nucl Mag Res Spectrosc 57:229–240

    Article  CAS  Google Scholar 

  • Huang T, Zhang W, Dai X, Zhang X, Quan C, Li H, Yang Y (2014) Precise measurement for the purity of amino acid and peptide using quantitative nuclear magnetic resonance. Talanta 125:94–101

    Article  PubMed  CAS  Google Scholar 

  • Josephs RD, Li M, Song D, Westwood S, Stoppacher N, Daireaux A, Choteau T, Wielgosz R, Xiao P, Liu Y, Gao X, Zhang C, Zhang T, Mi W, Quan C, Huang T, Li H, Flatschart R, Oliveira RB, Melanson JE, Ohlendorf R, Henrion A, Kinumi T, Wong L, Liu Q, Senal MO, Vatansever B, Ün I, Gören AC, Akgöz M, Quaglia M, Warren J (2017) Key comparison study on peptide purity—synthetic human C-peptide. Metrologia 54:08007

    Article  Google Scholar 

  • Kato M, Kato H, Eyama S, Takatsu A (2009) Application of amino acid analysis using hydrophilic interaction liquid chromatography coupled with isotope dilution mass spectrometry for peptide and protein quantification. J Chromatogr B 877:3059–3064

    Article  CAS  Google Scholar 

  • Pauli GF, Gödecke T, Jaki BU, Lankin DC (2012) Quantitative 1H NMR. development and potential of an analytical method: an update. J Nat Prod 75:834–851

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Singh S, Roy R (2016) The application of absolute quantitative 1H NMR spectroscopy in drug discovery and development. Expert Opin Drug Dis 11:695–706

    Article  CAS  Google Scholar 

  • Wu L, Yang B, Bi J, Wang J (2011) Development of bovine serum albumin certified reference material. Anal Bioanal Chem 400:3443–3449

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This work is financially supported by National Natural Science Foundation of China (No. 21275134) and National Key Technology Research and Development Program of the Ministry of Science and Technology of China (2013BAK10B01, 2011FY130100).

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Authors and Affiliations

  1. Division of Chemical Metrology and Analytical Science, National Institute of Metrology, Beijing, 100029, China

    Wei Zhang, Ting Huang, Hongmei Li & Dewei Song

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  1. Wei Zhang
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  2. Ting Huang
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  3. Hongmei Li
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  4. Dewei Song
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Correspondence to Ting Huang.

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Zhang, W., Huang, T., Li, H. et al. Purity Measurement of Human C-Peptide by High Performance Liquid Chromatography and Quantitative Nuclear Magnetic Resonance. Int J Pept Res Ther 24, 391–396 (2018). https://doi.org/10.1007/s10989-017-9620-6

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  • DOI: https://doi.org/10.1007/s10989-017-9620-6

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