Survey of peptide quantification methods and comparison of their reproducibility: A case study using oxytocin

USP’s peptide reference standards content is typically determined using an HPLC assay against an external standard for which the purity was determined by a mass balance approach. To explore the use of other analytical methods, the USP Biologics Department conducted a multi-laboratory collaborative s...

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Bibliographic Details
Published in:	Journal of pharmaceutical and biomedical analysis Vol. 166; pp. 105 - 112
Main Authors:	Li, Chensheng, Bhavaraju, Sitaram, Thibeault, Marie-Pier, Melanson, Jeremy, Blomgren, Andreas, Rundlöf, Torgny, Kilpatrick, Eric, Swann, Carolyn J., Rudd, Timothy, Aubin, Yves, Grant, Kevin, Butt, Margaret, Shum, WaiKei, Kerim, Tursun, Sherwin, William, Nakagawa, Yukari, Pavón, Sergi, Arrastia, Silvia, Weel, Tim, Pola, Arunima, Chalasani, Dinesh, Walfish, Steven, Atouf, Fouad
Format:	Journal Article
Language:	English
Published:	England Elsevier B.V 20-03-2019
Subjects:	Amino acid analysis (AAA) Amino Acids - analysis Chemistry Techniques, Analytical - methods Chromatography, High Pressure Liquid HPLC Magnetic Resonance Spectroscopy NMR Oxytocin - analysis Peptide Quantification Reference Standards Reproducibility of Results NMR Quantification Amino acid analysis (AAA) Peptide HPLC
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Summary:	USP’s peptide reference standards content is typically determined using an HPLC assay against an external standard for which the purity was determined by a mass balance approach. To explore the use of other analytical methods, the USP Biologics Department conducted a multi-laboratory collaborative study. The study determined the inter-laboratory variability for peptide quantitation using the following methods: HPLC assay, quantitative nuclear magnetic resonance (qNMR) spectroscopy, or amino acid analysis (AAA). The three methods were compared with regard to their suitability for quantitation of the nonapeptide oxytocin. In this study, the HPLC assay method using the same peptide bulk material as the standard showed the lowest inter-lab variability. The coefficient of variation (%CV) was calculated without counting the uncertainty associated with the purity assignment of the standard with mass balance. The proton qNMR method is a direct measurement of the peptide against an internal standard, which is not difficult to perform under common laboratory conditions. Because of the simpler operation and shorter analytical time, qNMR as a primary method for peptide reference standard value assignment deserves further exploration.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0731-7085 1873-264X
DOI:	10.1016/j.jpba.2018.12.028