Published: March 20, 2019
Chensheng Li, Sitaram Bhavaraju, Jeremy Melanson, Andreas Blomgren, Torgny Rundlof, Eric L. Kilpatrick, Timothy Rudd, Yves Aubin, Kevin Grant, Edward Saravolac, Tursun Kerim, William Sherman, Yukari Nakagawa, Sergi Pavon, Tim Weel, Arunima Pola, Dinesh Chalasani, Stephen Walfish, Fouad Atouf
USPs 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 analyti-cal methods, the USP Biologics Department conducted a multi-laboratory collaborative study. The study deter-mined the inter-laboratory variability for peptide quantitation using the following methods: HPLC assay, quanti-tative 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 as-signment of the standard with mass balance. The 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.
Citation: Journal of Pharmaceutical and Biomedical Analysis
Pub Type: Journals
Peptide, Quantitation, Quantification, HPLC, NMR, Amino acid analysis (AAA)
Created March 20, 2019, Updated January 30, 2019