1P-37 PDF
Development of human insulin solution certified reference material using a novel amino acid analysis method
We have developed human insulin solution certified reference material CRM 6209-a, which is intended for use as a primary reference material to enable the SI-traceable measurement of human insulin. The total protein concentration of human insulin in the present CRM was determined by amino acid analysis coupled to isotope-dilution mass spectrometry. We conducted two independent amino acid analyses in which the following independent hydrolysis and liquid chromatography-isotope dilution mass spectrometry were combined: one was gas-phase acid hydrolysis and pre-column derivatization liquid chromatography-tandem mass spectrometry method, and the other was liquid-phase acid hydrolysis and another pre-column derivatization liquid chromatography-tandem mass spectrometry method. The quantitative values of the two different amino acid analyses were in agreement within their uncertainties. The certified value was the weighted mean of the results of the two methods. The certified value and the expanded uncertainty (k = 2) of CRM 6209-a are (77.9 ± 2.3) mg/L.