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| Determination of the Mass Fraction of Pure Human Serum Albumin by HPLC-IDMS |
| LI Jia-le1,2,WU Li-qing2,JIN You-xun2,WANG Jing2,YANG Bin2,YANG Yi1 |
1. Beijing University of Chemical Technology, Beijing 100029, China
2. National Institute of Metrology, Beijing 100029, China |
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Abstract A mass balance method and a high performance liquid chromatography-isotope dilution mass spectrometry （HPLC-IDMS） method are established to determine the mass fraction of pure human serum albumin (HSA). With the mass balance method, the pure HSA is quantified by HPLC and its moisture and ash are quantified by Karl Fischer and ignition residue analysis, which give an average mass fraction of 0.861 g/g. By the HPLC-IDMS method, after the pure HSA is hydrolyzed by acid, the resulted Pro, Val and Phe are quantified. Consequently, the mass fraction of HSA is calculated by the average. Under the optimized conditions, the mass fraction of HSA is 0.853 g/g with a relative standard deviation (RSD) of 0.8% and an expanded uncertainty of 0.015 g/g (k=2). The limits of detection and quantification are 1.37×10-5 g/g and 4.55×10-5 g/g, respectively. Compared with the mass balance method, the relative bias of the HPLC-IDMS method is 0.9%.
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Received: 15 November 2014
Published: 22 March 2016
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2016, Vol. 37 
