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%.
李佳乐,武利庆,金有训,王晶,杨彬,杨屹. 人血清白蛋白纯品含量的同位素稀释质谱方法研究[J]. 计量学报, 2016, 37(3): 328-332.
LI Jia-le,WU Li-qing,JIN You-xun,WANG Jing,YANG Bin,YANG Yi. Determination of the Mass Fraction of Pure Human Serum Albumin by HPLC-IDMS. Acta Metrologica Sinica, 2016, 37(3): 328-332.
［1］赵颖怡, 汪嵘, 韦宇拓. 人血清白蛋白的研究进展［J］. 广西农业生物科学, 2002, 21(2): 126-128.
［2］吴婧, 文凌飞, 朱兴旺,等. 用铍试剂测定人血清白蛋白的分光光度法［J］. 分析测试学报, 2002,21(5): 54-56.
［3］Poór M, Li Y, Matisz G, et al. Quantitation of species differences in albumin-ligand interactions for bovine, human and rat serum albumins using fluorescence spectroscopy: A test case with some Sudlow's site I ligands［J］. Journal of Luminescence, 2014, 145: 767-773.
［4］Hong Y, Feng C, Yu Y,et al. Quantitation, Visualization, and Monitoring of Conformational Transitions of Human Serum Albumin by a Tetraphenylethene Derivative with Aggregation-Induced Emission Characteristics［J］. Analytical Chemistry, 2010, 82(16): 7035-7043.
［5］李先平, 王敏, 杨江玲. 用Helena SPIFE琼脂糖凝胶电泳系统进行血清蛋白电泳［J］. 实用医学杂志, 2003,(12): 1371-1373.
［6］Qian J, Tang Q, Cronin B, et al. Development of a high performance size exclusion chromatography method to determine the stability of human serum albumin in a lyophilized formulation of Interferon alfa-2b［J］. Journal of Chromatography A, 2008, 1194(1): 48-56.
［7］Chung M K, Riby J, Li H, et al. A sandwich enzyme-linked immunosorbent assay for adducts of polycyclic aromatic hydrocarbons with human serum albumin［J］. Analytical Biochemistry, 2010, 400(1): 123-129.
［8］蔡耘, 武力民, 杨松成. MALDI-TOF-MS法分析人血清白蛋白［J］. 分析测试学报, 1996,(4): 80-82.
［9］ISO 17511:2003, 体外诊断医疗器械—生物样品中量的测量—校准品和控制物质赋值的计量学溯源性［S］.
［10］常碧影, 梁冬生, 阎惠文. 氨基酸分析技术研究的进展［J］. 分析化学, 1993,(10): 1220-1227.
［11］戴新华, 暴海霞, 苏福海. 国际关键比对CCQM—K55C:L-缬氨酸纯度的测量［J］. 计量技术, 2013,(10): 15-19.
［12］黄挺, 张伟, 全灿. 定量核磁共振法研究进展［J］. 化学试剂, 2012, 34(4): 327-332.
［13］赵墨田. 同位素稀释质谱法特点［J］. 质谱学报, 2004, 25(z1): 167-168.
［14］Mun～oz A, Kral R, Schimmel H. Quantification of protein calibrants by amino acid analysis using isotope dilution mass spectrometry［J］. Analytical Biochemistry, 2011, 408(1): 124-131.
［15］JJG1036—2008, 电子天平检定规程［S］.
2016, Vol. 37 
