色谱法
胰蛋白酶
化学
人血清白蛋白
糜蛋白酶
白蛋白
检出限
血清白蛋白
人血浆
酶
生物化学
作者
Meiyun Shi,Xujian Duan,Xinyue Zheng,Di Lu,Yun-Cheng Ge,Ning Zhang,Yajun Liu,Jiansong You,Huadan Xue,Lei Yin
标识
DOI:10.1016/j.ab.2023.115316
摘要
The quantification of albumin is important in clinical medicine because the concentration of albumin in biological fluids is closely related to human health. In this study, we developed a highly selective and robust assay to determine human serum albumin (HSA) in human plasma by combining chymotrypsin/trypsin digestion coupled with targeted LC-MS/MS technique. Human plasma samples were denatured, reduced, alkylated, and digested with both chymotrypsin and trypsin to generate surrogate peptides. A unique chymotryptic peptide (NAETF) arising from human serum albumin was finally selected for targeted LC-MS/MS detection and quantification. Numerous parameters related to the targeted LC-MS/MS assay were evaluated, including lower limit of quantitation (LLOQ), linearity range, enzyme digestion efficiency, accuracy and precision. The LC-MS/MS assay was linear in the concentration range 0.05-1 mg/mL with intra-day and inter-day precision <10.2% and accuracy ranging from -3.94% to 4.89%. The assay was successfully applied to determine HSA in 148 human plasma samples.
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