RP-HPLC法测定血浆中聚乙二醇尿酸酶浓度的方法  

Determination of PEG-Uricase Concentration in Plasma by RP-HPLC

作　　者：范开[1] 马雪丰[1] 谢敏[1] 胡春兰[1] 谭克海[1] 

机构地区：[1]重庆理工大学药学与生物工程学院,重庆400050

出　　处：《重庆理工大学学报（自然科学）》2010年第5期29-32,共4页Journal of Chongqing University of Technology：Natural Science

基　　金：国家"重大新药创制"科技重大专项资助项目(2009zx09102-25)

摘　　要：建立一种快速、稳定、灵敏度高的测定血浆中聚乙二醇尿酸酶浓度的高效液相色谱方法。将血浆样品与尿酸反应5 min,加入高氯酸终止,经高速离心,取上清液20μL检测。色谱柱采用Waters symmetry C18(4.6×250 mm,5μm),流动相为磷酸-甲醇-超纯水(3∶10∶487),流速1.0 mL/min,检测波长293 nm。通过体系中尿酸的降低值间接测定血浆中聚乙二醇尿酸酶的浓度。结果表明,聚乙二醇尿酸酶在31.25 ng/mL^20μg/mL范围内呈线性关系,r=0.999 4,日内、日间精密度均小于10%,回收率在88%~105%。A rapid,stable and highly sensitive method for the determination of PEG-uricase concentration in plasma by RP-HPLC was established.The plasma samples reacted with uric acid for 5min,then the reaction was terminated by adding perchloric acid and high-speed centrifugate.The supernatant 20 μL injection was analyzed.The analysis was determined by RP-HPLC using C18 column（4.6×250 mm,5 μm） with a mobile phase consisting of phosphoric acid-methanol-water（3：10：487,v/v）.The flow rate was 1.0 mL/min and the wavelength was detected at 293nm.The concentration of PEG-uricase in plasma was calculated indirectly by lowering the value of versus uric acid digest.The calibration curve of PEG-uricase concentration displayed highly relative linearity over the range of 31.25ng/mL-20ug/mL.The intra-day and inter-day RSD were not more than 10%.The average recovery was 88%-105%.

关 键 词：聚乙二醇尿酸酶 RP-HPLCg 浓度 

分 类 号：R943[医药卫生—药剂学]