反相高效液相色谱紫外法检测人血清尿酸  

Determination of uric acid in human serum by reverse-phase high-performance liquid chromatography using ultraviolet detection

作　　者：江雅平[1] 刘欧[2] 徐国宾[1] 陈文祥[3] 张传宝[3] 张江涛[3] 

机构地区：[1]北京大学第一医院检验科,北京100034 [2]清华大学第一医院检验科,北京100016 [3]卫生部临床检验中心,北京100730

出　　处：《临床检验杂志》2012年第4期244-248,共5页Chinese Journal of Clinical Laboratory Science

基　　金："十一五"国家科技支撑计划资金(2007BA105B09);国家高技术研究发展计划(863计划)资金(2006AA020909);首都发展基金(2009-1012)

摘　　要：目的探讨日本临床化学会(JSCC)推荐的检测人血清尿酸(UA)的反相高效液相色谱紫外(HPLC-UV)法是否可作为候选参考方法来验证同位素稀释质谱法、评价常规方法。方法对反相HPLC-UV法进行复现并进行方法学评价。观察尿酸酶处理人血清样本后的色谱以评价其特异性;检测系列标准液的UA水平并绘制标准曲线,观察线性范围;用朗道公司质控品评价精密度;用UA标准液评价回收率;分别用有证参考物SRM 909b(Ⅰ和Ⅱ)、国家一级标准物质GBW 09176、GBW 09175、GBW 09174评价正确度,并与2008年国际临床化学与检验医学联合会(IFCC)参考实验室RELA(ring trials for reference labora-tories)结果进行比对;用改良Bland-Altman图评价5种常规检测系统与反相HPLC-UV法间的偏差。结果尿酸酶处理后UA色谱峰消失;本法的线性范围为2.08～1 785μmol/L;批内变异系数(CV)均<0.3%,批间CV均<0.4%,日间CV均<2.3%,总CV均<2.7%;平均相对回收率为96.0%～100.6%;与SRM909bⅠ和Ⅱ靶值的偏移分别为-2.5%、-2.3%;与GBW09176、GBW 09174、GBW 09175靶值的偏移分别为0.29%,-0.74%和0.06%;与参加RELA比对的另3家实验室的平均值进行比较,水平1偏移为0.35%,水平2为-0.69%;Hitachi、Beckman Coulter、Roche、Dade、Vitros常规检测系统检测人血清UA与反相HPLC-UA法的相关系数分别为0.998 9、0.996 5、0.999 2、0.999 2和0.998 7,与反相HPLC-UA法的偏差均小于5%的生物学变异。结论本法特异、简便、快速,精密度好,正确度高,与具有良好溯源的常规测定系统具有良好的相关性和一致性,可推荐作为人血清UA测定的候选参考方法。Objective To explore whether the reverse-phase high performance liquid chromatography assay coupled with ultra violet （HPLC-UV） detection recommended by Japanese Society of Clinical Chemistry （JSCC） may be used as the candidate reference meth- od, and verify the isotope dilution mass spectrometry （ID-MS）, evaluate the routine methods for the determination of uric acid in hu- man serum. Methods The reverse-phase HPLC-UV was reproduced and its methodology characteristics was evaluated. The chromato- grams of uric acid in the human serum sample treated by uricase were observed to evaluate the specificity. The linear range was ob- served by means of determining the concentration of a series of standard solutions of uric acid and drawing the standard curve. Randox quality control material was used to evaluate the precision of the assay. The standard solution of uric acid was used to evaluate the re- covery rate. The accuracy of the reverse-phase HPLC-UV was evaluated by using the certified reference materials （ SRM 909b） and the national primary reference materials GBW 09176, GBW 09175 and GBW 09174. The results determined by reverse-phase HPLC-UV were compared with those of RELA （ ring trials for reference laboratories） of 2008 IFCC （ International Federation of Clinical Chemistry and Laboratory Medicine Federation）. The modified Bland-Ahman graphical method was used to appraise the bias between the reverse- phase HPLC-UV method and 5 routine assay systems. Results After the serum samples were treated by the uricase, the chromato- graphic peak of uric acid disappeared. The linear range of reverse-phase HPLC-UV assay was from 2.08 to 1 785 ixmoL/L. The within- run, between-run, inter-day and total coefficients of variation （CV） of reverse-phase HPLC-UV were less than 0.3% , 0.4% , 2.0% and 2.7% respectively. The average relative recovery rate was from 96% to 100.6%. The proportional bias of target values of the cer- tified reference material SRM 909b-I and -1I was -2.5% and -2.3%, and the p

关 键 词：尿酸 高效液相色谱 有证参考物质 

分 类 号：R446.1[医药卫生—诊断学]