摘要
目的建立一种使用同位素稀释液相色谱串联质谱法(ID-LC/MS/MS)测定血清肌酐的候选参考方法。方法以[^2H3]肌酐为内标,用无水乙醇沉淀血清中的蛋白质,用氯仿纯化上清液,用液相色谱串联质谱分离测定,以包括法定量。结果血清肌酐测定的批内、批间和总变异系数的平均值分别为0.57%(范围0.52%-0.61%)、0.43%(范围0.11%~0.59%)和0.73%(范围0.62%~0.83%)。加样回收试验的回收率范围为99.09%~101.13%。分析两种参考物质SRM909b和SRM967b,测定结果与认定值的偏差小于0.4%。结论建立了ID-LC/MS/MS法测定血清肌酐的方法,方法准确、精密、简便,可望作为血清肌酐测定的参考方法。
Objective To develop a candidate reference method for the measurement of creatinine in human serum based on isotope dilution liquid chromatography tandem mass spectrometry ( ID-LC/MS/ MS). Methods An isotopically labeled internal standard [ ^2 H3 ] creatinine was added to the serum sample and equilibrated with the endogenous creatinine. The samples were treated with anhydrous ethanol to remove proteins by precipitation. After being washed with chloroform for further clean-up, the samples were analyzed by LC/MS/MS. Serum creatinine was quantified by a bracketing calibration. Results The within-run, between-run and total coefficients of variation ranged from 0. 52% to 0. 61%, 0. 11% to 0. 59% and 0. 61% to 0. 83% , and the averages were 0. 57% , 0.43% and 0. 73% , respectively. The analytical recoveries ranged from 99. 09% to 101.13% with an average of 100. 3%. The results of analyzing the certified reference material SRM 909b ( Level Ⅰ and Ⅱ ) and SRM 967b showed biases of less than 0. 4%. Conclusions An ID-LC/MS/MS method for measuring serum creatinine has been developed. The method is highly precise and accurate and may be used as a candidate reference method for serum ereatinine measurements.
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2009年第5期509-514,共6页
Chinese Journal of Laboratory Medicine
基金
国家科技支撑计划资助项目(2007BA105B09)
国家高技术研究发展计划(863)资助(2006AA020909)
关键词
肌酸酐
同位素标记
色谱法
液相
串联质谱法
Creatinine
Isotope labeling
Chromatography,liquid
Tandem mass spectrometry