Thienpont L M, Van Landuyt K G, Stöckl D, De Leenheer A P
Laboratoria voor Medische Biochemie en voor Klinische Analyse, University of Ghent, Belgium.
Clin Chem. 1995 Jul;41(7):995-1003.
We present a candidate Reference Method for determining creatinine in serum, based on isocratic HPLC. The chromatographic column, with alkaline-treated aluminum oxide as stationary phase, is eluted with an equivolume mixture of methanol/acetonitrile containing 70 mL/L aqueous NaOH (10 mmol/L), and ultraviolet absorbance is detected at 240 nm. We investigated the ruggedness of the method and validated its performance with isotope dilution gas chromatography-mass spectrometry (ID GC-MS) for a set of 22 patients' sera and 10 commercially available lyophilized control materials. The mean deviation from ID GC-MS was +0.1% (range, -2.2% to +2.2%). The between-day CV, calculated from six independent measurements performed on three different days, was 0.9% (range 0.2% to 1.9%); the within-run CV was 0.8%. The total error of the method was < 3%. These performance characteristics make the method suitable for target-setting of quality-control materials and accuracy assessment of routine test kits. For the latter application, done with split-sample measurements of a panel of 83 patients' specimens, we used the Boehringer Mannheim enzymatic creatinine PAP test on the Hitachi 911, the Kodak Ektachem single-slide enzymatic creatinine test on the Ektachem 700, the Merck Jaffé kinetic assay on the Mega analyzer, and the Roche Jaffé kinetic test on the Cobas Mira.
我们提出了一种基于等度高效液相色谱法测定血清中肌酐的候选参考方法。该色谱柱以碱处理的氧化铝为固定相，用含70 mL/L氢氧化钠水溶液（10 mmol/L）的甲醇/乙腈等体积混合物洗脱，并在240 nm处检测紫外吸光度。我们研究了该方法的耐用性，并使用同位素稀释气相色谱-质谱联用仪（ID GC-MS）对22例患者血清和10种市售冻干质控品进行了性能验证。与ID GC-MS的平均偏差为+0.1%（范围为-2.2%至+2.2%）。通过在三个不同日期进行的六次独立测量计算得出的日间变异系数为0.9%（范围为0.2%至1.9%）；批内变异系数为0.8%。该方法的总误差<3%。这些性能特征使该方法适用于质控品的靶值设定和常规检测试剂盒的准确性评估。对于后一种应用，通过对83例患者标本进行分样测量，我们在日立911上使用了勃林格殷格翰酶法肌酐PAP检测，在Ektachem 700上使用了柯达Ektachem单张酶法肌酐检测，在Mega分析仪上使用了默克Jaffé动力学分析，在Cobas Mira上使用了罗氏Jaffé动力学检测。
