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一种用于血清和唾液肌酐的简化、快速液相色谱-串联质谱分析法。

A simplified, rapid LC-MS/MS assay for serum and salivary creatinine.

作者信息

Bernstone Laura, Jayanti Anuradha, Keevil Brian

机构信息

Department of Clinical Biochemistry, Wythenshawe Hospital, Manchester University NHS Foundation Trust, Manchester, UK.

Department of Nephrology, Manchester Royal Infirmary, Manchester University NHS Foundation Trust, Manchester, UK.

出版信息

Clin Mass Spectrom. 2018 Nov 29;11:21-26. doi: 10.1016/j.clinms.2018.11.004. eCollection 2019 Jan.

Abstract

In routine clinical laboratories, serum creatinine is typically measured on automated analyzers using colorimetric or enzymatic assays, which are both susceptible to interferences that can lead to incorrect measurement. Here, we present a straightforward and rapid LC-MS/MS assay for serum creatinine using methanol extraction, with separation performed using a strong cation exchange column. Results from this newly developed method were compared against those from an automated Abbott Architect kinetic Jaffe method. We also assessed the effect of bilirubin and glucose, as interferants, on both methods. Our LC-MS/MS assay has a run time of 1.1 min, uses a relatively small sample volume of 10 µL and has a within-batch imprecision of 1.1-1.8% at the concentrations tested, which is within the range necessary for routine clinical use. Recovery from serum samples spiked with creatinine was >95%, and glucose and bilirubin were not found to interfere in the assay. Glucose was observed to significantly interfere in the kinetic Jaffe method, while bilirubin had a variable effect. We also determined that creatinine can be measured from saliva using our method, and that salivary concentrations are, on average, 15% of those in serum. This assay provides an alternative for patient sample analysis where interference is expected in routine creatinine methods.

摘要

在常规临床实验室中,血清肌酐通常使用比色法或酶法在自动分析仪上进行检测,这两种方法都容易受到干扰,从而导致测量结果不准确。在此,我们介绍一种使用甲醇萃取的直接快速的液相色谱-串联质谱(LC-MS/MS)法检测血清肌酐,采用强阳离子交换柱进行分离。将这种新开发方法的结果与自动雅培Architect动力学Jaffe法的结果进行了比较。我们还评估了胆红素和葡萄糖作为干扰物对这两种方法的影响。我们的LC-MS/MS法运行时间为1.1分钟,使用相对少量的10微升样本体积,在所测试的浓度下批内不精密度为1.1-1.8%,这在常规临床使用所需的范围内。加标血清样本中肌酐的回收率>95%,未发现葡萄糖和胆红素干扰该检测。观察到葡萄糖对动力学Jaffe法有显著干扰,而胆红素的影响则各不相同。我们还确定使用我们的方法可以从唾液中测量肌酐,唾液中的浓度平均为血清中的15%。该检测方法为预期常规肌酐检测方法存在干扰的患者样本分析提供了一种替代方法。

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