A simplified, rapid LC-MS/MS assay for serum and salivary creatinine
A simplified, rapid LC-MS/MS assay for serum and salivary creatinine
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DOI：
10.1016/j.clinms.2018.11.004
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发表时间：
2019-01-01
影响因子：
2.1
通讯作者：
Keevil, Brian
中科院分区：
文献类型：
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作者：
Bernstone, Laura;Jayanti, Anuradha;Keevil, Brian
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测量，这两种方法都容易受到干扰，导致测量结果不正确。在这里，我们提出了一种直接快速的LC-MS/MS检测方法，使用甲醇提取，使用强阳离子交换柱进行分离。这种新开发方法的结果与自动化Abbott Architect动态Jaffe方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素对两种方法的影响。我们的LC-MS/MS检测运行时间为1.1 min，使用相对较小的10 μ L样本量，在所测浓度下的批内不精密度为1.1-1.8%，在常规临床使用所需的范围内。加入肌酐的血清样品回收率为95%，葡萄糖和胆红素未发现干扰测定。葡萄糖对动力学Jaffe法有显著干扰，而胆红素对动力学Jaffe法有可变影响。我们还确定，使用我们的方法可以从唾液中测量肌酐，并且唾液浓度平均为血清浓度的15%。该试验提供了一种替代的病人样本分析，其中干扰是预期在常规肌酐方法。
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 mu L 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.