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首页> 外文期刊>Mass Spectrometry Letters >Simultaneous Determination of Five Porphyrins in Human Urine and Plasma Using High Performance Liquid Chromatography-Tandem Mass Spectrometry
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Simultaneous Determination of Five Porphyrins in Human Urine and Plasma Using High Performance Liquid Chromatography-Tandem Mass Spectrometry

机译：高效液相色谱-串联质谱法同时测定人尿和血浆中的五种卟啉

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A specific and sensitive liquid chromatography-electrospray ionization tandem mass spectrometry method (LC-ESI-MS/MS) was developed and validated for the simultaneous quantification of porphyrins (coproporphyrin, pentacarboxylporphyrin, hexacarboxylporphyrin, heptacarboxylporphyrin, and uroporphyrin) in human plasma and urine. Acidified plasma samples and urine samples were prepared by using liquid-liquid extraction using ethyl acetate and protein precipitation with acetonitrile, respectively. The separation was achieved onto a Synergi Fusion RP column ( , ) with a gradient elution of mobile phase A (0.1% formic acid in 2 mmol/L ammonium acetate, v/v) and mobile phase B (20% methanol in acetonitrile, v/v) at a flow rate of /min. Porphyrins and the internal standard (IS), coproporphyrin I- , were detected by a tandem mass spectrometer equipped with an electrospray ion source operating in positive ion mode. Multiple reaction monitoring (MRM) transitions of the protonated precursor ions and the related product ions were optimized to increase selectivity and sensitivity. The proposed method was validated by assessing selectivity, linearity, limit of quantification (LOQ), precision, accuracy, recovery, and stability. The calibration curves were obtained in the range of 0.1-100 nmol/L and the LOQs were estimated as 0.1 nmol/L for all porphyrins. Results obtained from the validation study of porphyrins showed good accuracy, precision, recovery, and stability. Finally, the proposed method was successfully applied to clinical studies on the autism spectrum disorder (ASD) diagnosis of 203 Korean children.
机译：建立了一种特异性灵敏的液相色谱-电喷雾串联质谱法（LC-ESI-MS / MS）并验证了该方法可同时定量测定人血浆和尿液中的卟啉（共卟啉，五羧基卟啉，六羧基卟啉，七羧基卟啉和尿卟啉）。通过分别使用乙酸乙酯进行液-液萃取和使用乙腈进行蛋白质沉淀，分别制备了酸化的血浆样品和尿液样品。使用流动相A（0.1％甲酸在2 mmol / L乙酸铵中的体积，v / v）和流动相B（20％甲醇在乙腈中的体积，v）的梯度洗脱，在Synergi Fusion RP柱（）上完成分离。 / v）。用配备有以正离子模式运行的电喷雾离子源的串联质谱仪检测卟啉和内标（IS）协同卟啉I-。优化了质子化前体离子和相关产物离子的多反应监测（MRM）跃迁，以提高选择性和灵敏度。通过评估选择性，线性，定量限（LOQ），精密度，准确性，回收率和稳定性来验证所提出的方法。获得的校准曲线范围为0.1-100 nmol / L，所有卟啉的LOQ估计为0.1 nmol / L。卟啉验证研究的结果显示出良好的准确性，精密度，回收率和稳定性。最后，该方法成功地应用于203例韩国儿童自闭症谱系障碍（ASD）诊断的临床研究。

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