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首页> 外文期刊>Journal of mass spectrometry: JMS >Quantification of L-ergothioneine in human plasma and erythrocytes by liquid chromatography-tandem mass spectrometry
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Quantification of L-ergothioneine in human plasma and erythrocytes by liquid chromatography-tandem mass spectrometry

机译：液相色谱-串联质谱法定量测定人血浆和红细胞中的L-麦角硫氨酸

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摘要

A sensitive analytical method has been developed and validated for the quantification of L-ergothioneine in human plasma and erythrocytes by liquid chromatography-tandem mass spectrometry. A commercially available isotope-labeled L-ergothioneine-d_9 is used as the internal standard. A simple protein precipitation with acetonitrile is utilized for bio-sample preparation prior to analysis. Chromatographic separation of L-ergothioneine is conducted using gradient elution on Alltime C18 (150 mm × 2.1 mm, 5 ?μ). The run time is 6 min at a constant flow rate of 0.45 ml/min. The mass spectrometer is operated under a positive electrospray ionization condition with multiple reaction monitoring mode. The mass transitions of L-ergothioneine and L-ergothioneine-d_9 are m/z 230 > 127 and m/z 239 > 127, respectively. Excellent linearity [coefficient of determination (r~2) ≥ 0.9998] can be achieved for L-ergothioneine quantification at the ranges of 10 to 10 000 ng/ml, with the intra-day and inter-day precisions at 0.9-3.9% and 1.3-5.7%, respectively, and the accuracies for all quality control samples between 94.5 and 101.0%. This validated analytical method is suitable for pharmacokinetic monitoring of L-ergothioneine in human and erythrocytes. Based on the determination of bio-samples from five healthy subjects, the mean concentrations of L-ergothioneine in plasma and erythrocytes are 107.4 ± 20.5 ng/ml and 1285.0 ± 1363.0 ng/ml, respectively.
机译：已经开发了一种灵敏的分析方法，并通过液相色谱-串联质谱法对人血浆和红细胞中的L-麦角硫氨酸进行定量验证。使用市售的同位素标记的L-麦角硫氨酸-d_9作为内标。在分析之前，用乙腈进行简单的蛋白质沉淀可用于生物样品的制备。 L-麦角硫氨酸的色谱分离是在Alltime C18（150 mm×2.1 mm，5 µμ）上进行梯度洗脱的。在0.45 ml / min的恒定流速下，运行时间为6分钟。质谱仪在正电喷雾电离条件下以多反应监测模式运行。 L-麦角硫氨酸和L-麦角硫氨酸-d_9的质量跃迁分别为m / z 230> 127和m / z 239> 127。 L-麦角硫氨酸定量在10至10000 ng / ml范围内可实现出色的线性[测定系数（r〜2）≥0.9998]，日内和日间精度为0.9-3.9％，分别为1.3-5.7％，所有质量控制样品的准确度在94.5和101.0％之间。这种经过验证的分析方法适用于人体和红细胞中L-麦角硫氨酸的药代动力学监测。根据对五个健康受试者的生物样品的测定，血浆和红细胞中L-麦角硫氨酸的平均浓度分别为107.4±20.5 ng / ml和1285.0±1363.0 ng / ml。

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