Determination of phenethyl isothiocyanate in human plasma and urine by ammonia derivatization and liquid chromatography-tandem mass spectrometry
Determination of phenethyl isothiocyanate in human plasma and urine by ammonia derivatization and liquid chromatography-tandem mass spectrometry
复制标题
DOI：
10.1016/j.ab.2003.08.011
复制
发表时间：
2003-12-01
影响因子：
2.9
通讯作者：
Morris, ME
中科院分区：
文献类型：
--
作者：
Ji, Y;Morris, ME
异硫氰酸苯乙酯（PEITC）是一种存在于十字花科蔬菜中的膳食化合物，具有预防癌症的特性。我们的目的是开发和验证一种新的液相色谱-串联质谱法分析PEITC在人血浆和尿液中的浓度。己烷萃取后，向样品中加入氨，使PEITC衍生为苯乙基硫脲。在C-18柱上以乙腈/5 mM甲酸（60：40，v/v）作为移动的流动相进行色谱分离，然后在多反应监测模式下进行串联质谱检测。氘标记的PEITC用作内标。检测限为2 nM，校准曲线在7.8至2000 nM范围内呈线性。日内和日间变异系数分别小于5%和10%。日内和日间准确度范围分别为101.0 - 104.2%和102.8 - 118.6%。加标人血浆和尿液的回收率范围分别为100.3 - 113.5%和98.3 - 103.9%。该测定用于测量从食用100 g豆瓣菜后的受试者获得的血浆和尿液样品中的PEITC。这种新的检测方法代表了第一种具有灵敏度和特异性的分析方法，可用于测定PEITC的血浆和尿液浓度。(C)2003年爱思唯尔公司All rights reserved.
Phenethyl isothiocyanate (PEITC) is a dietary compound present in cruciferous vegetables that has cancer-preventive properties. Our objective was to develop and validate a novel liquid chromatography-tandem mass spectrometry procedure to analyze PEITC concentrations in human plasma and urine. Following hexane extraction, ammonia was added to samples to derivatize PEITC to phenethylthiourea. Chromatographic separation was achieved on a C-18 column with acetonitrile/5 mM formic acid (60:40, v/v) as the mobile phase followed by tandem mass spectrometry detection in multiple reaction monitoring mode. Deuterium-labeled PEITC was used as the internal standard. The detection limit was 2 nM and calibration curves were linear from 7.8 to 2000 nM. The intra- and inter-day coefficients of variation were less than 5 and 10%; respectively. The intra- and inter-day accuracies ranged from 101.0 to 104.2% and from 102.8 to 118.6%, respectively. The recovery from spiked human plasma and urine ranged from 100.3 to 113.5% and from 98.3 to 103.9%, respectively. The assay was used to measure PEITC in plasma and urine samples obtained from subjects after consumption of 100 g of watercress. This novel assay represents the first analytical method with the sensitivity and specificity to determine plasma and urine concentrations of PEITC. (C) 2003 Elsevier Inc. All rights reserved.