High-performance liquid chromatography-based determination of total isothiocyanate levels in human plasma: Application to studies with 2-phenethyl isothiocyanate
High-performance liquid chromatography-based determination of total isothiocyanate levels in human plasma: Application to studies with 2-phenethyl isothiocyanate
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DOI：
10.1006/abio.2001.5030
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发表时间：
2001-04-15
影响因子：
2.9
通讯作者：
Chung, FL
中科院分区：
文献类型：
--
作者：
Liebes, L;Conaway, CC;Chung, FL
饮食和药物异硫氰酸酯（ITC）可能在降低某些癌症的风险方面发挥作用。对人体内ITCs的定量对于流行病学和药代动力学研究都很重要。我们描述了一种改进的基于HPLC的测定尿中ITCs的方法，用于人血浆。该方法利用1，2-苯二硫醇与人血浆中存在的ITCs的环化缩合反应，然后进行两步己烷提取，并通过HPLC使用365 nm处的UV检测进行分析。该方法在49-3003 nM苯乙基异硫氰酸酯（PEITC）范围内用人血浆显示出线性和重现性（r（2）= 0.996 +/- 0.003）。用PEITC的另外两种生物学上存在的缀合物：PEITC-N-乙酰半胱氨酸（PEITC-NAC）和PEITC-谷胱甘肽（PEITC-GSH）观察到类似程度的线性。多天评估的PEITC的回收率为96.6 +/- 1.5%，PEITC-GSH和PEITC-NAC的回收率为100%。多日取样的试验重现性显示，PEITC、PEITC-NAC和PEITC-GSH的平均%CV分别为6.5 +/- 0.3%、6.4 +/- 4.3和12.3 +/- 3.9。在临床研究中，对于23名受试者的非饮食对照组，确定平均血浆ITC水平为413 +/- 193 nM PEITC当量。3例接受40 mg PEITC单次给药的受试者的药代动力学血浆集的多日分析数据显示CV良好（范围：16-21%）。证明了该方法对PEITC在人体中的药代动力学研究的适用性。(C)北京：科学出版社.
Dietary and pharmacologic isothiocyanates (ITCs) may play a role in reducing the risk of certain cancers. The quantification of ITCs in humans is important both for epidemiological and pharmacokinetic studies. We describe a modification of an HPLC-based assay of urinary ITCs for use with human plasma The assay utilizes the cyclocondensation reaction of 1,2-benzenedithiol with ITCs present in human plasma, followed by a two-step hexane extraction and analysis by HPLC using W detection at 365 nm. The method shows linearity and reproducibility with human plasma over a range of 49-3003 nM phenethyl isothiocyanate (PEITC) (r(2) = 0.996 +/- 0.003). A similar degree of linearity was seen with two other biologically occurring conjugates of PEITC: PEITC-N-acetylcysteine (PEITC-NAC) and PEITC-glutathione (PEITC-GSH). The recovery of PEITC assessed on multiple days was 96.6 +/- 1.5% and was 100% for PEITC-GSH and PEITC-NAC. The reproducibility of the assay on multiday samplings showed a mean %CV of 6.5 +/- 0.3% for PEITC, 6.4 +/- 4.3 for PEITC-NAC and 12.3 +/- 3.9 for PEITC-GSH. In clinical studies, mean plasma ITC level of 413 +/- 193 nM PEITC equivalents was determined for a non-dietary-controlled group of 23 subjects. Multiday analysis data from pharmacokinetic plasma sets of 3 subjects taking a single dose of PEITC at 40 mg showed a good CV (range: 16-21%). The applicability of the methodology to pharmacokinetic studies of PEITC in humans is demonstrated. (C) 2001 Academic Press.