IDENTIFICATION OF COVALENT ADDUCTS TO PROTEIN SULFUR NUCLEOPHILES BY ALKALINE PERMETHYLATION
IDENTIFICATION OF COVALENT ADDUCTS TO PROTEIN SULFUR NUCLEOPHILES BY ALKALINE PERMETHYLATION
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DOI：
10.1006/abio.1993.1048
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发表时间：
1993-02-01
影响因子：
2.9
通讯作者：
HANZLIK, RP
中科院分区：
文献类型：
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作者：
SLAUGHTER, DE;ZHENG, J;HANZLIK, RP
我们最近报道了肝毒素溴苯与大鼠肝蛋白硫亲核试剂共价结合的代谢产物的鉴定。E. Slaughter和R. P. Hanzlik，Chem. Res. Toxicol. 4，349-359（1991）中描述的。该研究的核心是我们开发了一种称为碱性全甲基化的方法，该方法将异生物质亲电体的蛋白质-S加合物转化为稳定的可提取的苯甲醚衍生物。我们在这里报告的实质性改进，我们原来的碱性全甲基化方法，这将大大扩大其潜在的效用。具体而言，我们已经开发出显着温和的反应条件，消除副反应，提高了从各种巯基尿酸模型化合物的硫代苯甲醚产率的量和一致性，并增加了至少50倍的方法的整体灵敏度。使用本文所述的方法，常规上可以通过GC/MS产生、检测和鉴定少至2pmol的硫代苯甲醚衍生物。这种方法可能是相当普遍的，并应证明有用的反应代谢物的毒理学研究，工业卫生和生物监测，以及农业化学残留物分析。
We recently reported on the identification of metabolites of the hepatotoxin bromobenzene covalently bound to rat liver protein sulfur nucleophiles (D. E. Slaughter and R. P. Hanzlik, Chem. Res. Toxicol. 4, 349-359 (1991). Central to that study was our development of a method called alkaline permethylation which converts protein-S adducts of xenobiotic electrophiles to stable extractable thioanisole derivatives. We report here on substantial improvements to our original alkaline permethylation method which should greatly expand its potential utility. Specifically, we have developed significantly milder reaction conditions, eliminated side reactions, improved the amount and consistency of thioanisole yields from various mercapturic acid model compounds, and increased the overall sensitivity of the method at least 50-fold. Using the procedure described herein it is routinely possible to generate, detect, and identify by GC/MS as little as 2 pmol of a thioanisole derivative. This method is potentially quite general and should prove useful for studies in the toxicology of reactive metabolites, for industrial hygiene and biomonitoring, and for agrichemical residue analysis.