Lee Jin Young, Kim Changhwan, Lee Yong Han
Agency for Defense Development (ADD), PO Box 35-5, Yuseong-gu, Daejeon 305-600, Republic of Korea.
J Anal Toxicol. 2018 Jun 1;42(5):293-299. doi: 10.1093/jat/bkx066.
A sensitive method for the purification and determination of two protein adducts, organophosphorus (OP)-BChE and OP-albumin adducts, in a single sample using a simultaneous sample preparation method was developed and validated using liquid chromatography-tandem mass spectrometry. First, we isolated O-ethyl S-2-diisopropylaminoethyl methyl phosphonothiolate (VX) and O-pinacolyl methylphosphonofluoridate (soman, GD)-BChE adducts using an immunomagnetic separation (IMS) method and the HiTrap™ Blue affinity column was subsequently used to isolate and purify VX and GD-albumin adducts from the plasma of rhesus monkeys exposed to nerve agents. Additionally, we examined the time-concentration profiles of two biomarkers, VX and GD-nonapeptides and VX and GD-tyrosines, derived from OP-BChE and OP-albumin adducts up to 8 weeks after exposure. Based on the results, we determined that VX and GD-tyrosine is more suitable than VX and GD-nonapeptide as a biomarker owing to its longevity. This integrated approach is expected to be applicable for the quantification of other OP-BChE and OP-albumin adducts in human plasma, thus serving as a potential generic assay for exposure to nerve agents.
开发了一种灵敏的方法，可使用同步样品制备方法在单个样品中纯化和测定两种蛋白质加合物，即有机磷（OP）-丁酰胆碱酯酶（BChE）和OP-白蛋白加合物，并采用液相色谱-串联质谱法进行了验证。首先，我们使用免疫磁分离（IMS）方法分离了O-乙基-S-2-二异丙氨基乙基甲基硫代膦酸酯（VX）和O-频哪基甲基膦酸酯（梭曼，GD）-BChE加合物，随后使用HiTrap™ Blue亲和柱从暴露于神经毒剂的恒河猴血浆中分离并纯化VX和GD-白蛋白加合物。此外，我们研究了暴露后长达8周的两种生物标志物，即源自OP-BChE和OP-白蛋白加合物的VX和GD-九肽以及VX和GD-酪氨酸的时间-浓度曲线。基于这些结果，我们确定由于VX和GD-酪氨酸的持久性，它比VX和GD-九肽更适合作为生物标志物。这种综合方法有望适用于定量人血浆中其他OP-BChE和OP-白蛋白加合物，从而作为一种潜在的通用神经毒剂暴露检测方法。
