Development of Forensic Diagnosis of Acture Sarin Poisoning

合成了一种类似沙林的有机磷制剂——甲基膦酸异丙酯（BIMP）。该制剂具有与沙林相同的膦酸基团，也具有与沙林相同的抗乙酰胆碱酯酶活性效力。制备实验暴露bimp的人红细胞后，将bimp结合的乙酰胆碱酯酶（AChE）从红细胞膜上溶解，经免疫亲和层析纯化，胰酶消化，结合AChE的沙林水解产物经碱性磷酸酶消化释放。水解产物经三甲基硅基（TMS）衍生化，气相色谱-质谱联用（GC-MS）检测。在实验暴露于bimp的人红细胞中检测到沙林水解产物异丙基甲基膦酸和甲基膦酸。利用这种新方法，我们可以从东京地铁恐怖袭击受害者的红细胞和固定福尔马林的小脑中检测沙林水解产物。

A sarin-like organophosphorus agent, bis (isopropyl methyl) phosphonate (BIMP), was synthesized. This agent has the same phosphonate group as sarin and also has the same anti-acethylcholinesterase activity potency as sarin. After preparation of experimental BIMP-exposed human red blood cells, BIMP-bound acetylcholinesterase (AChE) was solubilized from erythrocyte membranes, purified by immunoaffinity chromatography, digested with trypsin, and the sarin hydrolysis products bound to AChE were released by alkaline phosphatasedigestion. The digested sarin hydrolysis products were subjected to trimethylsilyl (TMS) derivatization and detected by gas chromatography-mass spectrometry (GC-MS). Isopropyl methylphosphonic and methylphosphonic acids, which are the sarin hydrolysis products, were detected in experimental BIMP-exposed human red blood cells. Using this new method, we could detected the sarin hydrolysis products from erythrocytes and formalin-fixed cerebellums of victims of the Tokyo subway terrorist attack.