Department of Chemistry and Biochemistry , South Dakota State University , Box 2202, Brookings , South Dakota 57007 , United States.
Beckman Laser Institute and Medical Clinic , University of California , Irvine , California 92612 , United States.
Chem Res Toxicol. 2019 Apr 15;32(4):718-726. doi: 10.1021/acs.chemrestox.8b00384. Epub 2019 Feb 21.
The direct analysis of cyanide (HCN or CN inclusively symbolized as CN) to confirm exposure has major limitations due to cyanide's volatility, reactivity, and short half-life in biological fluids. These limitations have led to the exploration of cyanide detoxification products for indirect verification of cyanide exposure. Although cyanide interacts strongly with sulfur-containing molecules, to date, biomarkers resulting from the interaction of cyanide with glutathione (GSH; i.e., a biologically abundant sulfur-donating biomolecule) have yet to be discovered. In this study, we studied the interaction of CN and GSH to produce 2-aminothiazoline-4-oxoaminoethanioc acid (ATOEA). An LC-MS/MS method was developed and validated to analyze ATOEA from plasma, producing a linear range of 0.5-50 μM, a limit of detection of 200 nM, and excellent precision and accuracy. ATOEA concentrations were significantly elevated in the plasma of animals following cyanide exposure. Moreover, the production of ATOEA from cyanide exposure was confirmed by detection of both ATOEA and ATOEA-CN in rabbit plasma ( N = 11 animals) following administration of NaCN:KCN (1:1), with a similar amount of ATOEA and ATOEA-CN formed ( R = 0.9924, p < 0.05). The concentration of ATOEA increased with cyanide dose and then decreased rapidly when an antidote was administrated. This study definitively showed that ATOEA is produced from interaction of CN and GSH and can serve as a biomarker of cyanide exposure.
由于氰化物的挥发性、反应性和在生物体液中的半衰期短，直接分析氰化物（HCN 或 CN 统称 CN）来确认暴露情况存在重大限制。这些限制导致了对氰化物解毒产物的探索，以间接验证氰化物暴露情况。尽管氰化物与含硫分子强烈相互作用，但迄今为止，尚未发现由氰化物与谷胱甘肽（GSH；即生物体内丰富的硫供体生物分子）相互作用产生的生物标志物。在这项研究中，我们研究了 CN 与 GSH 相互作用生成 2-氨基噻唑啉-4-氧代乙氨基乙酸（ATOEA）的过程。开发并验证了一种 LC-MS/MS 方法来分析来自血浆的 ATOEA，产生了线性范围为 0.5-50 μM、检测限为 200 nM、精密度和准确度良好的结果。在动物暴露于氰化物后，其血浆中的 ATOEA 浓度显著升高。此外，通过检测 NaCN:KCN（1:1）给药后兔血浆中的 ATOEA 和 ATOEA-CN（N=11 只动物），证实了氰化物暴露会产生 ATOEA，两者的生成量相似（R=0.9924，p<0.05）。ATOEA 的浓度随氰化物剂量增加而增加，当给予解毒剂时迅速下降。这项研究明确表明，ATOEA 是由 CN 和 GSH 相互作用产生的，可作为氰化物暴露的生物标志物。
