VERIFIN, Finnish Institute for Verification of the Chemical Weapons Convention, Department of Chemistry, University of Helsinki, Helsinki, Finland.
J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Apr 15;879(13-14):908-14. doi: 10.1016/j.jchromb.2011.02.043. Epub 2011 Mar 5.
As a part of the project for screening unequivocal biomarkers after sulfur mustard exposure, a quantitative method for the determination of β-lyase metabolites 1,1'-sulfonylbis-[2-(methylsulfinyl)ethane] (SBMSE) and 1-methylsulfinyl-2-[2-(methylthio)ethylsulfonyl]ethane (MSMTESE) was validated. Full validation was conducted according to the FDA guidelines for method validation using pooled human urine as a sample matrix. The metabolites were extracted from urine with an optimized sample preparation procedure using ENV+ solid phase extraction cartridge with reduced volume of sample and solvents. Metabolites were detected by improved and faster liquid chromatography-heated electrospray ionization-tandem mass spectrometry (LC-HESI-MS/MS) method with two transitions of each chemical using non-buffered eluents, post-column splitter and higher flow-rate. These provided over five times faster analysis than previously published method providing 4.5 min/sample cycle time, to achieve up to 200 samples per day (24 h). Quantification was performed using deuterium labelled internal standard of SBMSE. The method was linear over the concentration range of 5-200 ng/ml (average correlation coefficients were R(2)=0.997 and R(2)=0.989) for both β-lyase metabolites, SBMSE and MSMTESE, respectively. The average retention times for SBMSE and MSMTESE were 1.96±0.01 min and 3.24±0.03 min (n=54). Calculated limits of detection were 4 ng/ml for both SBMSE and MSMTESE, respectively. Lower limits of quantification were 10 ng/ml and 11 ng/ml for SBMSE and MSMTESE, respectively. This validated method was successfully used in the First Confidence Building Exercise on Biomedical Samples Analysis organized by the Organisation for the Prohibition of Chemical Weapons (OPCW). Identification criteria for analysing unequivocal biomarkers of sulfur mustard with LC-MS/MS after alleged use is discussed and proposed based on the validation and exercise results.
作为筛选芥子气暴露后明确生物标志物项目的一部分，验证了定量测定β-裂合酶代谢物 1,1'-亚磺酰基双-[2-(甲亚磺酰基)乙烷]（SBMSE）和 1-甲亚磺酰基-2-[2-（甲硫基）乙磺酰基]乙烷（MSMTESE）的方法。根据 FDA 方法验证指南，使用混合人尿作为样品基质，对该方法进行了全面验证。通过使用 ENV+固相萃取小柱和减少样品和溶剂体积的优化样品制备程序，从尿液中提取代谢物。使用改进的更快的液相色谱-加热电喷雾串联质谱（LC-HESI-MS/MS）方法，使用每种化学物质的两个转换，在无缓冲洗脱液、柱后分流器和更高流速下检测代谢物。与之前发表的方法相比，该方法的分析速度提高了五倍以上，提供了 4.5 分钟/样品的循环时间，每天可处理多达 200 个样品（24 小时）。使用 SBMSE 的氘标记内标进行定量。该方法在 SBMSE 和 MSMTESE 两种β-裂合酶代谢物的浓度范围为 5-200ng/ml 时呈线性（平均相关系数分别为 R(2)=0.997 和 R(2)=0.989）。SBMSE 和 MSMTESE 的平均保留时间分别为 1.96±0.01 分钟和 3.24±0.03 分钟（n=54）。计算得出 SBMSE 和 MSMTESE 的检出限分别为 4ng/ml。SBMSE 和 MSMTESE 的定量下限分别为 10ng/ml 和 11ng/ml。该经过验证的方法成功用于禁止化学武器组织（OPCW）组织的第一次生物医学样品分析建立信心活动。根据验证和活动结果，讨论并提出了使用 LC-MS/MS 分析疑似使用芥子气后的明确生物标志物的鉴定标准。
Zotero 插件