Metabolite profiling can be used as a diagnostic measure for both short and long term co-exposure by individuals to benzene, toluene, ethylbenzene and xylenes (BTEX). A novel one pot derivatization in situ kit (OPDISK) was developed and optimized using a multivariate approach based on central composite design. The OPDISK was designed to simultaneously derivatize, in a urine sample matrix, a series of fourteen carboxylic acid and phenol-bearing urinary metabolites of BTEX to enhance their chromatographic analysis and sensitivity for detection by liquid chromatography – electrospray ionization – tandem mass spectrometry (LC-ESI-MS/MS). Using the reagent kit, the less responsive functional units on the molecules were converted to permanently positively-charged functional units. The kit was composed of three components, 2-fluoro-1-methylpyridinium p-toluenesulfonate (FMP), 3-carbinol-1-methylpyridinium iodide (CMP) and triethylamine (TEA) as a basic catalyst and, only after diluting a urine sample 20 fold with acetonitrile, was applied under mild conditions of room temperature and short reaction time of 20 min. The derivatized biomarkers were then directly analyzed using isotope dilution LC-ESI-MS/MS. The method was sensitive (limit of detection on column ranged from 1.4 pg to 3.1 ng), accurate (mean accuracy from 85% to 114%), and precise (mean coefficient of variation from 1% to 14%). The method results indicated a good linearity (R² ≥ 0.990) for all metabolites. ClinChek^® urine control samples were used successfully to demonstrate the accuracy of the method.

代谢物分析可作为个人短期和长期共同接触苯，甲苯，乙苯和二甲苯（BTEX）的诊断手段。一种新颖的原位锅衍生化套件（OPDISK）是基于中央复合设计的多变量方法而开发和优化的。OPDISK旨在在尿液样品基质中同时衍生一系列BTEX的十四种羧酸和含酚的尿代谢物，以增强其色谱分析和液相色谱–电喷雾电离–串联质谱（LC- ESI-MS / MS）。使用试剂盒，将分子上反应较弱的功能单元转换为永久带正电的功能单元。该试剂盒由三部分组成，即对甲苯磺酸2-氟-1-甲基吡啶鎓（FMP），碘化3-甲醇-1-碘化吡啶鎓（CMP）和三乙胺（TEA）作为碱性催化剂，并且仅在稀释尿液样品后才可使用。用乙腈20倍，在室温的温和条件下和20分钟的短反应时间下，使用硅藻土。然后使用同位素稀释LC-ESI-MS / MS直接分析衍生的生物标记。该方法灵敏（色谱柱的检出限为1.4 pg至3.1 ng），准确（平均准确度为85％至114％）和精确（平均变异系数为1％至14％）。方法结果表明线性良好（R²  ≥0.990）的所有代谢物。ClinChek ^®尿对照样品成功地用于说明该方法的精确度。