Ligocka D, Lison D, Haufroid V
Industrial Toxicology and Occupational Medicine Unit, Catholic University of Louvain, Brussels, Belgium.
J Chromatogr B Analyt Technol Biomed Life Sci. 2002 Oct 5;778(1-2):223-30. doi: 10.1016/s0378-4347(01)00441-8.
The aim of this work was to validate a sensitive method for quantitative analysis of 5-hydroxy-N-methylpyrrolidone (5-HNMP) in urine. This compound has been recommended as a marker for biological monitoring of N-methylpyrrolidone (NMP) exposure. Different solvents and alternative methods of extraction including liquid-liquid extraction (LLE) on Chem Elut and solid-phase extraction (SPE) on Oasis HLB columns were tested. The most efficient extraction of 5-HNMP in urine was LLE with Chem Elut columns and dichloromethane as a solvent (consistently 22% of recovery). The urinary extracts were derivatized by bis(trimethylsilyl)trifluoroacetamide and analysed by gas chromatography-mass spectrometry (GC-MS) with tetradeutered 5-HNMP as an internal standard. The detection limit of this method is 0.017 mg/l urine with an intraassay precision of 1.6-2.6%. The proposed method of extraction is simple and reproducible. Four different m/z signal ratios of TMS-5-HNMP and tetralabelled TMS-5-HNMP have been validated and could be indifferently used in case of unexpected impurities from urine matrix.
本研究的目的是验证一种用于尿液中5-羟基-N-甲基吡咯烷酮（5-HNMP）定量分析的灵敏方法。该化合物已被推荐作为N-甲基吡咯烷酮（NMP）暴露生物监测的标志物。测试了不同的溶剂和替代萃取方法，包括在Chem Elut柱上的液液萃取（LLE）和在Oasis HLB柱上的固相萃取（SPE）。尿液中5-HNMP的最有效萃取方法是使用Chem Elut柱和二氯甲烷作为溶剂的液液萃取（回收率始终为22%）。尿液提取物用双（三甲基硅基）三氟乙酰胺进行衍生化，并以四氘代5-HNMP作为内标，通过气相色谱-质谱联用（GC-MS）进行分析。该方法的检测限为0.017 mg/l尿液，批内精密度为1.6 - 2.6%。所提出的萃取方法简单且可重复。已验证了TMS-5-HNMP和四标记TMS-5-HNMP的四种不同m/z信号比，并且在尿液基质出现意外杂质的情况下可以无差别地使用。
