利用液相層析串聯質譜儀開發並建立Diisocyanate-related diamines之分析與應用

異佛爾酮二異氰酸酯（Isophorone diisocyanate，IPDI）、2,6-甲苯二異氰酸酯（2,6-toluene diisocyanate，2,6-TDI）、2,4-甲苯二異氰酸酯（2,4-toluene diisocyanate，2,4-TDI）、4,4′-二苯基甲烷二異氰酸酯（4,4′-methylene diphenyl diisocyanate，MDI）及1,5-萘二異氰酸酯（1,5-naphthalene diisocyanate，NDI）屬於Diisocyanate，是重要的聚氨酯（PU）工業原料，二異氰酸酯（Diisocyanate）已被研究與職業氣喘及呼吸道疾病有關，目前用以評估Isocyanate暴露量的最佳方法，除了對於空氣中濃度監測外，便是檢測人體尿液中二異氰酸酯轉換成胺類之代謝產物，其二異氰酸酯主要對應之胺類化合物分別為異佛爾酮二胺（Isophorone diamine，IPDA）、2,6-甲苯二胺（2,6-toluene diamine，2,6-TDA）、2,4-甲苯二胺（2,4-toluene diamine，2,4-TDA）、4,4′-二胺基二苯甲烷（4,4′-methylene diphenyl diamine，MDA）及1,5-萘二胺（1,5-naphthalene diamine，NDA）。本研究中利用微波用於尿液中代謝物的水解及搭配液相層析串聯質譜法，對於常用二異氰酸酯代謝物：IPDA、2,6-TDA、2,4-TDA、MDA及NDA建立一簡單且具高靈敏度的分析法。本研究中影響分析物在質譜分析下感度的因素很多，如溶劑中甲酸的添加，溶劑中甲醇與純水的混合比例等，影響尿液前處理的因素亦有水解酸鹼當量、微波水解能量與時間、萃取所需甲苯體積等，皆於本研究中有詳細探討。本方法可應用於人體尿液中對IPDA、2,6-TDA、2,4-TDA、MDA及NDA之分析，其尿液中定量範圍分別為1-50 ng/mL IPDA、2,6-TDA、2,4-TDA、NDA及0.5-25 ng/mL MDA，線性回歸係數皆大於0.999，其偵測極限分別為0.025 ng/mL IPDA, 0.4 ng/mL 2,6-TDA, 0.1 ng/mL 2,4-TDA, 0.025 ng/mL NDA 及0.025 ng/mL MDA (S/N=3，注射體積 10 μL)。在同日間及異日間分析結果中，相對標準差（R.S.D）分別小於6.5 %及11.7 %，相對誤差（R.E）分別小於7.1 %及7.0 %。

關鍵字

二異氰酸酯；二胺；液相層析串聯質譜

並列摘要

Isophorone diisocyanate(IPDI), 2,6-toluene diisocyanate(2,6-TDI), 2,4-toluene diisocyanate (2,4-TDI), 4,4’-methylene diphenyldiisocyanate (MDI) and 1,5-naphthalene diisocyanate (NDI) are important sources of polyurethane(PU) productions. Diisocyanates have been researched about occupational asthma and respiratory diseases. Currently, the main methods for determination of diisocyanate exposure are determination of the concentration of diisocyanate in air or the diisocyanate metabolites which are diisocyanate related diamines protein adduct in human urine. The main diisocyanate-related diamines are Isophorone diamine(IPDA), 2,6-toluene diamine(2,6-TDA), 2,4-toluene diamine(2,4-TDA), 4,4′-diaminodiphenylmethane(MDA) and 1,5-diamino naphtha-lene(NDA). In this study, we developed microwave method for diisocyanate related diamines protein adduct hydrolysis and a simple and sensitive method with liquid chromatography-tandem mass spectrometry to determine the diisocyanate-related diamines, IPDA, 2,6-TDA, 2,4-TDA, MDA and NDA. In the study, there are many effects of intensity for determination of IPDA, 2,6-TDA, 2,4-TDA, MDA and NDA, such as the formic acid addition in solvent, the solvent proportion of methanol and H2O. There are many effects of work-up procedure for urine sample, such as equivalent of acid and base, microwave power and irradiation time, volume of toluene. All were discussed in this study. The application of the method for human urine which determine IPDA, 2,6-TDA, 2,4-TDA, MDA and NDA were evaluated the concentrations over range of 1-50 ng/mL IPDA, 2,6-TDA, 2,4-TDA, NDA and 0.5-25 ng/mL MDA. The linear correlation coefficient are greater than 0.999. The detection limit are 0.025 ng/mL IPDA, 0.4 ng/mL 2,6-TDA, 0.1 ng/mL 2,4-TDA, 0.025 ng/mL NDA and 0.025 ng/mL MDA (S/N=3, injection volume 10 μL) The intra-day relative standard deviation (R.S.D) and the inter-day intra-day relative standard deviation (R.S.D) were below 6.5 % and 11.7 %, the relative error (R.E) were below 7.1 % and 7.0 %