利用液相層析串聯質譜儀結合4-二甲氨基苯甲酰氯(DMAB)化學衍生法偵測尿液中環己烷、環己酮及環己醇曝露指標成分

Determination of the Urinary Makers for the Occupational Exposure of Cyclohexane, Cyclohexanone and Cyclohexanol by using the DMAB Derivatization and LC-MS/MS

謝雅雯(Ya-Wen Hsien)

指導教授 ： 沈振峯

國立虎尾科技大學/文理學院/生物科技研究所/碩士(2014年)

摘要

環己醇、環己酮與環己烷為工業上常用溶劑，環己烷常用於製造環己醇與環己酮，而環己醇與環己酮常用於製造橡膠、尼龍與塗料工業上等。由文獻指出環己醇對眼睛、鼻子、喉嚨與皮膚有刺激性，而環己酮在動物實驗中會抑制中樞神經，長期吸入對人體造血、腎臟、中樞神經系統會有所為害，且已確定對動物會有致癌性，因此對環己酮、環己醇與環己烷進行生物偵測以了解人員的曝露程度是有其必要性的，由於環己醇、環己酮與環己烷在尿液中的曝露指標物為環己醇、1,2—環己二醇與1,4-環己二醇，本研究主要目的即為開發可同時偵測尿中環己醇、1.2-環己二醇與1,4-環己二醇的LC/MS/MS分析方法，研究中引用4-二甲氨基苯甲酰氯(DMAB)作為化學衍生劑，將三種曝露指標物與內標物，同時經衍生化後再以LC/MS/MS偵測所生成之DMAB-衍生物，除了在標準品可成功確定衍生物的生成之外，將指標物添加在人尿樣中，經衍生化後也可同時測得各DMAB-衍生物的訊號，在人尿樣中線性範圍為0.045-2.7 μg/ml，均有良好的線性(r2＞0.994)，三種衍生物的偵測極限濃度均可達0.045μg/ml。對本方法的精密度與準確度進行測試，在低、中及高三種濃度平均的%Error(RSD)分別為3% (13%), 7% (15%) 和 10 % (12%)。

關鍵字

環己醇 ； 環己二醇 ； 環己酮 ； 液相層析串聯式質譜儀 ； 化學衍生法 ； 生物偵測

並列摘要

Cyclohexanol, cyclohexanone and cyclohexane are wildly used organic solvents and/or chemical intermediates in industry. Cyclohexane is often used as solvent for cyclohexanol and cyclohexanone. Cyclohexanol and cyclohexanone are used for making rubber, nylon and the paint in industry. It has reported that cyclohexanol is an irritant of eyes, nose, throat and skin. Cyclohexanone could inhibit the central nervous system of animal and has carcinogenicity in animal experiments. Moreover, blood, kidney, central nervous system in human could be damaged by the long-term inhalation of cyclohexanone. To understand the exposure of factory workers, the biological monitoring of cyclohexanone, cyclohexanol and cyclohexane is necessary. At present, the corresponding urinary makers(cyclohexanol, 1,2-cyclohexanediol and 1,4-cyclohexanediol) of cyclohexane, cyclohexanone and cyclohexanol were analyzed with GC-MS (with chemical derivatization) and/or GC-FID. The prepartion of the urine sample was usually including acid hydrolysis, solvent extraction and/or chemical derivation, which was relative complicate. In this study, a bioanalytical methods was developed to determine the three makers in urine for monitoring the exposure of cyclohexane, cyclohexanone and cyclohexanol at the same time. The developed method applied the DMAB-derivatization to increase the LC-MS sensitivity of the markers. For the spiked urine sample, after acid hydrolysis, sample dilution and DMAB-derivatzation, the corresponding derivatives were analyzed by LC-ESI-MS/MS. The linearity of 0.045 to 2.72 μg/mL was obtained and the detection limits were measured to be about 0.045μg/ml for the three markers in spiked urine. The accuracy and precision were examined with urine samples spiked at L, M and H levels, where the average error (RSD) were obtained to be 3% (13%), 7% (15%) and 10 % (12%), respectively.

並列關鍵字

Cyclohexanone ； Cyclohexane ； 1,2-Cyclohexanediol ； 1,4-Cyclohexanediol ； Cyclohexanol ； LC-MS/MS ； DMAB-derivation

參考文獻

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