| 摘要: | 呋喃甲醛(furfural)被廣泛應用在化學石油工業上且為已知的食物性致突變物質，同時具有基因毒性，其暴露途徑包含吸入、皮膚接觸及食入。為了保護大眾的健康，降低勞工作業場所或一般環境下的致癌風險及基因毒性的影響，進行呋喃甲醛的暴露評估有其必要性。因此，本研究希望利用固相微萃取 (solid-phase microextraction, SPME) 與氣相層析/質譜儀（GC/MS）之結合，建立呋喃甲醛尿中代謝物與肌酸酐的分析技術，以發展方便、敏感且正確的生物偵測方法，進而提供呋喃甲醛暴露評估工具。 首先製備濃度為0.1～3倍furfural BEIs (Biological Exposure Indices)的添加樣本，再利用固相微萃取技術進行頂空吸附，並以GC/MS分析。萃取過程中，除了測試包括: PA、PDMS、PDMS/DVB、CAR/PDMS、CW/DVB等纖維的吸附效果外，其他如:溫度、攪拌速度、離子及含鹽量等可能影響纖維吸附量的因素，亦都是研究中探討的重點。肌酸酐的分析亦利用已知濃度的添加樣本進行測試，但進行分析之前，已吸附肌酸酐的SPME纖維需先以無水三氟醋酸（trifluoroacetic anhydride, TFAA）頂空裹附，經與肌酸酐反應生成衍生物(trifluoroacetamide)後，再以GC/MS進行分析。 試驗結果顯示使用65?m PDMS/DVB纖維最符合研究所需。關於肌酸酐的分析，可直接將纖維插入水樣本中，於室溫環境下配合1200 rpm的電磁攪拌速度，讓纖維在樣本中停留60sec，接著取出纖維並插入另一個置有100?L TFAA的4mL vial中並停留30sec，再取出纖維另插入一個空的4mL vial中並等待10min後，以GC/MS在注射口溫度230?C下進行分析(m/z=95、112)。至於呋喃甲酸的偵測，亦可使用 PDMS/DVB纖維在室溫下直接插入樣本並停留60sec進行吸附後(轉速 1200 rpm)，以GC/MS分析(m/z=95、112)。當纖維在注射口溫度230 ?C停留5min，其脫附效率>95%。本研究的呋喃甲酸的檢量線範圍: 0.03～0.89 g/L (R2=0.996)，約等於0.1 ~ 3倍的BEIs值，分析準確度在±10％以內。分析變異係數CVa％為2.40％。方法偵測極限為19.42 μg，儀器偵測極限為0.26μg；肌酸酐的檢量線範圍: 0.32～3.12 g/L (R2>0.995)，為有效尿液的肌酸酐濃度範圍。分析變異係數CVa％為3.5％。方法偵測極限為173.63μg，儀器偵測極限為0.26μg。不同條件對萃取呋喃甲酸的影響發現添加氯化鈉及亞硫酸鈉兩種鹽類及加溫、攪拌樣本均會增加纖維吸附呋喃甲酸的量；肌酸酐部分則發現添加氯化鈉、硫酸鈉及亞硫酸鈉會降低纖維萃取肌酸酐衍生物的量。 本研究發現固相微萃取技術可應用於尿液中呋喃甲酸及肌酸酐之分析。與現有方法相比較，本研究能有效縮短樣本前處理及分析時間並減少溶劑的使用，且與肌酸酐現有分析方法的平行比對具有一致性；然而在不更換衍生試劑下，本研究的方法並無法直接進行兩者的同步分析。可能的改善方式是先分析樣本中的呋喃甲酸濃度，再利用呋喃甲酸與肌酸酐混合經衍生後的檢量線來推算肌酸酐的濃度。; Furfural is widely used in petrochemical industry, and is a known dietary mutagens. The exposure routes of furfural include inhalation, skin contact and ingestion. Furfural is an irritant of the eyes, mucous membranes, and skin. Besides, furfural is a confirmed animal carcinogen. To perform the exposure assessment of furfural, biological monitoring is a better tool since inhalation is not the only route of exposure. However, there are drawbacks of the current methods. In recent years, a new analytical technique called solid-phase microextraction (SPME) has been developed which is favorable in terms of solvent-free and convenience. Therefore the purpose of this study was to develop a new analytical method for furoic acid and creatinine in urine based on SPME, to provide a better tool for the exposure assessment of furfural. Known concentrations of furoic acid equal 0.1～ 3 times BEIs (Biological Exposure Indices) of furfural were prepared in spiked urine samples. Headspace adsorptions were performed followed by the GC/MS analysis. Including 85?m PA, PDMS, PDMS/DVB, CAR/PDMS, and CW/DVB were tested for the abilities of adsorption. Besides, temperatures, speeds of magnetic stirring, and pH values were all the factors that needed to be investigated. For the analysis of creatinine, known concentrations of spiked urine samples were also prepared. However, trifluoroacetic anhydride (TFAA) must be loaded onto the fiber after the adsorption of creatinine. The derivatives from the reaction of TFAA and creatinine were then analyzed by GC/MS. The PDMS/DVB was selected for this research. For the analysis of creatinine, the SPME fiber was first directly immersed into a spiked water sample for 60 sec with 1200 rpm magnetic stirring. The fiber was then transferred to another 4 mL vial (which was filled with 100 ?L of TFAA) and stood for 30sec. Afterward the fiber was transferred to another blank 4 mL vial again and stood for 10 min before the analysis by GC/MS (DB-WAX column, injector temp.=230?C, m/z=95,112). The detection of furoic acid was also performed by the adsorption of PDMS/DVB with direct urine sample immersed for 30sec (1200 rpm) followed by the GC/MS analysis. The desorption efficiency > 95% when the SPME fiber was stayed in the injector for 5 min (230?C). The concentrations of the spiked furoic acid standards ranged from 0.03~0.89 g/L (R2=0.996) which was equal to 0.1~3 times BEIs. And the concentrations of the spiked creatinine standards ranged from |
