本研究中建立了水樣中磺胺類抗生素可靠的分析方法。水樣以固相萃取法進行前處理，首先固萃匣以5 mL甲醇活化，5 mL的去離子水平衡固萃匣，然後取水樣500 mL進樣，進樣後以5 mL去離子水清洗固萃匣，抽氣30分鐘，再以5 mL甲醇流析，隨後收集流析下來的溶劑吹氮至乾涸，最後加入1 mL甲醇回溶，利用液相層析串聯質譜儀(LC-MS/MS)分析10種磺胺類抗生素成分含量，分別為磺胺嘧啶(SDZ)、磺胺塞唑(STZ)、磺胺苯唑(SNZ)、磺胺氯噠嗪(SCP)、磺胺二甲嘧啶(SMZ)、磺胺二甲氧嘧啶(SDM)、磺胺喹噁啉(SQX)、磺胺間甲氧嘧啶(SMM)、磺胺甲基嘧啶(SMR)、磺胺甲二唑(SMT)。移動相使用甲醇與0.005 M的醋酸銨溶液進行梯度沖堤，各磺胺類抗生素的回收率介於52.1-86.8%，偵測極限為1.4-3.3 ng/L，定量極限為4.8-11.1 ng/L。
針對高屏溪沿岸水體在2013年10月2日和2014年2月6日採集六個測站進行環境水體中磺胺類抗生素成分含量分析，各測站均檢出磺胺類抗生素。SDZ濃度範圍在1.8-513.3 ng/L；STZ濃度範圍在ND-574.2 ng/L，SNZ濃度範圍在ND-6.8 ng/L，SCP濃度範圍在6.7-412.5 ng/L，SDM濃度範圍在2.6-77.3 ng/L，SQX濃度範圍在ND-28.0 ng/L，SMM濃度範圍在4.5-76.1 ng/L，SMR濃度範圍在ND-6.8 ng/L，SMZ和SMT各採樣點皆未檢出。從2013年10月2日採樣所測得磺胺類抗生素濃度略低於2014年2月6日之測得結果，應是降雨量不同影響環境中磺胺類抗生素含量。
高屏溪本流上的里嶺大橋、高屏大橋以及高屏溪出海口三測站，所測得的磺胺類抗生素濃度有漸增的趨勢，其支流上的河濱公園、萬大大橋下支流測站，所測得的磺胺類抗生素濃度偏高，排入高屏溪後導致本流磺胺類抗生素濃度升高。

A reliable analytical method for the determination of sulfonamide antibiotics in Gaoping River waters was developed. Solid phase extraction was used for sample pretreatment. First, the SPE cartridges were conditioned with 5 mL of methanol and balanced with 5 mL of ultrapure water, then 500 mL of water sample was loaded into the SPE cartridges. Next, the SPE cartridges were washed with 5 mL of ultrapure water and dried for 30 minutes in vacuum. Analytes were eluted from the cartridges with 5 mL of methanol, and then dried under a gentle flow of nitrogen gas, reconstituted by 1 mL of methanol, and transferred into a sample vial. Ten sulfonamide antibiotics including sulfadiazine(SDZ), sulfathiazole(STZ), sulfaphenazole(SNZ), sulfachloropyridazine(SCP), sulfamethazine(SMZ), sulfadimethoxine(SDM), sulfaquinoxaline(SQX), sulfamonomethoxine(SMM), sulfamerazine(SMR), sulfamethizole(SMT) were analyzed by LC-MS/MS equipped with a mobile phase composed of 0.005 M ammonium acetate and methanol to perform gradient elution. The recovery rates were 52.1-86.8 %, limit of detection were 1.4-3.3 ng/L and limit of quantitation were 4.8-11.14ng/L.
On October 2, 2013(wet season), and February 6, 2014(dry season), environmental water samples were collected from six sampling sites along the Gaoping River to investigate the contents of sulfonamide. Contents of SDZ, STZ, SNZ, SCP, SDM, SQX, SMM, SMR, SMZ and SMT were 1.8-513.3 ng/L, N.D.-574.2 ng/L, N.D.-6.8 ng/L, 6.7-412.5 ng/L, 2.6-77.3 ng/L, N.D.-28.0ng/L, 4.5-76.1 ng/L, N.D.-6.8 ng/L, N.D. and N.D. Concentrations of the measured sulfonamide antibiotics for water samples collected on October 2, 2013 were slightly lower than February 6, 2014, due to the difference in the amount of rainfall.
The concentrations of sulfonamide antibiotics at Gaoping Estuary were higher than the upper mainstream sampling sites Liling Bridge and Gaoping Bridge which is attributed to the inlet of two tributaries with high contents of sulfonamides.

論文審定書 i
中文摘要 ii
Abstract iii
壹、前言 1
一、研究緣起 1
二、研究目標 2
貳、文獻回顧 3
一、磺胺類藥物抗生素 3
(一)磺胺類抗生素簡介 3
(二)磺胺類抗生素的抑菌機制 3
(三)磺胺類抗生素的應用 4
(四)本研究使用磺胺類抗生素介紹 5
二、水樣萃取方法 8
(一)固相萃取法 8
(二)固相萃取吸附劑種類及其作用機制 9
三、分析儀器 11
參、材料與方法 14
一、實驗藥品與材料 14
二、分析方法 16
(一)標準品製備 16
(二)液相層析串聯式質譜儀參數條件 16
(三)採樣地點與水樣前處理 18
(四)水樣中分析物固相萃取法 21
肆、結果與討論 23
一、水樣萃取方法最佳化 23
(一)固相萃取匣之選擇 23
(二)流析溶劑對回收率之影響 25
二、液相層析移動相成分之探討 26
三、品保與品管 28
(一)檢量線 28
(二)最低偵測極限與最低定量極限 29
四、高屏溪溪水中磺胺類抗生素含量分析 30
(一)水文資料 30
(二)高屏溪水中磺胺類抗生素含量分析 31
伍、結論 37
參考文獻 38
附錄一、HLB、MCX、PCX三種固萃匣回收率(%) 41
附錄二、兩種流析溶劑對磺胺類抗生素的回收率(%) 42
附錄三、檢量線 43
附錄四、高屏溪各測站照片 48
附錄五、2005/1~2006/12 國貿局藥物進口量 54
附錄六、國內抗生素進口總量 54
附錄七、10月2日各測站磺胺類抗生素含量長條圖 55
附錄八、2月6日各測站磺胺類抗生素含量長條圖 58

(1) 經濟部水利署南區水資源局。http://www.wrasb.gov.tw/index5.aspx

(2) 行政院農業委員會動植物防疫檢疫局。https://www.baphiq.gov.tw/home.php

(3) 交通部中央氣象局。http://www.cwb.gov.tw/V7/

(4) 吳天賞，藥物化學，藝軒圖書出版社，2007。

(5) 林正芳，新興汙染物(抗生素與止痛藥)於特定汙染源環境之流佈，台灣大學環境工程研究所，2008年。

(6) 陳建輝，自來水源中殘留農藥檢驗方法之建立及水質安全之調查與評估，1998年。

(7) 劉耀馳，分子印跡技術在固相萃取中的應用與展望，湖南大學化學化工學院，2004年。

(8) 張慧兒，以電灑法串聯式液相層析質譜儀定量分析含heparin 之人體血漿中 的fentanyl，碩士論文，大同大學生物工程研究所，2010年。

(9) 楊易倫，利用微透析連結高效能液相層析法來檢測牛奶中的磺胺劑，碩士論文，高雄師範大學化學研究所，2002年。

(10) 莊崇昌，利用微透析連結高效能液相層析法來檢測雞蛋中的磺胺劑，碩士論文，高雄師範大學化學研究所，2002年。

(11) Assassi, N., Tazerouti, A., Canselier, J., P. 2005. Analysis of chlorinated, sulfochlorinated and sulfonamide derivatives of n-tetradecane by gas chromatography/mass spectrometry. J. Chromatogr. A., 1071(1-2), 71-80.

(12) Baran, W., Adamek, E., Ziemian´ ska, J., Sobczak, A. 2011. Effects of the presence of sulfonamides in the environment and their influence on human health. J. Hazard. Mater., 196, 1-15.

(13) Białk-Bielin´ ska, A., Kumirska, J., Palavinskas, R., Stepnowski, P. 2009. Optimization of multiple reaction monitoring mode for the trace analysis of veterinary sulfonamides by LC–MS/MS. Talanta., 80, 947-953.

(14) Connor, E. E. 1998. Sulfonamide antibiotics. Prim. Care Update Ob Gyns., 5, 32-35.

(15) Costanzo, S. D., Murby, J., Bates, J. 2005.Ecosystem response to antibiotics entering the aquatic environment. Mar. Pollut. Bull ., 51, 218-23.

(16) Dasenali, M. E., Thomaidis, N. S. 2010. Multi-residue determination of seventeen sulfonamides and five tetracyclines in fish tissue using a multi-stage LC-ESI-MS/MS approach based on advanced mass spectrometric techniques. Anal. Chim. Acta., 672, 93-102.

(17) Dı´az-Cruz, M. S., Garcı´a-Gala´n, M. J., Barcelo, D. 2008.Highly sensitive simultaneous determination of sulfonamide antibiotics and one metabolite in environmental waters by liquid chromatography–quadrupole linear ion trap–mass spectrometry. J. Chromatogr. A., 1193 , 50-59.

(18) Dinh, Q. T., Alliot, F., Moreau-Guigon, E., Eurin, J., Chevreuil, M., Labadie, P. 2011. Measurement of trace levels of antibiotics in river water using on-line enrichment and triple-quadrupole LC–MS/MS. Talanta ., 85 , 1238-1245.

(19) Gräslund, S., Bengtsson, B.-E. 2001. Chemicals and biological products used in south-east Asian shrimp farming, and their potential impact on the environment -- a review. Sci. Total Environ., 280, 93-131.

(20) Jury, K. L., Khan, S. J., Vancov, T., Stuetz, R. M., Ashbolt, N. J. 2011. Biogeochemistry and Water Quality of the Everglades: Symposium Overview. Crit. Rev. Env. Sci. and Technol., 41, 1–28.

(21) Le-Minh, N., Stuetz, R. M., Khan, S. J. 2012. Determination of six sulfonamide antibiotics, two metabolites and trimethoprim in wastewater by isotope dilution liquid chromatography/tandem mass Spectrometry. Talanta., 89 , 407-416.

(22) Matusik, J. E., Sternal, R. S., Barnes, C. J., Sphon, J. A. 1990. Confirmation of identity by gas chromatography/tandem mass spectrometry of sulfathiazole, sulfamethazine, sulfachloropyridazine, and sulfadimethoxine from bovine or swine liver extracts after quantitation by gas chromatography/electron-capture detection. J Assoc. Off Anal. Chem., 73(4), 529-533.

(23) Sharker, S. D., Latif, Z., Gray, A. I. 2006. Natural Products Isolation Second Edition. Humana Press Inc. Totowa, New Jersey.

(24) Shao, B., Dong, D., Wu, Y., Hu, J., Meng, J., Tu, X., Xu, S. 2005. Simultaneous determination of 17 sulfonamide residues in porcine meat, kidney and liver by solid-phase extraction and liquid chromatography-tandem mass spectrometry. Anal. Chim. Acta., 546, 174-181.

(25) Vega-Morales, T., Torres-Padron, M. E., Sosa-Ferrera, Z., Santana-Rodriguez, J.J. 2009. Determination of alkylphenol ethoxylates and their degradation products in liquid and solid samples. Trends Anal. Chem., 28(10), 1186-1200.

(26) Yang, J.-F. , Ying, G.-G., Zhao, J.-L., Tao, R., Su, H.-C., Chen, F. 2010. Simultaneous determination of four classes of antibiotics in sediments of the Pearl Rivers using RRLC–MS/MS. Sci. Total Environ., 408, 3424-3432.