臺灣博碩士論文加值系統

本論文為利用氣相層析法並配合火焰離子化檢測器對市售感冒糖漿製劑及飲品中之acetaminophen和caffeine作分析，建立一個簡單且快速的分析方法。將含有acetaminophen及caffeine之樣品，透過簡單的液相-液相萃取方式作前處理，直接以氣相層析儀分析，所使用的毛細管柱為VF-5ms (30 m × 0.32 mm, 0.25 μm)，管柱的溫度設在180 ℃，維持1分鐘，再以每分鐘20℃升溫至240℃，維持1分鐘。而注射部及檢測器的溫度分別設在250 ℃和 280 ℃。在基本分析方法，acetaminophen的檢量線範圍為100 - 2000 μg/mL，其偵測極限為2 μg/mL (S/N=3)；caffeine的檢量線範圍為50 - 1000 μg/mL，其偵測極限為5 μg/mL (S/N=3)。
利用本方法應用在市售感冒糖漿製劑及飲品的測定證實是可行的。Acetaminophen及caffeine在同日內分析 (n = 3) 其相對標準偏差 (R.S.D.) 皆小於0.8 %，而異日間分析 (n = 5) 的相對標準偏差分別小於0.9 % 及1.1 %，其萃取相對回收率在96.4 % ~ 102.2 %，說明本法具有良好之定量可靠性，可成功地應用在感冒糖漿和飲品中acetaminophen及caffeine之定量測定及品質管制。

A simple and rapid method for simultaneous analysis of acetaminophen and caffeine in commercial cold syrups and drinks is described. The method was based on direct gas chromatography with flame ionization detector. The samples were simply pretreated by liquid-liquid extraction and then analyzed by GC with a fused-silica VF-5ms capillary column (30 m × 0.32 mm, 0.25 μm). The temperatures of injector and detector were set at 250℃ and 280℃, respectively. The column oven was maintained at 180℃ for 1 min, ramped at a rate of 20℃/min to 240℃ and held for 1 min.
Several parameters affecting the extraction of acetaminophen and caffeine were investigated. The linear range of the method for the determination of acetaminophen and caffeine were 100 – 2000 μg/mL and 50 – 1000 μg/mL, respectively. The instrument detection limits (signal to noise ratio of 3) of acetaminophen and caffeine were 2.0 ?nand 5.0 μg/mL, respectively. Results of application of the method for the analysis of acetaminophen and caffeine in commercial syrup preparations and drinks were satisfactory. The intra-day relative standard deviation (n=3) for acetaminophen and caffeine were both less than 0.8 %. The inter-day relative standard deviation (n=5) for acetaminophen and caffeine were less than 0.9 % and 1.1 %, respectively. The assay should be useful for quality control of acetaminophen and caffeine in pharmaceutical syrup preparations and drinks.

目錄

中文摘要-------------------------------------------------i
英文摘要-------------------------------------------------ii
圖目錄---------------------------------------------------v
表目錄---------------------------------------------------vii
第一節：緒論---------------------------------------------1
第二節：實驗部份
壹、試藥及材料---------------------------------------16
貳、儀器---------------------------------------------17
參、試藥溶液之配製
(一) Acetaminophen標準溶液-----------------------18
(二) Caffeine標準溶液----------------------------19
(三) 內部標準品bibenzyl溶液----------------------19
肆、氣相層析之分析條件-------------------------------20
伍、基本分析條件之探討
(一) Acetaminophen和caffeine層析位置確認---------21
(二) 內部標準品之選擇----------------------------21
(三) 檢量線之製定法------------------------------22
(四) 精密度與準確度之探討------------------------24
(五) 滯留時間穩定度及安定性之探討----------------25
陸、感冒糖漿劑和飲品之應用分析
(一) 檢品前處理之探討----------------------------26
(二) 感冒糖漿劑及飲品之含量分析------------------28
(三) 感冒糖漿劑和飲品中添加回收率之探討----------30
第三節：結果與討論
壹、基本分析法之探討
(一) Acetaminophen和caffeine層析位置探討---------32
(二) 內部標準品之選擇----------------------------33
(三) Acetaminophen和caffeine之GC-FID層析圖-------36
(四) Acetaminophen和caffeine檢量線之建立---------37
(五) 精密度與準確度之測定------------------------40
(六) 滯留時間穩定度及安定性之探討----------------42
貳、感冒糖漿劑及飲品之應用分析
(一) 檢品前處理方法之選定--------------------44
(二) 感冒糖漿劑之GC-FID層析圖----------------50
(三) 飲品之GC-FID層析圖----------------------51
(四) 市售感冒糖漿與飲品之含量分析------------52
(五) 感冒糖漿劑和飲品中添加回收率之探討------55
第四節：結論---------------------------------------------57
第五節：參考文獻-----------------------------------------59

圖目錄
圖一：Structure of acetaminophen-------------------------1
圖二：Cyclooxygenase pathway ----------------------------3
圖三：Metabolism of acetaminophen------------------------5
圖四：Structures of caffeine-----------------------------7
圖五：Metabolism of caffeine in the liver----------------8
圖六：Acetaminophen和caffeine標準水溶液分析步驟圖--------22
圖七： Acetaminophen和caffeine標準氰甲烷溶液之氣相層析圖-32
圖八：Gas chromatograms for the determination of 4 different internal standard------------------------------35
圖九：Gas chromatograms for the determination of acetaminophen and caffeine in blank (A) and standard (B)-36
圖十：Calibration curves of acetaminophen standard in intra-day (A) and inter-day (B) analysis-----------------------38
圖十一：Calibration curves of caffeine standard in intra-day (A) and inter-day (B) analysis-----------------------39
圖十二：The stability of the retention time of acetaminophen, caffeine and bibenzyl standard------------43
圖十三：The stability of acetaminophen and caffeine standard-------------------------------------------------43
圖十四：Effects of the solvents on the extraction of the acetaminophen and caffeine from syrup sample solutions (A) and drinks (B)-------------------------------------------45
圖十五：Effects of the methods on the extraction of the acetaminophen and caffeine from syrup sample solutions (A) and drinks (B)-------------------------------------------46
圖十六：Effects of the temperature on the extraction of the
acetaminophen and caffeine from syrup sample solutions (A)
and drinks (B)-------------------------------------------47
圖十七：Effects of the time on the extraction of the acetaminophen and caffeine from syrup sample solutions (A) and drinks (B)-------------------------------------------48
圖十八：糖漿劑和飲品之分析步驟---------------------------49
圖十九：Gas chromatograms for the determination of acetaminophen and caffeine in syrup sample---------------50
圖二十：Gas chromatograms for the determination of caffeine in drinks------------------------------------------------51

表目錄
表一： 感冒糖漿劑和飲品之acetaminophen和caffeine標示含量-29
表二：Retention times of internal standard---------------34
表三：Precision and accuracy for the analysis of acetaminophen standard-----------------------------------41
表四：Precision and accuracy for the analysis of caffeine standard-------------------------------------------------41
表五：感冒糖漿劑中acetaminophen之含量分析結果------------52
表六：感冒糖漿劑中caffeine之含量分析結果-----------------53
表七：飲品中caffeine之含量分析結果-----------------------54
表八：感冒糖漿中添加acetaminophen和caffeine之
回收率分析結果-------------------------------------------56
表九：飲品中添加caffeine之回收率分析結果-----------------56

第五節：參考文獻

1. Anthony S. Paracetamol. Emerg. Nurse. 2007, 15 (3): 24-5.

2. Brodie B. B., Axelrod J. The fate of acetanilide in man. J. Pharmacol. Exp. Ther. 1948, 94 (1): 29–38.

3.http://www.chemheritage.org/EducationalServices/pharm/asp/asp08.htm A Festival of Analgesics. 2007.

4. http://vascular.free.fr/cascadaa.htm

5. Kis B., Snipes J. A., Busija D.W. Acetaminophen and the cyclooxygenase-3 puzzle: sorting out facts, fictions, and uncertainties. J. Pharmacol. Exp. Ther. 2005, 315 (1): 1–7.

6. Aronoff D. M., Oates J. A., Boutaud O. New insights into the mechanism of action of acetaminophen: Its clinical pharmacologic characteristics reflect its inhibition of the two prostaglandin H2 synthases. Clin. Pharmacol. Ther. 2006, 79 (1): 9–19.

7. Bertolini A., Ferrari A., Ottani A., Guerzoni S., Tacchi R., Leone S. Paracetamol: new vistas of an old drug. CNS Drug Rev. 2006, 12 (3-4): 250–75.

8. 吳玉琴. Acetaminophen於特殊病人群在建議劑量下引起肝毒性反應. 成醫藥誌. 2004, 14(5): 4-7.

9. Dart R. C., Erdman A. R., Olson K. R., Christianson G., Manoguerra A. S., Chyka P. A., Caravati E. M., Wax P. M., Keyes D. C., Woolf A. D., Scharman E. J., Booze L. L., Troutman W. G. American Association of Poison Control Centers. Acetaminophen poisoning: an evidence-based consensus guideline for out-of-hospital management. Clin. Toxicol. (Phila). 2006, 44 (1): 1–18.

10. Dong H., Haining R. L., Thummel K. E., Rettie A. E., Nelson S. D. Involvement of human cytochrome P450 2D6 in the bioactivation of acetaminophen. Drug Metab. Dispos. 2000, 28 (12): 1397–400.

11.http://upload.wikimedia.org/wikipedia/en/3/34/Paracetamol_metabolism.png

12. James A. N., Caffeine and related methylxanthines: possible naturally occurring pesticides. Science. 1984, 226 (4671): 184-7.

13. Zou J., Li N., Simple and environmental friendly procedure for the gas chromatographic-mass spectrometric determination of caffeine in beverages. J. Chromatogr. A. 2006, 1136 (1):106-10.

14. Bennett A. W., Bonnie K. B., The World of Caffeine. Routledge. 2001.

15. Fisone G., Borgkvist A., Usiello A., Caffeine as a psychomotor stimulant: mechanism of action. Cell Mol Life Sci. 2004, 61 (7–8): 857-72.

16. Grosso L. M., Bracken M. B., Caffeine Metabolism, Genetics, and Perinatal Outcomes: A Review of Exposure Assessment Considerations during Pregnancy. Ann. Epidemiol. 2005, 15 (6): 460-466.

17. Benowitz N. L., Peng M., Jacob P., Effects of cigarette smoking and carbon monoxide on chlorzoxazone and caffeine metabolism. Clin. Pharmacol. Ther. 2003, 74 (5): 468-474.

18. Diner H. C., Pfaffenrath V., Pageler L., Peil H., Aicher B., The fixed combination of acetylsalicylic acid, paracetamol and caffeine is more effective than single substances and dual combination for the treatment of headache: a multicentre, randomized, double-blind, single-dose, placebo-controlled parallel group study. Cephalalgia. 2005, 25: 776–787.

19. http://food.doh.gov.tw/Chinese/news_1.asp?idCategory=&idx=1710
行政院衛生署食品衛生處食品資訊網公告 (中華民國96年7月6日)

20. Smith A., Effect of caffeine on human behavior. Food Chem. Toxicol. 2002, 40: 1243–1255.

21. Kerrigan S., Lindsey T., Fatal caffeine overdose: Two case reports. Forensic Sci. Int. 2005, 153: 67–69.

22. Holmgren P., Norde´n-Pettersson L., Ahlner J., Forensic Sci. Int. 2004, 139: 71–73.

23. McCusker R. R., Goldberger B, A., Edward J. C., Caffeine content of specialty coffees. J. Anal. Toxicol. 2003, 27(7): 520-2.

24. McCusker R. R., Goldberger B, A., Edward J. C., Caffeine content of energy drinks, carbonated sodas, and other beverages. J. Anal. Toxicol. 2006, 30(2):112-4.

25. Rice S. L., Mitra S., Microwave-assisted solvent extraction of solid matrices and subsequent detection of pharmaceuticals and personal care products (PPCPs) using gas chromatography–mass spectrometry. Anal. Chim. Acta. 2007, 589: 125–132.

26. Lee H. F., Cheng C. C., Chou S. S., Determination and Investigation of Caffeine Content in Beverage. Ann. Rept. NLFD Taiwan R.O.C. 1993, 11: 257-264.

27. Wu H. Y., Cheng Y. N., Chang M. L., Lin Y. P., Quantitation of Active Ingredients in Cold Syrup-Collaborative Study. J. Food Drug Anal. 1994, 2(1):63-69.

28. Ku Y. R., Wen K. C., Ho L. K., Chang Y. S., Solid-phase extraction for the determination of caffeine in traditional Chinese medicinal prescriptions containing Theae folium by high performance liquid chromatography. J. Pharm. Biomed. Anal. 1999, 20: 351-356.

29. Martin M. J., Pablos F., Gonzalez A. G., Simultaneous determination of caffeine and non-steroidal anti-inflammatory drugs in pharmaceutical formulations and blood plasma by reversed-phase HPLC from linear gradient elution. Talanta. 1999, 49: 453–459.

30. Barbas C., Garcia A., Saavedra L., Castro M., Optimization and validation of a method for the determination of caffeine, 8-chlorotheophylline and diphenhydramine by isocratic high-performance liquid chromatography Stress test for stability evaluation. J. Chromatogr. A. 2000, 870: 97–103.

31. Chen Z., Pavelic P., Dillon P., Naidu R., Determination of caffeine as a tracer of sewage effluent in natural waters by on-line solid-phase extraction and liquid chromatography with diode-array detection. Water Res. 2002, 36: 4830–4838.

32. Horie H., Nesumi A., Ujihara T., Kohata K., Rapid determination of caffeine in tea leaves. J. Chromatogr. A. 2002, 942: 271–273.

33. Gardinali P. R., Zhaob X., Trace determination of caffeine in surface water samples by liquid chromatography–atmospheric pressure chemical ionization–mass spectrometry (LC–APCI–MS). Environ. Int. 2002, 28: 521– 528.

34. Minamisawa M., Yoshida S., Takai N., Determination of biologically active substances in roasted coffees using a diode-array HPLC system. Anal. Sci. 2004, 20: 325-328.

35. Jacob P., Haller C. A., Duan M., Yu L., Peng M., Benowitz N. L., Determination of Ephedra Alkaloid and Caffeine Concentrations in Dietary Supplements and Biological Fluids. J. Anal. Toxicol. 2004, 28: 152-159.


36. Pistos C., Stewart J. T., Assay for the simultaneous determination of acetaminophen–caffeine–butalbital in human serum using amonolithic column. J. Pharm. Biomed. Anal. 2004, 36: 737–741.

37. Tzanavaras P. D., Themelis D. G., Development and validation of a high-throughput high-performance liquid chromatographic assay for the determination of caffeine in food samples using a monolithic column. Anal. Chim. Acta. 2007, 581: 89–94.

38. Walker J. C., Zaugg S. E., Walker E. B., Analysis of beverages by capillary electrophoresis. J. Chromatogr. A. 1997, 781: 481-485.

39. Okamotoa H., Nakajimaa T., Itoa Y., Aketoa T., Shimadab K., Yamato S., Simultaneous determination of ingredients in a cold medicine by cyclodextrin-modified microemulsion electrokinetic chromatography. J. Pharm. Biomed. Anal. 2005, 37: 517–528.

40. Emre D., Ozaltın N., Simultaneous determination of paracetamol, caffeine and propyphenazone in ternary mixtures by micellar electrokinetic capillary chromatography. J. Chromatogr. B. 2007, 847: 126–132.

41.http://en.wikipedia.org/wiki/Image:Caffeine_metabolites.svg