臺灣博碩士論文加值系統

Prednisolone分析方法之建立及運動對prednisolone 藥物動力學之影響

摘 要

本實驗建立高效能液相層析法分析血漿及尿液中prednisolone(去氫皮質醇)的濃度，以RP-18e管柱在波長243nm的二極體陣列下偵測，此方法以betamethasone作為內標準品，血漿之檢量線範圍為15ng/mL – 250ng/mL；尿液之檢量線範圍為25ng/mL – 500ng/mL。此分析方法，血漿的回收率約69-74%，尿液的回收率約62-90%。
本研究給予八位健康受試者口服5mg的prednisolone後，分別進行不運動及次最大強度力竭性運動兩階段實驗，收取服藥後血液及尿液，測定prednisolone濃度以進行藥物動力學的分析。
在血液部份，血液中prednisolone的最高濃度(Cmax)運動組顯著低於不運動組(p<0.05)，而濃度-時間曲線下面積(AUC)、半衰期(t1/2)、清除率(CL)、擬分布體積(Vd)等兩組間無顯著差異；在尿液部份，排除速率常數(k)、排除半衰期(t1/2) 兩組間皆無顯著差異。結果顯示，運動介入並不會影響prednisolone的藥物動力學。

關鍵字：運動、藥物動力學、去氫皮質醇

Analytical Method Development for Prednisolone and Effect of Exercise on Prednisolone Pharmacokinetics

ABSTRACT

We describe a method for measuring concentrations of prednisolone in plasma and urine by high performance liquid chromatography. It was performed on a RP-18e column with diode array detection at 243nm. Betamethasone was uesd as the internal standard. The calibration curve is linear within the studied range from 15ng/mL – 250ng/mL in plasma and 25ng/mL – 500ng/mL in urine. The analytical recoveries are about 62-90% in urine and 69-74% in plasma.
In this study, eight healthy volunteers received 5mg prednisolone before a submaximal exhaustive exercise or a non-exercise session. Both plasma and urine drug concentrations for pharmacokinetic calculation were estimated.
In plasma samples peak plasma concentration of exercise group was significantly lower than non-exercise group (p<0.05). However, the AUC, t1/2 , CL, Vd were not significantly different among two groups. In urine samples, k and t1/2 were not significantly different among exercise and non-exercise group. We concluded that the pharmacokinetics of prednisolone did not affect by exercise.


Key Word：Exercise, Pharmacokinetics, Prednisolone

目 錄

第壹章 緒論

第一節 研究動機--------------------------------------------- 1
第二節 研究目的--------------------------------------------- 2

第貳章 文獻探討

第一節 Prednisolone簡介----------------------------------- 3
第二節 尿液及血液中prednisolone的分析方法---- 6
第三節 Prednisolone的藥物動力學性質------------- 7
第四節 運動對藥物動力學的影響---------------------- 10

第叁章 尿液中Prednisolone分析方法之建立

第一節 材料與方法---------------------------------- 14
第二節 結果與討論---------------------------------- 18
第三節 結論------------------------------------------ 25

第肆章 血液中Prednisolone分析方法之建立

第一節 材料與方法---------------------------------- 26
第二節 結果與討論---------------------------------- 31
第三節 結論------------------------------------------ 36

第伍章 運動對Prednisolone藥物動力學的影響
第一節 材料與方法--------------------------------- 37
第二節 數據分析及統計方法------------------------------ 43
第三節 結果與討論----------------------------------------- 45
第四節 結論與建議----------------------------------------- 55

參考文獻 56

附錄
一 受試者基本資料及最大攝氧量-------------------- 65
二-a 不運動組口服prednisolone 後之血中濃度- 66
二-b 運動組口服prednisolone後之血中濃度---- 67
三-1-a 編號1之受試者口服prednisolone後血中濃度曲線圖---------------------------------- 68
三-1-b 編號1之受試者prednisolone血中原始濃度及一室模式估計濃度曲線圖------------------------- 68
三-2-a 編號2之受試者口服prednisolone後血中濃度曲線圖---------------------------------- 69
三-2-b 編號2之受試者prednisolone血中原始濃度及一室模式估計濃度曲線圖-------------- 69
三-3-a 編號3之受試者口服prednisolone後血中濃度曲線圖------------------------------------------------------------------------ 70
三-3-b 編號3之受試者prednisolone血中原始濃度及一室模式估計濃度曲線圖-------------- 70
三-4-a 編號4之受試者口服prednisolone後血中濃度曲線圖----------------------------------mg的Prednisolone後之血中濃度曲線圖 71
三-4-b 編號4之受試者prednisolone血中原始濃度及一室模式估計濃度曲線圖-------------- 71
三-5-a 編號5之受試者口服prednisolone後血中濃度曲線圖---------------------------------- 72
三-5-b 編號5之受試者prednisolone血中原始濃度及一室模式估計濃度曲線圖-------------- 72
三-6-a 編號6之受試者口服prednisolone後血中濃度曲線圖---------------------------------- 73
三-6-b 編號6之受試者prednisolone血中原始濃度及一室模式估計濃度曲線圖-------------- 73
三-7-a 編號7之受試者口prednisolone後之血中濃度曲線圖---------------------------------- 74
三-7-b 編號7之受試者prednisolone血中原始濃度及一室模式估計濃度曲線圖-------------- 74
三-8-a 編號8之受試者口服prednisolone後血中濃度曲線圖----------------------------------------------------------------------- 75
三-8-b 編號8之受試者prednisolone血中原始濃度及一室模式估計濃度曲線圖------------- 75
四-a 不運動組口服prednisolone 後血中cortisol濃度--------------------------------- 76
四-b 運動組口服prednisolone 後血中cortisol濃度------------------------------------------- 77
五-a 不運動組口服prednisolone後，以非室體模式分析所計算出之藥物動力學參數------ 78
五-b 運動組口服prednisolone後，以非室體模式分析所計算出之藥物動力學參數--------- 78
五-c 不運動組口服pednisolone後，以一室模式分析所計算出之藥物動力學參數--------- 79
五-d 運動組口服pednisolone後，以一室模式分析所計算出之藥物動力學參數----------- 79
六-a 不運動組口服pednisolone後24小時內的尿液體積------------------------------------- 80
六-b 運動組口服pednisolone後24小時內的尿液體積------------------------------------- 80
七-a 不運動組口服pednisolone後24小時內的尿液濃度------------------------------------- 81
七-b 運動組口服pednisolone後24小時內的尿液濃度------------------------------------- 81
八-a 不運動組口服pednisolone後24小時內尿液中pednisolone的排泄量----------------- 82
八- b 運動組口服pednisolone後24小時內尿液中pednisolone的排泄量------------------ 82
九-a 不運動組口服prednisolone後尿液的藥物動力學參數----------------------------------- 83
九-b 運動組口服prednisolone後尿液的藥物動力學參數---------------------------------- 83
十 受試者須知---------------------------------- 84
十一 受試者同意書------------------------------- 85

表 目 錄

表 2-1 Prednisolone在尿液中的代謝物--------------- 8
表 2-2 在安靜、低強度、中等強度、高強度下血流的分佈------------------------------------------ 10
表 2-3 單次運動和連續運動介入對藥物動力學的------
影響----------------------------------------------------- 13
表 3-1 尿液分析方法之同日間精密度與準確度與回收率-------------------------------------------- 23
表 3-2 尿液分析方法之回收率----------------------- 23
表 4-1 梯度流析表(時間vs流速變化)---------------- 28
表 4-2 血液分析方法之同日間精密度、準確度與回收率--------------------------------------------- 34
表 4-3 血液分析方法之回收率--------------------- 35
表 5-1 受試者的基本資料及運動測試結果----------- 45
表 5-2 Prednisolone於一室及二室模式配適評估值-- 47
表 5-3 非室體模式及一室模式分析法計算所得之藥
物動力學參數---------------------------------- 48
表 5-4 尿液中Prednisolone的藥物動力學參數------ 54

圖 目 錄

圖 2-1 糖皮質類固醇的結構-------------------------- 5
圖 3-1 Prednisolone 波長200-300nm之UV吸收----------
光譜圖------------------------------------------ 18
圖 3-2 標準品於溶液中的層析圖(C8管柱)-------------------------- 19
圖 3-3 標準品於溶液中的層析圖(C18管柱)---------- 20
圖 3-4 空白尿液之層析圖----------------------------- 21
圖 3-5 尿液中含標準品層析圖----------------------- 21
圖 3-6 尿液中prednisolone 濃度的檢量線----------- 22
圖 4-1 血漿中含prednisolone、crtiol、betamethasone
之層析圖譜--------------------------------------------- 32
圖 4-2 血漿中prednisolone 濃度的檢量線----------- 33
圖 5-1 口服prednisolone之平均血漿濃度圖--------- 49
圖 5-2 不運動組和運動組cortisol的濃度變化------- 50
圖 5-3 尿液中prednisolone的排除累積量----------- 53

參考文獻

一、中文部分

方嘉佑(民91)。藥品分析。台北市﹕合記書局，44-45頁。
許美智(民89)。運動與禁藥。台北市﹕合記書局，38頁。
林敬二、楊美惠、楊寶旺、廖德章、薛敬和 (民82)。英中日化學大辭典。台北市﹕高立圖書有限公司，1250頁。
鄭瑞棠、陳文旭(民80)。藥理學。台北市：華杏出版社，853-868頁。


二、外文部分
Angeli, A., Frajria, R., De Paoli, R., Fonzo, D., & Ceresa, F. (1978). Diurnal variation of prednisolone binding to serum corticosteroid-binding globulin in man. Clinical Pharmacology and Therapeutics, 23(1), 47-53.
Arends, B. G., Bohm, R. O., van Kemenade, J. E., Rahn, K. H., & van Baak, M. A. (1986). Influence of physical exercise on the pharmacokinetics of propranolol. European Journal of Clinical Pharmacology, 31(3), 375-377.
Aslaksen, A., & Aanderud, L. (1980). Drug absorption during physical exercise. British Journal of Clinical Pharmacology, 10(4), 383-385.
Boudinot, F. D.,D’Ambrosio, R., & Jusko, W. J. (1986). Receptor-mediated pharmacodynamics of prednisolonein rat. Journal of Pharmacokinetics and Biopharmaceutics, 14, 469-493.
Boudinot, F. D., & Jusko, W. J. (1984). Fulid shifts and other factors affecting plasma protein binding of prednisolone by eequilibrium dialysis. Journal of Pharmaceutical Sciences, 73,774.
Brooks, P. M., Buchanan, W.W., Grove, M., & Downie, W. W. (1976). Effects of enzyme induction on metabolism of prednisolone. Clinical and laboratory study. Annals of the Rheumatic Diseases, 35(4), 339-343.
Brouns, F., Saris, W. H., & Rehrer, N. J. (1987). Abdominal complaints and gastrointestinal function during long-lasting exercise. International Journal of Sports Medicine, 8(3), 175-89.
Caldarella, A. M., Reardon, G. E, & Canalis, E. (1982). Analysis for cortisol in serum by liquid chromatography. Clinical Chemistry, 28(3), 538-43.
Causon, R. (1997). Validation of chromatographic methods in biomedical analysis viewpoint and discussion. Journal of chromatography. B, Biomedical applications,689, 175-180.
Cheng, M. H., Huang, W. Y., & Lipsey, A. I. (1988). Simultaneous liquid-chromatographic determination of prednisone and prednisolone in plasma. Clinical Chemistry, 34(9), 1987-1899
Costil, D. L. (1977). Sweating: its composition and effects on body fluids. Annals of the New York Academy of Sciences, 301, 160-174.
Deventer, K., & Delbeke, F. K. (2003). Validation of screening method for corticosteroids in doping analysis by liquid chromatography/tandem mass spectrometry. Rapid Cmmunication in Mass Spectrometry, 17, 2107-2114.
DiPiro, J. Y., Spruill, W. J., Blouin, R. A. & Pruemer, J. M. (2002). Concepts in Clincal Pharmacokinetics. (3rd)., Bethesda(MD), American Society of Health Systems Pharmacists
Dossing M. (1985) Effect of acute and chronic exercise on hepatic drug metabolism. Clincal Pharmacokinetics, 10(5), 426-431.
Frey, F. J. (1987). Kinetics and dynamics of prednisolone. Endocrine Reviews, 8(4), 453-473
Frey, B. M., & Frey, F. J. (1990). Clinical pharmacokinetics of prednisone and prednisolone. Clinical Pharmacokinetics, 19(2),126-146.
Frey, F. J., & Frey, B. M. (1983). Urinary 6 beta-hydroxyprednisolone excretion indicates enhanced prednisolone catabolism. The Journal of Laboratory and Clinical Medicine, 101(4), 593-604.
Frey, F. J., Frey, B. M., & Benet, L. Z. (1979). Liquid-chromatographic measurement of endogenous and exogenous glucocorticoids in plasma. Clinical Chemistry, 25(11), 1944-1947.
Gabrielsson, J. & Weiner, D. (2000). Pharmacokinetic and pharmacodynamic data analysis：concepts & applications. (3rd), Apotekarsocieten Inc.
Gambertoglio, J. G., Romac, D. R., Yong, C. L., Birnbaum, J., Lizak, P., & Amend, W. J. Jr. (1987). Lack of effect of sucralfate on prednisone bioavailability. The American Journal of Gastroenterology, 82(1), 42-45.
Gambertoglio, J. G., Amend, W. J. Jr, & Benet, L. Z., (1980). Pharmacokinetics and bioavailability of prednisone and prednisolone in healthy volunteers and patients: a review. Journal of Pharmacokinetics and Biopharmaceutics, 8(1), 1-52.
Grahame-smith, D. G. & Aronson, J. K. (1992). Oxford textbook of clinical pharmacology and drug therapy. Oxford, 584-585
Grille, W., Welter, U., Johnsen, K., Kolenda, K. D., & Loffler, H. (1996). Effect of physical activity on serum concentrations of digoxin and digitoxin. European Journal of Clinical Pharmacology, 50(3), 237-239.
Henry, J. A., Iliopoulou, A., Kaye, C. M., Sankey, M. G., & Turner, P. (1981). Changes in plasma concentrations of acebutolol, propranolol and indomethacin during physical exercise. Life Sciences, 28(17), 1925-1929.
Hirata, H., Kasama, T., Sawai ,Y., & Fike, R. R. (1994). Simultaneous determination of deflazacort metabolites II and III, cortisol, cortisone, prednisolone and prednisone in human serum. Journal of chromatography. B, Biomedical applications, 658(1), 55-61.
Hurwitz, G. A., Webb, J. G., Walle, T., Bai, S. A., Daniell, H. B., Gourley, L, Boyd, Loadholt, C., & Gaffney, T. E. (1983). Exercise-induced increments in plasma levels of propranolol and noradrenaline. British Journal of Clinical Pharmacology, 16(6), 599-608
Joreteg, T., & Jogestrand, T. (1984). Physical exercise and binding of digoxin to skeletal muscle--effect of muscle activation frequency. European journal of clinical pharmacology, 27(5), 567-570.
Jusko, W. J., Pyszczynski, N. A., Bushway, M. S., D'Ambrosio, R., & Mis, S. M. (1994). Fifteen years of operation of a high-performance liquid chromatographic assay for prednisolone, cortisol and prednisone in plasma. Journal of chromatography. B, Biomedical applications, 658(1), 47-54.
Kabra, P. M., Tsai, L. L., & Marton, L. J. (1979). Improved liquid-chromatographic method for determination of serum cortisol. Clinical Chemistry, 25(7), 1293-1296.
Kamimori, G. H., Smallridge, R. C., Redmond, D. P., Belenky, G. L., & Fein, H. G. (1990). The effect of exercise on atropine pharmacokinetics. European Journal of Clinical Pharmacology, 39(4), 395-397.
Keller, F., Maiga, M., Neumayer, H. H., Lode, H., & Distler, A. (1984). Pharmacokinetic effects of altered plasma protein binding of drugs in renal disease. European Journal of Drug Metabolism and Pharmacokinetics, 9(3), 275-282.
Khazaeinia, T., Ramsey, A. A., & Tam, Y. K. (2000). The effects of exercise on the pharmacokinetics of drugs. Journal of Pharmacy & Pharmaceutical Sciences, 3(3), 292-302.
Koivisto, V. A., & Felig, P. (1978). Effects of leg exercise on insulin absorption in diabetic patients. The New England Journal of Medicine, 298(2), 79-83.
Lenz, T. L., Lenz, N. J., & Faulkner, M. A. (2004). Potential Interactions between Exercise and Drug Therapy. Sports Medicine, 34(5), 293-306.
Majid, O., Akhlaghi, F., Lee, T., Holt, D. W., & Trull, A. (2001). Simultaneous determination of plasma prednisolone, prednisone, and cortisol levels by high-performance liquid chromatography. Therapeutic Drug Monitoring, 23(2), 163-168.
Marsh, T. D., Garnett, W. R., Poynor, W. J., & Pellock, J. M. (1983). Effects of exercise on valproic acid pharmacokinetics. Clinical Pharmacy, 2(1), 62-64.
Mason, W. D., Kochak, G., Winer, N., & Cohen, I. (1980). Effect of exercise on renal clearance of atenolol. Journal of Pharmaceutical Sciences, 69(3), 344-345.
McAllister, W. A., Winfield, C. & R., & Collins, J. V. (1981). Pharmacokinetics of prednisolone in normal and asthmatic subjects in relation to dose. European Journal of Clinical Pharmacology, 20(2), 141-145.
McArdle, W. D., Katch, F. I., & Katch, V. L. (1991). Exercise Physiology. 3rd ed. Philadelphia (PA), Lea & Febiger, 17
Mejer, L. E., & Blanchard, R. C. (1973). Fluorometric determination of plasma 11-hydroxycorticosteroids. I. Rapid procedure for clinical screening. Clinical Chemistry, 19(7), 710-717.
Mooy, J., Arends, B., van Kemenade, J., Boehm, R., Rahn, K.H., & van Baak, M. (1986). Influence of prolonged submaximal exercise on the pharmacokinetics of verapamil in humans. Journal of Cardiovascular Pharmacology, 8(5),940-942.
Park, S. J., Kim, Y. J., Pyo, H. S., Park, J. (1990). Analysis of corticosteroids in urine by hplc and thermospray LC/MS. J
Petereit, L. B., & Meikle, A. W., (1977). Effectiveness of prednisolone during phenytoin therapy. Clinical Pharmacology and Therapeutics, 22(6), 912-916.
Renner, E., Horber, F. F., Jost, G., Frey, B. M., & Frey, F. J. (1986). Effect of liver function on the metabolism of prednisone and prednisolone in humans. Gastroenterology, 90(4), 819-828.
Ryan, A. J, Chang, R.T., & Gisolfi, CV. (1996) Gastrointestinal permeability following aspirin intake and prolonged running. Medicine and Science in Sports and Exercise, 28(6), 698-705.
Sawrymowicz, M. (1997). The effect of exercise on the pharmacokinetics of acetaminophen and acetylsalicylic acid. Annales Academiae Medicae Stetinensis, 43, 57-66.
Schimmer, B. P., & Parker, K. L., (1995). Goodman&Gilman’s The Pharmacological Basis of Therapeutics, 9th ed., pp. 1465-1481 McGraw-Hill, New York.
Schlaeffer, F., Engelberg, I., Kaplanski, J., & Danon, A. (1984). Effect of exercise and environmental heat on theophylline kinetics. Respiration, 45(4), 438-442.
Shargel, L. Yu, A. B. C. (1999). Applied Biopharmaceutics & Pharmacokinetics. (4th ed.). Prentice Hall International, Inc. U.S.A. pp. 607-639.
Spector, A. A., & Santos, E. C. (1973). Influence of free fatty acid concentration on drug binding to plasma albumin. Annals of the New York Academy of Science, 226, 247-258
Stromberg, C., Vanakoski, J., Olkkola, K. T, Lindqvist, A., Seppala, T., & Laitinen, L. A. (1992). Exercise alters the pharmacokinetics of midazolam. Clinical Pharmacology and Therapeutics, 51(5), 527-32.
Stoschitzky, K., Zweiker, R., Klein, W., Zitta, S., Gruber, L. Lamprecht, G., & Lindner, W. (2002). Unpredicted lack of effect of exercise on plasma concentrations of carvedilol. Journal of Cardiovascular Pharmacology, 39(1), 58-60.
Suzuki, M., Sudoh, M., Matsubara, S., Kawakami, K., Shiota, M., & Ikawa, S. (1996). Changes in renal blood flow measured by radionuclide angiography following exhausting exercise in humans. European Journal of Applied Physiology and Occupational Physiology, 74(1-2), 1-7.
Uribe, M., Schalm, S. W., Summerskill, W. H., & Go, V. L. (1976). Effect of liquid diet on serum protein binding and prednisolone concentrations after oral prednisone. Gastroenterology, 71(2), 362-364.
Varis, T., Kivisto, K. T., & Neuvonen, P. J. (2000). The effect of itraconazole on the pharmacokinetics and pharmacodynamics of oral prednisolone. European Journal of Clinical Pharmacology, 56(1), 57-60.
van Baak, M. A. (1990). Influence of exercise on the pharmacokinetics of drugs. Clinical Pharmacokinetics, 19(1), 32-43.
van Baak, M. A, Mooij, J. M., & Schiffers, P. M. (1992). Exercise and the pharmacokinetics of propranolol, verapamil and atenolol. European Journal of Clinical Pharmacology, 43(5), 547-550.
Vermeulen, A. (1959). The metabolism of 4-C14 prednisolone. Journal of Endocrinology, 18, 278
Volin, P. (1995). Simple and specific reversed-phase liquid
chromatographic method with diode-array detection for simultaneous determination of serum hydroxychloroquine, chloroquine and some corticosteroids. Journal of chromatography. B, Biomedical applications, 666(2), 347-353.
WADA. (2004). International Standard for Laboratories, Version 3.0
WADA. (2004). The 2004 prohibited list international standard.
Ylitalo, P., & Hinkka, H. (1985). Effect of exercise on plasma levels and urinary excretion of sulphadimidine and procainamide. International Journal of Clinical Pharmacology Therapy and Toxicology, 23(10), 548-553.
Ylitalo, P., Hinkka, H., & Neuvonen, P. J. (1977). Effect of exercise on the serum level and urinary excretion of tetracycline, doxycycline and sulphamethizole. European Journal of Clinical Pharmacology, 12(5), 367-73.