利用衍生化反應搭配毛細管液相層析儀及紫外光偵測器偵測傳明酸

Determination of tranexamic acid coupled with chemical derivatization by capillary liquid chromatography with ultraviolet detection

廖芳儀(Fang-Yi Liao)

指導教授 ： 馮嘉嫻

高雄醫學大學/藥學院/香粧品學系碩士班/碩士(2013年)

摘要

傳明酸是賴胺酸的合成衍生物，藉由其相似賴胺酸的結構可以抑制纖維蛋白溶解酶原形成纖維蛋白溶解酶，進而減少酪胺酸酶活性和黑色素生成。除了美白用途之外，傳明酸在醫療上為抗纖維蛋白溶解劑，應用於出血性的情況和治療女性月經失血過多。
在傳明酸的分析方法中，因為其結構上缺少發光基團，常會使用衍生試劑來增加其偵測感度。本實驗利用dansyl chloride衍生傳明酸，與過去傳明酸文獻所使用的衍生試劑相比具有使用樣品量少和衍生物安定的優點，此外本實驗使用微波輔助反應進行來縮短反應時間，進而使操作更快速。為了延長分析管柱的使用壽命，本實驗利用分散液液微萃取法來萃取過量的衍生試劑，僅使用4 μL三氯甲烷進行萃取即完整移除過量的衍生試劑，與傳統的液液萃取法相比具有使用的有機溶媒量少和減少廢液產生的優點。
本實驗使用微小化液相層析儀和紫外光偵測器進行偵測，此方法的線性範圍為0.1-50 μM，相關係數為0.999，偵測極限為0.03 μM，同日內和異日間的相對標準差分別小於0.83%和3.08%。本實驗的分析方法已成功應用於偵測化粧品、藥劑、皮膚角質層萃取液及細胞樣品中的傳明酸含量。

關鍵字

傳明酸 ； 衍生反應 ； 毛細管液相層析儀

並列摘要

Tranexamic acid is a synthetic derivative of lysine. It is similar to lysine in structure that can use for the inhibition of plasminogen to plasmin, thereby reducing tyrosinase activity and melanin production. In addition to cosmetics uses, tranexamic acid is a medical antifibrinolytic drug used in haemorrhagic conditions and women menstrual blood loses.
Because tranexamic acid lacks the chromophore, the derivatization steps are needed for determination of tranexamic acid at low concentration by UV detector. Compared with the previous literature, we chose dansyl chloride as the derivatizing reagent in this study, which had the advantages of the less sample used and stable derivatives. Furthermore we used microwave-assisted reaction to reduce the reaction time which made the operation more quickly. In order to prolong the life time of analytical column, we used dispersive liquid-liquid microextraction (DLLME) to extract excess derivatizing reagent by 4 μL chloroform. Compared with the traditional liquid-liquid extraction, the advantages of DLLME in this study were the less organic solvent used and reduced the waste generated.
In this study, we were utilizing capillary liquid chromatography coupling with ultraviolet detector to monitor the dansyl derivative. The dynamic range was from 0.1 to 50 μM with a correlation coefficient 0.999 and the detection limit was 0.03 μM. The precision of intra- and interday were below 0.83 and 3.08%, respectively. Eventually, we were successfully applied this method to determine the content of tranexamic acid in pharmaceuticals, cosmetic products, the stratum corneum extractant and cell samples.

並列關鍵字

Tranexamic acid ； Derivatization method ； Capillary liquid chromatography

參考文獻

[1] S. Okamoto, U. Okamoto, Amino-methyl-cyclohexane-carboxylic acid: AMCHA a new potent inhibitor of the fibrinolysis, Keio Journal of Medicine, 11 (1962) 105-115.

[2] B.K. Fiechtner, G.A. Nuttall, M.E. Johnson, Y. Dong, N. Sujirattanawimol, W.C. Oliver, Jr., R.S. Sarpal, L.J. Oyen, M.H. Ereth, Plasma tranexamic acid concentrations during cardiopulmonary bypass, Anesthesia and Analgesia, 92 (2001) 1131-1136.

[3] D.A. Fergusson, P.C. Hebert, C.D. Mazer, S. Fremes, C. MacAdams, J.M. Murkin, K. Teoh, P.C. Duke, R. Arellano, M.A. Blajchman, J.S. Bussieres, D. Cote, J. Karski, R. Martineau, J.A. Robblee, M. Rodger, G. Wells, J. Clinch, R. Pretorius, A comparison of aprotinin and lysine analogues in high-risk cardiac surgery, The New England Journal of Medicine, 358 (2008) 2319-2331.

[4] G.A. Nuttall, M.C. Gutierrez, J.D. Dewey, M.E. Johnson, L.J. Oyen, A.C. Hanson, W.C. Oliver, Jr., A preliminary study of a new tranexamic acid dosing schedule for cardiac surgery, Journal of Cardiothoracic and Vascular Anesthesia, 22 (2008) 230-235.

[5] T.Y. Hsia, T.C. McQuinn, R. Mukherjee, R.L. Deardorff, J.E. Squires, R.E. Stroud, F.A. Crawford, S.M. Bradley, S.T. Reeves, F.G. Spinale, Effects of aprotinin or tranexamic acid on proteolytic/cytokine profiles in infants after cardiac surgery, The Annals of Thoracic Surgery, 89 (2010) 1843-1852.