應用高效液相色譜分析藥劑和皮膚樣品中的甲氨蝶呤   全文下載

Analysis of Methotrexate from Formulations and Skin Specimens by HPLC

本文介紹以高效液相色譜分析藥劑和透皮試驗後真皮及內皮中的甲氨蝶呤。採用MICRO BODAPAK C18, 20mm × 3.9mm 反相柱。藥量（範圍0.010至14.056 ug）和峰面積的標準曲線呈良好的直線關係。相關係數高達0.9999。用1 ug的甲氨蝶呤重覆分析七次，其峰面積為9627424+22111點（平均數+標準差；每點等於1／8毫伏特一秒，每毫伏特為2×10-6a.u.），其相對標準差為0.23%。甲氨蝶呤和葉酸的用量各為1 ug 時，其分離系數為33.54。此法可很好地測出凝膠和透皮試驗後真皮及內皮中的甲氨蝶呤。初步結果令人滿意，合乎第二十二版美國藥典的各項要求。本文提供新的皮膚分離方法（利用微波爐），皮膚抽取液的前處理，和在甲氨蝶呤的高效液相分析中保護柱的使用。"

A method for quantitative analysis of methotrexate (MTX) from formulated gels/creams and from tissue/solution samples after percutaneous permeation studies is described. High performance liquid chromatography (HPLC) with a MicroBondapak Cs, reverse-phase column was used. A calibration curve of doses versus peak areas for MTX showed linearity over the range of 0.010-14.056 µg with a very high correlation coefficient (r) of 0.99998. Replicate analysis of µg of MTX showed that the peak area was 9627424±22111 counts (mean ± SD; one count is 1/8 µV-sec, 1µV is 2 X 10-6a.u.) with a relative standard deviation of 0.23%. The resolution factor between MTX and folic acid was 33.35 when the applied doses of both were 1 µg each. Results demonstrated that the photolytic products of MTX, samples from gel and samples extracted from hairless guinea pig skin after percutaneous permeation and epidermis/dermis separation by microwave technique, could be well detected by this method. Preliminary results are satisfactory and meet the requirements of U.S. Pharmacopeia XXII(1). This report decribes novel approaches for determination of methotrexate content in the formulations and skin specimens. These approaches include the separation of epidermis and dermis by microwaving, the pretreatment procedures employed before analysis, and the use of guard column during HPLC analysis.