Martinavarro-Domínguez Adrià, Boseb Devasish, Durgbanshi Abhilasha, Gil-Agustí Mayte, Capella-Peiró Maria-Elisa, Broch Samuel Carda, Esteve-Romero Josep
Servei d'Anàlisis Clíniques, Consorci Hospital Provincial de Castelló, Av. Dr. Clara 19, 12002 Castelló, Spain.
J Chromatogr A. 2005 May 6;1073(1-2):309-15. doi: 10.1016/j.chroma.2004.10.069.
A procedure was developed for the determination of caffeine and theophylline using a C18 column (5 microm, 250 mm x 4.6 mm) and micellar liquid chromatography using hybrid mobile phases containing sodium dodecyl sulfate (SDS) and propanol, butanol or pentanol as modifiers. Detection was performed with a variable wavelength UV-vis detector at 272 nm. After the application of an interpretative strategy for the selection of the optimimum mobile phase, caffeine and theophylline can be resolved and determined in serum samples by direct injection, using a mobile phase made up of 50 mM SDS-2.5% (v/v) propanol-10 mM KH2PO4, pH 7, with an analysis time below 5 min. Calibration was linear in the range 0.05 to 50 microg mL(-1) with r > 0.999. The statistical evaluation of the method was examined by performing intra-day (n = 6) and inter-day calibration (n = 7) and was found to be satisfactory, with highly accurate and precise results. The proposed method was suitably validated and applied to the determination of caffeine and theophylline in serum samples of patients treated with bronchodilators.
开发了一种使用C18柱（5微米，250毫米×4.6毫米）和胶束液相色谱法测定咖啡因和茶碱的方法，该方法使用含有十二烷基硫酸钠（SDS）和丙醇、丁醇或戊醇作为改性剂的混合流动相。使用可变波长紫外可见检测器在272纳米处进行检测。在应用一种解释策略来选择最佳流动相后，通过直接进样，使用由50 mM SDS-2.5%（v/v）丙醇-10 mM KH2PO4（pH 7）组成的流动相，可在血清样品中分离并测定咖啡因和茶碱，分析时间在5分钟以内。校准在0.05至50微克/毫升（-1）范围内呈线性，r>0.999。通过进行日内（n = 6）和日间校准（n = 7）对该方法进行统计评估，结果令人满意，具有高度准确和精确的结果。所提出的方法经过了适当验证，并应用于测定接受支气管扩张剂治疗患者的血清样品中的咖啡因和茶碱。
