Soto-Otero R, Mendez-Alvarez E, Sierra-Marcuño G
J Clin Chem Clin Biochem. 1985 May;23(5):303-6. doi: 10.1515/cclm.1985.23.5.303.
A sensitive HPLC method is reported for the simultaneous determination of theophylline, caffeine, and phenobarbital in 100 microliters of plasma. After a single extraction of the drugs with chloroform/isopropanol (90 + 10 by volume) at low pH in the presence of an excess of ammonium sulphate they are resolved and quantified using a reversed-phase column (Spherisorb 5 ODS). The drugs are eluted with a binary-solvent gradient system (acetonitrile/phosphate buffer pH 4.6) at room temperature and monitored at 204 nm. Quantitation is based on peak-height ratio of analyte to interval standard (8-chlorotheophylline). Complete chromatographic resolution of all drugs is achieved within 15 min. The method is linear to 40 mg/l of theophylline and caffeine, and to 80 mg/l of phenobarbital. Numerous drugs and xanthine metabolites tested do not interfere.
报道了一种灵敏的高效液相色谱法，用于同时测定100微升血浆中的茶碱、咖啡因和苯巴比妥。在低pH值下，于过量硫酸铵存在的情况下，用氯仿/异丙醇（体积比90 + 10）对药物进行单次萃取后，使用反相柱（Spherisorb 5 ODS）对它们进行分离和定量。在室温下，用二元溶剂梯度系统（乙腈/ pH 4.6的磷酸盐缓冲液）洗脱药物，并在204 nm处进行监测。定量基于分析物与内标（8-氯茶碱）的峰高比。所有药物在15分钟内实现完全色谱分离。该方法对茶碱和咖啡因的线性范围为40 mg/l，对苯巴比妥的线性范围为80 mg/l。所测试的众多药物和黄嘌呤代谢物均不产生干扰。
