Athanikar N K, Peng G W, Nation R L, Huang S M, Chiou W L
J Chromatogr. 1979 Mar 1;162(3):367-76.
A method was developed for the rapid quantitative analysis of chlorpheniramine in plasma, saliva and urine using high-performance liquid chromatography. A diethyl ether or hexane extract of the alkalinized biological samples was extracted with dilute acid which was chromatographed on a reversed-phase column using mixtures of acetonitrile and ammonium phosphate buffer as the mobile phase. Ultraviolet absorption at 254 nm was monitored for the detection and brompheniramine was employed as the internal standard for the quantitation. The effects of buffer, pH, and acetonitrile concentration in the mobile phase on the chromatographic separation were investigated. A mobile phase 20% acetonitrile in 0.0075 M phosphate buffer at a flow-rate of 2 ml/min was used for the assays of plasma and saliva samples. A similar mobile phase was used for urine samples. The drug and internal standard were eluted at retention volumes of less than 17 ml. The method can also be used to quantify two metabolites, didesmethyl- and desmethylchlorpheniramine, in the urine. The method can accurately measure chlorpheniramine levels down to 2 ng/ml in plasma or saliva using 1 ml of sample, and should be adequate for biopharmaceutical and pharmacokinetic studies. Various precautions for using the assay are discussed.
开发了一种使用高效液相色谱法对血浆、唾液和尿液中的氯苯那敏进行快速定量分析的方法。将碱化生物样品的乙醚或己烷提取物用稀酸萃取，然后在反相柱上进行色谱分析，以乙腈和磷酸铵缓冲液的混合物作为流动相。监测254nm处的紫外吸收进行检测，并使用溴苯那敏作为定量内标。研究了流动相中缓冲液、pH值和乙腈浓度对色谱分离的影响。流动相为含20%乙腈的0.0075M磷酸盐缓冲液，流速为2ml/min，用于血浆和唾液样品的测定。尿液样品使用类似的流动相。药物和内标在保留体积小于17ml时被洗脱。该方法还可用于定量尿液中的两种代谢物，即去二甲基氯苯那敏和去甲基氯苯那敏。该方法使用1ml样品可准确测量血浆或唾液中低至2ng/ml的氯苯那敏水平，适用于生物制药和药代动力学研究。讨论了使用该测定法的各种注意事项。
