Ren Shuang, Chen Xiao-yan, Duan Xiao-tao, Zhong Da-fang
Laboratory of Drug Metabolism and Pharmacokinetics, Shenyang Pharmaceutical University, Shenyang 110016, China.
Yao Xue Xue Bao. 2006 Feb;41(2):188-92.
To develop and validate a liquid chromatography-tandem mass spectrometric (LC/MS/MS) method for the simultaneous quantification of ephedrine and chlorpheniramine in human plasma after oral administration of a compound preparation.
The analytes and the internal standard, diphenhydramine, were isolated from plasma by protein precipitation with methanol, then chromatographied on a Zorbax SB-C18 column (150 mm x 4.6 mm ID) using a mobile phase consisted of methanol-water-formic acid (80: 20: 0.5, v/v), at a flow rate of 0.5 mL x min(-1). A tandem mass spectrometer equipped with electrospray ionization source was used as detector and was operated in the positive ion mode. Selected reaction monitoring (SRM) using the precursor to produce ion combinations of m/z 166-->115, m/z 275-->230 and m/z 256-->167 were used to quantify ephedrine, chlorpheniramine and the internal standard, respectively. Results The linear concentration ranges of the calibration curves for ephedrine and chlorpheniramine were 0.50 - 200 microg x L(-1) and 0.050 - 20.0 microg x L(-1), respectively. The lower limits of quantification were 0. 50 microg x L(-1) for ephedrine and 0.050 microg x L(-1) for chlorpheniramine, individually. The intra- and inter-day relative standard deviation (RSD) across three validation runs over the entire concentration range was less than 9.3% for both ephedrine and chlorpheniramine. The inter-day accuracy (RE) was within +/- 3.4% for the analytes. Each sample was chromatographied within 3.3 min. The method was successfully used in pharmacokinetics study of ephedrine and chlorpheniramine in human plasma after oral administration of a compound preparation containing 5 mg ephedrine hydrochloride, 1 mg chlorpheniramine maleate, 50 mg phenytoin, 12.5 mg theophylline, 12.5 mg theobromine and 7.5 mg caffeine. No interaction among the six components was observed on their pharmacokinetic parameters.
The method was proved to be highly sensitive, selective, and suitable for pharmacokinetics investigations of different compound preparations containing low dosage of both ephedrine and chlorpheniramine.
建立并验证一种液相色谱 - 串联质谱（LC/MS/MS）法，用于同时定量口服复方制剂后人血浆中麻黄碱和氯苯那敏的含量。
采用甲醇蛋白沉淀法从血浆中分离分析物和内标苯海拉明，然后在Zorbax SB - C18柱（150 mm×4.6 mm内径）上进行色谱分离，流动相为甲醇 - 水 - 甲酸（80:20:0.5，v/v），流速为0.5 mL·min⁻¹。配备电喷雾电离源的串联质谱仪用作检测器，以正离子模式运行。分别采用m/z 166→115、m/z 275→230和m/z 2�6→167的前体离子产生离子对进行选择反应监测（SRM），以定量麻黄碱、氯苯那敏和内标。结果麻黄碱和氯苯那敏校准曲线的线性浓度范围分别为0.50 - 200 μg·L⁻¹和0.050 - 20.0 μg·L⁻¹。麻黄碱和氯苯那敏的定量下限分别为0.50 μg·L⁻¹和0.050 μg·L⁻¹。在整个浓度范围内，麻黄碱和氯苯那敏在三次验证实验中的日内和日间相对标准偏差（RSD）均小于9.3%。分析物的日间准确度（RE）在±3.4%以内。每个样品在3.3分钟内完成色谱分析。该方法成功用于口服含有5 mg盐酸麻黄碱、1 mg马来酸氯苯那敏、50 mg苯妥英、12.5 mg茶碱、12.5 mg可可碱和7.5 mg咖啡因的复方制剂后人血浆中麻黄碱和氯苯那敏的药代动力学研究。未观察到六种成分之间在药代动力学参数上的相互作用。
该方法被证明具有高灵敏度、高选择性，适用于含低剂量麻黄碱和氯苯那敏的不同复方制剂的药代动力学研究。
