Su Fenli, Wang Feng, Gao Wei, Li Huande
Clinical Pharmacy & Pharmacology Institute, Second Xiangya Hospital, Central South University, Changsha 410011, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Jun 15;853(1-2):364-8. doi: 10.1016/j.jchromb.2007.03.015. Epub 2007 Mar 24.
A rapid, sensitive and specific method to determination of ambroxol in human plasma using high performance liquid chromatography coupled with electrospray ionization mass spectrometry (HPLC-MS/ESI) was described. Ambroxol and the internal standard (I.S.), fentanyl, were extracted from plasma by N-hexane-diethyl ether (1:1, v/v) after alkalinized with ammonia water. A centrifuged upper layer was then evaporated and reconstituted with 100 microl mobile phase. Chromatographic separation was performed on a BDS HYPERSIL C18 column (250 mmx4.6 mm, 5.0 microm, Thermo electron corporation, USA) with the mobile phase consisting of 30 mM ammonium acetate (0.4% formic acid)-acetonitrile (64:36, v/v) at a flow-rate of 1.2 mL min(-1). The total run time was 5.8 min for each sample. Detection and quantitation was performed by the mass spectrometer using selected ion monitoring at m/z 261.9, 263.8 and 265.9 for ambroxol and m/z 337.3 for fentanyl. The calibration curve was linear within the concentration range of 1.0-100.0 ng mL(-1) (r=0.9996). The limit of quantification was 1.0 ng mL(-1). The extraction recovery was above 83.3%. The methodology recovery was higher than 93.8%. The intra- and inter-day precisions were less than 6.0%. The method is accurate, sensitive and simple for the study of the pharmacokinetics and metabolism of ambroxol.
描述了一种使用高效液相色谱-电喷雾电离质谱联用（HPLC-MS/ESI）测定人血浆中氨溴索的快速、灵敏且特异的方法。氨溴索和内标（I.S.）芬太尼在用氨水碱化后，通过正己烷-乙醚（1:1，v/v）从血浆中萃取。然后将离心后的上层清液蒸发，并用100微升流动相复溶。色谱分离在BDS HYPERSIL C18柱（250 mm×4.6 mm，5.0微米，美国热电公司）上进行，流动相由30 mM醋酸铵（0.4%甲酸）-乙腈（64:36，v/v）组成，流速为1.2 mL min⁻¹。每个样品的总运行时间为5.8分钟。通过质谱仪使用选择离子监测进行检测和定量，氨溴索的m/z为261.9、263.8和265.9，芬太尼的m/z为337.3。校准曲线在1.0 - 100.0 ng mL⁻¹浓度范围内呈线性（r = 0.9996）。定量限为1.0 ng mL⁻¹。萃取回收率高于83.3%。方法回收率高于93.8%。日内和日间精密度均小于6.0%。该方法对于氨溴索的药代动力学和代谢研究准确、灵敏且简单。
