Kim Hohyun, Yoo Jeong-Yeon, Han Sang Beom, Lee Hee Joo, Lee Kyung Ryul
Department of Pharmacokinetics, Seoul Clinical Laboratories, Seoul Medical Science Institute, 7-14 Dongbinggo-dong, Yongsan-gu, Seoul 140-809, South Korea.
J Pharm Biomed Anal. 2003 Jun 1;32(2):209-16. doi: 10.1016/s0731-7085(03)00059-1.
A sensitive and selective liquid chromatographic method coupled with tandem mass spectrometry (LC-MS/MS) was developed for the quantification of ambroxol in human plasma. Domperidone was used as internal standard, with plasma samples extracted using diethyl ether under basic condition. A centrifuged upper layer was then evaporated and reconstituted with 200 microl methanol. The reconstituted samples were injected into a C(18) XTerra MS column (2.1 x 30 mm) with 3.5 microm particle size. The analytical column lasted for at least 600 injections. The mobile phase was composed of 20 mM ammonium acetate in 90% acetonitrile (pH 8.8), with flow rate at 250 microl/min. The mass spectrometer was operated in positive ion mode using turbo electrospray ionization. Nitrogen was used as the nebulizer, curtain, collision, and auxiliary gases. Using MS/MS with multiple reaction monitoring (MRM) mode, ambroxol was detected without severe interferences from plasma matrix. Ambroxol produced a protonated precursor ion (M+H) at m/z 379 and a corresponding product ion at m/z 264. And internal standard (domperidone) produced a protonated precursor ion (M+H) at m/z 426 and a corresponding product ion at m/z 174. Detection of ambroxol in human plasma was accurate and precise, with quantification limit at 0.2 ng/ml. This method has been successfully applied to a study of ambroxol in human specimens.
建立了一种灵敏且具选择性的液相色谱-串联质谱联用（LC-MS/MS）方法，用于定量测定人血浆中的氨溴索。多潘立酮用作内标，在碱性条件下用乙醚萃取血浆样品。然后将离心后的上层清液蒸发，并用200微升甲醇复溶。将复溶后的样品注入粒径为3.5微米的C(18) XTerra MS柱（2.1×30毫米）。该分析柱至少可进行600次进样。流动相由90%乙腈中的20 mM醋酸铵（pH 8.8）组成，流速为250微升/分钟。质谱仪采用涡轮电喷雾电离在正离子模式下运行。氮气用作雾化气、帘气、碰撞气和辅助气。采用MS/MS的多反应监测（MRM）模式，氨溴索的检测不受血浆基质的严重干扰。氨溴索产生质荷比为379的质子化前体离子（M+H）和质荷比为264的相应产物离子。内标（多潘立酮）产生质荷比为426的质子化前体离子（M+H）和质荷比为174的相应产物离子。人血浆中氨溴索的检测准确且精密，定量限为0.2纳克/毫升。该方法已成功应用于一项关于人样本中氨溴索的研究。
