Li Yan, Sun Yuming, Dong Yu, Zhang Yifan, Zhong Dafang
Laboratory of Drug Metabolism and Pharmacokinetics, Shenyang Pharmaceutical University, Shenyang, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2006 Jan 2;830(1):71-80. doi: 10.1016/j.jchromb.2005.10.011. Epub 2005 Nov 8.
Two novel metabolites of benproperine (BPP), 1-[1-methyl-2-[2-(phenylmethyl)phenoxy]ethyl]-3-piperidinol (3-OH-BPP) and 1-[1-methyl-2-[2-(phenylmethyl)phenoxy]ethyl]-4-piperidinol (4-OH-BPP), were confirmed by comparison of retention times and mass spectra with those of synthetic standards using liquid chromatography-tandem mass spectrometry. Selective and sensitive procedures were developed for the simultaneous determination of BPP, 3-OH-BPP and 4-OH-BPP in human plasma and urine. The analytes were extracted from plasma sample and enzymatically hydrolyzed urine samples by liquid-liquid extraction, separated through a Diamonsil C(18) column (150 mm x 4.6 mm i.d.) and determined by tandem mass spectrometry with an electrospray ionization interface in selected reaction monitoring mode. Dextromethorphan was used as internal standard. The mobile phase consisted of acetonitrile-water-formic acid (34:66:1, v/v/v), and flow-rate was 0.5 ml min(-1). This method has a lower limit of quantification (LLOQ) of 60, 4.0 and 4.0 nmol l(-1)for BPP, 3-OH-BPP and 4-OH-BPP in plasma, 4.9, 4.7 and 2.4 nmol l(-1) in urine, respectively. The intra- and inter-run precision were measured to be below 9.2%, and the accuracy was within +/-4.3% for the analytes. The method was successfully used to determine BPP, 3-OH-BPP and 4-OH-BPP in plasma and urine for pharmacokinetic investigation. The results indicated residue of 3-OH-BPP in the body at least 192 h after an oral dose of BPP.
通过液相色谱 - 串联质谱法将保留时间和质谱与合成标准品进行比较，确认了苯丙哌林（BPP）的两种新型代谢产物，即1 - [1 - 甲基 - 2 - [2 - （苯甲基）苯氧基]乙基] - 3 - 哌啶醇（3 - OH - BPP）和1 - [1 - 甲基 - 2 - [2 - （苯甲基）苯氧基]乙基] - 4 - 哌啶醇（4 - OH - BPP）。开发了选择性和灵敏的方法，用于同时测定人血浆和尿液中的BPP、3 - OH - BPP和4 - OH - BPP。通过液 - 液萃取从血浆样品中提取分析物，并对尿液样品进行酶解，通过Diamonsil C(18)柱（150 mm×4.6 mm内径）进行分离，并在选择反应监测模式下使用电喷雾电离接口通过串联质谱法进行测定。右美沙芬用作内标。流动相由乙腈 - 水 - 甲酸（34:66:1，v/v/v）组成，流速为0.5 ml min(-1)。该方法对血浆中BPP、3 - OH - BPP和4 - OH - BPP的定量下限（LLOQ）分别为60、4.0和4.0 nmol l(-1)，尿液中分别为4.9、4.7和2.4 nmol l(-1)。批内和批间精密度测定低于9.2%，分析物的准确度在±4.3%以内。该方法成功用于血浆和尿液中BPP、3 - OH - BPP和4 - OH - BPP的药代动力学研究。结果表明，口服BPP后至少192小时，体内仍有3 - OH - BPP残留。
