Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, PR China.
J Pharm Biomed Anal. 2010 Apr 6;51(5):1134-41. doi: 10.1016/j.jpba.2009.11.024. Epub 2009 Dec 1.
(E)-4-hydroxydemethylbromhexine (E-4-HDMB) and (E)-3-hydroxydemethylbromhexine (E-3-HDMB) were found as major metabolites, while (Z)-4-hydroxydemethylbromhexine and (Z)-3-hydroxydemethylbromhexine as minor metabolites of bromhexine in human plasma. These compounds were identified in comparison with synthetic authentic samples. A sensitive and selective rapid resolution liquid chromatography tandem mass spectrometry (RRLC-MS/MS) method was developed to quantify the concentration of bromhexine and its two major metabolites (E-4-HDMB and E-3-HDMB) in human plasma. Following solid phase extraction, the analytes were separated on a Zorbax 1.8microm particle size reversed-phase C(18) column, using a gradient elution program with solvents consisting of 0.1% formic acid in acetonitrile and 0.1% formic acid in 5mM ammonium acetate at a flow rate of 0.7mL/min. Detection was carried out with an Agilent 6460 triple-quadrupole mass spectrometer operated with an electrospray ionization source mode operated in the positive ion mode. The recovery of bromhexine, E-4-HDMB, E-3-HDMB, and internal standard (IS) was 63.1-70.9%, 60.5-68.4%, 57.0-63.5%, and 87.8%, respectively. The matrix factors of bromhexine, E-4-HDMB, E-3-HDMB, and IS were 89.9-96.7%, 89.6-94.8%, 90.4-91.4%, and 103%, respectively. After an oral administration of 8.0mg bromhexine to five healthy male subjects, AUC(0-24h) values of bromhexine, E-4-HDMB, and E-3-HDMB were found to be 93.5+/-31.9, 34.0+/-14.5, and 15.8+/-6.89ngh/mL, respectively; while C(max) values were 24.6+/-5.16, 3.11+/-1.13, and 5.36+/-2.55ng/mL, respectively. Plasma concentration of bromhexine, E-4-HDMB, and E-3-HDMB declined with t(1/2) which gave 3.6+/-0.5, 8.4+/-2.7, and 6.4+/-2.5h, respectively.
（E）-4-羟去甲溴己新（E-4-HDMB）和（E）-3-羟去甲溴己新（E-3-HDMB）被发现为溴己新在人血浆中的主要代谢物，而（Z）-4-羟去甲溴己新和（Z）-3-羟去甲溴己新则为次要代谢物。这些化合物是通过与合成的纯正样品进行比较而鉴定的。开发了一种灵敏和选择性的快速分辨液相色谱串联质谱（RRLC-MS/MS）方法来定量人血浆中溴己新及其两种主要代谢物（E-4-HDMB 和 E-3-HDMB）的浓度。通过固相萃取后，分析物在 Zorbax 1.8μm粒径反相 C（18）柱上分离，使用包含 0.1%甲酸的乙腈和 0.1%甲酸的 5mM 乙酸铵的梯度洗脱程序，流速为 0.7mL/min。检测采用 Agilent 6460 三重四极杆质谱仪，采用电喷雾电离源模式，以正离子模式运行。溴己新、E-4-HDMB、E-3-HDMB 和内标（IS）的回收率分别为 63.1-70.9%、60.5-68.4%、57.0-63.5%和 87.8%。溴己新、E-4-HDMB、E-3-HDMB 和 IS 的基质因子分别为 89.9-96.7%、89.6-94.8%、90.4-91.4%和 103%。在 5 名健康男性受试者口服 8.0mg 溴己新后，发现溴己新、E-4-HDMB 和 E-3-HDMB 的 AUC（0-24h）值分别为 93.5+/-31.9、34.0+/-14.5 和 15.8+/-6.89ngh/mL，而 C（max）值分别为 24.6+/-5.16、3.11+/-1.13 和 5.36+/-2.55ng/mL。溴己新、E-4-HDMB 和 E-3-HDMB 的血浆浓度随 t（1/2）而下降，分别为 3.6+/-0.5、8.4+/-2.7 和 6.4+/-2.5h。
