Fujinami A, Miyazawa T, Tagawa N, Kobayashi Y
Clinical Chemistry Laboratory, Kobe Pharmaceutical University, Japan.
Biol Pharm Bull. 1998 Nov;21(11):1207-10. doi: 10.1248/bpb.21.1207.
We developed a method for simultaneous analysis of benzphetamine (BZ) and its metabolites, p-hydroxy-N-benzylamphetamine (pHBA), p-hydroxybenzphetamine (pHBZ), amphetamine (AP), methamphetamine and p-hydroxymethamphetamine by micellar electrokinetic chromatography (MEKC). Urine samples from 0-15 h (3-h intervals) after oral administration of BZ (10 mg) were hydrolyzed with beta-glucuronidase (EC 3.2.1.31) at 37 degrees C overnight. The treated urine was applied to a solid phase extraction column Bond Elut Certify. After sequentially washing the column with water, 0.1 mol/l acetic acid and methanol, the samples were eluted with dichloromethane:isopropanol:28% ammonium hydroxide=78.4:19.6:2.0 (v/v %). The eluate was evaporated and the residue dissolved in running buffer was analyzed by MEKC. In urine from 0-3 h, AP, pHBZ and pHBA were detected. After that, only pHBA, which is one of the major metabolites of BZ in human urine, could be detected in the urine by the present method. A method for quantitation of pHBA by MEKC is described here. The effects of acetonitrile and sodium dodecyl sulfate in the running buffer of MEKC on the separation of BZ and its metabolites are also reported.
我们开发了一种通过胶束电动色谱法（MEKC）同时分析苄非他明（BZ）及其代谢产物对羟基-N-苄基苯丙胺（pHBA）、对羟基苄非他明（pHBZ）、苯丙胺（AP）、甲基苯丙胺和对羟基甲基苯丙胺的方法。口服10mg BZ后0至15小时（间隔3小时）的尿液样本在37℃下用β-葡萄糖醛酸酶（EC 3.2.1.31）水解过夜。将处理后的尿液应用于固相萃取柱Bond Elut Certify。依次用水、0.1mol/L乙酸和甲醇冲洗柱子后，用二氯甲烷：异丙醇：28%氢氧化铵=78.4:19.6:2.0（v/v%）洗脱样品。将洗脱液蒸发，将残渣溶解在运行缓冲液中，通过MEKC进行分析。在0至3小时的尿液中检测到AP、pHBZ和pHBA。在此之后，通过本方法仅能在尿液中检测到pHBA，它是BZ在人尿液中的主要代谢产物之一。本文描述了一种通过MEKC定量pHBA的方法。还报道了MEKC运行缓冲液中的乙腈和十二烷基硫酸钠对BZ及其代谢产物分离的影响。
