液相色谱-串联质谱法分析樟柳碱大鼠肝匀浆代谢产物

Analysis of Anisodine and Its Metabolites in Homogenized Liver of Rat by Liquid Chromatography-Tandem Mass Spectrometry

运用液相色谱-电喷雾离子阱串联质谱(LC-MSn)联用技术研究了樟柳碱大鼠肝匀浆中的代谢物。采用离体实验的代谢研究方法,将樟柳碱与大鼠肝匀浆在富氧条件下温孵,代谢物以乙酸乙酯萃取,以ReliA-sil C18(5μm,2 mm×150 mm)为分离柱,甲醇/0.01%三乙胺溶液(用甲酸调节pH 3.5)(60∶40,V/V)为流动相,采用LC-MS及LC-MSn等方法进行分析。和原药相比较,根据温孵液中分析物分子量的变化(ΔM)及其多级质谱数据,在樟柳碱的大鼠肝匀浆培养液中鉴定出了2种代谢物,即脱水去甲基樟柳碱和N-氧化樟柳碱。实验结果表明,该液相色谱-串联质谱方法适合于药物代谢物的快速分析。

A method is described for the identification of anisodine and its metabolites in homogenized liver of rat by combining liquid chromatography and tandem mass spectrometry （LC-MS^n）. Anisodine was incubated with homogenized liver of rats in vitro under rich oxygen condition. Its metabolites were extracted with ethyl acetate and separated on a ReliAsil C l8 column（5μm, 2 mm × 150 mm） using methanol/0.01% trithylamine solution （adjusted to pH 3.5 with formic acid）（ 60：40, V/V） as mobile phase, then the metabolites were analyzed by LC-MS and LC-MS^n. Two metabolites （aponoranisodine and anisodine N-oxide） were found in the incubation solution by comparing their changes in molecular masses （△M） , full scan MS^n spectra with those of the parent drug and blank sample. The results indicate that the described HPLC-MS^n method can be effectively applied to identify the metabolites of drugs.