Chen Huaixia, Chen Yong, Du Peng, Han Fengmei
Ministry-of-Education Key Laboratory for the Synthesis and Application of Organic Functional Molecules & College of Chemistry and Chemical Engineering, Hubei University, Wuhan 430062, China.
J Pharm Biomed Anal. 2007 Jul 27;44(3):773-8. doi: 10.1016/j.jpba.2007.01.038. Epub 2007 Feb 1.
Liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESIMSn) was employed to investigate the in vivo and in vitro metabolism of anisodine. Feces, urine and plasma samples were collected after ingestion of 20 mg anisodine to healthy rats. Feces and urine samples were cleaned up by liquid-liquid extraction and solid-phase extraction procedures (C18 cartridges), respectively. Methanol was added to plasma samples to precipitate plasma proteins. Anisodine was incubated with homogenized liver and intestinal flora of rats in vitro, respectively, followed by extraction with ethyl acetate. LC-MSn was used for the separation and identification of the metabolites using C18 column with mobile phase of methanol/0.01% triethylamine solution (2 mM, adjusted to pH 3.5 with formic acid) (60:40, v/v). The results revealed that five metabolites (norscopine, scopine, alpha-hydroxytropic acid, noranisodine and hydroxyanisodine) and the parent drug existed in feces. Three new metabolites (dimethoxyanisodine, tetrahydroxyanisodine and trihydroxy-methoxyanisodine) were identified in urine. Four metabolites (norscopine, scopine, hydroxyanisodine and anisodine N-oxide) and the parent drug were detected in plasma. Two hydrolyzed metabolites (scopine and alpha-hydroxytropic acid) were found in rat intestinal flora incubation mixture, and two metabolites (aponoranisodine and anisodine N-oxide) were identified in homogenized liver incubation mixture.
采用液相色谱 - 电喷雾电离串联质谱法（LC - ESIMSⁿ）研究樟柳碱的体内外代谢情况。给健康大鼠灌胃20 mg樟柳碱后，收集粪便、尿液和血浆样本。粪便和尿液样本分别通过液 - 液萃取和固相萃取程序（C18柱）进行净化处理。向血浆样本中加入甲醇以沉淀血浆蛋白。樟柳碱分别与大鼠的肝匀浆和肠道菌群在体外孵育，随后用乙酸乙酯萃取。采用LC - MSⁿ，使用C18柱，以甲醇/0.01%三乙胺溶液（2 mM，用甲酸调节至pH 3.5）（60:40，v/v）作为流动相，对代谢产物进行分离和鉴定。结果显示，粪便中存在5种代谢产物（去甲东莨菪碱、东莨菪碱、α - 羟基托品酸、去甲樟柳碱和羟基樟柳碱）及母体药物。尿液中鉴定出3种新的代谢产物（二甲氧基樟柳碱、四羟基樟柳碱和三羟基甲氧基樟柳碱）。血浆中检测到4种代谢产物（去甲东莨菪碱、东莨菪碱、羟基樟柳碱和樟柳碱N - 氧化物）及母体药物。在大鼠肠道菌群孵育混合物中发现2种水解代谢产物（东莨菪碱和α - 羟基托品酸），在肝匀浆孵育混合物中鉴定出2种代谢产物（阿朴樟柳碱和樟柳碱N - 氧化物）。
