Determination of hypnotic benzodiazepines (alprazolam, estazolam, and midazolam) and their metabolites in rat hair and plasma by reversed-phase liquid-chromatography with electrospray ionization mass spectrometry.
Determination of hypnotic benzodiazepines (alprazolam, estazolam, and midazolam) and their metabolites in rat hair and plasma by reversed-phase liquid-chromatography with electrospray ionization mass spectrometry.
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DOI：
10.1016/s0731-7085(02)00520-4
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发表时间：
2003-01
影响因子：
3.4
通讯作者：
T. Toyo′oka;Y. Kumaki;M. Kanbori;Masaru Kato;Y. Nakahara
中科院分区：
文献类型：
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作者：
T. Toyo′oka;Y. Kumaki;M. Kanbori;Masaru Kato;Y. Nakahara
建立了反相高效液相色谱-电喷雾电离质谱法测定苯二氮卓类药物阿普唑仑(ALP)、艾司唑仑(EST)和咪达唑仑(MDZ)及其代谢物的方法。采用半微反相色谱柱(粒径3μm；10 0×2.0 mm，i.d.)以含1%冰醋酸的乙腈-水为洗脱液。质谱仪对母体药物及其代谢物的质子化分子离子[M+H]+进行选择离子监测。将该方法应用于黑腹大鼠(I.P.)毛干的测定。每日两次服用苯二氮卓类药物，连用5天。在毛干中发现了各种代谢物和母体药物，碱性磷酸酶给药的1-羟基阿普唑仑(1-HA)和4-羟基阿普唑仑(4-HA)，酯酶给药的8-chloro-6-phenyl-4H-[1，2，4]triazolo[4，3-a][1，4]benzodiazepine-4-one(K-EST)，MDZ给药的1-羟基咪唑仑(1-HM)和4-羟基咪唑仑(4-HM)。在头发样本中也检测到一些未知的代谢物。通过乙酸酐和吡啶的乙酰化反应确定了这些结构。单次腹腔注射后，还进行了原药及其在毛根和血浆中代谢物的时程研究。苯二氮卓类药物的给药。结果表明，毛发样品中母体药物和代谢物的浓度主要取决于血浆中的浓度。
Sensitive determination of benzodiazepines i.e., alprazolam (ALP), estazolam (EST), and midazolam (MDZ), and their metabolites, was carried out by reversed-phase liquid chromatography coupled with electrospray ionization mass spectrometry (HPLC-ESI-MS). The chromatography separations were achieved using a semi-micro HPLC column (3 μm particle size; 100×2.0 mm, i.d.) with acetonitrile–water containing 1% acetic acid as eluent. The mass spectrometer was operated in selected-ion monitoring mode at protonated-molecular ions [M+H]+of parent drugs and the metabolites. The proposed procedure was applied to the determination in hair shaft of Dark Agouti rats after intraperitoneal (i.p.) administration of benzodiazepines twice a day for 5 days. Various metabolites together with parent drugs were identified in the hair shaft, 1-hydroxyalprazolam (1-HA) and 4-hydroxyalprazolam (4-HA) from ALP administration; 8-chloro-6-phenyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine-4-one (K-EST) from EST administration; 1-hydroxymidazolam (1-HM) and 4-hydroxymidazolam (4-HM) from MDZ administration. A few unknown metabolites were also detected in the hair samples. These structures were elucidated with acetylation using acetic anhydride and pyridine. The time course studies of parent drugs and the metabolites in both hair root and plasma were also carried out after single i.p. administration of benzodiazepines. The results suggested that the concentrations of parent drugs and the metabolites in the hair samples were mainly dependent upon those in the plasma.