HPLC-MSn法鉴定葫芦巴碱及其在大鼠体内的主要代谢产物

目的建立快速灵敏的LC-MSⁿ检测葫芦巴碱及其在大鼠体内代谢物的分析方法。方法以葫芦巴碱对LC-MS²色谱及质谱条件进行优化，分析其电喷雾质谱的一级电离规律和多级质谱裂解规律，以此作为葫芦巴碱大鼠体内代谢物分析鉴定的依据。健康大鼠尾静脉注射8 mg·kg^-1葫芦巴碱，收集0～48 h的尿样，经C₁₈小柱固相萃取分离纯化后，直接采用LC-MSⁿ方法对尿样进行测定。结果根据生物体内药物代谢转化规律及母体药物的色谱-质谱行为规律，在尿样中鉴定出母药及其N-去甲基、N-去甲基环氧化产物，以及母药及其N-去甲基环氧化物的甘氨酸轭合物。结论本方法灵敏、快速、选择性高、专属性好，可用于葫芦巴碱的代谢产物研究。     

HPLC-ESI-ITMSn法鉴定麻黄碱及其大鼠体内主要代谢产物

目的建立快速灵敏的LC-ESI-ITMSⁿ分析检测麻黄碱及其大鼠体内代谢物的方法。方法以麻黄碱对照品对LC-ESI-ITMS²色谱及质谱条件进行了优化，分析总结其电喷雾质谱的一级电离规律和多级质谱裂解规律，以此作为麻黄碱大鼠体内代谢物分析鉴定的依据。健康大鼠空腹灌胃麻黄碱10 mg·kg^-1，收集0~48 h的尿样，经C₁₈小柱固相萃取分离纯化后，直接采用LC-ESI-ITMSⁿ方法对尿样进行测定。结果根据生物体内药物代谢转化规律及母体药物的色谱-质谱行为规律，在尿样中鉴定出3个第I相代谢产物，未发现第II相代谢产物。结论本方法灵敏、快速、选择性高、专属性好，可用于麻黄碱的代谢产物研究。     

LC/MS分析大鼠体内氧化苦参碱及其主要代谢物LC/MS分析大鼠体内氧化苦参碱及其主要代谢物

目的研究氧化苦参碱在大鼠体内的主要代谢产物。方法以氧化苦参碱和苦参碱为对象优化液相色谱/电喷雾离子阱质谱(LC/ESI-ITMSⁿ)实验条件，分析总结其电喷雾质谱的一级电离规律和二级质谱裂解规律，作为氧化苦参碱大鼠体内代谢物结构分析的依据。健康大鼠腹腔肌注40 mg·kg^-1氧化苦参碱，收集0~24 h的尿样，尿样中的代谢物经C₁₈小柱进行富集与纯化后，在优化的LC/ESI-ITMSⁿ条件下进样分析。代谢物的结构推导主要依据代谢物的色谱保留时间及其电喷雾离子阱质谱(ESI-ITMSⁿ)电离规律。结果在大鼠尿样中有原药及其6种I相氧化及还原代谢产物，且主要代谢物为苦参碱。未发现II相代谢物。结论本法不仅操作简便、快速，而且灵敏度高、专属性强。该分析技术是研究药物代谢最有效的方法之一。     

液相色谱-串联电喷雾离子阱质谱法鉴定大鼠尿液中药根碱代谢物

目的研究药根碱在大鼠体内的主要代谢产物。方法健康大鼠尾静脉注射12 mg·kg^-1药根碱，收集0～72 h的尿样，尿样经C₁₈小柱固相萃取分离纯化后，经液相色谱-串联电喷雾离子阱质谱(LC-ESI/ITMSⁿ)分析鉴定其中的代谢物。代谢物的结构鉴定主要依据各代谢物与原药的一级质谱电离规律和二级或三级质谱裂解规律间的关联性。结果在大鼠尿样中检测到7种I相代谢产物(如原药的脱氢、脱甲基、羟基化代谢物)及11种II相代谢产物(如甲基化轭合物和葡糖醛酸轭合物)。结论本方法用于大鼠尿样中药根碱的代谢物研究不仅操作简单、快速，而且灵敏度高、专属性强。     

罂粟碱的体内与体外代谢物研究

利用HPLC-MSⁿ检测罂粟碱及其在大鼠体内(大鼠粪便)和体外(肝微粒体, 肠道菌)代谢物。体内与体外的代谢物经C₁₈小柱进行富集和纯化后, 直接采用优化的HPLC-ESI/ITMSⁿ方法对样品进样分析。以罂粟碱标准品为对象优化高效液相色谱/电喷雾离子阱质谱(HPLC-ESI/ITMSⁿ )实验条件, 分析总结其电喷雾质谱的一级电离规律和多级质谱裂解规律, 作为罂粟碱大鼠体内与体外代谢物结构分析的依据。代谢物的结构推导主要依据代谢物的色谱保留时间及其电喷雾离子阱质谱HPLC-ESI/ITMSⁿ电离规律。在大鼠粪便中有原药及其8种代谢产物。体外代谢物检测到原药的脱甲基和羟基化。     

液相色谱-串联电喷雾离子阱质谱法鉴定大鼠尿液中大豆黄素的羟基化代谢产物及其硫酸酯轭合物

目的鉴定大豆黄素在大鼠体内的羟基化及其结合形代谢产物。方法SD大鼠分别单剂量给药500 mg·kg^-1，收集0～24 h尿样。尿样经SPE ODS C₁₈固相萃取柱纯化后，用LC-ESI/MSⁿ对尿样中的代谢物分别进行选择离子监测(SIM)和多级质谱(MSⁿ)分析。结果在大鼠尿中检测到几种尚未在国内外报道过的羟基化代谢产物及其硫酸酯轭合物。结论LC-ESI/MSⁿ法可以快速、简捷、准确地鉴定大豆黄素在大鼠体内羟基化及其结合形代谢产物。     

五味子酯甲在大鼠体内代谢产物的鉴定

目的 考察五味子酯甲在大鼠体内的代谢转化。方法 采用超高效液相色谱串联四级杆飞行时间质谱(UPLC-TOF-MS/MS)分析鉴定大鼠灌胃五味子酯甲后,其在尿样中的代谢产物。Phenomenex UPLC C₁₈色谱柱,流动相为乙腈-1‰甲酸水,梯度洗脱,质谱仪离子源为电喷雾离子源(ESI),正离子方式检测。结果 经代谢物软件处理后,根据MS/MS给出质谱碎片信息对五味子酯甲代谢产物进行结构推测,共检测到5种代谢产物。结论 五味子酯甲在大鼠体内的代谢途径主要为氧化反应和还原反应。     

HPLC-ESI/ITMSn法分析大鼠体内4-甲基哌嗪-1-二硫代甲酸-(3-氰基-3，3-二苯基)丙酯盐酸盐及其主要代谢物

目的研究新化合物4-甲基哌嗪-1-二硫代甲酸-(3-氰基-3，3-二苯基)丙酯盐酸盐(TM208)在大鼠体内的主要代谢产物。方法大鼠ig TM208 500 mg·kg^-1后，收集粪样、尿样和血样，用液相色谱-电喷雾离子阱质谱法测定。根据TM208及其代谢产物的色谱保留时间和电喷雾离子阱质谱(ESI-ITMSⁿ )电离规律及生物体内药物代谢转化规律，推导代谢物的结构。结果在粪样中发现8种I相代谢产物，在尿样和血样中发现5种I相代谢产物，未发现II相代谢产物。结论本法操作简便、快速、灵敏度高、专属性强，是一种研究TM208体内代谢产物的有效方法。     

新型抗炎镇痛剂SFZ-47在家兔体内羟基化及其结合型代谢产物的鉴定

目的：鉴定新型抗炎镇痛剂SFZ-47［3H-1,2-二氢-2-(4-甲基苯胺基)甲基-1-吡咯里嗪酮］在家兔体内的羟基化及其结合型代谢产物。方法：选择4只健康家兔单剂量口服100 mg SFZ-47,收集0～10 h的尿样。将未经或经过β-D-葡萄糖苷酸酶/硫酸酯酶水解的尿样以Sep-Pak C₁₈固相萃取柱纯化后,采用LC/MSn方法对尿中推测的代谢物分别进行选择离子监测(SIM)和多级全扫描质谱(MSⁿ)分析。结果：在尿中首次检测到SFZ-47羟基化及其β-D-葡糖苷酸与硫酸结合型代谢物。结论：SFZ-47在家兔体内主要代谢途径应为苯环上的甲基先氧化成羟甲基和羧基,再分别与β-D-葡糖醛酸或硫酸结合。     

牡荆素在大鼠体内的代谢研究

目的 研究黄酮碳苷牡荆素在大鼠体内的代谢产物,并推测其代谢途径。方法 SD大鼠灌胃给予5 mg·kg^-1牡荆素,收集0~3 h,3~6 h,6~12 h的尿液,采用UPLC-Q-TOF检测尿样中代谢产物。结果 采用Metabolynx XS代谢物分析软件,根据质谱碎片信息对代谢物进行结构鉴定,最终得到3个代谢产物。结论 牡荆素在大鼠尿液中检测得到1个一相代谢产物,2个二相代谢产物,推测牡荆素在大鼠体内主要发生氧化、甲基化和葡萄糖醛酸结合反应,其中葡萄糖醛酸化反应是较强的代谢种类。     

Characterization of new metabolites from in vivo biotransformation of norisoboldine by liquid chromatography/mass spectrometry and NMR spectroscopy

Norisoboldine (1,9-dihydroxy-2,10-dimethoxynoraporphine) is one of the major bioactive isoquinoline alkaloids in Linderae Radix, a commonly used Chinese herbal medicine. The aim of this study is to isolate and characterize metabolites of norisoboldine after gavage feeding in rats. High-performance liquid chromatography coupled with electrospray ionization and ion-trap mass spectrometry (HPLC-ESI/MSⁿ) was used to identify metabolites of norisoboldine in rat urine and bile samples. A total of five metabolites of norisoboldine were characterized by comparing retention time and UV absorption in HPLC, and by molecular mass and fragmentation pattern of the analytes by mass spectrometry with those of norisoboldine. Two new glucuronide conjugates of norisoboldine, noriosboldine-1-O-β-d-glucuronide and norisoboldine-9-O-α-d-glucuronide, were isolated from rat urine samples and their structures were confirmed by NMR spectroscopy (¹H, ¹³C, HMBC and HSQC) for the first time. The results suggested that glucuronidation and sulfation were involved in metabolic pathways of norisoboldine in rat.     

Identification of metabolites of tectoridin in-vivo and in-vitro by liquid chromatography-tandem mass spectrometry

In this work, liquid chromatography-electrospray ionization tandem ion-trap mass spectrometry (LC-MS(n)) was used to investigate the in-vivo and in-vitro metabolism of tectoridin. After oral administration of a single dose (100 mg kg(-1)) of tectoridin to healthy rats, faeces and urine samples were collected for 0-48 h and 0-24 h, respectively. Tectoridin was also incubated with rat intestinal flora and rat liver microsomes. Samples from in-vivo and in-vitro metabolism studies were purified using a C(18) solid-phase extraction cartridge, then separated using a reverse-phase C(18) column with methanol/ water (30:70, v/v, adjusted to pH 10.0 with ammonia water) as mobile phase and detected by an on-line MS(n) system. The structure of the metabolites was elucidated by comparing their molecular weights, retention times and full-scan MS(n) spectra with those of the parent drug. The results revealed six metabolites of tectoridin in urine (tectorigenin, hydrogenated tectorigenin, mono-hydroxylated tectorigenin, di-hydroxylated tectorigenin, glucuronide-conjugated tectorigenin and sulfate-conjugated tectorigenin); three metabolites in faeces (tectorigenin, di-hydroxylated tectorigenin and sulfateconjugated tectorigenin); one metabolite in the intestinal flora incubation mixture (tectorigenin), and four in the liver microsomal incubation mixture (tectorigenin, hydrogenated tectorigenin, mono-hydroxylated tectorigenin and di-hydroxylated tectorigenin). Except for tectorigenin, all other metabolites of tectoridin are reported for the first time.     

左旋一叶碱的代谢转化

目的研究一叶碱［securinine,(-)SE］在大鼠体内外的代谢转化。方法采用大鼠肝微粒体体外温孵法对(-)SE的代谢转化进行了研究,优化了代谢体系,建立了反相HPLC法同时分离检测(-)SE及其体外代谢产物的分析方法。用液液萃取,制备TLC及半制备HPLC分离纯化了4个代谢产物并进行了光谱鉴定。在此基础上,建立了生物体液中(-)SE及其代谢物的反相HPLC分析方法,并用该法检测了ip给药后大鼠的胆汁、尿样及其经β-葡糖醛酸苷酶水解后的样品。结果代谢物分别鉴定为6-位羟基,6-位羰基及5-位α及β羟基取代的(-)SE,还证实了体内6-位羟基代谢物进一步形成了二相结合型产物。结论基本阐明(-)SE在大鼠体内外代谢转化的途径。     

Identification of propentofylline metabolites in rats by gas chromatography/mass spectrometry

Propentofylline (PPF, 3-methyl-1-(5-oxohexyl)-7-propylxanthine) has been reported to be a compound for treatment of both vascular dementia and dementia of the Alzheimer type. The short half-life (about 15 min) of PPF at the terminal elimination phase and poor bioavailability after oral administration of PPF to rabbits (Kim et al., 1992) suggest in part that this drug takes the extensive first-pass metabolism in the liver. In addition, the metabolic pathway for PPF remains unclear. The objective of this experiment is to identify urinary metabolites of PPF in rats. For the identification of the metabolites, rat urine was collected after oral administration of 100 mg/kg PPF. PPF metabolite, 3-methyl-1-(5-hydroxyhexyl)-7-propylxanthine, was synthesized and confirmed by gas chromatography/mass spectroscopy (GC/MS) and 1H nuclear magnetic resonance spectroscopy. The urinary metabolites of PPF were extracted with diethyl ether and identified by electron impact and chemical ionization GC/MS. One urinary metabolite was confirmed to be 3-methyl-1-(5-hydroxyhexyl)-7-propylxanthine by synthesized authentic compound. Several metabolites of monohydroxy- and dihydroxy-PPF were identified based on mass fragmentation of both intact and trimethylsilylated derivatives of PPF metabolites and the novel structure of these metabolites is suggested based on mass spectra.