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1.
Sci Rep ; 8(1): 537, 2018 01 11.
Artigo em Inglês | MEDLINE | ID: mdl-29323165

RESUMO

In this study, the biotransformation in the plasma, urine and feces of rats following oral administration of protopine (PRO) and allocryptopine (ALL)were explored using HPLC-QqTOF MS. An HPLC-MS/MS method for the determination of tissues was developed and applied to the tissue distribution study in rats following intragastric administration of Plume Poppy Total Alkaloid for 3 weeks. A total of ten PRO metabolites and ten ALL metabolites were characterized in rats in vivo. Among these metabolites, six PRO metabolites and five ALL metabolites were reported for the first time. The predicated metabolic pathways including ring cleavage, demethylation following ring cleavage, and glucuronidation were proposed. The low-concentration residue of PRO and ALL in various tissues was detected at 24 h and 48 h after dosing, which indicated that both compounds could be widely distributed in tissues and exist as low levels of residue. The activities of erythromycin N-demethylase, aminopyrine N-demethylase and NAD (P)H quinone oxidoreductase in female rats can be induced post-dose, but these activities were inhibited in male rats. The proposed biotransformation and residues of PRO and ALL and their effects on enzymes may provide a basis for clarifying the metabolism and interpreting pharmacokinetics.


Assuntos
Benzofenantridinas/farmacocinética , Alcaloides de Berberina/farmacocinética , Fígado/metabolismo , Aminopirina N-Desmetilase/metabolismo , Animais , Benzofenantridinas/sangue , Benzofenantridinas/urina , Alcaloides de Berberina/sangue , Alcaloides de Berberina/urina , Citocromo P-450 CYP3A/metabolismo , Feminino , Inativação Metabólica , Fígado/enzimologia , Masculino , Papaveraceae/química , Quinona Redutases/metabolismo , Ratos , Ratos Sprague-Dawley , Distribuição Tecidual
2.
J Colloid Interface Sci ; 498: 144-152, 2017 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-28324720

RESUMO

To date, the natural alkaloids are mostly used in the field of pharmacological applications and the active substance of palmatine was extensively used in cancer therapy and other biomedical applications. Hence, in this study we report a simple preparation of poly-l-lysine (PLL) electro-polymerized on the surface of functionalized multiwalled carbon nanotubes (f-MWCNT) for electrochemical detection of palmatine content in human serum and urine samples. The active amino group of PLL plays a vital role towards the oxidation palmatine and exhibits superior electrocatalytic activity. Under optimum conditions, the prepared f-MWCNT/PLL composite shows a wide linear response range over the palmatine concentration ranging from 0.5µM to 425µM, and a detection limit (LOD) of 0.12µM based on S/N =3 (signal to noise ratio). The real time monitoring of palmatine content in serum and urine samples displays an appropriate recoveries and excellent performance for the practical analysis. The advantage of this developed system was simple, higher electrocatalytic activity, long-term stability and low cost. We hope that the prepared composite opens a new way for the fabrication of different biosensors in the field of biomedical application.


Assuntos
Alcaloides de Berberina/sangue , Alcaloides de Berberina/urina , Nanotubos de Carbono/química , Polilisina/química , Materiais Biocompatíveis/química , Técnicas Biossensoriais/métodos , Catálise , Técnicas Eletroquímicas , Eletrodos , Humanos , Limite de Detecção , Microscopia Eletrônica de Varredura , Nanocompostos/química , Oxirredução , Tamanho da Partícula , Espectroscopia Fotoeletrônica , Polimerização , Espectroscopia de Infravermelho com Transformada de Fourier , Propriedades de Superfície
3.
Rapid Commun Mass Spectrom ; 31(6): 523-537, 2017 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-28044413

RESUMO

RATIONALE: Palmatine (PAL), a protopalmatine alkaloid, is an active constituent in a number of medicinal plants. In order to obtain a comprehensive and systematic metabolic profile of PAL, we investigated its metabolites in plasma, liver tissue, bile, urine, and feces samples after intragastrical administration to Sprague-Dawley rats with a dose of 100 mg/kg/day. METHODS: In this study, a rapid and sensitive method by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC/QTOF-MS), and Metabolynx™ software with the mass defect filter (MDF) technique was developed for screening and identification of the metabolites. The structural elucidation of the metabolites was performed by comparing their molecular weights and fragment ions with those of the parent drug. RESULTS: As a result, a total of 58 metabolites were identified in rat biological samples including 46 metabolites in urine, 18 metabolites in plasma, 34 metabolites in bile, 26 metabolites in liver tissue, and 10 metabolites in feces. Among them, six major metabolites were fully confirmed using reference standards and others were identified by retention time, accurate mass and fragment ions. CONCLUSIONS: These results indicated that phase I reactions (demethylation and hydroxylation) and phase II reaction (glucuronidation and sulfation) were the main metabolic pathways of PAL in vivo. This research enhances our understanding of metabolism of PAL in rats, and provides useful information on the action mechanism of PAL. Copyright © 2017 John Wiley & Sons, Ltd.


Assuntos
Alcaloides de Berberina/química , Alcaloides de Berberina/metabolismo , Animais , Alcaloides de Berberina/sangue , Alcaloides de Berberina/urina , Cromatografia Líquida de Alta Pressão , Masculino , Metaboloma , Estrutura Molecular , Ratos , Ratos Sprague-Dawley , Espectrometria de Massas em Tandem
4.
J Pharm Biomed Anal ; 128: 371-381, 2016 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-27343900

RESUMO

Levo-tetrahydropalmatine (l-THP) is a tetrahydroprotoberberine isoquinoline alkaloid and has been used as an analgesic agent in China for over 50 years. Recent studies revealed that l-THP was effective in the treatment of drug addiction. However, the plasma metabolic profile, mass balance and clearance pathways of l-THP in human remain unknown. In the present study, an analytical strategy was developed for qualitative and quantitative investigation of metabolism and disposition of l-THP in human. Detection and structural characterization of l-THP metabolites were performed using liquid chromatography-quadrupole time-of-flight mass spectrometry. Selected major metabolites in plasma, urine and feces determined by liquid chromatography with UV detection were further quantified using a triple quadruple mass spectrometry and reference standards. A total of 20 metabolites were identified, most of which were formed via demethylation, mono-hydroxylation, and glucuronidation and sulfonation of desmethyl metabolites. Five major metabolites accounted for over 10% of the parent drug concentration in plasma. Major urinary and fecal metabolites and the parent drug that were monitored for 72h accounted for 46.3% of the dose, while only 0.16% of the dose was the unchanged drug. Multiple demethylations followed by glucuronide and sulfate conjugations and renal excretion were the major drug clearance pathways of l-THP in human.


Assuntos
Alcaloides de Berberina/farmacocinética , Adulto , Alcaloides de Berberina/sangue , Alcaloides de Berberina/urina , Cromatografia Líquida de Alta Pressão , Fezes/química , Humanos , Masculino , Espectrometria de Massas em Tandem
5.
Artigo em Inglês | MEDLINE | ID: mdl-26882127

RESUMO

Corynoline and acetycorynoline, the major active components derived from Corydalis bungeana Herba, showed multiple pharmacological activities. However, quantification of the two compounds in human urine has not been reported. A simple liquid chromatography with tandem mass spectrometry method for the simultaneous determination of corynoline and acetycorynoline in human urine has been developed and fully validated. The analytes were extracted from urine samples by simple liquid-liquid extraction. Chromatographic separation was achieved on a Hedera ODS-2C18 column with the mobile phase of water (containing 0.5% formic acid) and acetonitrile (28:72, v/v) at a flow rate of 0.4mL/min. A tandem mass spectrometric detection was conducted using multiple reaction monitoring via an electrospray ionization source in positive mode. The monitored ion transitions were m/z 368.1→289.1 for corynoline, m/z 410.2→289.2 for acetycorynoline and m/z 380.2→243.2 for donepezil (internal standard), respectively. The calibration curves were linear (correlation coefficients>0.9970) over the concentration ranges of 3.0-3000pg/mL for corynoline and 3.0-1000pg/mL for acetycorynoline. The established method was highly sensitive with the lower limit of quantification (LLOQ) of 3.0pg/mL for both analytes. The intra- and inter-day precision was lower than 10% in terms of relative standard deviation for the low, medium, and high quality control samples, and lower than 16% for the LLOQ samples of the analytes. The accuracy was within ±10% in terms of relative error for both analytes. The method was successfully applied to a urinary excretion study after oral administration of the Chinese medicine formula Shuanghua Baihe tablets in healthy volunteers. The urinary excretion profiles of corynoline and acetycorynoline in human were first reported. The results of this study suggest that renal excretion was not the main excretion pathway of corynoline and acetycorynoline in humans.


Assuntos
Alcaloides de Berberina/urina , Espectrometria de Massas em Tandem/métodos , Administração Oral , Alcaloides de Berberina/administração & dosagem , Cromatografia Líquida/métodos , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/farmacocinética , Feminino , Humanos , Limite de Detecção , Extração Líquido-Líquido/métodos , Masculino
6.
Drug Metab Dispos ; 43(10): 1491-8, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26261285

RESUMO

Corynoline is a 1,3-benzodioxole-containing isoquinoline alkaloid isolated from Corydalis bugeana Turcz., a traditional herbal medicine. Corynoline has reportedly demonstrated multiple pharmacologic properties. Previous studies have also shown that corynoline induced cytotoxicity and inhibited cytochrome P450 (CYP) enzymes, but the mechanisms of the adverse effects remain unknown. The major objective of the present study was to identify reactive metabolites of corynoline responsible for the cytotoxicity and enzyme inhibition. Three oxidative metabolites (M1-M3) were detected by liquid chromatography-tandem mass spectrometry in rat liver microsomal incubations after exposure to corynoline. M1 and M2 were two isomers of catechol derivatives, and M3 was a di-catechol. The M1-M3 metabolites were also observed in urine of rats given corynoline. A total of four N-acetylcysteine (NAC) conjugates (M4-M7) were detected in microsomes containing corynoline, NAC, and NADPH. Apparently, M4 and M5 were derived from M1, M6 resulted from M2, and M7 was a M3-derived NAC conjugate. This indicates that corynoline was bioactivated to ortho-quinone derivatives. No corynoline-derived NAC conjugates (M4-M7) were detected in urine of rats given corynoline; however, three corresponding cysteinylglycine conjugates (M8-M10) were observed instead. Recombinant P450 enzyme incubations demonstrated that the CYPs 2C9, 3A4, and 2C19 were mainly involved in metabolic activation of corynoline. The metabolism study facilitates the understanding of corynoline-induced cytotoxicity and P450 enzyme inhibition.


Assuntos
Alcaloides de Berberina/metabolismo , Alcaloides de Berberina/farmacologia , Microssomos Hepáticos/metabolismo , Animais , Alcaloides de Berberina/urina , Inibidores das Enzimas do Citocromo P-450/metabolismo , Inibidores das Enzimas do Citocromo P-450/farmacologia , Sistema Enzimático do Citocromo P-450/metabolismo , Humanos , Masculino , Microssomos Hepáticos/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Espectrometria de Massas em Tandem/métodos
7.
J Pharm Biomed Anal ; 94: 152-62, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24598170

RESUMO

The characterization and quantification of the metabolites of chiral drugs still remain a great challenge due to the complexity of the metabolites and most of them are not commercially available. In this study, a practical approach based on the combinatory liquid chromatography-mass spectrometric techniques has been proposed for the evaluation of metabolism profiles and urinary excretion kinetics of chiral drugs and their metabolites. Racemic tetrahydropalmatine (rac-THP), a biologically active ingredient isolated from a traditional Chinese herb Rhizoma Corydalis, was chosen as the model chiral drug. Ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) was applied to characterize the metabolites of THP enantiomers in rat urine after administration of (+)-THP or (-)-THP. Accurate mass measurement was used to determine the elemental composition of metabolites and thus to confirm the proposed structures of these metabolites. More than 30 potential metabolites were found in rat urine, most of which were identified for the first time, and the metabolic pathways in vivo were involved in demethylation, oxidation, glucuronide conjugation and sulfation, etc. And the tridesmethlyzed metabolite and didesmethlyzed coupled with oxidation metabolite were found only in (+)-THP treated rats. Afterwards, a liquid chromatography tandem mass spectrometry (LC-QqQ/MS) assay was developed and validated for the determination of the urine level of THP enantiomers and their metabolites. Semi-quantification of three phase I metabolites and two phase II metabolites were performed. Enantiomeric (-/+) cumulative urinary excretion ratios of THP and its five metabolites were obtained, which indicated the stereoselective aspects of metabolites of THP enantiomers in vivo. The study demonstrated the enormous potential of this strategy for the qualitative characterization, quantitative assay and the stereoselectivity of chiral drugs and their metabolites.


Assuntos
Alcaloides de Berberina/química , Alcaloides de Berberina/metabolismo , Animais , Alcaloides de Berberina/urina , Cromatografia Líquida de Alta Pressão/métodos , Corydalis/química , Medicamentos de Ervas Chinesas/química , Masculino , Desintoxicação Metabólica Fase I/fisiologia , Desintoxicação Metabólica Fase II/fisiologia , Ratos , Ratos Sprague-Dawley , Espectrometria de Massas em Tandem/métodos
8.
J Sep Sci ; 37(6): 696-703, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24453165

RESUMO

l-tetrahydropalmatine (l-THP) is a tetrahydroprotoberberine isoquinoline alkaloid that has been used as an analgesic agent in China for more than 40 years. Recent studies indicated its potential application in the treatment of drug addiction. In this study, a sensitive and rapid method using ultra high performance liquid chromatography with MS/MS was developed and validated for simultaneous quantitation of l-THP and its desmethyl metabolites. Enzymatic hydrolysis was integrated into sample preparation to enable the quantitative determination of both free and conjugated metabolites. Chromatographic separation was achieved on an Agilent Poroshell 120 EC-C18 column. Detection was performed by MS in the positive ion ESI mode. The calibration curves of the analytes were linear (r(2) > 0.9936) over the concentration range of 1-1000 ng/mL with the lower limit of quantification at 1 ng/mL. The precision for both intra- and interday determinations was <8.97%, and the accuracy ranged from -8.74 to 8.65%. The recovery for all the analytes was >70% without significant matrix effect. The method has been successfully applied to the urinary excretion study of l-THP in rats. The conjugates were found to be the major urine metabolites of the drug.


Assuntos
Alcaloides de Berberina/análise , Alcaloides de Berberina/urina , Alcaloides de Berberina/metabolismo , Cromatografia Líquida de Alta Pressão , Humanos , Estrutura Molecular , Espectrometria de Massas em Tandem
9.
Spectrochim Acta A Mol Biomol Spectrosc ; 122: 198-203, 2014 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-24316533

RESUMO

This report presents a highly sensitive, poly(A)-stabilized gold nanoparticle-based assay with dual readouts (resonance light scattering and colorimetric) for detecting palmatine hydrochloride (PaH) in real samples. The detection mechanism is based on the fact that palmatine hydrochloride has strong affinity to poly(A), which can stabilize gold nanoparticles at high ionic strength, and cause the aggregation of poly(A)-stabilized AuNPs, resulting in the enhanced resonance light scattering (RLS). At the same time, the color change of poly(A)-stabilized AuNPs solution is from red to blue via purple. Thus a highly sensitive RLS assay for PaH has been developed with a linear range of 0.023-2.5 µg/mL. The limit of detection (LOD, 3σ) is 2.3 ng/mL. In this work, the reaction mechanism of this system was investigated by scanning electron microscope (SEM), dark-field light scattering images (DLSI), dynamiclight scattering (DLS) and circular dichroism (CD). This proposed method was also applied successfully for the determination of PaH in pharmaceutical preparations and urine samples with RSD⩽4.0%. The results are in good agreement with those from the official method.


Assuntos
Alcaloides de Berberina/análise , Bioensaio/métodos , Ouro/química , Nanopartículas Metálicas/química , Poli A/química , Adulto , Alcaloides de Berberina/química , Alcaloides de Berberina/urina , Cápsulas , Dicroísmo Circular , Colorimetria , Humanos , Luz , Nanopartículas Metálicas/ultraestrutura , Espalhamento de Radiação , Espectrofotometria Ultravioleta , Comprimidos
10.
Appl Spectrosc ; 67(7): 747-51, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23816126

RESUMO

A post-chemiluminescence (PCL) phenomenon was observed when palmatine was injected in a mixture of N-bromosuccinimide (NBS) and alkaline dichlorofluorescein (DCF) after the chemiluminescence (CL) reaction of NBS-alkaline DCF had finished. Based on the PCL reaction, a rapid and sensitive method for the determination of palmatine was established. Under optimum conditions, the CL intensity was linear, with the concentration of palmatine in the range of 5.0 × 10(-9) to 1.0 × 10(-6) M. The detection limit was 6.0 × 10(-10) M for palmatine. The relative standard deviation for 11 parallel measurements of 1.0 × 10(-7) M palmatine was 1.5%. This method was applied to the determination of palmatine in pharmaceutical samples and biological fluids, with satisfactory results. The possible reaction mechanism is discussed briefly.


Assuntos
Alcaloides de Berberina/sangue , Alcaloides de Berberina/urina , Misturas Complexas/sangue , Misturas Complexas/urina , Análise de Injeção de Fluxo/instrumentação , Medições Luminescentes/instrumentação , Alcaloides de Berberina/química , Misturas Complexas/química , Desenho de Equipamento , Análise de Falha de Equipamento , Análise de Injeção de Fluxo/métodos , Medições Luminescentes/métodos
11.
J Sep Sci ; 34(22): 3279-86, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22028314

RESUMO

Dispersive liquid-liquid microextraction (DLLME) coupled with high-performance liquid chromatography (HPLC)-UV detection was applied in rat urine for the extraction and determination of tetrahydropalmatine (THP) and tetrahydroberberine (THB), both active components in Rhizoma corydalis. Various parameters affecting the extraction efficiency, such as the type and volume of extraction and dispersive solvent, pH, etc. were evaluated. Under the optimal conditions (extraction solvent: 37 µL of chloroform, dispersive solvent: 100 µL of methanol, alkaline with 100 µL of 1 mol/L NaOH, and without salt addition), the enrichment factors of THP and THB were more than 30. The extraction recoveries were 69.8-75.8% and 72.7-77.6% for THP and THB in rat urine, respectively. Both THP and THB showed good linearity in the range of 0.025-2.5 µg/mL, and the limit of quantification was 0.025 µg/mL (S/N=10, n=6). The intra-day and inter-day precision of THP and THB were <12.6%. The relative recoveries ranged from 95.5 to 107.4% and 96.8 to 100.9% for THP and THB in rat urine, respectively. The method has been successfully applied to rat urine samples. The results demonstrated that DLLME is a very simple, rapid and efficient method for the extraction and preconcentration of THP and THB from urine samples.


Assuntos
Alcaloides de Berberina/isolamento & purificação , Alcaloides de Berberina/urina , Berberina/análogos & derivados , Cromatografia Líquida de Alta Pressão/métodos , Medicamentos de Ervas Chinesas/análise , Medicamentos de Ervas Chinesas/isolamento & purificação , Microextração em Fase Líquida/métodos , Animais , Berberina/isolamento & purificação , Berberina/urina , Ratos
12.
Planta Med ; 76(16): 1859-63, 2010 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-20549593

RESUMO

Coptidis Rhizoma has been used as a traditional Chinese herbal medicine to treat typhoid, pharyngolaryngitis, diabetes mellitus, gastroenteritis and secretory diarrhea for more than a thousand years in China. However, there is little information on the IN VIVO chemical constituents of Coptidis Rhizoma following oral administration. In this paper, the alkaloid constituents in urine were studied in humans following oral administration of Coptidis Rhizoma decoction. Using macroporous adsorption resin chromatography, open ODS column chromatography, and preparative high-performance liquid chromatography, twelve protoberberine alkaloid constituents were isolated. Their structures were elucidated by chemical evidence, enzymatic deconjugation and analyses of mass, (1)H-NMR and NOESY spectra. The identified alkaloid constituents include berberine ( P1), groenlandicine 3-O- ß-D-glucuronide (M1), dehydrocheilanthifoline 2-O-ß-D-glucuronide (M2), thalifendine 10-O-ß-D-glucuronide (M3), jatrorrhizine 3-O-ß-D-glucuronide (M4), columbamine 2-O-ß-D-glucuronide (M5), berberrubine 9-O-ß-D-glucuronide (M6), jatrorrhizine 3-O-sulfate (M7), demethyleneberberine 2-O-sulfate (M8), dehydrocorydalmine 10-O-sulfate (M9), 3,10-demethylpalmatine 10-O-sulfate (M10) and 2,3,10-trihydroxyberberine 2-O-sulfate ( M11). No other parent protoberberine alkaloids from Coptidis Rhizoma except for a trace of berberine were found in the urine. These findings suggested that the protoberberine alkaloids, which were absorbed in vivo following oral administration of Coptidis Rhizoma decoction, were mainly conjugated with glucuronic acid or sulfuric acid to form phase II metabolites directly or after biotransformation to phase I metabolites, and finally excreted in urine.


Assuntos
Alcaloides de Berberina/metabolismo , Coptis/química , Extratos Vegetais/metabolismo , Administração Oral , Adulto , Alcaloides de Berberina/urina , Biotransformação , Humanos , Inativação Metabólica , Masculino , Estrutura Molecular , Extratos Vegetais/química , Extratos Vegetais/urina , Rizoma , Adulto Jovem
13.
Chirality ; 22(3): 355-60, 2010 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-19544352

RESUMO

The objective of this work was to study the stereoselectivity in excretion of tetrahydropalmatine (THP) enantiomers by rats and identify the metabolites of racemic THP (rac-THP) in rat urine. Urine and bile samples were collected at various time intervals after a single oral dose of rac-THP. The concentrations of THP enantiomers in rat urine and bile were determined using a modification of an achiral-chiral high-performance liquid chromatographic (HPLC) method that had been previously published. The cumulative urinary excretion over 96 h of (-)-THP and (+)-THP was found to be 55.49 +/- 36.9 microg and 18.33 +/- 9.7 microg, respectively. The cumulative biliary excretion over 24 h of (-)-THP and (+)-THP was 19.19 +/- 14.6 microg and 12.53 +/- 10.4 microg, respectively. The enantiomeric (-/+) concentration ratios of THP changed from 2.80 to 5.15 in urine, and from 1.36 to 1.80 in bile. The mean cumulative amount of (-)-THP was significantly higher than that of (+)-THP both in urine and bile samples. However, the enantiomeric (-/+) concentration ratios in rat urine and bile were significantly lower than those ratios in rat plasma. These findings suggested the excretion of THP enantiomers was stereoselective rather than a reflection of chiral pharmacokinetic aspects in plasma and (-)-THP was preferentially excreted in rat urine and bile. Three O-demethylation metabolites and the parent drug rac-THP were detected by liquid chromatography-tandem mass spectrometry in rat urine. One metabolite was obtained by preparative HPLC and identified as 10-O-demethyl-THP.


Assuntos
Alcaloides de Berberina/urina , Bile/química , Medicamentos de Ervas Chinesas/química , Espectrometria de Massas/métodos , Animais , Cromatografia Líquida de Alta Pressão/métodos , Ratos , Ratos Sprague-Dawley , Estereoisomerismo
14.
J Pharm Pharmacol ; 61(5): 647-52, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-19406004

RESUMO

OBJECTIVES: Despite its important therapeutic value, the metabolism of palmatine is not yet clear. Our objective was to investigate its in-vivo and in-vitro metabolism. METHODS: Liquid chromatography-tandem electrospray ionization mass spectrometry (LC-ESI/MSn) was employed in this work. In-vivo samples, including faeces, urine and plasma of rats, were collected after oral administration of palmatine (20 mg/kg) to rats. In-vitro samples were prepared by incubating palmatine with intestinal flora and liver microsome of rats, respectively. All the samples were purified via a C18 solid-phase extraction procedure, then chromatographically separated by a reverse-phase C18 column with methanol-formic acid aqueous solution (pH 3.5, 70:30 v/v) as mobile phase, and detected by an on-line MSn detector. The structure of each metabolite was elucidated by comparing its molecular weight, retention time and full-scan MSn spectra with those of the parent drug. KEY FINDINGS: The results revealed that 12 metabolites were present in rat faeces, 13 metabolites in rat urine, 7 metabolites in rat plasma, 10 metabolites in rat intestinal flora and 9 metabolites in rat liver microsomes. Except for six of the metabolites in rat urine, the other in-vivo and in-vitro metabolites were reported for the first time. CONCLUSIONS: Seven new metabolites of palmatine (tri-hydroxyl palmatine, di-demethoxyl palmatine, tri-demethyl palmatine, mono-demethoxyl dehydrogen palmatine, di-demethoxyl dehydrogen palmatine, mono-demethyl dehydrogen palmatine, tri-demethyl dehydrogen palmatine) were reported in this work.


Assuntos
Alcaloides de Berberina/análise , Animais , Alcaloides de Berberina/sangue , Alcaloides de Berberina/urina , Cromatografia Líquida , Fezes/química , Feminino , Masculino , Microssomos Hepáticos/metabolismo , Ratos , Ratos Wistar , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em Tandem
15.
Electrophoresis ; 30(4): 661-7, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19156769

RESUMO

New methods based on MEEKC coupling with field-amplified sample injection (FASI) induced by ACN were proposed for five isoquinoline alkaloids (berberine, palmatine, jatrorrhizine, sinomenine and homoharringtonine) in no salt and high salt sample solution (HS). For the separation of five isoquinoline alkaloids, a running buffer composed of 18 mM sodium cholate, 2.4% v/v butan-1-ol, 0.6% v/v ethyl acetate, 10% v/v (or 30% v/v) methanol and 87.0% v/v (or 67% v/v) 5 mM Na2B4O7~10 mM NaH2PO4 buffer (pH 7.5) was developed. In order to improve the sensitivity, FASI induced by ACN was applied to increase the detection sensitivity. The detection limit was found to be as low as 0.0002 microg/mL in no salt sample solution and 0.062 microg/mL in HS. The method has been applied for the analysis of human urine spiked with analytes, and the assay results were proved to be satisfactory, and also the determination of berberine in urine sample after oral administration berberine.


Assuntos
Acetonitrilas/química , Alcaloides/urina , Cromatografia Capilar Eletrocinética Micelar/métodos , 1-Butanol/química , Acetatos/química , Alcaloides de Berberina/urina , Harringtoninas/urina , Mepesuccinato de Omacetaxina , Humanos , Concentração de Íons de Hidrogênio , Metanol/química , Morfinanos/urina , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Dodecilsulfato de Sódio/química , Água/química
16.
Biopharm Drug Dispos ; 28(9): 511-6, 2007 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17891749

RESUMO

A high-performance liquid chromatographic method with ultraviolet detection was established and validated for quantification of three alkaloids (coptisine, palmatine and berberine) in rat urine. Following a single-step liquid-liquid extraction, the analytes were separated on a reversed-phase C(18) column with water-formic acid-triethylamine-methanol as the mobile phase at a flow rate of 1 ml/min. The linear ranges of the calibration curves were 1.6-160 ng/ml for all three alkaloids. The lower limit of quantification was 1.6 ng/ml for all three alkaloids. The within-batch accuracy was 90.4-108.3% for coptisine, 88.6-107.8% for berberine and 88.4-110.1% for palmatine. The between-batch accuracy was 99.3-100.3% for coptisine, 94.3-100.6% for berberine and 93.7-100.0% for palmatine. The within-batch and between-batch precisions were

Assuntos
Alcaloides de Berberina/urina , Cromatografia Líquida de Alta Pressão/métodos , Coptis/química , Plantas Medicinais/química , Administração Oral , Animais , Berberina/análogos & derivados , Berberina/urina , Medicamentos de Ervas Chinesas/química , Feminino , Masculino , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Rizoma
17.
Rapid Commun Mass Spectrom ; 21(13): 2019-22, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17534853

RESUMO

Palmatine is an isoquinoline alkaloid that has been widely used in China for the treatment of various inflammatory diseases such as gynecological inflammation, bacillary dysentery, enteritis, respiratory tract infection, urinary infection, etc. In the study reported in this paper, a simple and rapid high-performance liquid chromatography/electrospray ionization (ESI) tandem mass spectrometric method (MS/MS) was developed for elucidation of the structures of metabolites of palmatine in rat urine after administration of a single dose (20 mg/kg). The rat urine samples were collected and purified through C18 solid-phase extraction cartridges, and then injected onto a reversed-phase C18 column with 60:40 (v/v) methanol/0.01% triethylamine solution (2 mM, adjusted to pH 3.5 with formic acid) as mobile phase and detected by on-line MS/MS. Identification of the metabolites and elucidation of their structures were performed by comparing changes in molecular masses (DeltaM), retention times and spectral patterns of product ions with those of the parent drug. As a result, six phase I metabolites, the parent drug palmatine and two phase II metabolites were identified in rat urine for the first time.


Assuntos
Alcaloides de Berberina/metabolismo , Alcaloides de Berberina/urina , Cromatografia Líquida/métodos , Redes e Vias Metabólicas/fisiologia , Espectrometria de Massas em Tandem/métodos , Animais , Alcaloides de Berberina/análise , Alcaloides de Berberina/química , Estrutura Molecular , Ratos , Ratos Wistar , Espectrometria de Massas por Ionização por Electrospray , Fatores de Tempo
18.
Biomed Chromatogr ; 20(1): 95-100, 2006 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-15959869

RESUMO

L-tetrahydropalmatine (L-THP) is an active alkaloid from Stephania ainiaca Diels. In order to compare the similarities and differences of microbial and mammalian metabolisms of L-THP, the microbial transformation by Penicillium janthinellum and metabolism in rats were investigated. Biotransformation of L-THP by Penicillium janthinellum AS 3.510 resulted in the formation of three metabolites. Their structures were identified as L-corydalmine, L-corypalmine and 9-O-desmethyl-L-THP, respectively, by comprehensive nuclear magnetic resonance and mass spectrometry (MS) analysis. Six metabolites (M1-M6) were detected from the in vivo study in rats and three of which (L-corydalmine, L-corypalmine and 9-O-desmethyl-L-THP) were identified as the same compounds as those obtained from microbial metabolism by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis and comparison with reference standards obtained from microbial metabolism. The structures of the additional three metabolites were tentatively deduced as 2-O-desmethyl-L-THP and two di-O-demethylated L-THP by LC-MS/MS analysis. Time courses of microbial and rat metabolisms of L-THP were also investigated.


Assuntos
Alcaloides de Berberina/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Espectrometria de Massas/métodos , Penicillium/metabolismo , Animais , Alcaloides de Berberina/urina , Biotransformação , Fezes/química , Espectroscopia de Ressonância Magnética , Masculino , Ratos , Ratos Sprague-Dawley , Espectrofotometria Ultravioleta
19.
Artigo em Inglês | MEDLINE | ID: mdl-15458726

RESUMO

Protopine was extracted from Fumaria officinalis and purified by column chromatography. Urine samples were collected from horses and a human volunteer that had been administered either F. officinalis or protopine free base. Plant and urine samples were acetylated and analysed by GCMS after solid-phase extraction (SPE). The urinary metabolites of protopine were identified as 4,6,7,13-tetrahydro-9,10-dihydroxy-5-methyl-benzo[e]-l,3-benzodioxolo [4,5-1][2] benzazecin-12(5H)-one, 4,6,7,13-tetrahydro-10-hydroxy-9-methoxy-5-methyl-benzo[e]-1,3-benzodioxolo[4,5-1][2] benzazecin-12(5H)-one and 4,6,7,13-tetrahydro-9-hydroxy-10-methoxy-5-methyl-benzo[e]-1,3-benzodioxolo[4,5-l][2] benzazecin-12(5H)-one, chelianthifoline, isochelianthifoline and 2-O-desmethylchelianthifoline. The metabolic formation of the tetrahydroprotoberberines by closure of the bridge across N5 and C13 is rate limited and protopine-like metabolites accumulate only when the route is overloaded. Metabolism was qualitatively similar in the horse and human.


Assuntos
Alcaloides de Berberina/urina , Animais , Benzofenantridinas , Cromatografia Gasosa-Espectrometria de Massas , Cavalos
20.
Artigo em Inglês | MEDLINE | ID: mdl-12726842

RESUMO

Eschscholtzia californica preparations are in use as phytopharmaceuticals and as herbal drugs. Studies are described on the metabolism and the toxicological analysis of the Eschscholtzia californica alkaloids californine and protopine in rat urine using gas chromatography-mass spectrometry. The identified metabolites indicated that californine is extensively metabolized by N-demethylation and/or single or double demethylenation with consecutive catechol-O-methylation of one of the hydroxy groups. Protopine, however, only undergoes extensive demethylenation of the 2,3-methylenedioxy group followed by catechol-O-methylation. All phenolic hydroxy metabolites were found to be partly conjugated. The authors' systematic toxicological analysis procedure using full-scan gas chromatography-mass spectrometry after acid hydrolysis, liquid-liquid extraction and microwave-assisted acetylation allowed the detection of the main metabolites of californine and protopine in rat urine after a dose which should correspond to that of drug users. Therefore, use of Eschscholtzia californica preparations should also be detectable in human urine by the authors' systematic toxicological analysis procedure.


Assuntos
Alcaloides de Berberina/toxicidade , Eschscholzia/química , Cromatografia Gasosa-Espectrometria de Massas/métodos , Isoquinolinas/toxicidade , Animais , Benzofenantridinas , Alcaloides de Berberina/urina , Isoquinolinas/urina , Masculino , Ratos
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