RESUMEN
AIM: To identify the structure of unknown metabolites of berberine (Ber) in human urine after oral administration. METHODS: Urine samples were obtained from 5 volunteers after they orally took Ber chloride 0.9 g per day for three days. Metabolites in urine samples were isolated and purified by polyporous resin column chromatography. The individual metabolites were identified mainly using electrospray ionization mass spectroscopy (ESI-MS) and proton nuclear magnetic resonance (1H NMR) spectroscopy. RESULTS: Three unknown metabolites (M1, M2, and M3) were isolated. They were susceptible to arylsufatase. ESI-MS measurements of M1, M2, and M3 produced quasimolecular ions [M+H]+, m/z 17.9, 404.0, and 402.0 respectively. Especially, each of them produced a characteristic protonated ion [M-80+H]+, which can be ascribed as quasimolecular ions lost a SO3 fragment. 1H NMR spectra of the metabolites were also obtained and each of 1H signals was assigned. CONCLUSION: Structures of M1, M2, and M3 were firmly identified as jatrorrhizine-3-sulfate, demethyleneberberine-2-sulfate, and thalifendine-10-sulfate, and the major metabolite was M2.
Asunto(s)
Berberina/análisis , Berberina/metabolismo , Berberina/orina , Sulfatos/metabolismo , Adulto , Antidiarreicos/metabolismo , Antidiarreicos/orina , Berberina/análogos & derivados , Berberina/farmacocinética , Humanos , Espectroscopía de Resonancia Magnética , Masculino , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
It has been demonstrated that low concentrations of basic lipophilic drugs in biological fluids may be extracted and concentrated 10(4)-10(6) times by a series of extraction procedures in which the ratio of the extracting solvent to that of the solution to be extracted is of the order of 1:100 (microphase extraction procedure). Typically, basic drugs (atropine, aminopentamide, hyoscine, chloroquine, pyrimethamine) were extracted and concentrated sufficiently for direct GC analysis from 24-hour urine samples by a procedure involving three simple consecutive extraction steps. Using this procedure, it was demonstrated that after administration of aminopentamide (300 micrograms) to patients in the form of anti-diarrhoeal tablets, measurable quantities of the free, unchanged drug can be demonstrated in 24-hour urine samples. The main advantages of the method are simplicity, rapidity and sensitivity due to the low background interference in the GC separations. The principle involved can be extended to the analysis of acidic drugs with suitable solubility properties.