RESUMO
In this work, a new sample-preparation method based on hollow-fiber liquid-phase microextraction (HF-LPME) was developed for analysis of magnoflorine in rat plasma. Analysis was accomplished by reversed-phase high-performance liquid chromatography (HPLC), with ultraviolet detection by use of a photodiode-array detector. An orthogonal array design (OAD) was found to be effective for optimization of major conditions which may affect the efficiency of HF-LPME. Under the optimized conditions (pH of donor and acceptor phases 12 and 2.0, respectively; extraction time 20 min; stirring speed 800 rpm; and addition of 10 % (w/v) salt), the preconcentration factor for magnoflorine was 355. Calibration curves with reasonable linearity (r(2)≥0.9994) were obtained in the range 10-1000 ng mL(-1). Intra-day and inter-day precision (RSD) were <5.5 % and the limit of detection (LOD) for the analyte was 3.0 ng mL(-1) (S/N=3). The validated method was successfully used for pharmacokinetic studies of magnoflorine in rat plasma after intravenous administration.
Assuntos
Aporfinas/sangue , Cromatografia Líquida de Alta Pressão/métodos , Microextração em Fase Líquida/métodos , Animais , Aporfinas/farmacocinética , Calibragem , Limite de Detecção , Masculino , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos TestesRESUMO
A simple, rapid and specific method based on cloud-point extraction (CPE) was developed to determine ampelopsin in rat plasma after oral administration by reversed-phase high-performance liquid chromatography. The non-ionic surfactant Genapol X-080 was chosen as the extract solvent. Some important parameters affecting the CPE efficiency, such as the nature and concentration of surfactant, extraction temperature and time, centrifuge time and salt effect, were investigated and optimized. Separation was accomplished using a C(18) column by gradient elution with a acetonitrile-phosphate buffer solution as the mobile phase. The detection wavelength was set at 290 nm. Under optimum conditions, the linear range of ampelopsin in rat plasma was 20-2000 ng/mL (r(2)=0.9996). The limit of detection was 6 ng/mL (S/N=3) with the limit of quantification being 20 ng/mL (S/N=10). The proposed method has been successfully applied for pharmacokinetic studies of ampelopsin from rat plasma after oral administration.
Assuntos
Ampelopsis/química , Fracionamento Químico/métodos , Cromatografia Líquida de Alta Pressão/métodos , Medicamentos de Ervas Chinesas/isolamento & purificação , Medicamentos de Ervas Chinesas/farmacocinética , Flavonoides/isolamento & purificação , Flavonoides/farmacocinética , Animais , Flavonoides/sangue , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
A simple, inexpensive and efficient method based on the mixed cloud point extraction (MCPE) combined with high performance liquid chromatography was developed for the simultaneous separation and determination of six flavonoids (rutin, hyperoside, quercetin-3-O-sophoroside, isoquercitrin, astragalin and quercetin) in Apocynum venetum leaf samples. The non-ionic surfactant Genapol X-080 and cetyl-trimethyl ammonium bromide (CTAB) was chosen as the mixed extracting solvent. Parameters that affect the MCPE processes, such as the content of Genapol X-080 and CTAB, pH, salt content, extraction temperature and time were investigated and optimized. Under the optimized conditions, the calibration curve for six flavonoids were all linear with the correlation coefficients greater than 0.9994. The intra-day and inter-day precision (RSD) were below 8.1% and the limits of detection (LOD) for the six flavonoids were 1.2-5.0 ng mL(-1) (S/N=3). The proposed method was successfully used to separate and determine the six flavonoids in A. venetum leaf samples.
Assuntos
Apocynum/química , Flavonoides/química , Extratos Vegetais/química , Folhas de Planta/química , Cetrimônio , Compostos de Cetrimônio/química , Cromatografia Líquida de Alta Pressão/métodos , Concentração de Íons de Hidrogênio , Quempferóis/química , Polietilenoglicóis/química , Quercetina/análogos & derivados , Quercetina/química , Rutina/química , Tensoativos/químicaRESUMO
A simple, rapid and specific method was developed to separate as well as to determine the four phenylethanoid glycosides (PhGs) (echinacoside, tubuloside B, acteoside and isoacteoside) in rat plasma after oral administration of Cistanche salsa extract by reversed phase high performance liquid chromatography using a microemulsion as the mobile phase. The separations were performed on a Zorbax Extend-C18 column at 25°C. Photodiode-array detector was conducted at 322nm and with a flow rate of 0.8mLmin(-1). The optimized microemulsion mobile phase consisted of 0.3% triethylamine in 20mM phosphoric acid at pH 6.0, 0.8% (v/v) ethyl acetate as oil phase, 1.5% (v/v) Genapol X-080 as surfactant, 2.5% (v/v) n-propanol as co-surfactant. Under the optimal conditions, the calibration curve for four PhGs was linear in the range of 10-1000ngmL(-1) with the correlation coefficients greater than 0.9994. The intra-day and inter-day precision (RSD) were below 8.64% and the limits of detection (LOD) for the four PhGs were 0.4-1.3ngmL(-1) (S/N=3). The microemulsion liquid chromatography (MELC) method was successfully applied to separate and determine the four PhGs in rat plasma after oral administration of C. salsa extract.
Assuntos
Cromatografia Líquida/métodos , Cistanche/química , Glicosídeos/sangue , Extratos Vegetais/administração & dosagem , Administração Oral , Animais , Cromatografia Líquida de Alta Pressão , Estabilidade de Medicamentos , Glicosídeos/química , Glicosídeos/isolamento & purificação , Concentração de Íons de Hidrogênio , Masculino , Extratos Vegetais/química , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos TestesRESUMO
A simple and solvent-minimized sample preparation technique based on two-phase hollow fiber liquid phase microextraction (HF-LPME) was developed and used to quantify the echinacoside in Parkinson's disease rat plasma following oral administration. Analysis was accomplished by reversed-phase high performance liquid chromatography (HPLC) with ultraviolet detection. The influence factors relevant to the HF-LPME processes were optimized. Under the optimized conditions, the preconcentration factor for echinacoside was 337. Calibration curves with reasonable linearity (r(2) ≥ 0.9998) were obtained in the range of 5-750 ng mL(-1). Intra-day and inter-day precision (RSD) were ≤ 5.43% and the limit of detection (LOD) for the analyte was 2.0 ng mL(-1) (S/N=3). The validated method has been successfully applied for pharmacokinetic studies of echinacoside in Parkinson's disease (PD) rat plasma after oral administration.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Glicosídeos/farmacocinética , Microextração em Fase Líquida/métodos , Transtornos Parkinsonianos/tratamento farmacológico , Administração Oral , Animais , Calibragem , Cromatografia de Fase Reversa/métodos , Glicosídeos/administração & dosagem , Limite de Detecção , Masculino , Transtornos Parkinsonianos/fisiopatologia , Ratos , Ratos WistarRESUMO
A simple and solvent-minimized sample preparation technique based on two-phase hollow fiber liquid phase microextraction has been developed and used to quantify the osthole in cerebral ischemia reperfusion rat plasma following oral administration. The analysis was performed by reversed phase high performance liquid chromatography with fluorescence detection. Extraction conditions such as solvent identity, agitation rate, salt concentration, extraction time, and length of the hollow fiber were optimized. Under the optimized conditions, the linear range of osthole in rat plasma was 5-500 ng mL(-1) (r(2) = 0.9997). The limit of detection (LOD) was 2 ng mL(-1) (S/N = 3) with limit of quantification (LOQ) being 5 ng mL(-1). The validated method has been successfully applied for pharmacokinetic studies of osthole from cerebral ischemia reperfusion rat plasma after oral administration.