Simultaneous determination of active xanthone glycosides, timosaponins and alkaloids in rat plasma after oral administration of Zi-Shen Pill extract for the pharmacokinetic study by liquid chromatography-tandem mass spectrometry.

A sensitive and reliable liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) has been developed and validated for simultaneous determination of active components, i.e., xanthone glycosides (neomangiferin and mangiferin), timosaponins (timosaponin E1, timosaponin B-II and timosaponin B) and alkaloids (palmatine and berberine) in rat plasma after oral administration of Zi-Shen Pill extract. Plasma samples were pretreated by protein precipitation with acetonitrile containing the internal standards ginsenoside Re (for xanthone glycosides and timosaponins) and tetrahydroberberine (for alkaloids). LC separation was achieved on a Zorbax SB-C(18) column (150 mm x 2.1 mm I.D., 3.5 microm) with gradient elution using a mobile phase consisting of acetonitrile-0.1% formic acid in water at a flow rate of 0.25 mL/min. The detection was carried out by a triple-quadrupole tandem mass spectrometer in multiple reaction monitoring (MRM) mode via polarity switching between negative (for xanthone glycosides and timosaponins) and positive (for alkaloids) ionization mode. Linear calibration curves were obtained over the concentration range of 5-1000 ng/mL for mangiferin, 0.5-100 ng/mL for neomangiferin, timosaponin E1, timosaponin B-II and timosaponin B, and 0.05-10 ng/mL for palmatine and berberine. The mean recovery of all the analytes ranged from 64.7 to 93.8%. The intra- and inter-day precision (% R.S.D.) was within 11.7% and accuracy (% bias) ranged from -9.0 to 10.9%. This fully validated method was successfully applied to pharmacokinetic study of the above seven compounds in rats.

Cai F ，Xu W ，Wei H ，Sun L ，Gao S ，Yang Q ，Feng J ，Zhang F ，Chen W ... -  《-》

Simultaneous determination of timosaponin B-II and A-III in rat plasma by LC-MS/MS and its application to pharmacokinetic study.

A rapid, specific and sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed and validated for the simultaneous determination of timosaponin B-II (TB-II) and A-III (TA-III) in rat plasma. Plasma samples were pretreated via simple protein precipitation with acetonitrile and ginsenoside Rg2 was used as internal standard. Chromatographic separation was carried out on an Agilent XDB-C8 (150 mm × 2.1mm i.d., 5 μm) column by isocratic elution with acetonitrile-2 mmol/L ammonium acetate (55:45, v/v). The detection was performed on a Sciex API 4000(+) triple-quadrupole tandem mass spectrometer with TurboIonSpray ionization (ESI) inlet via the negative ion multiple reaction monitoring (MRM) mode. The results showed that the calibration curve was linear in the concentration range of 3-3,000 ng/mL for TB-II and 0.3-3,000 ng/mL for TA-III, respectively. The intra- and inter-day precisions were less than 13.25%, and the accuracy ranged from 100.88% to 104.07% at three QC levels for both. The pharmacokinetic profiles of TB-II and TA-III in timosaponins (total timosaponin) at three dose levels (TB-II 150, 300, 600 mg/kg and TA-III 0.59, 1.17, 2.34 mg/kg, respectively) and in timosaponins-Huangbai alkaloids mixtures (1:1, 1:3, w/w, TB-II 300 mg/kg and TA-III 1.17 mg/kg) were studied for the first time in rats by this LC-MS/MS method. After single oral administration of timosaponins, mean Cmax and AUC0-t of TB-II and TA-III increased but non-proportional to the oral doses. When timosaponins-Huangbai alkaloids (1:1, 1:3, w/w) mixtures were administered, Cmax and AUC0-t of TB-II in the mixtures were obviously higher than the corresponding values in timosaponins at the same dose level.

Feng Y ，Chen B ，Lin A ，Liu Y ... -  《-》

A rapid and sensitive liquid chromatography-tandem mass spectrometric method for the determination of timosaponin B-II in blood plasma and a study of the pharmacokinetics of saponin in the rat.

Cai F ，Sun L ，Gao S ，Yang Y ，Yang Q ，Chen W ... -  《JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS》

Simultaneous determination of active flavonoids and alkaloids of Tang-Min-Ling-Pill in rat plasma by liquid chromatography tandem mass spectrometry.

A rapid, sensitive and reliable liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed and validated for simultaneous determination of five active flavonoids (wogonin, chrysin, oroxylin A, naringenin, hesperetin) and four major alkaloids (berberine, coptisine, jatrorrhizine, palmatine) from Tang-Min-Ling-Pill in rat plasma. Plasma samples (100 μL) were spiked with internal standards daidzein (for flavonoids) and tetrahydropalmatine (for alkaloids), acidified with HCl and extracted by liquid-liquid extraction with acetone. Chromatographic separation was performed on a Zorbax SB-C(18) column with the mobile phase of water (containing 0.1% of formic acid)-acetonitrile (30:70, v/v) at a flow rate of 0.3 mL/min in a run time of 7.0 min. Detection was performed by multiple reaction monitoring mode using electrospray ionization in the positive ion mode. All analytes showed good linearity over the investigated concentration range (r>0.9900). The validated lower limit of quantification was 1.01 ng/mL for wogonin and oroxylin A, 0.238 ng/mL for chrysin, 1.01 ng/mL for naringenin, 0.998 ng/mL for hesperetin, 0.0505 ng/mL for berberine, 0.0996 ng/mL for coptisine, 0.0501 ng/mL for jatrorrhizine, 0.0889 ng/mL for palmatine, respectively. Intra- and inter-day precision (RSD%) was less than 15% and accuracy (RE%) ranged from -7.5% to 4.5%. The validated method was successfully applied to investigate the pharmacokinetics of the major flavonoids and alkaloids of Tang-Min-Ling-Pill after oral administration to rats.

Zhu Y ，Tong L ，Zhou S ，Sun H ，Bi K ，Zhang B ... -  《-》

Simultaneous determination of six alkaloids and one monoterpene in rat plasma by liquid chromatography-tandem mass spectrometry and pharmacokinetic study after oral administration of a Chinese medicine Wuji Pill.

A simple and sensitive method for simultaneous determination of seven active constituents, jatrorrhizine, berberine, coptisine, palmatine, evodiamine, rutacarpine and paeoniflorin, from a Chinese medicine Wuji Pill in rat plasma was developed based on a liquid chromatography and tandem mass spectrometry method. The separation of these seven compounds was carried out on a Shiseido CAPCELL PAK C(18) column using a mobile phase consisting of acetonitrile (containing 0.1% formic acid and water (containing 0.1% formic acid and 10 mmol/L ammonium acetate) and carbamazepine as an internal standard. Electrospray ionization in positive-ion mode and multiple reaction monitoring was used to identify and quantitate active components. All calibration curves gave good linearity (r>0.993) over the concentration range from 0.42-208.0 ng/mL to 4.18-418.0 ng/mL for all components. The precision of the in vivo study was evaluated by intra- and inter-day assays and the percentages of relative standard deviation were all within 15%. The method was successfully applied to pharmacokinetic study of all six alkaloids and one monoterpene in rat plasma after oral administration of the Wuji Pill.

Yuan J ，Wang Y ，An R ，Wang S ，Li SJ ，Jia JY ，Bligh SW ，Wang XH ，Ma YM ... -  《-》