Effect of Gegen Qinlian decoction on hepatic cytochrome CYP450 isozymes in rats by HPLC-MS/MS / 中国中药杂志

To study the effect of Gegen Qinlian decoction and its major effective components on five hepatic microsomal CYP450 isozymes in rats. The in vitro hepatic microsomal incubation technique was used to co-culture Gegen Qinlian decoction and its major effective components together with each probe substrate. HPLC-MS/MS was used to establish the analytical method for metabolites of the five isoform probe substrates of CYP450 isozymes, detect the linearity among micoromal protein concentration, incubation time and metabolite formation amount. And HPLC-MS/MS was applied to determine the formation rate (V) of corresponding metabolites (acetaminophen, 4-OH-chlorzoxazone, dextrophan, 6-OH-chlorzoxazone and 6β-hydroxytestosterone) specific probe substrates of the five isoform probe substrates of CYP450 isozymes (phenacetin, polbutamide, dextromethorphan, chlorzoxazone, testosterone), in order to determine the activity of each isozyme. The result showed good linearity among acetaminophen, 4-OH-tolbutamide, dextrophan, 6-OH-chlorzoxazone and 6β-hydroxytestosterone, satisfactory precision, stability and average recovery, suggesting the method was feasible. The optimized in vitro microsomal incubation conditions conformed to the requirements in the guideline of drug-drug interaction. Gegen Qinlian decoction showed different degrees of inhibitor effect on 5 CYP450 isoforms (CYP1A2, CYP2C11, CYP2D2, CYP2E1, CYP3A1/2). Its major effective component berberine could inhibit each CYP450 isoform at high concentrations (except for CYP1A2, CYP3A1/2).

Analysis and comparison of intestinal absorption of components of Gegenqinlian decoction in different combinations based on pharmacokinetic parameters / 药学学报

To analyse and compare the characteristics of the intestinal absorption of puerarin, baicalin, berberine and liquiritin in different combinations of Gegenqinlian decoction based on pharmacokinetic parameters, a sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was applied for the quantification of four components in rat's plasma. And pharmacokinetic parameters were determined from the plasma concentration-time data with the DAS software package. The influence of different combinations on pharmacokinetics of four components was studied to analyse and compare the absorption difference of four components, together with the results of the in vitro everted gut model and the rat single pass intestinal perfusion model. The results showed that compared with other combinations, the AUC values of puerarin, baicalin and berberine were increased significantly in Gegenqinlian decoction group, while the AUC value of liquiritin was reduced. Moreover, the absorption of four components was increased significantly supported by the results from the in vitro everted gut model and the rat single pass intestinal perfusion model, which indicated that the Gegenqinlian decoction may promote the absorption of four components and accelerate the metabolism of liquiritin by the cytochrome P450.

Rapid determination of acteoside, lobetyolin and another eleven effective components of Shengan Guanxin Mixture by LC-MS/MS / 中国药学杂志

OBJECTIVE: To establish an LC-MS/MS method to determine eleven components(paeoniflorin, acteoside, ferulic acid, naringin, salvianolic acid B, lobetyolin, hesperidin, liquiritin, isoliquiritoside, liquiritigenin, isoliquiritigenin) in Shengan Guanxin Mixture(traditional Chinese medicines) simultaneously. METHODS: The HPLC analysis was performed on an Agilent SB-C18 column(2.1 mm × 50 mm, 1.8 μm) with gradient elution of 0.1% formic acid and methanol containing 0.1% formic acid, the flow rate was 0.3 mL · min, and the column temperature was room temperature. Electrospray ionization (ESI) source was used. Quantitation was performed by using selected multiple reaction monitoring (MRM) mode. RESULTS: The method had good linearity in the ranges of 0.7580-30.32 μg · mL-1 (r =0.9949) for paeoniflorin, 0.3040-12.16 μg · mL-1 (r =0.9995) for acteoside, 0.2480-9.920 μg · mL-1 (r = 0.9969) for ferulic acid, 0.5035-20.14 μg · mL-1 (r = 0.9983) for naringin, 0.9100-36.40 μg · mL-1 (r = 0.9994) for salvianolic acid B, 0.0975-3.900 μg · mL-1 (r = 0.9989) for lobetyolin, 0.9965-39.86 μg · mL-1(r = 0.9982) for hesperidin, 0.2535-10.14 μg · mL-1 (r =0.9975) for liquiritin, 0.2480-9.920 μg · mL-1 (r = 0.9967) for isoliquiritoside, 0.2770-11.08 μg · mL-1 (r = 0.9950) for liquiritigenin, and 0.1530-6.120 μg · mL-1 (r = 0.9928) for isoliquiritigenin. The average recoveries of the eleven active components were 98.47%-101.91% with RSDs of less than 2.5% (n=6). CONCLUSION: The method is convenient, rapid, sensitive and reliable. It may provide a basis for the quality control of Shengan Guanxin Mixture.

Intestinal absorption of different combinations of active compounds from Gegenqinlian decoction by rat single pass intestinal perfusion in situ / 药学学报

The aim is to study the intestinal absorption of different combinations of active compounds out of Gegenqinlian decoction. Rat single pass intestinal perfusion model with jugular vein cannulated was used. Samples were obtained continuously from the outlet perfusate and the mesenteric vein. The levels of puerarin, daidzin, liquilitin, baicalin, wogonoside, jatrorrhizine, berberine and palmatine were determined by LC-MS/MS and their permeability coefficients were calculated. The results showed that Glycyrrhiza could promote the absorption of the active ingredients in Pueraria which is the monarch herb; meanwhile, Pueraria also played a role in promoting the absorption of liquilitin. Based on the Gegenqinlian decoction and the different combinations experiments, the results concerning the absorption of baicalin and wogonoside were as follows. For baicalin, Pueraria and Glycyrrhiza could promote its absorption and the effect of Pueraria was more obvious. For wogonoside, Pueraria could also promote its absorption, while Glycyrrhiza played a opposite role. Pueraria and Glycyrrhiza both played a part in promoting the absorption of jateorhizine, berberine and palmatine, the effective compounds in Coptis.

A new anthraquinone glycoside from the root of Rheum officinale Baill / 药学学报

A new and a known anthraquinone glycosides were isolated from the ethanol extract of the roots of Rheum officinale Baill. The extract was purified by various chromatographies, such as silica gel, Sephadex LH-20, RP-C18 column chromatography and HPLC. Two compounds were identified by the spectroscopic techniques of NMR, MS, and chemical method. In addition, they were tested for their cytotoxic effects against HepG2 cell. Unfortunately, they showed no or weak activity.