OBJECTIVE:To improve the method for the determination of 2 main components in Ticarcillin disodium and potas-sium clavulanate for injection. METHODS:HPLC was performed on the column of Waters XBridgeTM C18 with mobile phase of 0.01 mol/L ammonium dibasic phosphate solution(pH 7.0)-menthol(80:20,V/V)at a flow rate of 1.0 mL/min,the detection wave-length was 220 nm, column temperature was 30 ℃, and injection volume was 20 μL. RESULTS:The linear range was 1.95-195.22 μg/mL for ticarcillin (r=0.9999) and 0.12-12.18 μg/mL for clavulanate(r=0.9999);RSDs of precision,stability (under 4 ℃) and reproducibility tests were lower than 1.0%;recoveries were 99.3%-100.5%(RSD=0.4%,n=9) and 99.2%-101.0%(RSD=0.7%,n=9). CONCLUSIONS:The method is rapid,accurate and reliable,and can be used for the determination of 2 main components in Ticarcillin disodium and potassium clavulanate for injection.

OBJECTIVE:To study the inhibition effect of spinosin on 7 subtypes (CYP2B6,CYP2C8,CYP2C9,CYP2D6, CYP1A1,CYP2C19 and CYP3A4)of cytochrome P450(CYP450)enzymes from human liver microsomes in vitro. METHODS:Tak-ing 200.00,100.00,50.00,25.00,12.50,6.25,3.13,1.56,0.78,0.39 μmol/L spinosin and human liver microsomes for incuba-tion,using daktarin,bupropion,amodiaquine hydrochloride,diclofenac sodium,mephenytoin,dextromethorphan hydrobromide and midazolam as the specific probe drugs for above-mentioned 7 subtypes of CYP450 enzymes. UPLC-Q-TOF-MS was conducted to detect generation amount of 7 probe drug metabolites,and the half inhibitory concentration (IC50) of spinosin on 7 subtypes of CYP450 enzymes from human liver microsomes was calculated. RESULTS:IC50 of spinosin on 7 subtypes of CYP450 enzymes from human liver microsomes were 1714,1158,226.1,2288,80.59,101.1,1119 μmol/L,respectively,which were higher than 50μmol/L. CONCLUSIONS:Spinosin has no inhibition effect on above-mentioned 7 subtypes of CYP450 enzymes from human liver microsomes,with very low probability of inducing metabolic drug interactions.

OBJECTIVETo establish a HPLC-DAD method for determining concentrations of betulic acid and phenol red in intestinal circulation liquid, and probe into the absorption kinetic characteristics of betulic acid at different intestine segments in rats and the effect of different drug concentrations on absorption.

METHODThe rat intestinal absorption model was established to detect the impact of absorption site, drug concentration and pH value on drug absorption.

RESULTWithin the range from 75-125 mg x L(-1), the absorption rate and the quality concentration of betulic acid had a linear relation, with Ka value keeping unchanged. The absorption rate for each intestinal segment showed no remarkable difference, with Ka values in duodenum, jejunum, ileum and colon being (0.151 +/- 0.0049), (0.159 +/- 0.0056), (0.156 +/- 6.0083), (0.149 +/- 0.0041) h(-1), respectively.

CONCLUSIONBetulic acid is proved to be well absorbed in intestines marked by no specific absorption site in the intestine. The absorption mechanism of the drug conforms to passive transport mechanism and first-order kinetics. The bioavailability of betulic acid preparation can be increased by enhancing the dissolution rate and the solubility.

Animals ; Anti-HIV Agents ; pharmacokinetics ; Antineoplastic Agents, Phytogenic ; pharmacokinetics ; Chromatography, High Pressure Liquid ; Intestinal Absorption ; Intestines ; metabolism ; Kinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Reference Standards ; Reproducibility of Results ; Sensitivity and Specificity ; Triterpenes ; pharmacokinetics

Objective To develop a sensitive, simple, and accurate method for the determination of shionone in rat plasma after ig administration of Asteris Radix petroleum ether extract (RAPE). Methods The separation was achieved by HPLC on a RP18 column (150 mm × 3.9 mm, 5μm) with a mobile phase composed of acetotitrile-0.05% phosphoric acid water (98: 2) at a flow rate of 1.0 mL/min. UV Detector was set at 200 nm and friedelin was chosen as an internal standard. Results The linear range of the standard curves was (0.3443-22.0) μg/mL with the correlation coefficient of 0.9968. The intra- and inter-day precisions were all below 10% and the relative error was -3.5%-1.1%.Conclusion The developed method can be successfully applied to the pharmacokinetic study. After ig administration of RAPE, T1/2(ka) is (33.09 ± 7.32) min and T1/2(ke) is (84.95 ± 22.34) min.

OBJECTIVETo develop and validate a HPLC-UV-ELSD method for the simultaneous determination of ginsenosides and epimedium flavonoids in rat urine after intravenous administration of Jiweiling freeze-dried powder.

METHODChromatographic separation was performed on a C18 HPLC column, with gradient elution of acetonitrile and water as mobile phase. An UV detector was used at detection wavelength of 220 nm. An evaporative light scattering detector (ELSD) was used at drift tube temperature of 80 degrees C and gas pressure of 172.4 kPa.

RESULTThe calibration curves were linear over the investigated concentration ranges with all correlation coefficients higher than 0.998. The a intra- and inter-day RSD were less than 9.1% and the relative errors were verage extraction recoveries for all compounds were between 88.67% and 101.2%. The within the range of -11.58% to 10.89%.

CONCLUSIONThe proposed method showed appropriate accuracy and selectivity and was successfully applied to the rat urine samples analysis of saponins and flavonoids after intravenous administration of Jiweiling freeze-dried powder, which may provide some references to the apprehension of the action mechanism and clinical application.

Animals ; Calibration ; Chromatography, High Pressure Liquid ; methods ; Epimedium ; chemistry ; metabolism ; Flavonoids ; urine ; Ginsenosides ; urine ; Phytotherapy ; Plant Preparations ; Rats ; Saponins ; analysis ; Ultraviolet Rays

OBJECTIVETo develop a HPCE analysis method for fingerprints of Forsythia suspensa from Hebei province, get reference fingerprint and compare the fingerprints of F. suspensa collected from different producing areas and different parts of the plant.

METHODElectrophoresis was performed on a fused silica capillary column (75 microm x 60 cm, 30 cm). The running buffer was composed of 50 mmol x L(-1) borax (adjust to pH 9.90 with 0.1 mol x L(-1) NaOH). The applied voltage was 15 kV and the temperature was 20 degrees C. The detection wavelength was 214 nm. The semblances to the crude drugs of different producing areas were compared.

RESULTThe mutual mode of HPCE fingerprints was set up with 12 common peaks. The fingerprints of F. suspensa from Hebei province had high similarity, F. suspensa from Shanxi and Henan were also of good quality. The chemical composition in different parts of the herb had big differences.

CONCLUSIONThe method is simple, quick, accurate and can be used as a new means for the quality control of F. suspensa.

China ; Drugs, Chinese Herbal ; analysis ; Electrophoresis, Capillary ; methods ; Forsythia ; chemistry

A micellar electrokinetic capillary chromatography method with ultraviolet detection was developed for the simultaneous determination of psoralen, xanthotoxin, isoimpinellin, bergapten and scopoletin in Radix Glehniae. The separation was performed on an uncoated fused silica capillary column (50.2 cm x 75 microm x 40 cm) with 20 mmol x L(-1) borax solution (pH 9.6) containing 16 mmol x L(-1) sodium dodecylsulfate (SDS) and 15% acetonitrile as running buffer at applied voltage of 22 kV. The detection wavelength was 214 nm. The effects of concentrations of borax solution, sodium dodecylsulfate (SDS), and organic modifier, voltage, temperature on the separation and sensitivity were investigated. The five active constituents were completely separated within 7 min. The linear ranges of psoralen, xanthotoxin, isoimpinellin, bergapten and scopoletin were 9.91-82.6, 37.2-162, 2.23-18.6, 2.73-22.3 and 2.89-20.1 mg x L(-1), respectively. And the average recoveries were 98.9%, 98.4%, 101.3%, 99.1% and 98.0%, respectively. This simple and rapid method provided a new basis for assessment on quality of Radix Glehniae.
Apiaceae ; chemistry ; Chromatography, Micellar Electrokinetic Capillary ; methods ; Coumarins ; analysis ; isolation & purification ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Plant Roots ; chemistry

Stilbene glycoside (TSG) has been shown to have many beneficial properties. It is therefore essential to understand the absorption and metabolism of TSG in detail. We determined the recovery of TSG and its metabolites (TSG sulfate/glucuronides) in rat gastric contents, gastric mucosa, portal vein plasma, celiac arterial plasma, bile, and urine after administration of 15mg of TSG in 0.5mL physiological saline or incubation for 20min in situ in the stomach of rats. Within 20min, (64.0±9.8)% of the administered TSG disappeared from the stomach; later, TSG was recovered in both free and conjugated forms in plasma and bile, but not in urine. On the other hand, only free TSG was detected in the gastric contents and mucosa; it was also detected in the portal vein plasma as (48.1±3.5)% of the total TSG (all forms of TSG). However, the proportion of free TSG in the celiac arterial plasma and bile decreased to 4%-10%. In addition, the proportion of free TSG to total TSG in the liver microsome incubation mixture after TSG was incubated in liver microsome at 37℃ for 30min was very low [(10.6 ± 2.6)%]. These results indicate that TSG could be quickly absorbed from the rat stomach, conjugated in liver and excreted in bile. Such novel information would be helpful for the use of TSG as a beneficial natural product which may improve its proposed efficacy in preventing chronic diseases.

Objective To study the pharmacokinetics of aristolochic acid A in Radix Aristolochiae and the compound preparation of Guanxinsuhe Capsule in mice in vivo after single-dose oral administration and observe the difference of aristolochic acid A absorption and distribution. Methods Aristolochic acid A assay was performed by RP-HPLC on a Waters apparatus with a DiamonsilTM C18 column (250 mm × 4.6mm, 5 μm), a mobil phase: a mixture of methanol-water-acetic acid (72: 27 : 1), flow rate: 1.0 mL/min, detection wavelength: 315 nm, and column temperature: 20 ℃. Results Mice were given Radix Aristolochiae and Guanxinsuhe Capsule by ig at the same level of 2. 5 mg/kg of aristolochic acid A, respectively, which were suspended in 0. 3% CMC-Na solution. Plasma concentrations were determined by RPHPLC. After single-dose ig administration of Radix Aristolochiae or Guanxinsuhe Capsule to mice, the mean plasma concentration-time courses of aristolochic acid A obtained fitted the one-compartment model.The main pharmacokinetic parameters of aristolochic acid A in Radix Aristolochiae, t1/2ka, t1/2 ke, tmax,AUC, Cmax are 5. 103 min, 43. 63 min, 17.89 min, 80. 45 (μg · min)/mL, and 0. 916 8 μg/mL; the rela tive pharmacokinetic parameters in Guanxinsuhe Capsule are 5. 294 min, 43.50 min, 18. 32 min, 33.08(μg · min)/mL, and 0. 381 8 μg/mL. Conclusion The Cmax of aristolochic acid A in Guanxinsuhe Capsule is significantly less than that in Radix Aristolochiae, which indicates that the compound compability could decrease the absorption of aristolochiae acid A.

OBJECTIVE:To establish a RP-HPLC method for the assay of geniposide in Gardenia jasminoides Ellis and its preparations.METHODS:The determination was carried out with RP-HPLC on Kromasil C 18 column(4.6mm?250mm,5?m)with acetonitrile-water(10∶90)as the mobile phase,and the geniposide was detected at a UV-wavelength of238nm.RESULTS:The linear range of geniposide was0.596?g～2.98?g(r=0.9997).It was found that the contents of geni?poside in Gardenia jasminoides Ellis,its fruit,peel,seed,Jiaweixiaoyao pill and Qingkailing injection were41.8mg/g,12.3mg/g,67.3mg/g,4.26mg/g and0.316mg/ml,respectively.The average recoveries of geniposide in Gardenia jasminoides Ellis,Ji?aweixiaoyao pill and Qingkailing injection were99.35%,99.51%and99.57%and the relative standard deviations(RSD)were0.52%,0.72%and0.73%,respectively.CONCLUSION:The method is simple,accurate,sensitive and stable.It can be used for the determination of geniposide in Gardenia jasminoides Ellis and its preparations.