Inhibition Effect of Spinosin on Cytochrome P450 Enzymes from Human Liver Microsomes in vitro / 中国药房

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.

Simultaneous Determination of 2 Main Components in Ticarcillin Disodium and Potassium Clavulanate for Injection by Improved HPLC / 中国药房

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.

Study on absorption kinetics of betulic acid in rat's intestines / 中国中药杂志

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHOD</b>The rat intestinal absorption model was established to detect the impact of absorption site, drug concentration and pH value on drug absorption.</p><p><b>RESULT</b>Within 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.</p><p><b>CONCLUSION</b>Betulic 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.</p>

Determination of Shionone in Rat Plasma by HPLC and Its Pharmacokinetic study / 中草药·英文版

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.

In situ absorption and metabolism of stilbene glycoside in rat / 药物分析学报

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.

Determination of five coumarins in radix glehniae by micellar electrokinetic capillary chromatography / 中国中药杂志

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.

HPCE fingerprints of Forsythia suspensa from Hebei province / 中国中药杂志

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHOD</b>Electrophoresis 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.</p><p><b>RESULT</b>The 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.</p><p><b>CONCLUSION</b>The method is simple, quick, accurate and can be used as a new means for the quality control of F. suspensa.</p>

Simultaneous determination of ginsenosides and epimedium flavonoids in rat urine by HPLC-UV-ELSD / 中国中药杂志

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHOD</b>Chromatographic 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.</p><p><b>RESULT</b>The 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%.</p><p><b>CONCLUSION</b>The 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.</p>

Pharmacokinetics of aristolochic acid A in Radix Aristolochiae and Guanxinsuhe Capsule / 中草药

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.

Determination of the Contents of Geniposide in Gardenia Jasminoides Ellis and Its Preparations by RP-HPLC / 中国药房

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.

Studies on antiarrhythmic portion in Cornus officinalis extract / 中草药

Object To study the effective portion in Cornus officinalis Sieb. et Zucc. extract (COE) for the treatment of arrhythmia. Methods Effect of COE on chloroform induced ventricular fibrillation in mice and electrophysiology of isolated guinea pig papillary muscle were studied. Results Antiarrhythmic effect of COE may be related to its prolongation of action potential duration, increase of the absolute value of resting potential and a decrease of autonomy of sinus node. The effective portion in COE may be its total organic acid and a certain yet unknown trace substance, whereas its total glycosides were devoid of such activities. Conclusion Pharmacodynamic and myocardial electrophysiologic studies showed that the total organic acid and a certain unknown trace substance possessed the obvious antiarrhythmic activity.

Preparation of loganin and morroniside reference substances from Fructus Corni / 中草药

Objective To establish a separation method of loganin and morroniside reference substance from Fructus Corni. Methods After extracted with hot water and precipitated with alcohol, the extract of Fructus Corni was isolated and purified by macroporous resin, silica gel column chromatography, and preparative HPLC. Loganin and morroniside were identified by UV, 1H-NMR, 13C-NMR, and MS. Results The contents of loganin and morroniside was higher than 98% by normalization method of HPLC. Conclusion The developed method is simple with lower cost, by which loganin and morroniside can be used as reference substances for the qualitative and quantitative analysis of Chinese herbal medicine.

Pathway and method in studies on potential basis of Chinese herbal formula / 中草药

To study the potential basis of Chinese herbal formula is very important for elucidating their compatible mechanism and promoting the modernization of Chinese materia medica. The pathway and method in studies on the potential basis of Chinese herbal formula, such as altering prescriptions, serum pharmacology, serum pharmacochemistry, fingerprints, pharmacokinetics, and metabonomics, etc. are summed up in this paper. The above-mentioned pathway and method will provide the significant basis in studying the effective components and fractions in Chinese herbal formula.

Simultaneous determination of chlorogenic acid,puerarin and paeoniflorin in Kangganning Mixture by multiple wavelength RP-HPLC / 中成药

AIM: To establish a method for simultaneously determining chlorogenic acid,puerarin and paeoniflorin in Kangganning Mixture(Flos Lonicerae japonicae,Radix Puerariae Lobatae,Radix Paeoniale rubra,Fructus Forsythiae,etc.). METHODS: A multiple wavelength HPLC method was devoloped.The analysis was performed on an Agilent Zorbax SB C_(18) column(250 mm?4.6 mm,5 ?m) with acetonitrile-0.1%H_3PO_4(11∶89) as the mobile phase.The detection wavelengthes were monitored at 324 nm,254 nm and 230 nm for chlorogenic acid,puerarin and paeoniflorin,respectively.The flow rate was 1.0 mL/min and the column temperature was at 30 ℃.(RESULTS:) The calibration curves of chlorogenic acid,puerarin and paeoniflorin showed good linearity at the ranges of 0.179-2.864 ?g,0.071 55-1.144 8 ?g and 0.372 5-5.96 ?g,respectively.The average recoveries were(100.7%,) 103.3%,102.6% with RSD of 2.2%,2.4%,1.9%,respectively. CONCLUSION: The method is simple,accurate and reproducible for determining chlorogenic acid,puerarin and paeoniflorin in Kangganning Mixture.

Quality standard for Guishao Soft Capsules / 中成药

AIM: To establish the quality standard for Guishao Soft Capsules(Radix Paeoniae Alba,Radix Angelicae Sinensis,Rhizoma Chuanxiong, etc.) METHODS: Radix Paeoniae Alba,Rhizoma Atractylodis,Rhizoma Chuanxiong,Rhizoma Alismatis and Radix Angelicae Sinensis were identified by TLC.The contents of albiflorin and paeoniflorin in Guishao Soft Capsules were determined by HPLC.HPLC analytical method was carried out(using) a C_(18) column and a mixture containing 17 volume of acetonitrile and 83 volume of 0.05% acetic acid as the mobile phase.The detection wavelength was set at 230 nm. RESULTS: Spots obtained from the test solutions had the same color in reference solution and medical material in the same location,and the blank solution had no interfernece.The linear range of albiflorin was from 0.53 ?g to 1.06 ?g(r=0.999 8),the average recovery was(100.3%.) The linear range of paeoniflorin was from 0.29 ?g to 2.00 ?g(r=0.999 9),the average recovery was(99.49%.) CONCLUSION: In TLC,the spots are very clear and specific to identify the herbal medicine in Guishao Soft Capsules.It is simple,quick,high precise and accurate for HPLC to control the quality of Guishao Soft Capsules.