| Weiyanling granule is a kind of preparation in zhangjiagang hospital of traditional Chinese medicine,in which the main constituents were gardenia,radices paeoniae alba,fructus aurantii immaturus,radix curcumae and so on.It has the effect of soothing liver,regulating qi,clearing liver,relieving heat,relieving spasm and relieving pain.It is definite in clinical effect,often with some enhanced gastric motivation or protecting gastric mucosa drug combination in the treatment of bile reflux gastritis,such as doperidone,etc.At present,a lot of the related research based on clinical observation and application.Considering the safety of clinical medication,the article is based on the content determination of the effective components and the protein binding rate of weiyanling granule and attempts to study the effect of weiyanling granule on the activity of cytochrom P450 and the drug-drug interaction of weiyanling granule.To investigate the safety and rationality of weiyanling granule when combined with western medicine and traditional Chinese medicine,so as to provide basis for clinical rational and safe medication of weiyanling granule.1.Simultaneous Determination of Gardenoside,Paeoniflorin and Naringin in Weiyanling granule by HPLCThe contents of gardenoside,paeoniflorin and naringin in weiyanling granules were determined simultaneously by HPLC,and the above three components in three batches of weiyanling granule were quantitatively analyzed.The chromatographic seperations were on Agilent Zorbax SB-C18 column(4.6 mm × 150 mm,5μm)with the mobile phase by gradient elution of acetonitrile-0.1%phosphoric acid aqueous solution.The column temperature was set at 30℃ and the detection wavelength was 234 nm.The flow rate was 1.0 mL·min-1 and the injection volume was 10μL.The results showed that the linear range of gardenoside,paeoniflorin and naringin were 0.515～20.6 μg·mL-1(r=0.9997)for gardenoside,0.516～20.64μg·mL-1(r=0.9999)for paeoniflorin and 0.243～9.72 μg·mL-1(r=0.9998)for naringi.The recovery of the samples were 98.76%,98.07%,99.15%with RSD 0.90%,1.29%,1.25%,respectively.The method is simple,accurate and reproducible,which can provide basis for quality control of weiyanling granule.2.Determination of the protein binding rate of gardenioside,paeoniflorin and naringin in weiyanling granule in different species of plasmaDetection methods were established to determinate the concentrations of geniposide,paeoniflorin and naringin by HPLC,and to measure the protein binding rates of the three components in bovine serum albumin,rat plasma and human plasma by equilibrium dialysis method.The results showed that the linear relationship,precision,extraction recovery and stability of gardenoside,paeoniflorin and naringin all met the methodological.Within the low,middle and high concentration(5.084,10.04 and 20.08 mg·mL-1)of weiyanling granule,the protein-binding rate of gardenioside and bovine serum albumin were(23.77±13.39)%,(10.75± 1.34)%,(5.82±6.53)%respectively;the protein binding rate of paeoniflorin were(51.81 ±6.53)%,(30.12±6.61)%,(21.39±9.45)%,respectively;the protein binding rate of naringin were(70.21 ±3.31)%,(67.61 ± 1.31)%,(56.00±3.46)%respectively;the protein binding rate of gardenioside and rat plasma were(71.56±3.00)%,(43.56± 12.22)%,(40.63±4.73)%,respectively;the protein binding rate of paeoniflorin and rat plasma were(55.72±1.60)%,(47.59±6.33)%,(40.07±0.72)%respectively;the protein binding rate of naringin and rat plasma were(85.85±3.53)%,(85.38±0.99)%,(79.99±2.57)%respectively;the protein binding rate of gardenioside and human plasma were(13.56±2.40)%,(3.02±2.41)%,(5.82±2.08)%respectively;the protein binding rate of paeoniflorin and human plasma were(9.49±5.23)%,(5.01 ± 1.10)%,(3.98± 1.54)%respectively;the protein binding rate of naringin and human plasma were(25.04±2.61)%,(26.09± 13.82)%,(20.20±2.24)%,respectively.Within 5.084,10.04 and 20.08 mg·mL-1,the protein binding-rate of gardenioside,paeoniflorin and naringin were low binding type in human plasma.The protein binding rates of the three components were shown as rat plasma>bovine serum albumin>human plasma.There were significant differences in species.3.Evaluation of influence of weiyanling granule on activities of cytochrome P450 in rats by Cocktail probe drugsTo study the effect of Weiyanling granule on the activities of six CYP450 isoforms in rats by coctail probe drug.Dextromethorphan,midazolam,omeprazole,theophylline,chlorzoxazone and tolbutamide as the probe drug for CYP2D6,CYP3A2,CYP2C19,CYP2E1 and CYP2C9,respectively.We choose six rats which were intragastric administered with weiyanling granule(4 g·kg-1)daily,and the blood concentrations of 6 probe drugs in rats before and after administration were determined by the liquid chromatography mass spectrometry(LC-MS/MS).Results show that after treated with Weiyanling granules for 10 days,Compared with predose,the metabolism of dextromethorphan(p<0.01)and tolbutamide(p<0.01)decreased,the metabolism of chlorzoxazone(p<0.01)increased,but no significantly difference in metabolism of omeprazole,midazolam and theophylline.Weiyanling granule has a moderate strong inhibitory on CYP2D6 in rats,and has a weak inhibitory effect on CYP2C9,and has a weak induction effect on CYP2E1.It is indicated that gastritis particles have little effect on CYP3 A2,CYP2C19 and CYP1A2.4.Effect of weiyanling granule on pharmacokinetics of domperidone in ratsIn order to study the effect of weiyanling granule on pharmacokinetics of domperidone in rat.We choose six rats which were intragastric administered with weiyanling granule for 10 consecutive days,and domperidone(4 mg·kg-1)was oral administered to all the rats before the first time and after the last time giving weiyanling granule.To investigate the effect of combined application of weiyanling granule on the plasma concentration of domperidone.We used the liquid chromatography mass spectrometry(LC-MS/MS)to determinate the blood concentration of domperidone in rats.The results showed that weiyanling granule had no effect on the metabolism of domperidone,so it was verified that weiyanling granule had no effect on CYP3A2 in rats(human CYP3A4). |
PDF Full Text Request