| Selaginella moellendorfii Hieron is the dry whole herbs of Selaginella Spring, Selaginellaceae. It is distributed in areas to the south of the Changjiang River and also areas in the south of Shaanxi. It can clear heat-toxin, promote blood circulation and remove edema. It is used for acute infectious hepatitis, contusion of chest and waist, anasarca, thrombocytopenia and so on.The main efficacy component of Selaginella moellendorfii Hieron is flavonoids, including one effective constituent, i.e. amentoflavone, which has many kinds of biological activities, such as anti-inflammatory, anti-oxidation, antiviral, antitumor, reducing blood sugar and dilating blood vessels. However, there are a few researches on its pharmacokinetics at the present.In order to clearly explain the pharmacokinetics of amentoflavone in vivo, we detected the concentration of rabbits’serum by HPLC and therefore found out its pharmacokinetics through gavage with flavonoid extract which contained amentoflavone. The research could be used for improving the new drug design and providing basis for optimal dosing, so as to give full play to its curative effect in clinical medication.1. Preparation of the flavonoid extract of Selaginella moellendorfii HieronPulverize and sieve the crude drugs of Selaginella moellendorfii Hieron; then use85%ethanol for reflux extraction, in twice, one hour each time; degrease3times by petroleum ether, and then precipitate by low concentrated ethanol. The powder of flavonoids extract was gotten by decompression drying at last.As a result, there was23.41%amentoflavone in the flavonoid extract according to the determination of HPLC.2. The method of determining amentoflavone in rabbit serum by HPLCPretreatment of serum sample:saturated NaH2PO4was used to deposit protein and ethyl acetate was used to extract the sample. Centrifuged for10min at4000rpm, repeated the extraction one more time, then used nitrogen to blow and dry it. Residue was dissolved with500μL methanol and filtrated by0.45μm filter membrane.20μL solution was taken for liquid phase analysis.The analytical column was Cosmosil5C18-MS-II waters (4.6mm×250mm,5μm), the mobile phase consisted of acetonitrile (A) and water (including2%THF and0.1%TFA)(B) with gradient elution, pH value was2.2, the flow rate was1.0mL·min-1, column temperature was30℃, detection wavelength was335nm, the sample size was20μLIt was turned out that, the linear limits of amentoflavone ranged from1.024ng to41ng (r=0.9995,n=5). The absolute recovery for low, medium and high concentration was (59.17±0.41)%,(54.51±0.65)%and (51.52±0.69)%. The relative recovery for low, medium and high concentration was (83.98±0.98)%,(96.24±1.22)%and (100.50±1.38)%, respectively RSD≤2%(n=6). The lowest detection limit was0.41ng.3. Pharmacokinetics of amentoflavone in rabbit serum0.30g/kg flavonoid extract (amount to amentoflavone70.23mg/kg) was dissolved by0.5%tween-80and gavaged to rabbits; then took2ml blood from the auricular vein at the30th,45th,60th,75th,90th,120th,150th,180th,240th,360th,480th and720th minutes. After pretreating the sample, determinated it by HPLC. We got that the pharmacokinetics of amentoflavone was fitted one-compartment model. And t1/2was (125.015±71.326)min, Ke was (0.009±0.008)/min, V1/F was (99.586±59.97)L/kg, CL/F was (0.583±0.157) L/min/kg, AUC(0-t) was (125.521±34.387)mg/L·min, AUC(0-∞) was (136.803±36.995)mg/L·min, Ka was (0.142±0.19)/min, t1/2ka was (13.56±11.138)min,and Tlag was(24.771±3.147)min。... |
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