| Danshen (Salvia miltiorrhiza Bunge) is one of the traditional Chinese medicinal herbs widely used for the treatment of various Cardiovascular and cerebrovascular diseases. The pharmacologically active compounds of danshen comprise two fractions: lipophilic diterpenoid tanshinones and water-soluble phenolic acids. Danshensu, chemical name D-(+)-β-(3,4-dihydroxyphenyl) lactic acid, is the major water-soluble components of Danshen. Danshensu has significant pharmacological activities such as - antioxidative activity, cardioprotection against ischemia reperfusion injury, inhibit myocardial cell apoptosis and anti-tumor activity. Sodium Danshensu, [C9H10O5Na, MW 220], is a sodium salt of danshensu. In recent years, the incidence of cardiovascular disease increased year by year, a serious threat to human health, but also because of its high safety Danshensu, a significant effect more and more people of all ages. Because of low bioavailability, limiting the development of oral formulations Danshensu, so preparation of high bioavailability of oral formulations of sodium danshensu imperative.The present study was aimed to estiblish a method for determining Sodium Danshensu in rats plasma by and evaluate the pharmacokinetics of Sodium Danshensu after oral administration in rats. In addition, we added absorption enhancers into Sodium Danshensu to increase bioavailability.1. The estiblishment of analytical method of Sodium Danshensu in rats plasmaObjective To estiblish a method for determining Sodium Danshensu in rat plasma by UHPLC-QTOF-MS. Methods Sodium Danshensu was extrated for determination by UHPLC-QTOF-MS. Analytical column was Agilent ZORBAX SB-C18 (100×2.1 mm, 3.5μm). The mobile phase was Acetonitrile: water (0. 1% formic acid )= 10:90. The flow rate was 0.3 mL/min, injection volume was 10μL. capillary 4500 V, nebulizer 50 psig, drying gas 11 L/min. Mass spectrum coditions was ESI performing in the MRM mode using target ions m/z. Results The calibration curve was liner over the range of 10.0~2000.0 ng/ml. The LLOQ of Sodium Danshensu in plasma was 10.0 ng/ml. The intra- and inter-day RSD were < 10.0%. The extracted recovery was ranged from 98.0% to 110.0%. Conclusion The method is sensitive, rapid, simple and accurate to Sodium Danshensu plasma concertration and suitable to study preclinical test of Sodium Danshensu.2. The pharmacokinetics study of Sodium Danshensu in ratsFollowing a single in intravenous administration with different doses of Sodium Danshensu to rats(15, 30 and 60 mg/kg), the Blood samples were withdrawn from the hind limbs vein into heparinized glass tubes before and at 0, 0.083, 0.167, 0.25, 0.5, 1.0, 1.5, 2.0, 3.0, 4.0, 6.0, 8.0, 12.0 h after intravenous administration of Sodium Danshensu. All collected blood samples centrifuged to obtain plasma and the concentrations of Sodium Danshensu in plasma were determined by UHPLC-QTOF-MS method described as above. Pharmacokinetics parameter calculations were carried out using non-compartmental analysis method. The peak plasma concetration were 12.58 mg×h/ml, 34.35 mg×h/ml and 58.23 mg×h/ml, which increased with the dose, showing apparent dose-dependentship (r=0.987). The estimated elimination half-life were 3.39 h, 2.51 h and 2.67h. The MRT were 0.46, 0.41 and 0.28 h.Following a single oral administration with different doses of Sodium Danshensu to rats(30 and 180 mg/kg), the Blood samples were withdrawn from the hind limbs vein into heparinized glass tubes before and at 0.083, 0.17, 0.25, 0.5, 0.75, 1.0, 1.5, 2.0, 3.0, 4.0, 6.0, 8.0 and 12.0 h after oral administration of Sodium Danshensu. All collected blood samples centrifuged to obtain plasma and the concentrations of Sodium Danshensu in plasma were determined by UHPLC-QTOF-MS method described as above. Pharmacokinetics parameter calculations were carried out using non-compartmental analysis method. It was shown by the plasma concentration-time data that the concentarations of Sodium Danshensu in rat plasma were reduced slowly after oral administration and the difference of the rats was evident. The peak plasma concetration were 1.39±0.05 and 8.76±0.05 mg/ml, which were all obtained at 1.5 h and the estimated elimination half-life were 3.99±0.15mg×h/ml and 27.37±4.54 mg×h/ml for 30 and 180 mg/kg doses respectively.SD rats were administered a single dose of 30 mg/kg after intravenous and 180 mg/kg after oral sodium danshensu oral administration administration of Sodium Danshensu for the stduy of bioavailability study. Pharmacokinetics parameter calculations were carried out using non-compartmental analysis method. The peak plasma concetration were 60.36±16.14 mg×h/ml, 27.37±4.54 mg×h/ml, for 30 and 180 mg/kg doses respectively, which were positive correlatated with the doses and the correlation coeffcients was 0.987. Absolute bioavailability was 9.84%.3. bioavailability improvement of Sodium DanshensuSD rats were randomly divided into two groups, oral administration of Danshen sodium (30 mg/kg), the experimental group danshensu oral administration of sodium (30 mg/kg) and borneol (60 mg/kg). Borneol cholate group can not increase the bioavailability. SD rats were randomly divided into two groups, oral administration of Danshen sodium (30 mg/kg), the experimental group danshensu oral administration of sodium (30 mg/kg) and sodium cholate (30 mg/kg). The peak plasma concetration increased from 3.89±0.13 mg×h/ml to 5.85±0.24 mg×h/ml. The Cmax increased from 1.39±0.07μg/ml to 2.10±0.12μg/ml. Sodium cholate group can increase the bioavailability, the effect is significant. Sodium Danshensu combined with borneol, a reference to Compound Danshen Tablets, the results can not achieve the desired results, there is no increase in bioavailability. It will be conducted to further explore the reasons; We expected through adding absorption enhancers to promote absorption of the gastrointestinal by Sodium Danshensu combination of sodium cholate. The results of the experimental group Salvia sodium Cmax and AUC0~∞were increased by about 50%, bioavailability was significantly improved. The results of this study improve the bioavailability of Sodium Danshensu and played a facilitating role for the development of medical industry. |
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