ABSTRACT
Objective:To compare the anti-thrombin activity and the effects on the coagulation pathway between Whitmania pigra Whitman and Hirudinaria manillensis to provide scientific reference for the anticoagulation mechanism revelation for Hirudo. Methods:Anti-thrombin titration and chromogenic substrate assay-extraction-HPLC were applied to study the anti-thrombin activity of Whitmania pigra Whitman and Hirudinaria manillensis. APTT, PT and TT were determined by a clotting assay to compare the effects on the path-way of blood clotting. Results:The anticoagulation activity order measured by anti-thrombin titration was living Hirudinaria manillensis> dried Hirudinaria manillensis > > dried Whitmania pigra Whitman. The results of chromogenic substrate assay-extraction-HPLC in-dicated that the low dose of aqueous extract promoted the thrombin activity, while the high dose inhibited the thrombin activity. Hirudi-naria manillensis significantly inhibited the activity of thrombin, while Whitmania pigra Whitman showed weak anti-thrombin activity only at the higher dose. All leeches could prolong APTT, PT and TT. However, living Hirudinaria manillensis mainly affected TT, and dried Hirudinaria manillensis mainly affected APTT. Dried Whitmania pigra Whitman dramatically influenced APTT and TT. All the results indicated that the anticoagulant activity of Whitmania pigra Whitmanis was significantly higher than that of Hirudinaria manillen-sis. Conclusion:There are notable differences in the anti-thrombin activity and the effect on the pathway of blood clotting between Whitmania pigra Whitman and Hirudinaria manillensis.
ABSTRACT
Objective:To investigate the enantiomer separation for atropine by capillary electrophoresis .Methods:Capillary elec-trophoresis was used with an elastic quartz capillary column (60 cm ×75 mm, effective length of 40 cm).The concentration of phos-phate buffer was 30 mmol· L-1 .The high and time of injection was 10 cm and 5 s, respectively.The detection wavelength was 225 nm.The best separation conditions were selected including the type and concentration of chiral resolving agent , pH of the buffer solu-tion, operating voltage and organic solvent.Results:The optimum conditions of separation were as follows:the pH of buffer solution was 7.0, the concentration of S-β-CDP was 10 mg· ml-1 , and the operating voltage was 12 kV.Conclusion: The method is simple and fast, which can be used to se parate the optical isomers of atrpo ine.
ABSTRACT
Objective To establish a capillary electrophoresis method to separate ephedrine and psedudoephedrine. Methods RM-β-CD and HP-β-CD were set as additives. A capillary electrophoresis method was set up. The effects of types and concentrations of additives, the concentrations and pH values of buffered solution, running voltage and organic solvent on the separation of ephedrine and psedudoephedrine were investigated.Results Ephedrine and pseudoephedrine could be successfully separated by using either RM-β-CD or HP-β-CD as additives. When RM-β-CD was used as additive, the best separation conditions were as follows: separation voltage 10 kV, 25 mmol/L Tris-H3PO4 (pH 2.42), 20 mg/mL of RM-β-CD. Under the conditions, the resolution of ephedrine and pseudoephedrine was 1.56 and they were separated successfully within 13 min. When HP-β-CD was used as additive, the best separation conditions were as follows: separation voltage 10 kV, 25 mmol/L Tris-H3PO4 (pH 3.00), 50 mg/mL of HP-β-CD. Under the conditions, the resolution of ephedrine and pseudoephedrine was 2.73 and they were separated successfully within 15 min.ConclusionThis method is reliable, rapid and repeatable. It can be used as separation determination method for ephedrine and pseudoephedrine.
ABSTRACT
Objective To establish a pre-column derivation HPLC-UV method for the content determination of hyodeoxycholic acid in artificial calculus bovis. Methods The hyodeoxycholic acid was derived by 2-bromo-2’-acetonaphthoneat using triethylamine as the catalyst in 60 ℃ water bath. After that, a HPLC-UV method was established to determine the content of hyodeoxycholic acid in artificial calculus bovis. Results When the derivatising time at 60 ℃ water bath was 50 min, the radio of the molar amount of derived reagents and hyodeoxycholic was over 20:1 and the radio of catalyst and hyodeoxycholic was over 15:1; the reaction was completed. The calibration curve was linear within the range of 0.1–2 μg for hyodeoxycholic acid (r=0.999 7), and the average recovery was 97.85% (RSD=1.6%). In this sample, the content of hyodeoxycholic is 4.12%. Conclusion The method is with high sensitivity, highly reproducible, reliable and accurate for the content determination of hyodeoxycholic acid in artificial calculus bovis.