ABSTRACT
Objective:To further confirm the anti-endotoxin active parts of Lidanpaidu prescription through observing the anti-en-dotoxin intensity based on a half-in vivo method. Methods:The extracting solution of Lidanpaidu prescription was extracted by the sol-vents with different polarity to obtain the active part. The anti-endotoxin activity of the samples was determined by using the Azo color matrix method, and the amount of inactivated endotoxin was compared among the various extraction parts with different dosages. Re-sults:Compared with that of the control group, the amount of inactivated endotoxin by the rat plasma in each administration group was all higher, and the differences were statistically significant (P0. 05). The rat plasma of the water soluble fraction administration groups also showed endotoxin inactivation, and the difference be-tween the high dosage group and the low dosage group was statistically significant (P<0. 05). Conclusion:The results of the half-in vivo method showed the consistent anti-endotoxin effect of the whole prescription and the water soluble fraction, furthermore, the dose-effect relationship is preliminarily appeared, suggesting that the water soluble fraction remains the effective components of the whole prescription to a great extent. Therefore, the water soluble fraction can be further confirmed as the active anti-endotoxin part of Lidan-paidu prescription.
ABSTRACT
Objective: To analyze the influencing factors in thrombin titration for the determination of anticoagulant activity of Whitmania Pigra Whitman. Methods: The white porcelain plates were used as the titration carriers instead of tubes in the titration ( called white porcelain method for short) . The effect of different carriers, interval time of titration and thrombin concentration on the results of anticoagulant activity test was studied. Results:Under the same conditions, the anticoagulant activity was more accurate and stable using white porcelain method. Using white porcelain method with 20 u·ml-1 or 10 u·ml-1 as the thrombin concentration and titrating 5μl each time, once every minute, the thrombin consumption volume was linear with the sample concentration within the range of 0. 125-0. 333 g·ml-1(r20 =0. 961 and r10 =0. 992), and the anticoagulant activity respectively was (33. 08 ± 2. 64) and (31. 24 ±1.32) u·g-1(RSD20 =8.0% and RSD10 =4.2%). As for a certain sample concentration (0.333 g·ml-1), the theoretical error of determination was not more than 10% and 5%. Conclusion:The improved white porcelain method is more suitable for determining anticoagulant activity of Whitmania Pigra Whitman with more stable results and accurate end point states than tube method. Under the conditions of 10 u·ml-1 thrombin concentration, titrating 5μl each time, once every minute, the linearity, accuracy and precision are all promising.