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China Journal of Chinese Materia Medica ; (24): 5798-5808, 2023.
Article in Chinese | WPRIM | ID: wpr-1008777


Based on the concept of quality by design(QbD), the Box-Behnken design-response surface methodology combined with standard relation(SR) and analytic hierarchy process(AHP)-entropy weight method(EWM) was applied to optimize the extraction process of the classic prescription Yihuang Decoction. The content of geniposidic acid, phellodendrine hydrochloride, and berberine hydrochloride in Yihuang Decoction, the extract yield, and fingerprint similarity were used as the critical quality attributes(CQAs) of the extraction process. The extraction time, water addition, and extraction times were used as the critical process parameters(CPPs). After determining the levels of each factor and level through single-factor experiments, response surface experiments were designed according to the Box-Behnken principle, and the experimental results were analyzed. The SR between each sample and the reference sample under various evaluation indicators of different extraction parameters was calculated. The weights of the five evaluation indicators were determined using AHP-EWM, followed by comprehensive evaluation. A function model between CPPs and CQAs characterized by comprehensive scores was established to predict the optimal extraction process parameters. In the final comprehensive weight coefficients, the yield rate accounted for 43.1%, and the content of berberine hydrochloride, phellodendrine hydrochloride, and geniposidic acid accounted for 35.1%, 6.3%, and 15.5%, respectively. After comprehensive score analysis with SR, the established second-order polynomial model was statistically significant(P<0.01, and the lack of fit was not significant). The predicted optimal extraction conditions for Yihuang Decoction were determined as follows: 8-fold volume of water, extraction time of 1.5 h, and extraction once. The mean comprehensive score of the validation experiment was 85.77, with an RSD of 0.99%, and it met the quality control stan-dards for the reference sample of Yihuang Decoction. The results indicate that the optimized extraction process for Yihuang Decoction is stable and reliable, and the water extract is close in quality attributes to the reference sample. This can serve as a foundation for the research and development of granules in the future. Box-Behnken design-response surface methodology combined with SR and AHP-EWM can provide references for the modern extraction process research of other classic prescriptions.

Drugs, Chinese Herbal , Analytic Hierarchy Process , Berberine , Entropy , Water
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 24-31, 2021.
Article in Chinese | WPRIM | ID: wpr-906139


Objective:To explore the quality transmitting relationship between decoction pieces and substance benchmarks with the fingerprint, index component content and dry extract rate as evaluation indexes, and investigate the key quality attributes of 15 batches of substance benchmarks of Yihuangtang, and establish the quality standard of this substance benchmarks. Method:Fifteen batches of Yihuangtang substance benchmarks freeze-dried powder samples were prepared, the fingerprint and index component content of 15 batches of decoction pieces and substance benchmarks were determined by high performance liquid chromatography (HPLC), the mobile phase was acetonitrile (A)-0.1% phosphoric acid aqueous solution (B) for gradient elution (0-6 min, 97%B; 6-12 min, 97%-92%B; 12-25 min, 92%-90%B; 25-35 min, 90%-89%B; 35-50 min, 89%-82%B; 50-75 min, 82%-72%B; 75-85 min, 72%-35%B), the detection wavelength was set at 230 nm, combined the dry extract rate to clarify the attribution of characteristic peaks and the range of similarity with the control chromatogram, the content range and transfer rate range of geniposidic acid and berberine hydrochloride, the dry extract rate range and the variation range of the substance benchmarks. Result:The established HPLC fingerprint had good precision, repeatability and stability, and could be used for the simultaneous determination of decoction pieces and substance benchmarks of Yihuangtang. The similarities between the control chromatogram and fingerprint of substance benchmarks were >0.99. A total of 15 characteristic peaks were assigned, and 8 characteristic peaks were identified by the reference substances, of which 6 were from Phellodendri Chinensis Cortex processed with salt, 1 was from Plantaginis Semen processed with wine, and 1 was from stir-fried Dioscoreae Rhizoma. The content ranges of geniposidic acid and berberine hydrochloride in 15 batches of substance benchmarks of Yihuangtang were 0.10%-0.16% and 0.63%-1.05%, the transfer rate ranges of them were 20.91%-32.65% and 19.60%-29.59%, respectively. The dry extract rate range of the substance benchmarks was 8.45%-9.92%. Conclusion:The quality standard of Yihuangtang substance benchmarks can be preliminarily formulated by the combination of fingerprint, dry extract rate and determination of index component, which can provide the basis for the quality control of Yihuangtang and the development of related preparations.

Chinese Journal of Experimental Traditional Medical Formulae ; (24): 166-171, 2021.
Article in Chinese | WPRIM | ID: wpr-905941


Objective:A strong antithrombotic protein component, named PvQ, was purified and enriched from total protein of <italic>Pheretima vulgaris</italic>,<italic> </italic>a<italic> </italic>traditional Chinese medicine. Moreover, we evaluated its fibrinolytic and anticoagulant activity, and expected to provide reference for the research on antithrombotic substances of Pheretima. Method:A rapid <italic>in</italic> <italic>vitro</italic> activity-oriented separation combined with the AKTA-Pure protein purification system conducted on <italic>P. vulgaris</italic>. Meanwhile, the fibrinolytic and anticoagulant activities of PvQ were measured by fibrin plate method and fibrinogen-thrombin time (Fibg-TT) method. And the <italic>in vitro</italic> thrombolysis assay was used for evaluating the lysis ability of PvQ to thrombus. Then the stability of PvQ was also analyzed for its anticoagulant activity at different pH and temperature. Result:The PvQ was successfully enriched and its activity was determined to have significant fibrinolytic and anticoagulant activities. And the result of <italic>in vitro</italic> thrombolysis assay revealed that PvQ could hydrolyze more than 80% of thrombus after 5 h of incubation at 37 ℃. In addition, the changes of temperature and pH had significant effects on antithrombotic activity, and this study showed that PvQ was rapidly inactivated at ≥60 ℃ or in acidic conditions (pH<7). While, the activity of PvQ was unaffected or less affected at ≤50 ℃ and under alkaline conditions. Conclusion:A feasible preparation method of PvQ is established, and it can affect fibrin and fibrinogen at the same time, thus exerting a dual fibrinolytic effect and possessing significant fibrinolytic and anticoagulant activities. It provides a scientific interpretation for the treatment of thrombotic diseases by PvQ and a reference for the development of antithrombotic protein products of Pheretima.

China Journal of Chinese Materia Medica ; (24): 4410-4416, 2021.
Article in Chinese | WPRIM | ID: wpr-888140


This study was mainly based on the compatibility of Puerariae Lobatae Radix and Chuanxiong Rhizoma to prepare submicron emulsion and evaluated its physical and pharmaceutical properties. Firstly, pseudo-ternary phase diagrams were drawn by dripping method which took Chuanxiong oil as the oil phase and the area of microemulsion region as the index. On this basis, suitable emulsifier and co-emulsifier were screened for the preparation of Chuanxiong oil submicron emulsion. Then, the formula realizing the largest oil loading was selected. Finally, puerarin substituted part of emulsifier and co-emulsifier to lower their content, so as to form puerarin-Chuanxiong oil submicron emulsion featuring the combination of medicine and adjuvant. Its particle size, zeta potential, centrifugal stability and storage stability were determined, and the in vitro drug release behavior was investigated by dialysis bag method, based on which the quality of the as-prepared submicron emulsion was evaluated comprehensively. The proposed method was proved feasible for the preparation of Chuanxiong oil submicron emulsion, which adopted polyoxyethylene castor oil(EL-40) as the emulsifier and was free from co-emulsifier. The formula of the maximum oil loading was found as Chuanxiong oil∶EL-40∶water 3∶7∶90. Further, puera-rin successfully replaced up to 10% of the emulsifier in submicron emulsion. Eventually, the optimal drug-loading formula was determined as puerarin∶Chuanxiong oil∶EL-40∶water 7∶30∶63∶900. The quality evaluation results of the as-prepared submicron emulsion demonstrated that the average emulsion droplet size was 333.9 nm, the PDI 0.26, and the zeta potential-10.12 mV. The submicron emulsion had a good centrifugal stability and did not present any instable phenomena such as delamination and precipitation during its standing still for 50 days. The evaluation of in vitro drug release behavior indicated that the submicron emulsion was capable of releasing the drug completely. The puerarin-chuanxiong oil submicron emulsion prepared in this study possessed a stable quality and to some extent increased the solubility of puerarin along with a sustained-release effect. This study provided ideas for the clinical application of puerarin.

Emulsions , Isoflavones , Particle Size , Solubility