Optimization of high pressure machine decocting process for Dachengqi Tang using HPLC fingerprints combined with the Box-Behnken experimental design.

Using Dachengqi Tang (DCQT) as a model, high performance liquid chromatography (HPLC) fingerprints were applied to optimize machine extracting process with the Box-Behnken experimental design. HPLC fingerprints were carried out to investigate the chemical ingredients of DCQT; synthetic weighing method based on analytic hierarchy process (AHP) and criteria importance through intercriteria correlation (CRITIC) was performed to calculate synthetic scores of fingerprints; using the mark ingredients contents and synthetic scores as indicators, the Box-Behnken design was carried out to optimize the process parameters of machine decocting process under high pressure for DCQT. Results of optimal process showed that the herb materials were soaked for 45 min and extracted with 9 folds volume of water in the decocting machine under the temperature of 140 °C till the pressure arrived at 0.25 MPa; then hot decoction was excreted to soak Dahuang and Mangxiao for 5 min. Finally, obtained solutions were mixed, filtrated and packed. It concluded that HPLC fingerprints combined with the Box-Behnken experimental design could be used to optimize extracting process of traditional Chinese medicine (TCM).

Study on spectrum-effect relationship of rhizoma Rhei, cortex Magnoliae Officinalis, fructus Aurantii Immaturus and their formula.

Rhizoma Rhei, cortex Magnoliae Officinalis and fructus Aurantii Immaturus compose dachengqi tang (DCQT), a classical formula of traditional Chinese medicine (TCM) that is used for acute intestinal obstruction and has been proven to be effective and economic. However, the ingredients of TCM are complicated, and it is unclear which ingredients are the most important for its effects. In this paper, the relationship between the spectra and effects is discussed to provide a powerful method and some insights into the quality control of the herbs and their formula. High-performance liquid chromatographic (HPLC) fingerprint analysis was performed to investigate the chemical structures in different batches of rhizoma Rhei, cortex Magnoliae Officinalis, fructus Aurantii Immaturus and DCQT. Hierarchical clustering analysis was employed to evaluate the similarities between fingerprints. Animal model of small intestinal propulsion was established to study the purgative functions of the herbs and DCQT. The relationship between the chemical ingredients and the effects was explored by regression analysis. HPLC fingerprint analysis results demonstrated variations between ingredients in different batches of rhizoma Rhei, cortex Magnoliae Officinalis, fructus Aurantii Immaturus and DCQT. The origin, collection time and preparation process may have contributed to these differences. Small intestinal propulsion results showed that, compared with the control group, the positive and therapeutic groups including single herbs and formula were significantly effective (P < 0.05). Spectrum-effect relationship results indicated that seven peak ingredients, hesperidin, aloe-emodin, honokiol, rhein, magnolol, emodin and sennoside A, were inducted in the regression equation, among which, the influence of sennoside A was the largest and most positively associated with the effects. The data analysis results indicated that many ingredients contributed to the purgative effects, among which, sennoside A might be the most important effective component; therefore, sennoside A should be determined for quality control. Furthermore, the spectrum-effect relationship is simple, operative and suitable for the quality evaluation of TCM.

Bufothionine, a possible effective component in cinobufocini injection for hepatocellular carcinoma.

ETHNOPHARMACOLOGICAL RELEVANCE: Cinobufacini has been traditionally used in China for the treatment of tumor since hundreds years ago. For recent years, its modern preparation,cinobifucini injection has also obtained satisfactory therapeutic functions for cancer. MATERIALS AND METHODS: High performance liquid chromatography (HPLC) analysis was applied to determine the content of cinobufagin, resibufogenin and bufothionine in cinobufacin extract liquid and injection; MTT assay and flow cytometric analysis were also respectively used to study the effect of cinobufacini extract liquid, injection and three chemical structures on cells and cell cycles. RESULTS: HPLC results demonstrated that in cinobufacini extract liquid three ingredients (cinobufagin, resibufogenin and bufothionine) were all monitored while in cinofacini injection only bufothinone was detected; MTT assays showed bufothionine could obviously inhibit the proliferation of human hepatocellular carcinoma cell lines such as SMMC-7721 and BEL-7402 in a dose- and time-dependent manner as well as cinobufagin and resibufogenin; further flow cytometric analysis indicated obvious increases in G2/M phase and decrease in G0/G1 phase when SMMC-7721 cell line exposure to bufothionine (480 µg/ml). CONCLUSIONS: These results suggested bufothionine could be involved in treatment of human cancer for cinobufacini injection and the mechanism might be relative to induce G2/M phase cell cycle arrest.