| Periploca forrestti Schltr. belongs to Asclepiadaceae family, Periploca Linn. genus, the rhizome of the southwest Periploca sepium. It is the characteristic medicine of Miao. It was commonly used in the treatment of rheumatic arthritis, traumatic injury, irregular menses, fracture. Recently, the researches about this medicine mostly focus on chemical constitutent and pharmacological activities. According to the previous study, the saponins constituents in this medicine has close relationship with the anti-inflammatory effect. The current research on saponins include pharmacological, biological transformation and biogenetic synthesis. The biological transformation of saponins is mainly focused on the structural modification and extraction of the target compounds. The application of biological technology in the transformation of saponins in Periploca forrestti Schltr. is conducive to the development and utilization of resources of this medicine.Combined with the preliminary study of Periploca forrestti Schltr., this paper using enzyme assisted extraction method increased the extraction rate of saponins components in Periploca forrestti Schltr.. At the same time, the enzyme hydrolysis process was optimized and validated. Then, the anti-inflammatory activity was investigated. And the characteristic peak of chlorogenic acid in Periploca forrestti Schltr. was identified and the HPLC determination method for it was established. The results were as follows.1 By the UV spectrophotometry technology established the determination method of the saponins compounds in Periploca forrestti Schltr. and through nineteen enzymes it was found that the mix-enzyme(cellulase and pectinase) which can significantly improve the extraction rate of saponins compounds. On the basis of single factor investigation, the Box-Behnken response surface methodology was used to optimize the enzymatic extraction process. The optimal enzymatic extraction conditions are 51.97 oC, 3.5 of pH, 1.55% of enzyme dosage, 8: 2 of enzyme ratio(pectinase and cellulase). And the experiment results showed that the extraction rate was 55% more than the traditional extract method. The verification and amplification experiment were done later. The method provided reference for the further utilization of the active components in the Periploca forrestti Schltr.2 Adjuvant arthritis rat model was established to investigate the pharmacodynamic effects of the extract which was got by the enzyme extraction method from the Periploca forrestti Schltr. The experiments with diameter of the ankle joint and foot palm thickness of rats, weight, content of PGE2, organ coefficient of thymus, renicapsule, spleen, the change degree of ankle joint pathological as evaluation indexes to observed and compared effect of traditional extraction and enzymatic extraction of saponins compounds on AA rats. Ultimately it was confirmed that the enzyme-assisted extract could effectively improve the anti rheumatoid arthritis pharmacodynamic activity of Periploca forrestti Schltr.. It provided scientific basis for expanding the application of Periploca forrestti Schltr. on the anti rheumatoid arthritis.3 It was confirmed that chlorogenic acid was an absorbed component in prototype in Periploca forrestti Schltr.. With the content of chlorogenic acid as index, the orthogonal design method was used to optimize the pretreatment process of Periploca forrestti Schltr. The selected extraction conditions were 70 oC of reflux temperature, 1 h of extract time twice, 60% methanol of solvent, 10 mL of solvent volume. Then, the C18 column was used for separation of chlorogenic acid in Periploca forrestti Schltr. Results showed that under the conditions of flow rate of 1mL·min-1, the mobile phase consisted of 0.5% formic acid: water =9: 91, the recovery rate could reach 98.569%. The method was convenient, rapid and had good recovery rate and precision, which provides a basis for the research of the quality control of the Periploca forrestti Schltr. |
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