Study On Extraction Process And Preliminary Pharmacodynamics Of Fugui Gutong Decotion

Fugui Gutong Decotion is made of 8 kinds of Chinese herbs, including aconite root, monkshood root, cinnamon, lanceolata, angelica, white peony root, epimedy and olibanum. The prescription was an effective recipe-Wutou Decotion documented in《Synopisis of the Golden chamber》written by doctor Zhang Zhongjing in Han dynasty.The efficiencies of Fugui Gutong Decotion are cleaning away prolonged cold-damp obstruction, expelling cold and damp pathogen, warming meridian to activate, subduing swelling and alleviating pain, which is used to treat knee osteoarthritis. It was reported that the clinical effective rate of Fugui Gutong Decotion is above 95 %.At present, Fugui Gutong Decotion is extracted by conventional method. Though it is the ideal drug for the disease, patients have to take a large dose due to the low content of effective components in Fugui Gutong Decotion. In order to overcome the above shortcomings, this study makes a more thorough study, including the follows:(1)The establishment of analytical methods: Aconitine-type alkaloids in Fugui Gutong decotion is determined by the acid dye colorimetry based on the spectrometric features of ion pair compound, which is extracted by chloroform, produced in the reaction between aconitine-type alkaloids and bromcresol green in citric acid-disodium hydrogen phosphate buffer solution. The absorbance of ion pair compound was detected at the wavelength of 412.5nm. The method is accurate, reliable, feasible, and has good exclusivity for determining the content of aconitine-type alkaloids in Fugui Gutong decotion.Paeoniflorin in Fugui Gutong Decotion is determined by HPLC method and the mobile phase was acetonitrile-0.1 % phosphoric acid (14:86). The results showed that Paeoniflorin peak was separated from other impurities with the sample prepared simply. The method was simple, accurate and reproducible, and suitable for the determination-(2) Optimization of extraction process for Fugui Gutong Decotion: This study investigates three extraction methods, including yellow rice wine, semi-bionic, and microbial fermentation. Orthogonal design was used to optimize the extraction conditions of the above extraction methods with the evaluation indexes such as aconitine-type alkaloids, paeoniflorin and dried extraction. The optimized conditions for yellow rice wine extraction comprise the follows: the extraction solvent was 85% yellow rice wine; material granularity was decoction pieces; total time of extraction was 4 h. When semi-bionic extraction was as follows: the extraction pH was 2.2, 6.0, 9.0 respectively, and total time of extraction was 4 h. The results of comparing the extraction methods by comprehensive value Y showed that: the microbial fermentation was the highest, semi-bionic extraction followed, and the yellow rice wine extraction water extraction was the lowest.(3) Study on primary pharmacodynamics for Fugui Gutong Decotion fermented. The oral acute toxicity study revealed that the LD₅₀ of fermented extract was 133.70g·kg^-1 in mice, which was higher than that of conventional extract LD₅₀(118.16g·kg^-1). The results suggested that the fermented Fugui Gutong Decotion was safer than conventional extract. Fermented extract could inhibit ear inflammation induced by xylene in mice and the increase of abdominal cavity capillary permeability in mice caused by acetic acid. It was demonstrated that Fugui Gutong Decotion fermented had anti-inflammatory effects. Analgesic tests showed that the fermented extract could significantly improve the pain threshold of mice, and effectively reduce writhing response frequency induced by acetic acid. The results showed that anti-inflammatory and analgesic effect of Fugui Gutong Decotion fermented was obviously better than that of conventional extraction.