| Chinese medicine played a significant role in fighting against plague over long history in our country. In addition, it has not only summarized a huge amount of classical prescriptions but also enriched and developed many excellent Chinese medicine theories such as Defense-Qi-Nutrient-Blood Syndrome and Triple Energizer Pattern Identification. In recent years, SARS and Influenza A virus subtype H1N1 spread severely all over the world, which led to an urgent need for anti-bacteria and anti-viruses hand sanitizer in market. In this study, Xiang Xu" Couple Herbs in Lonicera japonica-Forsythia suspense(LF) were chosen as the main subject, which was derived from "Yin qiao powder"--a well known powder composed of Forsythia suspense, Lonicera japonica, Platycodon grandiflorus, Herba Menthae Heplocalycis, Zanthoxylum armatum, Radix Rhapontici Rhaponticum uniflorum, Schizonepeta tenuifolia, Fructus Arctii. The main objective of this study is to investigate the synergistic effect of active components from LF, develop Yinghua-lianqiao hydrogel formulation with the combination of traditional Chinese clinical experience and modern dose technology, establish complete quality controlling system, declare and register the disinfection product and further promote it to market. The content and results are shown as follows:1. Chlorogenic acid,forsythoside A were chosen as the antibacterial index components in LF. Loganin and anti-inflammatory effective parts of Forsythia suspense were selected as the anti-inflammatory index components. The difference between the antibacterial and anti-inflammatory effect before and after the compatibility of active components was also investigated.The results indicated that:①Forsythia shell decoction was the main anti-inflammatory part of Forsythia suspense and it showed better anti-inflammatory effect than essential oil and decoction without volatile oil of Forsythia seeds; The cooperated loganin and Forsythia shell decoction displayed significantly different anti-inflammatory effect; forsythoside A did not have remarkable anti-inflammatory effect, however, this effect was greatly improved after forsythoside A was cooperated with chlorogenic acid or loganin. Therefore LF had synergistic effect in anti-inflammatory.②The cooperated chlorogenic acid and forsythoside A had synergistic effect in anti E.coli, which proved they had synergistic antibacterial effect.2. With chlorogenic acid and forsythoside A as index components, single decoction and mixed decoction of LF cooperation from different producing areas were analyzed by HPLC. The extraction of LF was optimized by orthogonal design. In addition, single factor and orthogonal experiment were employed to select favorable hydrogel forming agent and optimize the description and technology of Yinghua-lianqiao hydrogel formulation.The results indicated①There were totally 15 peaks of single and mixed decoction of LF. In addition, the peak1,3,4,5,6,7,8,9,11,13,14 of the mixed decoction was apparently higher than those of single extraction. The peak number and area of the mixed decoction of LF produced in henan province were much higher than those produced in other places.②The optimal orthogonal condition for the extraction of LF was A1B2C3D1,that is ultrosonic extraction time of 10 min,50% ethanol, extraction of 3 times and solvent of 20 ml.③Carbopol U20 is better than other supplementary materials in the hydrogel shape and transparency. The prescription of Yinghua-lianqiao hydrogel formulation was A3B1C1D1,namely the contents of ethanol, extraction agent and humectant were 60%,0.1%,0.8% and 1.0% respectively.3. The quality standard introduction of Yinghua-lianqiao hydrogel formulation was drafted and its inspection items were investigated:property, identification, determination of active components content, initial stability, penetration degree, bacteriostatic effect and repeated test of skin stimulation.The quality standard of Yinghua-lianqiao hydrogel formulation was established. The results indicated that:①The spots of TLC were clear, undisturbed and easy to distinguish.②In the meanwhile, a HPLC method was also established to quantify chlorogenic acid,forsythin and forsythoside A in hydrogel formulation simultaneously. The investigation of methodology showed the calibrated curves of chlorogenic acid,forsythin and forsythoside A exhibited excellent linear relationship between 8.0～48.0,1.0～100.0 and 5.0～1000.0μg/mL (r=0.9992,0.9996,0.9996)and the average recovery rate were 99.23±2.26%,99.48±2.29% and 99.21±1.77%, respectively. In addition, the contents of chlorogenic acid,forsythin and forsythoside A in hydrogel formulation were 0.093±1.28%,0.020±2.61% and 0.058±0.98%;③Yinghua-lianqiao hydrogel formulation was able to inhibit Staphylococcus aureus,Escherichia Coli and Candida albican;④The active components of Yinghua-lianqiao hydrogel formulation cannot pass through skin. Yinghua-lianqiao hydrogel formulation displayed light stimulation.⑤The decreasing rates of ethanol and active components were lower than 4% and 2% respectively showing a good stability.4. The amplifying pilot production of Yinghua-lianqiao hydrogel formulation, product positioning and production were carried out. The results indicated Yinghua-lianqiao hydrogel formulation was well produced in pilot plant and it could also be further promoted to market by designing the logo introduction of product as well as declaring disinfection product. |
