Development Of A Hemostatic Disinfectant From Callicarpa Nudiflora Hook.et Arn.in Li Nationality

Callicarpa nudiflora Hooket Am.is a kind of folk medicine used in Li nationality area of Hainan Province.In this paper,the hemostatic disinfectant was developed by the leaves of nudiflora which was served as raw materials.Then,we have performed the extraction process,content determination,selection of auxiliary materials,preparation technology and the function test.The main contents are as follows:Take the total flavonoids as the index,then the comparison of ASE,ultrasound,microwave and multi-function concentration extraction for total flavonoids from the leaf of nudiflora have been performed.Ultimately,the ASE extraction rate of total flavonoid was the highest about 1.26%,ultrasonic and microwave extraction method following by approximately 1.03%and 0.98%,and the multi-function concentrated extraction was the lowest about 0.36%.In the optimization of solvent selection,resazurin microplate screening experiment has been tested.We obtained that ethanol and n-butanol had lower concentrations with 0.022 g.mL-1 and 0.033 g.mL-1 respectively comparing with other 3 kinds of extraction liquid.However,the n-butanol has higher toxicant than ethanol,so it was selected as agent extract of nudiflora leaf.Verbascoside content in nudiflora is high as a hemostatic active ingredient was consided as the index for its detection.When HPLC was carried out with chromatographic conditions as follows:Hypersil Gold column(4.6 mm × 250 mm,5μm),the mobile phase of acetonitrile-methanol-water(20:5:75),flow rate at 1 mL·min-1,and the detection wavelength at 334 nm,the retention time of nudiflora was 6 min,the separation degree of principal component and other peaks was more than 1.5 and the number of pedal was greater than 5000,the linearity of verb ascoside was good in the range of 0.05～0.2mg/mL(r2 = 0.9909),the average recovery was 99.41%,while RSD was 1.44%.There were good precision,repeatability and stability,and the RSD respectively was 1.34%,1.24%,1.62%(n =6).Finally,,the method was simple,feasible and reproducible and was used to control the quality of nudiflora.On the basis of the above experiments,the optimum extraction process of Rulein was optimized by orthogonal method,and then MIC test was carried out to determine the concentration of the drug for A disinfectant with good bactericidal and hemostatic effects.Then,due to consider enlarge production,so the most commonly method of water extraction alcohol precipitation was used.The results showed that when the extraction temperature was 120 ℃,times was 1,the concentration of ethanol was 50%and the time was 12 min,the extraction rate of Verbascoside was 2.3%and the MIC was 0.02 g/mL.Ultimately,we found Verbascoside concentration at least 2 mg/mL,total flavonoids concentration at least 0.2 mg/mL so that it can play the hemostatic and antibacterial effect.In the choice of excipients and process optimization,glycerin,propylene glycol,salicylic acid and borax were selected for adjunct dosage and sequencing test.And the centrifugal speed,the added amount test of activated carbon and choice of dosage form was tested.Finally,the centrifuged speed of extractive liquid was 5000 rad/min,without the use of activated carbon,propylene glycol first,then add salicylic acid to adjust the pH to 4.5±0.5～6±0.5.Because the spray is evenly sprayed and consumed less,used without contact with the skin,so,the dosage form of spray was selected.In order to determine the quality and efficacy of the new disinfectant samples,this experiment has tested the sensory properties of samples,heavy metal content,rapid killing of microorganisms,skin and eye irritation,the long-term bacteriostasis and the hemostatic property,as well Analgesic and anti-inflammatory,stability.The results show that the sensory traits had no obvious change in three months.Pb,As,Hg were lower than the requirements.The rapid bacteriostatic test was meted the requirements.The sample did not irritate to the skin and eyes of the rabbits.Both sterilization and hemostatic effects of the samples were stronger than Yunnan Baiyao.The positive drug had the similar analgesic effect with the sample.54 ℃ stability trial showed the original liquid and sample pH changes were stable and the total flavonoids absorbance was become larger.The original liquid Verbascoside peak area significantly decreased compared to the sample,which may be the sample has added stability adjuncts.4 ℃ and 25 ℃ have no significant effect on the sample and the original liquid.However,light has a significant effect on Verbascoside and flavonoid of the original liquid,so it is suggested to keep the sample in dark with low temperature.