Preparation And Evaluation And Its Hemostatic Mechanism Of Chlorine Dioxide Spray System

Objective: For preparation binary aerosol, stable chlorine dioxide solution wasregarded as main drug, suitable polymer materials as its carrier, which can overcomethe instability of chlorine dioxide. In this paper, about above-mentioned aerosol,making quality evaluation and screening of antibacterial activity. Experimenting forskin irritation and safety evaluation with animal subjects, making preliminaryresearch for hemostatic mechanism.Methods:The iodometric method was used to determine the contents of self-madestable chlorine dioxide solution. To establish a determine method about contents ofself-made chlorine dioxide aerosol through adopting a reversed phase, highefficiency，liquid ion to chromatography. The release degree is regarded as theevaluation index. UV spectrophotometric method was used to set up a method ofdetermining contents. Making screen respectively prescription composition of sodiumalginate and use level of glycerol,etc. The orthogonal test has been adopted toascertain prescription composition.According to the provisions of ChinesePharmacopoeia2010edition about characters of aerosol, pH value, centrifugal testetc.,the quality evaluation has been finished.In order to making evaluate the safety ofchlorine dioxide aerosol, rabbit skin was regarded as the research object to doexperiment of skin irritation. Staphylococcus aureus, Escherichia coli and Candidaalbicans were regarded as testing fungus, to have been tested for antibacterialproperty of chlorine dioxide aerosol. It has been measured by a semi automatic bloodcoagulation analyzer for the plasma thrombin time (TT), prothrombin time (PT),activated partial thromboplastin time (APTT), fibrinogen (FIB), that rabbits acted asanimal subjects. To measuring the rat platelet aggregation rate under acting ofexutory of two-adenosine-monophosphate(ADP), peanut four acid (AA), plateletactivating factor (PAF), etc., whose animal subjects were rats. Making preliminaryresearch for hemostatic mechanism of chlorine dioxide.Results:The three batch of self-made stable chlorine dioxide solution concentrationwere21.96mg/mL,23.77mg/mL,20.60mg/mL respective that has been measured with iodometric method. The values of RSD were respective1.4%,0%,1.7%. Tomeasure contentss of the chlorine dioxide aerosol through adopting a reversed phase,high efficiency， liquid ion to chromatography. The average percent recoveryof sample is98.6%, the results of the RSD value is2.4%, which indicates thismethod being accurate and reliable. To measure contentss of stable chlorine dioxidesolution through adopting UV spectrophotometry. The average percent recoveryof sample is97.9%, RSD value is1.7%. The ultimate determined formulation ofchlorine dioxide aerosol is: sodium alginate was0.25g, glycerol is1.0g, polyacrylicacid is200μL, ethanol is4.0mL, stable chlorine dioxide solution is24.72mg, that hasbeen ascertained through orthogonal method. The self-made aerosol is a kind ofcolorless transparent solution and its pH value is about4.6. There is no stratificationphenomenon after being centrifugated. Its stability test has showed that thepreparation should be stored in a dark place and about20℃. The safety of preparationis reliable. There is no redness swelling and plaque eschar on the skin of rabbits,which was done on the intact or damaged skin of rabbits after single and multiplestimulation testing. After making the minimum bacteriostatic and bactericidal testwith Staphylococcus aureus, Escherichia coli, Candida albicans, we have known thatminimum bacteriostatic concentrations are:18.6,18.6,37.1μg/mL; minimumbactericidal concentrations are:148.6,148.6,1188μg/mL, their preparationconcentrations correspond on three kinds of above bacteria respectively; When theconcentration of chlorine dioxide is in the range of0.05mg/mL-0.20mg/mL,thechange percentage value of thrombin time (TT)and in the range of0.08mg/mL-0.15mg/mL, the average value of partial activated thrombin time (APTT)were better than result of Yunnan Baiyao (in the positive contrast group) respective;Following the increase of concentration of chlorine dioxide, the average value ofprothrombin time (PT) changed irragularily, meanwhile, the coagulation timeof contentss of fibrinogen (FIB) decreased, which was compared with the blank;However its tendency showed that decreased at first and then increased; When thedose of test group equals to positive contrast group, the result of average valuechange percentage of thrombin time (TT) indicated that chlorine dioxide has a slight advantage over Yunnan Baiyao about coagulation time. The result indicated thatthrombin time (APTT)of partial activated test group excels obviorsly the YunnanBaiyao group; The test result of prothrombin time (PT) is slightly higher than thepositive control group, however, the corresponding results of the contentss offibrinogen (FIB) closes to the Yunnan Baiyao group; Through test of plateletaggregation rate, it was known that aggregation rate of Yunnan Baiyao group was40%-50%. While it was also40%-50%that aximum aggregation rate of test group ofchlorine dioxide solution which was induced by two adenosine monophosphate(ADP). It was20%-30%that aximum aggregation rate of test group of chlorinedioxide solution which was induced by peanut four acid (AA) and platelet activatingfactor (PAF), and aximum aggregation rate of saline was0.Conclusions: In this paper, it was stated how to prepare a kind of chlorine dioxidebinary aerosol which can stop bleeding rapidly and is external use on skin. Thequality standard of preparation conforms to the rule inside the ChinesePharmacopoeia of2010edition. Experimental results show that the preparation haskilling effect on Staphylococcus aureus, Escherichia coli, Candida albicans, and therewere no irratation to intact or damaged skin.So it can be used for local rapidhemostasis on skin. It can show through measuring four items indexof Coagulation, the average value of partial activated thromboplastin time (APTT)was degression. According to it, we can infer preliminary that chlorine dioxide maybe a kind of thrombin activator which can promote activation of coagulation factor XIand factor XII, speed up starting the pathway of the endogenous coagulation,thus which can acts as rapid coagulation. Meanwhile, there is no thrombosis tofollow during using it. According to irregular change of average value of theprothrombin time (PT), to infer preliminary that chlorine dioxide induce hardly orpromote the pathway of the extrinsic coagulation. According to degression of wholeaverage value of the thrombin time (TT), when the concentration of chlorine dioxideis in the range of0.01mg/mL-0.20mg/mL. it actson promotion ability of turning plasma fibrinogen into fibrin and quickens thischange, to form more fibrin in a unit of time, thereby, it can act on stopping bleeding quickly. Platelet occursdifferent activating reaction under the actionof different inducer.The result of vitro experiment showed that chlorine dioxide couldcause different degree aggregation of rat vitro platelet under the inducement actionof ADP, PAF and AA. Furthermore, platelet aggregation falls after rising. The dataalso indicate that chlorine dioxide can stop bleeding rapidly, however, there occurnopotential factors of bring thrombosis during stopping bleeding. Among three kindsof inducer which are ADP, PAF and AA, it was expressed that ADP has astronger inducement action on chlorine dioxide, which induce platelet aggregation ofrat. The chlorine dioxide has inducement action on three ways of ADP, PAF and AA,it induces platelet to activate, may be through a common part. At present, itis accepted that platelet activation can pass three ways, they are ADP,AA and PAF.However, wholeprocess aboutplatelet activation aggregation is becauseof dropping cAMP level in platelet and increasing concentration of intracellular freecalcium ion finally. This provides the theory basis for chlorine dioxide becoming aking of waiting to choice hemostatic drug.