| Objective:purpose:When using Gamuzhuer powder for external use,it has poor adhesion,easy to fall off,not used in special locations,and low bioavailability.Therefore,modern methods and materials are used to study the dosage form change of Gamuzhuer powder to provide patients with more aspect.Method:(1)Taking the uniformity of the solution,the shedding phenomenon of the powder,the color and weight difference of the patch as the inspection indicators,the Gamuzhuer band-aids were prepared through single factor and orthogonal experiments.(2)Refer to“Chinese Pharmacopoeia”(2015)and related literature to determine the shape,identification,content and other aspects of Gamuzhuer Band-Aid.Appearance is identified by color and sense of smell:chemical method is used to identify the main component of red gypsum,CaS04·2H2O,mainly to observe whether a white precipitate is formed;whether the precipitate is dissolved in hydrochloric acid.Identification of cinnabar and silver vermilion:add hydrochloric acid and nitric acid solution and observe whether there will be precipitation decomposition in the last step of titration.The borneol content in the preparation was determined by gas chromatography(GC),TG-WAXMS capillary column(30×0.25mm,0.25μm),FID detector,the inlet temperature was 220℃,and the detection temperature was 250℃.The column temperature is 100℃ as the initial temperature,keep for 2min,increase to 110℃ with 2℃·min-1,keep 3min,increase to 205℃with 50℃·min-1,keep 7min,increase with 20℃·min-1 Keep it at 250℃ for 2 minutes;the carrier gas is nitrogen,the flow rate is 1ml·min-1,the injection method is split injection,the split ratio is 10:1;the injection volume is lul,and naphthalene is used as the internal standard.The content of calcium sulfate in water in red gypsum was determined by coordination titration with disodium ethylenediaminetetraacetate.The content of arsenic disulfide in realgar was determined by titration with iodine titrant.(3)The 72 Wistar rats were divided into a drug treatment group and a model group,with 12 rats in each group.The 6 treatment groups were high-dose Gamuzhuer wound patch group,medium-dose Gamuzhuer wound patch group,low-dose Gamuzhuer wound patch group,Gamuzhuer powder group,and blank matrix group.A round skin full-thickness incision wound of about 0.6cm×0.6cm wound was made on the back of each rat.All wounded rats were treated immediately after preparing the skin damage wounds for 9 consecutive days.After 3、5、7 and 9 days after the operation,the wounds of each group were photographed,and the data of the residual wound area of the rats were recorded.And 3、5、7、9 days after the operation,tissue samples were taken for HE staining.Observe the histopathological features under an ordinary optical microscope.(4)Skin irritation and allergy test:Before the experiment 4-24h,the rabbit back hair was depilated with sodium sulfide,and the skin was wiped with distilled water at 40℃.After the sodium sulfide was washed and removed,the experiment was carried out.Single contact test:fix the patch on the depilatory part for 4h.After the contact period is over,remove the applicator,wash it with warm water or other suitable non-irritating solvent,and dry it carefully.Respectively after removing the patch 4h,record the situation of each contact part.Repeated exposure experiment:repeat this experiment after the single exposure test is completed,4 hours a day,continuous 3d observation and recording.Record the contact area after each removal of the patch and before contacting again.After the last contact,record the conditions of each contact site after removing the patch.Observation indicators:Observe the local skin erythema,edema,congestion and extent of rabbits with naked eyes,and express them by scoring;at the same time,observe the general status,behavior and physical signs of rabbits.After the experiment,HE staining was performed.Observe the morphological changes of the skin tissue after administration under a microscope.Allergic experimental study:sensitization exposure:stick the Gamuzhuer patch group,matrix group,and positive control(1%2,4-dinitrochlorobenzene)on the left side of the back of the guinea pig for 8 hours,on the 7th day and The same method was repeated once on the 14th day.Provocation contact:14 days after the last sensitization of the test substance,the test substance was applied to the depilatory area on the right side of the corresponding animal.After 8 hours,the test substance was removed for observation,and the skin allergy was observed again at 24h、48h、72h Reflect the situation and score according to the "Scoring Standard for the Degree of Skin Allergic Reaction",and calculate the average score of the animal reaction in each group.Result:Gamuzhuer patch was prepared with 20ml of 60%ethanol as the dispersion medium,0.4g of flax oil as the oil phase,and 0.4g of glycerin as the humectant.(2)Gamuzhuer Band-Aid has both the light yellow and aromatic flavor of Gamuzhuer powder.The identification reaction of red gypsum produces a white precipitate;the precipitate does not dissolve in hydrochloric acid.Identification of cinnabar and cinnabar:add a mixed solution of hydrochloric acid and nitric acid to dissolve,evaporate to dryness,add water to dissolve,filter,add lead acetate test solution to the filtrate to produce white precipitate,separate,and the precipitate is dissolved in sodium hydroxide test solution.Content result:The order and level of the influence of various factors on the content of borneol and isoborneol are analyzed through orthogonal experiment results.A>B>C,A1>B1>C1 that is,the best extraction process is 20 patches and 10ml acetic acid.Ethyl,ultrasound at 200 C for 5 min.The peak areas of borneol and isoborneol have a good linear relationship in the ranges of 3.66～234.800μg/ml and 2.228～146.400μg/ml,respectively.The linear equations of borneol and isoborneol are Y=0.055x+0.057(R2=0.999),Y=0.089x-0.023(R2=0.999),the average recoveries were 89.7%and 93.8%,RSD were 1.4%and 2.8%,respectively;the average content of borneol and isoborneol in the three batches of samples was 49.6 μg/tablet,35μg/tablet.The linear equation of red gypsum is that the recovery rate is 91.55%,RSD=2.29%,the average content of red gypsum in the three batches of samples is 89.9 u g/piece,and RSD=1.15%.The realgar recovery rate was 93.9%,RSD=2.4%,the average content of realgar in the three batches of samples was 49.9 u g/piece,and RSD=1.6%.(3)On the 3rd and 5th day after the operation,the medium-dose Gamuzhuer band-aid group was compared with the high-dose Gamuzhuer band-aid group,and the low-dose Gamuzhuer band-aid group was compared with the high-dose Gamuzhuer band-aid group.There was a statistically significant difference between the powder group and the high-dose Gamuzhuer band-aid group(P<0.05).There was a statistically significant difference between the low-dose Gamuzhuer band-aid group on the 7th and 9th day compared with other groups(P<0.05).Observed under an optical microscope,it was found that the low-dose Gamuzhuer band-aid group had less inflammation and more new granulation tissues than the model group.Compared with the high-dose Gamuzhuer band-aid group,the inflammation reduction time was earlier,the granulation tissue appeared earlier,and the skin recovery time was shorter than that of the high-dose Gamuzhuer band-aid group.The low-dose Gamuzhuer bandage group had the smallest residual wound area and the best healing effect.(4)No erythema and edema were observed by naked eyes.Under the HE staining light microscope,the drug group had no damage to the epidermis and dermis of the rabbits,the thickness did not change,and there was no inflammatory exudation.There was no redness,swelling,water spots,etc.on the guinea pigs.Gamuzhuer patch is not irritating and allergic.Conclusion:1.0.4g flax oil,0.4g glycerin,60%20ml ethanol is the best process for preparing Gamuzhuer Band-Aid.2.Identification of red gypsum,cinnabar and silver vermilion by physical and chemical methods.The content of borneol is 84.6 u g/piece.The content of red gypsum is 89.9 μg/piece.The realgar content is 49.9μg/piece.3.The low-dose Gamuzhuer bandage group has the best efficacy.4.Gamuzhuer Band-Aid has no irritation and allergic reactions. |
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