Study On The Quality Standard Setting,stability And Antibacterial And Anti-inflammatory Effects Of Horse Oil Arnebia Euchroma Cream

The horse oil contains oleic acid,linoleic acid and palmitto acid,in which the content of unsaturated fatty acids is more than 60%.The horse oil has the effects of diminishing skin inflammation and enhancing permeability.It will be a potential market prospect if the horse oil is used for materials of products.For above reasons,the horse oil was combined with Arnebia euchroma to play important roles in anti-inflammatory and antibacterial effects.Firstly,the compatibility between raw material and excipients in the horse oil Arnebia euchroma cream was investigated.Then,the quality standard of horse oil Arnebia euchroma cream,stability,anti-inflammatory and antibacterial effects were studied for providing a basis of the further development and application of horse oil functional products.The main results are as follows:(1)The compatibility was investigated between Arnebia euchroma oil raw material and excipients such as horse oil,glycerol diesters horse oil,white beeswax,petroleum jelly,vitamin E and lavender essential oil by the influence factor experiments using the retention rate of total hydroxynaphthoquinone as measurement indicator,which is the active ingredient in Arnebia euchroma.Glycerol diesters horse oil,petrolatum,white beeswax,vitamin E and lavender essential oil were optimized as excipients in the horse oil Arnebia euchroma cream.Furthermore,when the horse oil Arnebia euchroma cream was prepared,before Arnebia euchroma oil was added into,the temperature should be fallen below 55℃ and protected from light to ensure the stability of the total hydroxynaphthoquinone pigment in Arnebia euchroma.The products should be stored and transported in low temperatures,light-proof and dry conditions.(2)The quality standards of horse oil Arnebia euchroma cream were established in the reference of the Q/320500 SZAY-002(emollient ointment),the corresponding indexes of the China Pharmacopoeia(2020 edition),and the test results of three batches of samples.The results showed that three batches of horse oil Arnebia euchroma cream were purplish red,uniform and exquisite,a light lavender odour,peroxide values were less than 10 mmol/kg.In thin-layer chromatograms,the position and color of the spots of the samples were consistent with those of the Arnebia euchroma control herbs and reference standard of levotaxanin,while the negative control didn’t interfere.The content of total hydroxynaphthoquinone in all three batches of horse oil Arnebia euchroma cream was greater than 0.35mg/g,which met the requirements of the quality standards of horse oil Arnebia euchroma cream.The stability trial of horse oil Arnebia euchroma cream was conducted in the reference of guidelines for drug stability test in China Pharmacopoeia(2020 edition).The results showed that horse oil Arnebia euchroma cream should be stored in light-proof,seal,and low temperature(5℃)conditions.In these situations,the expiration date of products is tentatively fixed for twelve months.(3)The Oxford cup method was used to study the inhibitory effects of horse oil Arnebia euchroma cream on Staphylococcus aureus,Pseudomonas aeruginosa,and Escherichia coli in vitro.The effects of horse oil Arnebia euchroma cream on auricle swelling degree,auricle histomorphology and skin capillary permeability in mice were investigated through the models of auricle swelling and skin capillary permeability increasing caused by xylene in mice,in order to investigate the ameliorative effect of horse oil Arnebia euchroma cream on skin inflammation.Results showed that the high dose group of horse oil Arnebia euchroma cream was medium sensitive to Staphylococcus aureus and Escherichia coli and low sensitive to Pseudomonas aeruginosa.Furthermore,the antibacterial effects of horse oil Arnebia euchroma cream were comparable to those of Jingwanhong Ointment(positive control).The middle and high dose groups of horse oil Arnebia euchroma cream could significantly reduce auricle swelling degree in mice(P<0.05,P<0.01).Notably,the auricle swelling degree in the high dose group of horse oil Arnebia euchroma cream was significantly lower(P<0.05)than that of Jingwanhong Ointment(positive control).Meanwhile,horse oil Arnebia euchroma cream could obviously help improve symptoms caused by inflammatory cell infiltration such as auricl epidermal thickening,epidermal sponge-like edema,and vascular congestion.The middle and high dose groups of horse oil Arnebia euchroma cream could significantly reduced capillary permeability in the abdomen of mice(P<0.05,P<0.01).Most of all,the capillary permeability in the high dose group of horse oil Arnebia euchroma cream was significantly lower(P<0.05)than that of Jingwanhong Ointment(positive control).It is shown that the horse oil Arnebia euchroma cream not only inhibits the pathogenic bacteria causing inflammation,but also improve the symptoms induced by inflammatory such as swelling of skin tissue,increasing capillary permeability and tissue fluid exudation.(4)The anti-inflammatory active components,targets and mechanisms of Arnebia euchroma were explored through network pharmacology.The results could predict that the anti-inflammatory active components in Arnebia euchroma are mainly 11 chemical components such as Des-O-methyllasiodiplodin,Mandenol,Ethyl oleate,Shikonin and Acetylshikonin,etc.Meanwhile,three proteins such as PTGS2,ALB and MMP9 are main targets.Moreover,the mechanism of anti-inflammatory may be the cancer pathway,lipid and atherosclerosis,arachidonic acid metabolism,TNF signaling pathway,IL-17 signaling pathway,and AGE-RAGE signaling pathway,etc.