Study On The Inhalation Toxicity,Mechanism Microcapsules Predicition,Purification And Controlled Release Of Essential Oil From Acorus Tatarinowii

Acorus tatarinowii is the dried rhizome of Acorus tatarinowii Schott of Araceae.It has the effects of resolving phlegm and opening orifices,awakening mind and improving intelligence,resolving dampness and harmonizing stomach.It is known as“aromatic depression”.Essential oil is one of the main active components of Acorus tatarinowii.Studies have shown that nasal administration of essential oil from Acorus tatarinowii is more likely to penetrate the blood-brain barrier and play a good role in the prevention and treatment of mental diseases,such as epilepsy,depression,post-traumatic stress disorder（PTSD）,etc.However,the lack of attention and research on the safety of essential oil inhalation administration of Acorus tatarinowii,coupled with the shortcomings of essential oil,such as poor stability,low solubility and volatilization,have severely limited its drug development and clinical application.Therefore,this topic chooses to study the inhalation safety of essential oil from Acorus tatarinowii,predict the toxic mechanism,explore the purification process,and prepare it into microcapsules to provide scientific basis for subsequent clinical drug use and product development.1.Study on the safety of essential oil inhalation from Acorus tatarinowii.Through acute and subacute toxicity tests,the general performance,blood routine,blood biochemistry,organ coefficient and other indexes of animals were observed to evaluate the safety of inhalation administration of essential oil from Acorus tatarinowii.The results showed that the LD₅₀ of Acorus tatarinowii essential oil on mice was 38.05m L/m³,with 95%confidence interval of 35.23～41.12 m L/m³.The 12.68 and 44.39m L/m³ essential oil of Acorus tatarinowii significantly decreased the food intake of female and male rats and inhibited the weight gain（P<0.05）;RBC,HGB,HCT,PLT,AST,BUN,CR and T-CHO were significantly increased in male rats,and MPV,PDW and GLU were decreased,while HGB,HCT,PLT,PDW,AST,BUN,T-CHO and CR were significantly increased in female rats.Reduced WBC,Mon,MCHC,GLU,TP,and ALB（P<0.05）;Significantly increased brain,liver,lung and kidney indexes of female and male rats（P<0.05）;Histopathologic sections showed that the liver,kidney,lung,brain and nose of female and male rats showed obvious pathological injuries,with varying degrees of inflammatory cell infiltration and cell necrosis.2.Network toxicology of essential oil from Acorus tatarinowii.The toxic components,action targets and pathways of essenyial oil from Acorus tatarinowii,were predicted by GC-MS combined with network toxicology.The results showed that 28components were identified from the essential oil of Acorus tatarinowii.It is possible that major toxic components such as methyleugenol andα-pinene act on key targets such as EP300,EGFR,CREBBP,RELA and ESR1,and regulate HIF-1 signaling pathway,JAK-STAT signaling pathway,and TH17 cell differentiation to produce liver and renal toxicity.It is harmful to nervous system,respiratory system and reproductive development system.3.Preliminary study on purification technology of essential oil from Acorus tatarinowii.Taking the toxic components methyleugenol and pharmacodynamic componentsβ-asarone as the target components,the initial separation and purification process parameters were determined by changing the evaporation intensity of molecular distillation and taking the total recovery and the relative contents of the two components as the objects of investigation.The results showed that the total recovery reached 90.0%when the feed liquid flow rate was 2～2.5 m L/min,the film scraping rate was 250 r/min,the evaporation temperature was 60℃,the vacuum degree was 1 mbar,the primary condensation temperature was 5℃,the secondary condensation temperature was-15℃,and theβ-asarone was mainly concentrated in the heavy fraction,and the methyleugenol content decreased by 10.63%,while the middle fraction only contained methyleugenol,and noβ-asarone was detected,which initially realized the removal of toxic components and enrichment of effective components.4.Study on essential oil microcapsules from Acorus tatarinowii.The essential oil microcapsules of Acorus tatarinowii were prepared by spray drying method and the encapsulation technology was optimized by single factor analysis.The morphology,size and properties of microcapsules were studied by means of SEM,laser particle size analyzer,FTIR,TG and XRD.The odor and in vitro drug release characteristics of toxic/active components after microencapsulation of essential oil from Acorus tatarinowii were analyzed and evaluated by electronic nose and in vitro release experiments,and the stability of high temperature and high humidity were measured.The results showed that when the dosage of solid material was 10%,the ratio of gelatin to Arabic was 1:1,the ratio of capsule core to capsule material was 1:4,the stirring temperature was 60℃,the stirring speed was 1000 r/min,the air inlet temperature of spray dryer was 155℃,and the injection rate was 10%,the encapsulation rate and drug loading were the highest,and the process was stable and reliable.The surface of the essential oil microcapsules prepared under the optimal prescription was spherical with no cracks or holes,and the particle size was 12.64±0.24μm.RTIR,TG and XRD confirmed that the essential oil was successfully embedded in the microcapsules,and the embedding process was only a simple physical embedding process.The results of electronic nose showed that the essential oil of Acorus tatarinowii improved its bad odor obviously after microencapsulation.The in vitro release results showed that the microencapsulation technology could effectively reduce the release of toxic/active components of essential oil from Acorus tatarinowii,and the release process was consistent with the first order equation.The results of high temperature and high humidity test showed that the stability of the essential oil from Acorus tatarinowii was greatly improved and the change of its quality was reduced after microencapsulation.