Preparation,Quality Evaluation And Safety Study Of Nasal Mucosa Moisturizing Microemulsion

Objective:In this paper,nasal mucosal moisturizing microemulsion was prepared with peppermint oil as the main drug and glycerol as the moisturizer.The formulation and preparation process were optimized,and the quality evaluation,stability,efficacy,toxicity and irritation of the microemulsion were studied.Methods:?1?Preparation of nasal mucosa moisturizing microemulsion:?1?Prescription screening:The oil?O?,surfactant?SF?,cosurfactant?CoSF?,the mass ratio of surfactant to cosurfactant?K_m?,glycerol content,each component and dosage of nasal mucosa moisturizing microemulsion were studied by HLB,single factor test and pseudo-ternary phase diagram.?2?Preparation process:The single factor experiment was conducted to investigate the influence of stirring time,stirring speed and sample adding mode on the process.The preparation process of nasal mucosa moisturizing microemulsion was optimized through the multi-index orthogonal test design.?2?Quality evaluation of nasal mucosa moisturizing microemulsion:?1?The quality of nasal mucosa moisturizing microemulsion was evaluated by appearance,centrifugation test,turbidity,viscosity,particle size,Zeta potential,pH,content and other indicators.?2?A determination method of menthol in nasal mucosa moisturizing microemulsion based on gas chromatography?GC?to establish and verify the methodology.?3?According to the 2015 verison of Chinese Pharmacopoeia"Guidelines for the Stability Test of Raw Materials and Preparations"in the Fourth Edition,the stability and storage conditions of preparation were initially investigated.?3?Study on the efficacy,toxicity and irritation of nasal mucosa moisturizing microemulsion:?1?The microscopic observation of mouse nasal mucosal epithelial cell morphology,and the adhesion test of the maxillary mucosa of toad in vivo were used to evaluate the efficacy of nasal mucosal moisturizing microemulsion.?2?The toxicity of nasal mucosal moisturizing microemulsion was evaluated by acute oral toxicity test in mouse and ciliary duration test isolated toad.?3?The irritation of nasal mucosa moisturizing microemulsion was evaluated by rat nasal mucosa irritation test and rabbit eye mucosa irritation test.Results:?1?The selected oil phase was peppermint oil with the optimum HLB was 15.The surfactant was polyoxyethylene hydrogenated castor oil?CO-40?,the cosurfactant was anhydrous ethanol,and the mass ratio of surfactant and cosurfactant?K_m?was 1:1.Its prescription composition:1.5%for peppermint oil,18%for CO-40,18%for anhydrous ethanol,9%for glycerin,and 53.5%for distilled water.The optimal preparation process was as follows:firstly add peppermint oil to CO-40 and stir evenly,then add other components of the prescription amount,and stir for 2 h at a stirring speed of 1200 r/min.?2?The prepared nasal mucosa moisturizing microemulsion was a colorless transparent,uniform and stable liquid dispersion system with a mint fragrance.The particle morphology was observed by transmission electron microscopy and the particle size distribution was between 10¹00 nm.There was no delamination after centrifugation at 10000 r/min for 15 min.The average turbidity was 1.16±0.09 NTU,the average viscosity was 34.6±6.75 mPa·s,and the pH was 6.3⁶.5.The staining method showed that the water-soluble dye spreads faster;the conductivity test showed that the microemulsion was a bicontinuous?B.C?microemulsion.When the water content of the system was more than 69.70%,O/W microemulsion was formed.Thus,the formulation was characterized by an oil-in-water type and contains partially bicontinuous particles.Menthol was determined by GC in nasal mucosa moisturizing microemulsion:The linear range of menthol was 0.183×10^-32.745×10^-3 g/mL,the intra-day precision RSD was1.34%².85%,the daytime precision RSD was 1.19%².96%,the recovery result was between 94.44%¹04.14%,the average recovery rate was 98.73%,and the RSD was3.26%.The average content of peppermint oil calculated by menthol was 2.544 mg/mL in the moisturizing microemulsion,the RSD was 2.89%,and the RSD of the content determination between batches was between 0.58%².33%.The stability test results showed that the total RSD were 3.79%,3.69%and 4.18%in the strong light exposure test,high temperature test and acceleration test,respectively.In-10?freeze-thaw,-80?freeze-thaw and low temperature test,the total content of RSD was 3.35%,2.85%and1.38%.And the particle sizes were all less than 100 nm,other indicators were no significant change.?3?Pharmacodynamic test:In the microscopic observation of mouse nasal mucosa cells morphological,the results showed that the cells treated with hypertonic saline could quickly rehydrate and restore their original full morphology after being treated with nasal mucosa moisturizing microemulsion.In the mucoadhesive test,the relative transmission rate of the high,meddle,and low dosegastrodin groups of self-made moisturizing microemulsion were 0.55,0.66,and 0.78 times of the saline group,with statistical difference?P<0.05?.Toxicity test:In the ciliary toxicity test,there was no significant difference in the duration of ciliary exercise between the high,meddle and low dosegastrodin nasal mucosa moisturizing microemulsion group compared with the saline group?P>0.05?.And the results of mucosal section indicated that it was basically non-toxic.In the acute oral toxicity test,the mouse given the maximum concentration and the maximum gavage dose were all healthy and survived in the test group,and their main organs was no obvious pathological changes by naked eyes observing.Stimulating test:In the rat nasal mucosa irritation test,there was no significant effect on the body weight and daily behavior of the rat.At the same time,there was no significant difference in the average daily number of nose scratching and head shaking compared with the saline group?P>0.05?.In the eye mucosal irritation test of rabbit,the nasal mucosa moisturizing microemulsion group had shorter time of corneal turbidness,conjunctival congestion,a small amount of secretions and slight edema,but less irritation than compound menthol nasal drops.Conclusion:The peppermint oil nasal mucosa moisturizing microemulsion was innovatively prepared through prescription screening,preparation process optimization and quality control,and nano-loaded drug release technology used to encapsulate the peppermint oil to form a liquid-liquid nano-dispersion system,which reduced the volatility and irritation of peppermint oil.The results of efficacy test,toxicity test and irritant test indicated that the microemulsion had superior moisture retention,excellent air permeability and no toxicity or irritability.The preparation was expected to provide a scientific basis for improving the dryness of the nasal mucosa,reducing and preventing the occurrence of respiratory diseases.To provide a new,safe and effective approach for the clinical treatment of the diseases caused by nasal dryness,such as dryness,atrophic,allergic rhinitis and upper respiratory tract infection.It had very important practical application value to protect the first pass of human respiration and improve people's life quality.