Study On Quality Control And Stability Of Caryophylli Flos Extract Microemulsion

Caryophylli Flos?CF?is the dried flower bud of Myrtle family plant Eugenia caryophyllata Thunb.It has the effect of warming middle energize and descending adverse-rising,invigorating kidney and strengthening Yang.Studies have shown that CF has the functions of antioxidant,antimicrobial and anti-mites.It contains 14%²0%volatile oil,in which contains 78%⁹5%eugenol.The present study investigated executed quality control of CF,CF extract,CF extract microemulsion and the stability of CF extract microemulsion,on the basis of previous studies.The following results were observed:?1?Quality control of CFA:Identification:The method was in accordance with the CF identification method mentioned in 2015 Edition 2^nd section of Chinese Veterinary Pharmacopoeia.The sample of CF solution,the control herbs of CF solution and eugenol solution 5?L were dipped on the same silica gel G plate,separately.The plate was developed by petroleum ether?60⁹0??-ethyl acetate?9:1?.After drying in the air,5%vanillin in thick sulfuric acid solution was sprayed to the plate.The plate was heated over 105?until bands clearly appeared.The results showed that the chromatogram of the herbs of CF,control herbs of CF and eugenol showed the same color spots in the corresponding position.B:Content determination:The method to determine CF eugenol content with HPLC was established.Chromatographic condition was:Welchrom C₁₈?4.6×250nm,5?m?;methanol-water?55:45?as the mobile phases;flow rate was 1 mL/min;the column temperature was 40?;the detection wavelength was 282 nm;injection quantity was 20?L.Eugenol concentration and peak area had a good liner relationship within the range of0.01994⁰.13958 mg/mL.Regression equation was y=23045083.4647x+23918.2857,r=0.9998,the average recovery rate was 101.46%,with RSD of 0.60%.The results showed that the content of eugenol in CF is 15.52%.The above results demonstrated the method is specific and reproducible.?2?Quality control of CF extractA:Identification:TLC identification method of CF extract was established.Briefly,the method was in accordance with the CF identification method mentioned in 2015 Edition 2ⁿdd section of Chinese Veterinary Pharmacopoeia.CF extract solution?10?L?,control herbs of CF solution?5?L?and eugenol solution?5?L?were dipped on the same silica gel G plate separately.The plate were developed by petroleum ether?60⁹0??-ethyl acetate?5:1?,dried in the air,colored by 10%ethanol sulfate solution spray,heating over 105?and observed under UV lamp?365 nm?.The chromatogram of the CF extract had the same color spots in the corresponding position with the control herbs of CF solution and eugenol solution.The method has strong durability.B:Content determination:Accurately weighed CF extract 0.06 g,added 50 mL methanol,and ultrasonic demulsification for 20 min.Then leave it to room temperature,filtered.Determined by HPLC method which was established for the quality control of CF.The results showed that the content of eugenol in CF extract was 5.87%.?3?Quality control of CF extract microemulsionA:Identification:20.0 g of CF extract microemulsion was weighed,in which anhydrous sodium sulfate 2.0 g was added,and ultrasonic demulsification for 30 min.Then,20 mL petroleum ether?30⁶0??was added,extracted for 3 times,combined and dried in room temperature.CF extract microemulsion solution was obtained by adding 2 mL ether and dissolve.Blank microemulsion solution was prepared in the same way.Took 20?L CF extract microemulsion and blank microemulsion and identified by TLC method established for the quality control of CF extract.The chromatogram of the CF extract microemulsion had the same color spots in the corresponding position with the control herbs of CF solution and eugenol solution,and the negative control had no spot.B:Content determination:Accurately weighed CF extract microemulsion 3 g,added methanol to 50 mL and determined by HPLC method which was established for the quality control of CF.The results showed that the blank microemulsion had no spot and the content of eugenol in CF extract microemulsion was 0.05%.?4?The stability of CF extract microemulsionAfter highlight exposure test,high temperature test,accelerated test and long-term stability test showed that the CF extract microemulsion is tawny,homogeneous and transparency,no layering phenomenon,and the main content of eugeno l had no significant change.The results represented that CF extract microemulsion is stable for 6 months under conditions of room temperature in dark.