Study On The Extraction And Determination Of Fatty Acids And The Antibacterial Activity In Maggots

Object:This work is aimed at the optimization of not only the extraction of maggot oils but also the precolumn derivatization HPLC methods for the of the oil content.Through the detail studies of related antibacterial activities of the extraction of fatty acid of maggots,we are looking forward to obtain further fundamental theoretic for the application of maggots and to provide a new sight for the development of new drugs.Methods:1.The best extraction time,extraction times and reasonable liquid-to-material ratio of the extraction of maggot oil were optimized by the response surface method?RSM?on the basis of the screening of the extraction methods and solvents.2.With linoleic acid,palmitic acid,oleic acid,stearic acid as the reference,seventeen alkyl acid as the internal standard,the contents of fatty acids in maggots were determined by the precolumn derivatization of high performance liquid chromatography?HPLC?,and the contents of fatty acids were also determined by the internal standard method3.Taking Staphylococcus aureus?SA?and Escherichia coli?EC?as the research object,the minimum inhibitory concentration?MIC?of fatty acid from maggot is determined by microdilution broth method and the effect was determined by crystal violet staining method.Results:1.The optimized method of maggot oil extraction was ultrasonic extraction,the best solvent was CH₂Cl_2,the best extraction conditions were ultrasonic extraction 4 hours,solvent amount 15 times and 2 times of extraction.2.By the precolumn derivatization of HPLC method for the determination of fatty acid content of maggots,the content of linoleic acid,palmitic acid,oleic acid and stearic acid were,0.0955 mg/mL?0.1864 mg/m L?0.1658 mg/m L?0.0550 mg/m L,respectively.3.Microdilution broth measured fatty acid extract from maggots on Staphylococcus aureus the MIC was 0.250 mg/mL,1/2 MIC showed that the concentration of crystal violet staining method of Staphylococcus aureus biofilm formation was inhibited,the MIC drug concentration damaged Staphylococcus aureus mature biofilm,drug concentration showed a dose dependent,fatty acid extract from maggots on Escherichia coli had no obvious inhibitory effect,the drug also had no obvious effect on Escherichia coli biofilm formation and mature biofilm.Conclusions:1.Compared with the traditional orthogonal experiment,the response surface method is used to optimize the extraction process of Fructus Glutamate oil,and the result is more accurate and higher accuracy.2.The pre-column derivatization of HPLC method is stable and reproducible,and can be used for determination of fatty acid content of maggot.The method was employed to determine the content of fatty acid in maggots.3.Fatty acid extract of Maggot inhibits Staphylococcus aureus obviously,the drug also inhibit the biofilm formation of Staphylococcus aureus and its mature biofilm,but the drug had no obvious effect on Escherichia coli biofilm formation and mature biofilm.