Study On Optimization Of Extraction Process Of Blumea Balsamifera Oil And Its Antibacterial Activity

Blumea balsamifera oil is a subsidiary product in the process of extracting borneol from Blumea balsamifera?L.?DC.,and it is widely concerned because of its effective antibacterial,anti-inflammatory,antioxidant and other biological activities.Due to the complexity of the composition of Blumea balsamifera oil and the lack of unified standard monitoring,the content of each component of Blumea balsamifera oil on the market is uneven and activity is uncertain.The chemical composition and composition of drugs are the material basis for their efficacy,while the quality of the new chinese medicine drugs is based on the premise of material basis.Therefore,this paper used in vitro bacteriostatis test to screen the antibacterial activities of different sources of Blumea balsamifera oil and to search for the antibacterial active substance.The heating reflux method was used to extract Blumea balsamifera oil through single factor test and orthogonal design.Combined with the comprehensive scores of extraction rate,bacteriostatic zone and anti-inflammatory activity—sensitized guinea pig ileum test to determine the inhibitory effect of Blumea balsamifera oil on the release of slow reactive substances?SRS-A?from guinea pig lung tissue—to screen the best extraction process of Blumea balsamifera oil.The gas chromatography-mass spectrometry?GC-MS?was used to analyze the composition of three Blumea balsamifera oil—A9 with good antibacterial effect,A5 with good anti-inflammatory effect and B screened by comprehensive score.the protective effect of Blumea balsamifera oil on mice infected with Staphylococcus aureus?S.aureus?was investigated by in vivo experiment.the antibacterial mechanism of S.aureus was studied by measuring the growth,electrical conductivity,macromolecular substance content and morphological structure,nucleic acid and soluble protein content.The results showed that?1?Blumea balsamifera oil from different sources had different antibacterial effects on the tested strains,in which the diameter of bacteriostatic zone of S.aureus and Pseudomonas aeruginosa was 11.69±0.51mm?16.07±0.29 mm and 11.57±0.40 mm?14.41±0.16 mm,the maximum value of MIC was 54.69?g/m L,MBC was 218.75?g/m L.The chemical components with better antibacterial activity were pinene and linalool,and the MIC values for the tested strains were?156.25?g/m L and?312.5?g/m L.?2?According to the comprehensive score,the best extraction process of Blumea balsamifera oil was selected as A₃B₂C₁D₁?B?.that is,the ratio of material to liquid was 1?14,ultrasonic-assisted soaking 30min at 30?,and then soaked for 1.5 hours,Borneol was collected after reflux extraction at 100?for 60min,and then continuing to extract for 4 hours,the extraction rate of Blumea balsamifera oil collected was0.175%.the diameter of bacteriostatic zone for S.aureus and Pseudomonas aeruginosa was 20.2±0.18 mm and 20.34±0.33 mm,and the MIC values were 9.77?g/m L and 13.68?g/m L,MBC values were 19.54?g/m L and 27.35?g/m L,the inhibition rate of SRS-A production was 59.2%.36 common chemical components were analyzed from the Blumea balsamifera oil extracted by three processes?A5,A9,B?.There are 14 chemical components with more than 1%content in A5 and A9,and13 chemical components with more than 1%content in B.Among them,the percentage content of?-?-Borneol in A5,A9 and B was all more than 10%,and the proportion trend of other chemical components in Blumea balsamifera oil extracted by different processes was different.?3?Different concentrations of Blumea balsamifera oil?B?inhibited the death of mice infected with S.aureus in different degrees.Among them,20mg/m L Blumea balsamifera oil significantly inhibited the death of mice?P<0.5?,and significantly reduced the splenomegaly and white spots in kidney and renal tissue necrosis in mice?P<0.01?,When S.aureus was treated with?0.5MIC B solution,the growth of the strain was inhibited within 9h,while that of?1MIC B solution was not obvious within24h.The concentration of 2MIC B solution caused the change of bacterial conductivity,but did not cause the leakage of macromolecules.The morphology of the bacteria was destroyed and irregular folds and fragments appeared.The content of nucleic acid decreased and the content of soluble protein decreased obviously with the increase of concentration,in which the protein band became shallower when the molecular weight was about 75 KDa,135 KDa and 245 KDa.The results suggest that the antibacterial effects of Blumea balsamifera oil from different sources are different,Pinene and Linalool compounds may be its antibacterial substances,and the proportion of chemical components in Blumea balsamifera oil will its afficacy,it is necessary to establish a quality standard combined with the effectiveness of Blumea balsamifera oil.High quality Blumea balsamifera oil was obtained by combining with the comprehensive scores of extraction yield,antibacterial and anti-inflammatory activity,which reduced the content of?-?-Borneol in the oil and made it have high antibacterial and anti-inflammatory effect.The Blumea balsamifera oil extracted by the best process has a significant effect on the growth rate of S.aureus and has a significant protective effect on S.aureus infected mice.and destroys the integrity of the cell by affecting the membrane permeability of S.aureus,and finally leads to cell death by affecting the synthesis of nucleic acid and protein,but does not directly act on the cell DNA.This will provide a theoretical basis for the establishment of the quality standard of Blumea balsamifera oil and further targeting research.