| Objecive:Lonicera japonica,dry bud or first flower of Lonicera japonica Thunb.,has the pharmaceutical effect of heat-clearing,detoxification and antibacterial.The essential oil extracted from L.japonica has antibacterial,and people usually put it into medicine,health products,drinks,food and daily chemical industry.However,the low production of essential oil limits its development.It has confirmed that the supercritical carbon dioxide could extract efficiently L.japonica essential oil.And yet,there were lacking of the research on composition,flavor and antibacterial of the L.japonica essential oil extracted by supercritical carbon dioxide extraction,we have no enough proof on how to use the essential oil.If we want to use the essential oil,the composition,flavor and biological activity must be considered.They can provide,respectively,the reference for medicinal ingredients,quality control and application.Also,there are three causes that variety,locality and bloom can affect the composition,flavor,and antibacterial of the L.japonica essential oil,and interfere with its use.To give references for the industrial extraction and products development of L.japonica essential oil,we have done some researches as follows.Firstly,this paper optimized extraction technique of the L.japonica essential oil extracted by the supercritical carbon dioxide extraction,then,applied the technique on extracting "varieties-localities-bud and flower stage" L.japonica essential oil,finally,studied the composition,flavor,and antibacterial of the essential oilsMethods:1.The first part of the paper applied supercritical carbon dioxide extraction on extracting the essential oil from dry L.japonica.Then,the study investigated the effect of causes that crushing degree,static extraction time,dynamic extraction time,extraction temperature,extraction pressure and gas flow of carbon dioxide on the extraction rate of L japonica essential oil,Based on the results,the study selected causes that static extraction time,dynamic extraction time and extraction temperature,and carried on a response surface experiment for ideal extracting technique.2.The second part analyzed the composition of the essential oils which were extracted from "varieties-localities-bud and flower stage" L japonica by the ideal extracting technique.Then,this part analyzed the effect of variety,locality and bloom on the essential oil composition.And by Orthogonal partial least-squares discrimination analysis(OPLS-DA),the study identified differential components among the essential oils.3.The third part gained chromatograms of "varieties-localities-bud and flower stage"L.japonica essential oil from Flash gas chromatography electronic nose.By the results,the study analyzed the flavor of the essential oil,and discriminated the category of the essential oil by iscriminant factor analysis(DFA).4.The fourth part adopted the dilution separation to get the result that "varieties-localities-bud and flower stage,L.japonica essential oil inhibiting the growth of Staphylococcus aureus.Also,to quantify the antibacterial of the essential oil,the study calculated the concentration of the L.japonica essential oils needed to inhibit 50%and 90%S.aureus,called IC50 and IC90,respectively.Results and conclusion:1.The first part has gotten the ideal extracting technique for the L.japonica essential oil.The technique was as follows:extraction pressure was 46 MPa,extraction temperature was 47?,the gas flow of carbon dioxide was 2.2 L/min,crushing degree was 100 to 120 mesh,static extraction time was 10min,dynamic extraction time was 40min.The extraction process is reliable,and can provide a reference for extracting L.japonica essential oil in industry.2.The second part has identified 74 compounds from "varieties-localities-bud and flower stage" L.japonica essential oil.The compounds belongs to aliphatic acid,alkane,aldehyde,ester,alcohol,olefin,and ketone.Among them,aliphatic acid and alkane were abundant.Further,these compounds that decane,undecane,dodecane,methyl laurate,methyl myristate,pentadecanoic acid,palmitoleic acid,palmitic acid,heptadecanoic acid,linoleic acid,linolenic acid,stearic acid,arachidic acid,heneicosan oic acid,behenic acid,tricosoic acid,heptacosane,lignoceric acid,triacontane and cerotic acid were common components of the L.japonica essential oil.The study has also distinguished the category of "varieties-localities" L.japonica essential oils by OPLS-DA,but without "bud and flower stage".These components that lignoceric acid,3,3,5,5-tetramethylcyclohexanol,cyclobutyloctadecyl oxalate,trans-oleic acid,stearic acid,cerotic acid,behenic acid,tricosane,heptacosane,linoleic acid,lignocerane,methyl laurate,linolenic acid,capric acid and caproic acid were discriminating components among the "varieties" L.japonica essential oils.The components that dimethoxyhexane,heptadecanone,lignoceric acid,tetratriacontane,stearic acid,docanoic acid,carboceric acid,decane,undecane,(E)-9-dodecenoic acid,dicetyl,dodecane,pentacosoic acid,3,3,5,5-tetramethylcyclohexanol,octane,palmitic acid,2-methyl octacosane,caproic acid,1,1-dimethylhexadecane,15-octadecenoic acid,squalene,1,3-di-tert-butylbenzene,triundane and dioctyl terephthalate were discriminating components among the "localities" L.japonica essential oils.The results can provide a reference for adding the L.japonica essential oil into the products.3.The third part has gotten the flavor of "varieties-localities-bud and flower stage" L.japonica essential oil.Moreover,the components contributing to flavor were as follows:methyl isobutyrate,2,3-butanedione,(5Z)-1,5-octadien-3-one,myristyl ether,methyl laurate,propionaldehyde,n-butyraldehyde,valeraldehyde,2,3-pentanedione,isoamyl alcohol,2-methylbutanol,trans-2-pentanol,3-hexanone,n-hexanal,isoamyl acetate,hexane methyl ester,2-heptanol,6-methyl-2-heptanone,n-pentyl propionate,mushroom alcohol,methyl heptanoate,trans-2-octenal,4-nonanol,nonanal,acetic acid phenylethyl,L-carvone and cinnamaldehyde.To sum up,the L.japonica essential oil has pungent,caramel,creamy,sweet and sour flavors.These flavors were related to many objects that fruits,butter,cheese,chocolate,malt,sesame oil,wine,mushroom,cinnamon,herbs,butter,almond,lavender,honey,rose and tobacco.In addition,the L.japonica essential oil was also waxy and pungent.Among of them,the waxy was related to trans-2-octenal,nonal and methyl laurate,and the pungent related to propionaldehyde.The study has also obtained the DFA models based on sensors,then,distinguished the category of the L.japonica essential oils divided by varieties,localities and bloom.The sensors of identifying"varieties" L.japonica essential oil were as follows:585.79-1-RH,697.89-1-RH,863.45-1-RH,1107.41-1-RH,789.89-2-RH,1210.54-2-RH,1274.24-2-RH,1311.11-2-RH and 1419.37-2-RH.The sensors of identifying "localities" L.japonica essential oil were as follows:16.20-1-RH,17.60-1-RH,28.99-1-RH,41.41-1-RH,54.24-1-RH,16.74-2-RH,18.96-2-RH,20.32-2-RH,29.59-2-RH,31,15-2-A and 45.30-2-A.The sensors of identifying "bud and flower stage" L.japonica essential oil were as follows:113.32-1-RH,902.62-1-RH,1078.57-1-RH,582.63-2-RH,929.89-2-RH,1052.05-2-RH and 1274.24-2-RH.The results can play an important role in quality control of the L.japonica essential oil.4.The fourth part has resulted that the L.japonica essential oil can inhibit the growth of S.aureus within the administration of 0.01?10mg/ml.The key components of bacteriostasis were as follows:palmitoleic acid,linoleic acid,linolenic acid,palmitoleic acid,stearic acid and arachidic acid.However,to kill S.aureus,the administration of the L.japonica essential oil should be exceeded 10mg/ml.The study has also gotten the bactericidal curves of the L.japonica essential oil,then,calculated IC50 and IC90.IC50 were as follows:"A" was 31.95?g/ml,"B" was 41.67 ?g/ml,"C" was 72.20 ?g/ml,"D" was 75.89 ?g/ml,"E" was 74.37?g/ml,"F" was 91.37 ?g/ml,"G" was 139.50 ?g/ml,"H" was 117.60 ?g/ml,"I" was 92.90?g/ml,"J" was 77.43 ?g/ml,"K" was 57.20 ?g/ml,"L" was 41.16?g/ml.IC90 were as follows:"A" was 1.37mg/ml,"B" 0.73 mg/ml,"C" was 1.41 mg/ml,"D" was 1.44 mg/ml,"E"was 3.33 mg/ml,"F" was 5.31 mg/ml,"G" was 4.94 mg/ml,"H" was 3.33 mg/ml,"I" was 4.72 mg/ml,"J" 6.51 mg/ml,"K" was 2.34 mg/ml,"L" was 4.21 mg/ml.The results can provide evidence of efficacy for developing products with the L.japonica essential oil. |
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