CN114698843B - Perilla seed oil microemulsion-hydrogel system and preparation method and application thereof - Google Patents
Perilla seed oil microemulsion-hydrogel system and preparation method and application thereof Download PDFInfo
- Publication number
- CN114698843B CN114698843B CN202210302211.5A CN202210302211A CN114698843B CN 114698843 B CN114698843 B CN 114698843B CN 202210302211 A CN202210302211 A CN 202210302211A CN 114698843 B CN114698843 B CN 114698843B
- Authority
- CN
- China
- Prior art keywords
- seed oil
- perilla seed
- hydrogel
- oil microemulsion
- microemulsion
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 239000001335 perilla frutescens leaf extract Substances 0.000 title claims abstract description 149
- 239000000017 hydrogel Substances 0.000 title claims abstract description 105
- 238000002360 preparation method Methods 0.000 title claims abstract description 32
- 239000004530 micro-emulsion Substances 0.000 claims abstract description 61
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 48
- 229920002148 Gellan gum Polymers 0.000 claims abstract description 36
- 239000000216 gellan gum Substances 0.000 claims abstract description 35
- 235000010492 gellan gum Nutrition 0.000 claims abstract description 35
- WMBWREPUVVBILR-WIYYLYMNSA-N (-)-Epigallocatechin-3-o-gallate Chemical compound O([C@@H]1CC2=C(O)C=C(C=C2O[C@@H]1C=1C=C(O)C(O)=C(O)C=1)O)C(=O)C1=CC(O)=C(O)C(O)=C1 WMBWREPUVVBILR-WIYYLYMNSA-N 0.000 claims abstract description 27
- WMBWREPUVVBILR-UHFFFAOYSA-N GCG Natural products C=1C(O)=C(O)C(O)=CC=1C1OC2=CC(O)=CC(O)=C2CC1OC(=O)C1=CC(O)=C(O)C(O)=C1 WMBWREPUVVBILR-UHFFFAOYSA-N 0.000 claims abstract description 27
- 239000005018 casein Substances 0.000 claims abstract description 27
- 238000002156 mixing Methods 0.000 claims abstract description 26
- 239000003921 oil Substances 0.000 claims abstract description 25
- 235000019198 oils Nutrition 0.000 claims abstract description 25
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 claims abstract description 17
- 235000021240 caseins Nutrition 0.000 claims abstract description 17
- 230000008014 freezing Effects 0.000 claims abstract description 16
- 238000007710 freezing Methods 0.000 claims abstract description 16
- 238000010257 thawing Methods 0.000 claims abstract description 16
- 238000004108 freeze drying Methods 0.000 claims abstract description 7
- 239000002245 particle Substances 0.000 claims description 28
- 150000002632 lipids Chemical class 0.000 claims description 25
- 238000000265 homogenisation Methods 0.000 claims description 23
- 230000000694 effects Effects 0.000 claims description 20
- 238000000034 method Methods 0.000 claims description 16
- 229920001661 Chitosan Polymers 0.000 claims description 7
- 150000003626 triacylglycerols Chemical class 0.000 claims description 6
- 239000003814 drug Substances 0.000 claims description 5
- 238000010438 heat treatment Methods 0.000 claims description 5
- 235000013376 functional food Nutrition 0.000 claims description 4
- 238000002844 melting Methods 0.000 claims description 4
- 230000008018 melting Effects 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims 1
- 238000011161 development Methods 0.000 abstract description 5
- 238000012545 processing Methods 0.000 abstract description 4
- 230000001766 physiological effect Effects 0.000 abstract description 2
- 235000013402 health food Nutrition 0.000 abstract 1
- 239000000839 emulsion Substances 0.000 description 76
- 241000244206 Nematoda Species 0.000 description 57
- 239000000243 solution Substances 0.000 description 34
- 239000000203 mixture Substances 0.000 description 29
- 239000003925 fat Substances 0.000 description 28
- 235000019197 fats Nutrition 0.000 description 28
- 239000012071 phase Substances 0.000 description 22
- 238000009825 accumulation Methods 0.000 description 19
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 18
- 238000010186 staining Methods 0.000 description 18
- NPGIHFRTRXVWOY-UHFFFAOYSA-N Oil red O Chemical compound Cc1ccc(C)c(c1)N=Nc1cc(C)c(cc1C)N=Nc1c(O)ccc2ccccc12 NPGIHFRTRXVWOY-UHFFFAOYSA-N 0.000 description 14
- 238000002474 experimental method Methods 0.000 description 14
- 239000000463 material Substances 0.000 description 14
- 230000003647 oxidation Effects 0.000 description 13
- 238000007254 oxidation reaction Methods 0.000 description 13
- 238000003860 storage Methods 0.000 description 13
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 12
- 241000244203 Caenorhabditis elegans Species 0.000 description 11
- 239000007788 liquid Substances 0.000 description 11
- 235000013305 food Nutrition 0.000 description 10
- 239000011780 sodium chloride Substances 0.000 description 9
- 239000007908 nanoemulsion Substances 0.000 description 8
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 8
- 239000008367 deionised water Substances 0.000 description 7
- 229910021641 deionized water Inorganic materials 0.000 description 7
- 238000005538 encapsulation Methods 0.000 description 7
- 239000006228 supernatant Substances 0.000 description 7
- 229960000583 acetic acid Drugs 0.000 description 6
- DTOSIQBPPRVQHS-PDBXOOCHSA-N alpha-linolenic acid Chemical compound CC\C=C/C\C=C/C\C=C/CCCCCCCC(O)=O DTOSIQBPPRVQHS-PDBXOOCHSA-N 0.000 description 6
- 230000008859 change Effects 0.000 description 6
- 239000004519 grease Substances 0.000 description 6
- 150000002500 ions Chemical class 0.000 description 6
- 238000005259 measurement Methods 0.000 description 6
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 6
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 6
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 6
- 238000005406 washing Methods 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 241000588724 Escherichia coli Species 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 4
- 239000003995 emulsifying agent Substances 0.000 description 4
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 230000001590 oxidative effect Effects 0.000 description 4
- 239000001301 oxygen Substances 0.000 description 4
- 229910052760 oxygen Inorganic materials 0.000 description 4
- 230000009467 reduction Effects 0.000 description 4
- 238000000518 rheometry Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 230000009471 action Effects 0.000 description 3
- 235000020661 alpha-linolenic acid Nutrition 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 230000007423 decrease Effects 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 238000004043 dyeing Methods 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 239000000796 flavoring agent Substances 0.000 description 3
- 235000019634 flavors Nutrition 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 229960004488 linolenic acid Drugs 0.000 description 3
- 230000007935 neutral effect Effects 0.000 description 3
- VOFUROIFQGPCGE-UHFFFAOYSA-N nile red Chemical compound C1=CC=C2C3=NC4=CC=C(N(CC)CC)C=C4OC3=CC(=O)C2=C1 VOFUROIFQGPCGE-UHFFFAOYSA-N 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 229920001282 polysaccharide Polymers 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 238000009210 therapy by ultrasound Methods 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- 101100499232 Caenorhabditis elegans dhs-3 gene Proteins 0.000 description 2
- 101100136092 Drosophila melanogaster peng gene Proteins 0.000 description 2
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- 229930040373 Paraformaldehyde Natural products 0.000 description 2
- 235000004347 Perilla Nutrition 0.000 description 2
- 241000229722 Perilla <angiosperm> Species 0.000 description 2
- 102000009105 Short Chain Dehydrogenase-Reductases Human genes 0.000 description 2
- 108010048287 Short Chain Dehydrogenase-Reductases Proteins 0.000 description 2
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 2
- 235000010419 agar Nutrition 0.000 description 2
- MBMBGCFOFBJSGT-KUBAVDMBSA-N all-cis-docosa-4,7,10,13,16,19-hexaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCC(O)=O MBMBGCFOFBJSGT-KUBAVDMBSA-N 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 150000001840 cholesterol esters Chemical class 0.000 description 2
- 239000002131 composite material Substances 0.000 description 2
- 230000006866 deterioration Effects 0.000 description 2
- 235000014113 dietary fatty acids Nutrition 0.000 description 2
- 239000006185 dispersion Substances 0.000 description 2
- 235000013601 eggs Nutrition 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 239000000194 fatty acid Substances 0.000 description 2
- 229930195729 fatty acid Natural products 0.000 description 2
- 150000004665 fatty acids Chemical class 0.000 description 2
- 210000001035 gastrointestinal tract Anatomy 0.000 description 2
- 239000012362 glacial acetic acid Substances 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 238000000227 grinding Methods 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- 229920002521 macromolecule Polymers 0.000 description 2
- 229940118019 malondialdehyde Drugs 0.000 description 2
- 210000003463 organelle Anatomy 0.000 description 2
- 230000004792 oxidative damage Effects 0.000 description 2
- 229920002866 paraformaldehyde Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000012460 protein solution Substances 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 239000001100 (2S)-5,7-dihydroxy-2-(3-hydroxy-4-methoxyphenyl)chroman-4-one Substances 0.000 description 1
- RVBUGGBMJDPOST-UHFFFAOYSA-N 2-thiobarbituric acid Chemical compound O=C1CC(=O)NC(=S)N1 RVBUGGBMJDPOST-UHFFFAOYSA-N 0.000 description 1
- 244000153158 Ammi visnaga Species 0.000 description 1
- 235000010585 Ammi visnaga Nutrition 0.000 description 1
- 238000009020 BCA Protein Assay Kit Methods 0.000 description 1
- 101100124874 Caenorhabditis elegans hsf-1 gene Proteins 0.000 description 1
- 241000233779 Cyclocarya paliurus Species 0.000 description 1
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- QUQPHWDTPGMPEX-UHFFFAOYSA-N Hesperidine Natural products C1=C(O)C(OC)=CC=C1C1OC2=CC(OC3C(C(O)C(O)C(COC4C(C(O)C(O)C(C)O4)O)O3)O)=CC(O)=C2C(=O)C1 QUQPHWDTPGMPEX-UHFFFAOYSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 1
- 241000207923 Lamiaceae Species 0.000 description 1
- 239000006137 Luria-Bertani broth Substances 0.000 description 1
- 229910019093 NaOCl Inorganic materials 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 241001028048 Nicola Species 0.000 description 1
- 244000124853 Perilla frutescens Species 0.000 description 1
- 235000004348 Perilla frutescens Nutrition 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 241000790234 Sphingomonas elodea Species 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 238000005411 Van der Waals force Methods 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- PNNNRSAQSRJVSB-BXKVDMCESA-N aldehydo-L-rhamnose Chemical compound C[C@H](O)[C@H](O)[C@@H](O)[C@@H](O)C=O PNNNRSAQSRJVSB-BXKVDMCESA-N 0.000 description 1
- JAZBEHYOTPTENJ-JLNKQSITSA-N all-cis-5,8,11,14,17-icosapentaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O JAZBEHYOTPTENJ-JLNKQSITSA-N 0.000 description 1
- AEMOLEFTQBMNLQ-WAXACMCWSA-N alpha-D-glucuronic acid Chemical compound O[C@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-WAXACMCWSA-N 0.000 description 1
- 125000003368 amide group Chemical group 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 238000003149 assay kit Methods 0.000 description 1
- QUQPHWDTPGMPEX-UTWYECKDSA-N aurantiamarin Natural products COc1ccc(cc1O)[C@H]1CC(=O)c2c(O)cc(O[C@@H]3O[C@H](CO[C@@H]4O[C@@H](C)[C@H](O)[C@@H](O)[C@H]4O)[C@@H](O)[C@H](O)[C@H]3O)cc2O1 QUQPHWDTPGMPEX-UTWYECKDSA-N 0.000 description 1
- 210000000467 autonomic pathway Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 210000004958 brain cell Anatomy 0.000 description 1
- 239000012267 brine Substances 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 239000009096 changqing Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- APSNPMVGBGZYAJ-GLOOOPAXSA-N clematine Natural products COc1cc(ccc1O)[C@@H]2CC(=O)c3c(O)cc(O[C@@H]4O[C@H](CO[C@H]5O[C@@H](C)[C@H](O)[C@@H](O)[C@H]5O)[C@@H](O)[C@H](O)[C@H]4O)cc3O2 APSNPMVGBGZYAJ-GLOOOPAXSA-N 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 239000004064 cosurfactant Substances 0.000 description 1
- 229920006037 cross link polymer Polymers 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 235000013325 dietary fiber Nutrition 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 235000020669 docosahexaenoic acid Nutrition 0.000 description 1
- 229940090949 docosahexaenoic acid Drugs 0.000 description 1
- 235000020673 eicosapentaenoic acid Nutrition 0.000 description 1
- 229960005135 eicosapentaenoic acid Drugs 0.000 description 1
- JAZBEHYOTPTENJ-UHFFFAOYSA-N eicosapentaenoic acid Natural products CCC=CCC=CCC=CCC=CCC=CCCCC(O)=O JAZBEHYOTPTENJ-UHFFFAOYSA-N 0.000 description 1
- 230000005489 elastic deformation Effects 0.000 description 1
- 230000009881 electrostatic interaction Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 235000021323 fish oil Nutrition 0.000 description 1
- 238000002073 fluorescence micrograph Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000000451 gelidium spp. gum Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- VUYDGVRIQRPHFX-UHFFFAOYSA-N hesperidin Natural products COc1cc(ccc1O)C2CC(=O)c3c(O)cc(OC4OC(COC5OC(O)C(O)C(O)C5O)C(O)C(O)C4O)cc3O2 VUYDGVRIQRPHFX-UHFFFAOYSA-N 0.000 description 1
- QUQPHWDTPGMPEX-QJBIFVCTSA-N hesperidin Chemical compound C1=C(O)C(OC)=CC=C1[C@H]1OC2=CC(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO[C@H]4[C@@H]([C@H](O)[C@@H](O)[C@H](C)O4)O)O3)O)=CC(O)=C2C(=O)C1 QUQPHWDTPGMPEX-QJBIFVCTSA-N 0.000 description 1
- 229940025878 hesperidin Drugs 0.000 description 1
- 239000007970 homogeneous dispersion Substances 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 230000001418 larval effect Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 229940031182 nanoparticles iron oxide Drugs 0.000 description 1
- 239000005445 natural material Substances 0.000 description 1
- ARGKVCXINMKCAZ-UHFFFAOYSA-N neohesperidine Natural products C1=C(O)C(OC)=CC=C1C1OC2=CC(OC3C(C(O)C(O)C(CO)O3)OC3C(C(O)C(O)C(C)O3)O)=CC(O)=C2C(=O)C1 ARGKVCXINMKCAZ-UHFFFAOYSA-N 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 239000007764 o/w emulsion Substances 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 229920001277 pectin Polymers 0.000 description 1
- 239000001814 pectin Substances 0.000 description 1
- 235000010987 pectin Nutrition 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 238000005502 peroxidation Methods 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000035935 pregnancy Effects 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- -1 salt ions Chemical class 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 230000002522 swelling effect Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 229920001187 thermosetting polymer Polymers 0.000 description 1
- 230000009974 thixotropic effect Effects 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
- 239000008852 wen-xin Substances 0.000 description 1
- 230000037303 wrinkles Effects 0.000 description 1
- 229920001285 xanthan gum Polymers 0.000 description 1
- 239000000230 xanthan gum Substances 0.000 description 1
- 235000010493 xanthan gum Nutrition 0.000 description 1
- 229940082509 xanthan gum Drugs 0.000 description 1
- 238000000733 zeta-potential measurement Methods 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/115—Fatty acids or derivatives thereof; Fats or oils
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/03—Organic compounds
- A23L29/035—Organic compounds containing oxygen as heteroatom
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/03—Organic compounds
- A23L29/045—Organic compounds containing nitrogen as heteroatom
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
- A23L29/269—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of microbial origin, e.g. xanthan or dextran
- A23L29/272—Gellan
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
- A23L29/275—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of animal origin, e.g. chitin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
- A23L29/294—Inorganic additives, e.g. silica
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
- A61K36/535—Perilla (beefsteak plant)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The invention provides a perilla seed oil microemulsion-hydrogel system, a preparation method and application thereof, belonging to the technical field of health food processing, wherein the preparation method comprises the following steps: 1) Mixing casein with water to obtain a water phase, mixing EGCG with perilla seed oil to obtain an oil phase, mixing the water phase with the oil phase, and homogenizing to obtain a perilla seed oil microemulsion; 2) Dropping gellan gum solution into chitosan-MgCl 2 Repeatedly freezing and thawing for 3-5 times in the solution to obtain hydrogel; 3) And mixing the hydrogel with the perilla seed oil microemulsion for 10-14 hours after freeze-drying to obtain a perilla seed oil microemulsion-hydrogel system. The preparation method provided by the invention utilizes the perilla seed oil to prepare the oil-in-water type EGCG perilla seed oil microemulsion, and prepares the oil-in-water type EGCG perilla seed oil microemulsion into a hydrogel system, so that the stability and the physiological activity of the perilla seed oil are further improved, and a new thought is provided for solving the current situation that the development of the perilla seed oil is limited.
Description
Technical Field
The invention belongs to the technical field of health-care food processing, and particularly relates to a perilla seed oil microemulsion-hydrogel system and a preparation method and application thereof.
Background
The perilla seed oil is obtained from mature seed of Perilla frutescens (Perilla frutesccns) belonging to Labiatae. The content of unsaturated fatty acid and polyunsaturated fatty acid in the purple perilla seed oil is 91.1 to 93.8 percent and 70.7 to 83.4 percent respectively. Wherein the highest proportion of alpha-linolenic acid is 58.8-70.9%. Alpha-linolenic acid is a necessary fatty acid for human body, and can be converted into eicosapentaenoic acid and docosahexaenoic acid in human body, which are effective active ingredients in fish oil. Alpha-linolenic acid has the functions of resisting tumor, resisting thrombus, reducing blood fat, nourishing brain cells, regulating autonomic nerves and the like and receives much attention.
On the other hand, the perilla seed oil contains a large amount of unsaturated fatty acid, the content of the unsaturated fatty acid is extremely easy to be influenced by factors such as illumination, temperature and the like, the problem of oxidative deterioration is generated, and the loss of nutrient components and the deterioration of flavor are caused, so that the application of the perilla seed oil in foods is limited; and unsaturated fatty acid is a macromolecular substance, so that the physiological absorptivity is poor, and the unsaturated fatty acid cannot be fully absorbed in a human body, so that the loss of nutritional ingredients is caused.
Microemulsions are thermodynamically stable, isotropic, transparent or translucent homogeneous dispersions composed of spontaneous formation of water, oil, surfactants, cosurfactants, and the like. The oil-in-water emulsion takes oil as a disperse phase and is embedded in water under the action of an emulsifier, so that oxygen is isolated, oxidation is delayed, and stability is improved.
Gellan gum is an extracellular polysaccharide of pseudomonas dysenteriae (Sphingomonas elodea (ATCC 31461)). The main components of the gellan gum are complex linear high molecular saccharide polymers formed by connecting glucose, D-glucuronic acid and L-rhamnose according to the proportion of 2:1:1 and four sugar molecules sequentially through glycosidic bonds. Gellan gum has good flavor release, high transparency, strong stability, strong gel ability, wide acid resistance range and other good characteristics, gradually starts to replace pectin, agar and xanthan gum, and plays an important role in the fields of chemical industry, food, medicine and the like.
Hydrogels (hydrogels) are a new class of polymeric materials composed of a three-dimensional network of chemically or physically crosslinked polymers and a large amount of water. The main chain or side chain of the three-dimensional network polymer constituting the hydrogel generally contains a large number of hydrophilic groups such as hydroxyl groups, carboxyl groups, amino groups, amide groups, etc., so that the hydrogel exhibits extremely strong hydrophilicity, can absorb a large amount of moisture and maintains a certain shape. The hydrogel has certain mechanical strength, high permeability to small molecules and various stimulus responsivity, and the performance and unique structure can meet the application requirements in various aspects. So in recent years, efficient novel hydrogels have been the hot spot of research. The hydrogel is used as a structural or functional material, has wide application in various fields of food engineering, agricultural production, medical and health, sewage treatment and the like, and is a material with great development prospect.
The perilla seed oil is easily oxidized due to the high content of unsaturated fatty acid, and has unstable property and is not shelf-stable. The oil substances have the characteristics of poor water solubility, low oral administration utilization rate and the like, and limit the application of the oil substances in food processing. Although the microemulsion can improve the stability of the perilla seed oil, the microemulsion is limited in a plurality of application scenes, and emulsion breaking is easy to occur under the conditions of long-time storage, different pH values and ion concentrations. It is important to find a more stable system to protect the perilla seed oil.
In recent years, research on the composition of perilla seed oil fatty acid and the measurement and extraction process at home and abroad is more, and reports are made on the application and processing of the perilla seed oil, especially, the effective delivery carrier mainly taking the perilla seed oil is processed and constructed through a new food technology, for example, research on preparing the perilla seed oil emulsion is not reported, and related products of the perilla seed oil emulsion are not available in the market. In recent years, food-grade emulsions and hydrogels have been widely used as some of the existing forms of fats and oils in foods for embedding and delivering functional fats and oils, fat-soluble nutrients and flavor substances, preventing oxidation of functional nutrients and improving their water solubility and bioavailability. Therefore, the preparation of the food-grade nanoemulsion and the hydrogel by using the perilla seed oil has great significance.
Conventional emulsion systems still have problems such as instability in long-term storage, different pH, ion concentration, easy loss of encapsulated materials, etc.
Disclosure of Invention
In view of the above, the invention aims to provide a perilla seed oil microemulsion-hydrogel system, a preparation method and application thereof; the oil-in-water type EGCG perilla seed oil microemulsion is prepared by using the perilla seed oil, and is prepared into a hydrogel system, so that the stability and the physiological activity of the perilla seed oil are further improved, and a new thought is provided for solving the current situation that the development of the perilla seed oil is limited.
The invention provides a preparation method of a perilla seed oil microemulsion-hydrogel system, which comprises the following steps:
1) Mixing casein with water to obtain a water phase, mixing EGCG with perilla seed oil to obtain an oil phase, mixing the water phase with the oil phase, and homogenizing to obtain a perilla seed oil microemulsion;
2) Dropping gellan gum solution into chitosan-MgCl 2 Repeatedly freezing and thawing for 3-5 times in the solution to obtain hydrogel;
3) Mixing the hydrogel after freeze-drying with the perilla seed oil microemulsion for 10-14 hours to obtain a perilla seed oil microemulsion-hydrogel system;
there is no time sequence limitation between step 1) and step 2).
Preferably, the mass percentage of the casein in the perilla seed oil microemulsion is 0.4-1.5%, the mass percentage of the EGCG in the perilla seed oil microemulsion is 0.008-0.05%, and the mass percentage of the perilla seed oil in the perilla seed oil microemulsion is 5-15%.
Preferably, the homogenizing includes sequentially performing a first homogenizing and a second homogenizing; the rotation speed of the first homogenization is 8000-12000 rpm, the repetition time of the first homogenization is 2-4 times, and the time of each time is 0.5-1.5 min; the second homogenization is high-pressure homogenization, the pressure of the second homogenization is 450-550 Mpa, the repetition number of the second homogenization is 4-6, and the time of each time is 20-40 s.
Preferably, the preparation method of the gellan gum solution in the step 2) comprises mixing gellan gum with water, and heating at 85-95 ℃ for 15-25 min.
Preferably, the chitosan-MgCl 2 The pH value of the solution is 3.4-3.6, and the chitosan-MgCl 2 MgCl in solution 2 The mass percentage of the chitosan-MgCl is 0.10-0.15 percent 2 The mass percentage of chitosan in the solution is 2-4%.
Preferably, the mass percentage of gellan gum in the hydrogel obtained in the step 2) is 1% -3%.
Preferably, the freezing temperature of the repeated freezing and thawing is-3 to-6 ℃ and the freezing time is 7 to 9 hours; the melting temperature of the repeated freezing and thawing is 10-30 ℃.
The invention also provides a perilla seed oil microemulsion-hydrogel system prepared by the preparation method.
The invention also provides application of the perilla seed oil microemulsion-hydrogel system in preparation of functional foods or medicines with lipid-lowering efficacy.
Preferably, the lipid-lowering comprises lowering the content of triglycerides and/or the particle size of lipid droplets.
Compared with the prior art, the invention has the following beneficial effects: the invention mixes the purple perilla seed oil microemulsion and the hydrogel, and uses gellan gum and Mg 2+ Cross-linking between and Chitosan-MgCl 2 The stability of the perilla seed oil is obviously improved through the hydrogel formed by electrostatic interaction with gellan gum, and the stability of the perilla seed oil and the perilla seed are improved through the hydrogelThe stability study of the oil microemulsion and the perilla seed oil microemulsion-hydrogel system shows that the perilla seed oil microemulsion-hydrogel system prepared by the invention can better protect the perilla seed oil and prevent oxidation, and can effectively protect grease. In addition, experiments show that compared with emulsion, the perilla seed oil microemulsion-hydrogel system prepared by the invention has more remarkable lipid-lowering effect and better lipid-lowering effect.
The perilla seed oil microemulsion-hydrogel system prepared by the invention has good swelling property, good viscoelasticity, obvious thixotropic property and certain memory property under the action of different stresses; has good embedding slow release effect on water-soluble and fat-soluble functional active substances, and can delay the release rate of functional components.
The preparation method for the purple perilla seed oil microemulsion-hydrogel system has the advantages of simple process and low cost, and can adapt to different application scenes. The raw materials used in the invention are natural substances, are convenient to obtain, and have the environment-friendly concept of sustainable development of food.
Drawings
FIG. 1 shows the particle size, potential and PDI changes of the perilla seed oil microemulsion under different pH conditions;
FIG. 2 shows the particle size, potential and PDI changes of the perilla seed oil microemulsion under different NaCl concentration conditions;
FIG. 3 shows MDA content change of the perilla seed oil microemulsion at 37 ℃ for 1-5 days, wherein PSO is perilla seed oil, PSO-Casein is mixture of the perilla seed oil and Casein, and PSO-Casein-EGCG is perilla seed oil nanoemulsion;
FIG. 4 shows fat accumulation of nematodes after treatment with perilla seed oil microemulsion;
FIG. 5 is a graph showing nematode triglyceride content after treatment with perilla seed oil microemulsion;
FIG. 6 is the effect of perilla seed oil microemulsion on the average size of the transgenic line worm drops ZXW 618;
FIG. 7 is a graph showing malondialdehyde content of various emulsions and perilla seed oil microemulsion-hydrogel systems as a function of days;
FIG. 8 is the appearance and microstructure of 1%, 2%, 3% and 2% GG-PSO emulsion-hydrogel systems;
FIG. 9 is the storage modulus (G') of a 1% GG, 2% GG, 3% GG, 2% GG-PSO emulsion-hydrogel system and gellan gum, wherein the gellan gum is a hydrogel prepared using only gellan gum, without other materials;
FIG. 10 is a quantitative graph of nematode oil red O staining; wherein: blank (OP 50), PSO-Casein-EGCG emulsion, 2% GG (hydrogel with 2% gellan gum), 2% GG-PSO complex (hydrogel with 2% gellan gum and PSO-Casein-EGCG emulsion complex).
Detailed Description
The invention provides a preparation method of a perilla seed oil microemulsion-hydrogel system, which comprises the following steps: 1) Mixing casein with water to obtain a water phase, mixing EGCG with perilla seed oil to obtain an oil phase, mixing the water phase with the oil phase, and homogenizing to obtain a perilla seed oil microemulsion; 2) Dropping gellan gum solution into chitosan-MgCl 2 Repeatedly freezing and thawing for 3-5 times in the solution to obtain hydrogel; 3) And mixing the hydrogel with the perilla seed oil microemulsion for 10-14 hours after freeze-drying to obtain a perilla seed oil microemulsion-hydrogel system.
In the invention, casein is mixed with water to obtain a water phase, and EGCG is mixed with perilla seed oil to obtain an oil phase. In the invention, the mass percentage of the casein in the perilla seed oil microemulsion is preferably 0.4-1.5%, and more preferably 0.5-1.0%. In the present invention, the mass ratio of casein to water is preferably 1g:160 to 200ml, more preferably 1g:180ml; in the invention, the casein is preferably subjected to ultrasonic treatment after being mixed with water, and the ultrasonic treatment time is preferably 3-8 min, and more preferably 5min; the power of the ultrasonic treatment is not particularly limited, and the casein can be completely dissolved. In the invention, the EGCG and the perilla seed oil are mixed to obtain an oil phase, and the mass percentage of the EGCG in the perilla seed oil microemulsion is 0.008-0.05%, preferably 0.01-0.04%; the mass percentage of the perilla seed oil in the perilla seed oil microemulsion is preferably 5% -15%, more preferably 8% -12%, and even more preferably 10%. The invention does not limit the sequence of the preparation of the water phase and the oil phase.
After the water phase and the oil phase are obtained, the water phase and the oil phase are mixed and homogenized to obtain the perilla seed oil microemulsion. In the present invention, the homogenization includes a first homogenization and a second homogenization which are sequentially performed; the rotation speed of the first homogenization is preferably 8000 to 12000rpm, more preferably 9000 to 11000rpm, still more preferably 10000rpm; the number of repetitions of the first homogenization is preferably 2 to 4, more preferably 3, and the time for each is preferably 0.5 to 1.5min, more preferably 0.8 to 1.2min, more preferably 1min. In the present invention, the first homogenization is preferably performed by using a homogenizer, and the present invention is not limited to the homogenizer, and a homogenizer conventional in the art may be used. In the present invention, the second homogenization is preferably high-pressure homogenization, and the pressure of the second homogenization is preferably 450 to 550Mpa, more preferably 480 to 520Mpa; the number of repetitions of the second homogenization is preferably 4 to 6, and the time for each is preferably 20 to 40s, more preferably 25 to 35s. In the present invention, the second homogenization is preferably performed using a high-pressure homogenizer.
In the present invention, gellan gum solution is added dropwise to chitosan-MgCl 2 And repeatedly freezing and thawing for 3-5 times in the solution to obtain the hydrogel. In the invention, the mass percentage of gellan gum in the hydrogel is preferably 1% -3%, and more preferably 2%. In the invention, the preparation method of the gellan gum solution comprises the steps of mixing gellan gum with water, and heating for 15-25 min at 85-95 ℃; the heating temperature is preferably 88 to 92 ℃, and more preferably 90 ℃; the heating time is 18 to 22 minutes, and more preferably 20 minutes. In the invention, the mass percentage of gellan gum in the gellan gum solution is 2% -6%, preferably 4%. After the gellan gum is prepared and obtained, the gellan gum solution is dripped into chitosan-MgCl 2 In solution. In the present invention, the chitosan-MgCl 2 The pH of the solution is preferably 3.4 to 3.6, more preferably 3.5; the chitosan-MgCl 2 MgCl in solution 2 The mass percentage of (a) is preferably 0.10-0.15%, more preferably 0.11-0.14%, even more preferably 0.13%; the chitosan-MgCl 2 The mass percentage of chitosan in the solution is preferably 2% -4%, more preferably 2.5% -3.5%, and even more preferably 3%. In the present invention, the chitosan-MgCl 2 The pH of the solution is preferably adjusted by glacial acetic acid. In the present invention, the chitosan-MgCl 2 By passing said MgCl 2 Mixing the aqueous solution with chitosan solution to obtain MgCl 2 The volume ratio of the aqueous solution to the chitosan-acetic acid solution is preferably 1:1; the chitosan-acetic acid solution uses water as a solvent, the volume percentage of acetic acid is 3%, the mass percentage of chitosan is 4-8%, more preferably 5% -7%, still more preferably 6%.
In the invention, the freezing temperature of the repeated freezing and thawing is preferably-3 to-6 ℃, and is further preferably-4 to-5 ℃; the freezing time is preferably 7 to 9 hours, more preferably 8 hours; the melting temperature of the repeated freeze thawing is 10-30 ℃, and the invention has no limit on the melting time of the repeated freeze thawing, and is suitable for complete thawing. In the present invention, the repeated freeze thawing functions to improve the stability of the hydrogel.
After the hydrogel is obtained, the hydrogel is freeze-dried. In the present invention, the temperature of the lyophilization is preferably-45 to-35 ℃, more preferably-40 ℃, and the time of the lyophilization is preferably 20 to 30 hours, more preferably 24 hours.
In the invention, the freeze-dried hydrogel and the perilla seed oil microemulsion are mixed for 10 to 14 hours to obtain a perilla seed oil microemulsion-hydrogel system. In the present invention, the mixing time is preferably 11 to 13 hours, more preferably 12 hours; the mixing time is within the above-defined range, so that the hydrogel can fully absorb the perilla seed oil microemulsion.
The water involved in the above preparation method of the present invention is preferably deionized water.
The invention also provides a perilla seed oil microemulsion-hydrogel system prepared by the preparation method.
The invention also provides application of the perilla seed oil microemulsion-hydrogel system in preparation of functional foods or medicines with lipid-lowering efficacy. In the present invention, the lipid-lowering efficacy preferably includes lowering the content of triglycerides and/or the particle size of lipid droplets. The specific composition and preparation method of the functional food and drug are not particularly limited.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Preparation of purple perilla seed oil microemulsion
0.5g casein was dissolved in 90mL deionized water at room temperature of 25℃and sonicated for 5min as the aqueous phase. 0.01g EGCG is dissolved in 10g perilla seed oil and is used as oil phase after being mixed evenly. Mixing the water phase and the oil phase, homogenizing for 1min at 10000rpm with a homogenizer for 3 times to form coarse emulsion, homogenizing under high pressure for 500Mpa for 5 times, and 30s each time; and preparing the perilla seed oil microemulsion.
Preparation of hydrogels
0.26% (w/v) MgCl 2 The aqueous solution was added drop wise in equal volumes to a chitosan solution (6 g chitosan in 100ml of 3% acetic acid solution). The pH of the above solution was adjusted to 3.5 with glacial acetic acid. Gellan gum powder was stirred in deionized water as a 4% (w/v) solution at 90 ℃ for 20min until completely dissolved to promote hydration of gellan gum. Continuously and gently stirring at room temperature, dropwise adding 10mL of gellan gum solution to 10mL of chitosan-MgCl 2 In solution. Freezing at-4deg.C for 8 hr, thawing at room temperature, repeating freeze thawing for 4 times to obtain hydrogel, and lyophilizing to obtain lyophilized hydrogel.
Preparation of purple perilla seed oil microemulsion-hydrogel system
Adding the perilla seed oil microemulsion into hydrogel, and waiting for 12 hours according to the ratio of the perilla seed oil microemulsion to the hydrogel being 1:1, so that the hydrogel fully absorbs the perilla seed oil microemulsion to obtain a perilla seed oil microemulsion-hydrogel system.
Experimental example 1
The particle size, potential and PDI change of the perilla seed oil microemulsion prepared in the example 1 under different pH conditions, the particle size, potential and PDI change under different NaCl concentration conditions and the MDA content change for 1-5 days at 37 ℃ are detected.
According to the method described in example 1, a CK mixture was prepared without addition of EGCG.
The specific method for parameter measurement comprises the following steps:
determination of the average particle size of the emulsion
1mL of freshly prepared emulsion is taken into a 10mL centrifuge tube, 5mL of distilled water is added for dilution, the mixture is uniformly shaken, 2mL of the mixture is taken out and put into a particle size analyzer for measuring the average particle size.
Emulsion Zeta potential measurement
1mL of freshly prepared emulsion is taken out in a 10mL centrifuge tube, 5mL of distilled water is added for dilution, the mixture is uniformly shaken, 2mL of the mixture is taken out and put into a particle size analyzer for measurement of Zeta potential.
Determination of the polydispersity index of an emulsion
1mL of freshly prepared emulsion is taken into a 10mL centrifuge tube, 5mL of distilled water is added for dilution, the mixture is uniformly shaken, 2mL of the mixture is taken out and put into a particle size analyzer for measuring the polydispersity index.
Encapsulation efficiency measurement
The encapsulation efficiency (EE,%) of the CK mixture and the perilla seed oil emulsion was determined using hexane extraction of the unencapsulated perilla seed oil (Palchoudhury Soubantika, lead Jamie R.Aface and cost-effective method for separation ofoil-water mixtures using polymer-coated iron oxide nanoparticles [ J ]. Environmental science & technology,2014,48 (24)).
The materials were vortexed for 1min by taking 5mL each of the liquid to be tested and mixing with 2.5mL of hexane. The mixture was then centrifuged at 959×g for 3min and the supernatant liquid was transferred to a new clean conical tube. The extraction process was repeated 2 times and the 2 times of supernatant liquid was combined to give 5mL of extract per sample. The content of perilla seed oil in the organic phase was measured at 320nm by ultraviolet-visible spectrophotometry (Shimadzu corporation).
Experimental results
TABLE 1 particle size, potential, encapsulation efficiency, PDI, encapsulation efficiency (%)
The particle sizes of the CK mixture and the perilla seed oil microemulsion are about 360nm, the particle sizes are smaller, the particle size ranges of the nanoemulsion are met, and the stability is achieved. Compared with the CK mixture, the particle size of the emulsion is not changed significantly after the EGCG tea polyphenol is added, which indicates that the addition of EGCG has no significance on the particle size of the emulsion.
The higher the absolute value of Zeta potential, the greater the repulsive force between particles due to the effect of repulsive force with electric charge, the less likely the system will aggregate, i.e. the more stable the colloid will be. The potential of the CK mixture and the perilla seed oil microemulsion in the experiment are respectively-20.5+/-0.96 and-20.77+/-0.68, which shows that the CK mixture and the perilla seed oil microemulsion have good stability.
The PDI values are both small (< 0.3) from the polydispersity index point of view, again indicating good stability. In addition, compared with the CK mixture prepared directly by casein, the encapsulation rate of the perilla seed oil microemulsion added with EGCG is as high as 94.3%. The fact that the perilla seed oil is embedded in the perilla seed oil to a great extent is explained, and compared with the perilla seed oil microemulsion, the encapsulation rate is better, probably because EGCG is used as an antioxidant to play a role in protecting an oil-water interface layer.
In general, from the basic physical characterization, the stability of the perilla seed oil microemulsion prepared by adding EGCG is good.
Physical stability of perilla seed oil microemulsion
pH stability
By means of dipotassium hydrogen phosphate (K) 2 HPO 4 ) Monopotassium phosphate (KH) 2 PO 4 ) And disodium hydrogen phosphate (Na) 2 HPO 4 ) CK mixtures and perilla seed oil microemulsions were prepared at different pH (3, 4, 5, 6, 7, 8, 9), and their average particle size, dispersion coefficient PDI, and Zeta potential were determined by reference to the above methods.
Ion stability
CK mixtures and perilla seed oil microemulsions with different ion concentrations (0.1 mmol/L,1mmol/L,10mmol/L,100mmol/L,1 mol/L) were prepared by using sodium chloride (NaCl), and after 1 day of storage at room temperature, the average particle size, the dispersion coefficient PDI and the Zeta potential were determined by referring to the above methods.
Oxidative stability of perilla seed oil microemulsion
MDA content determination
Malondialdehyde MDA is a lipid oxidation end product, commonly used to evaluate the degree of peroxidation and oxidative damage of body lipids. The higher the MDA content, the more serious the oxidative damage of the organism. The oxidation product (MDA) of the emulsion was measured by measuring the thiobarbituric acid conjugate (TBARS) content of the emulsion, and the degree of oxidation of the oil in the reaction emulsion (Ali Ali, ghozlene, mekhlufi, nicolas Huang, florence Agnely,. Beta. -lactoglobulin stabilized nanemulsions-Formulation and process factors affecting droplet size andnanoemulsion stability. International Journal ofPharmaceutics;2016.500 (1-2): 291-304.).
The results are shown in FIGS. 1, 2 and 3.
As can be seen from A in FIG. 1, the CK mixture (labeled PSO-Casein in the figure) and the perilla seed oil microemulsion (labeled PSO-Casein-EGCG in the figure) both maintain a stable particle size at pH 3-9, indicating their stability at different pH's. From B in FIG. 1, it can be seen that the absolute value of the potential of the perilla seed oil microemulsion is higher at pH 5-7, which indicates that the emulsion has good stability at pH 5-7. At pH 3, the emulsion potential is positive; the potential of the emulsion at pH 5-7 is negative and the absolute value of the potential increases with increasing pH. According to the change in Zeta potential, it is shown that the surface charge of both emulsions is provided by casein, because nanoemulsions with casein as emulsifier are stabilized by electrostatic repulsion, which tends to form aggregates by hydrophobic attraction and van der waals interactions when the pH approaches isoelectric point (pI) 4.8, when the electrostatic repulsion is insufficient to overcome these attractive forces. In the measurement of emulsion particle size, the net charge of the emulsion is small at pI, but highly positive or highly negative at pH values below or above pI. The polydispersity index (PDI) can be used as a measure of the homogeneity of the nanoemulsion, as can be seen from C in FIG. 1, the PDI of PSO-Casein-EGCG emulsion is relatively low at pH around 5-7, indicating that at pH 5-7 the PSO-Casein-EGCG emulsion is homogeneous, and that the addition of EGCG makes the emulsion more homogeneous compared to CK mixtures.
Thus, both emulsions were stable over a pH range of 5.0 to 7.0.
As is clear from FIG. 2, at 0.1-1 mmol/L NaCl, the particle size and PDI of PSO-Casein-EGCG are smaller, the absolute value of potential is larger, and both emulsions are not layered and are more stable. When the ion concentration of the emulsion is 10-1000 mmol/L NaCl, obvious layering is generated, PDI and particle size are larger, and emulsion particles are aggregated at the moment. The absolute value of the potential of the emulsion decreases with the increase of the ion concentration of the emulsion, which indicates the instability of the nano emulsion under high salt concentration. Presumably due to shielding of electrostatic repulsive forces of salt ions to emulsion interfaces. In high concentration brine (10-1000 mmol/L NaCl), the electrostatic repulsion is no longer strong enough and hydrophobic interactions dominate, leading to droplet aggregation. In contrast, at relatively low salt concentrations (0.1-1 mmol/L NaCl), electrostatic repulsion is sufficient to overcome the interaction between Van der Waals forces and hydrophobicity, and the emulsion is stable, and the PSO-Casein-EGCG emulsion can maintain good stability at low salt concentrations (0.1-1 mmol/L NaCl).
As shown in fig. 3, the amount of the antioxidant EGCG added had a significant effect on the oxidation degree of the grease under the same emulsifier conditions. The MDA content increased significantly over time during storage for 1-5 days, indicating lipid oxidation of both emulsions. At the same storage time, the MDA content in the PSO-Casein-EGCG emulsion is significantly lower than that of the CK mixture without EGCG, for example, the MDA content in the PSO-Casein-EGCG emulsion is increased from 0.03mg/mL to 0.11mg/mL, and the MDA content in the CK mixture is increased from 0.06mg/mL to 0.16mg/mL. Thus, PSO-Casein-EGCG emulsions have better oxidative stability than CK mixtures without EGCG. This is probably because EGCG exerts its oxidation resistance, which prolongs the stabilization time of the grease and reduces the oxidation degree of the grease; in addition, casein is used as an emulsifier, so that oxygen can be prevented from being generated, and an oil-water system is more stable.
Experimental example 2
Effect of perilla seed oil microemulsion prepared in example 1 on nematode feeding
Nematode culture and synchronization
Generalized culture of nematodes
For all nematodes, the experiment was performed using solid medium. The caenorhabditis elegans grows on a NGM (Nematode growth medium) solid medium, and takes E.coli OP50 as a food source, and the temperature of the growing environment is constantly set to 20 ℃ and the humidity is 40-60%.
Preparation of culture Medium and related solution reference Geng Yiwen method (Geng Yiwen. Investigation of modified apple pomace dietary fiber and lipid-lowering function by Hydrogen peroxide method [ D)]Beijing, national academy of agricultural sciences, 2015.) E.coli OP50 was first inoculated into LB broth liquid medium and shaken in a shaker at 37℃for 12h. When the absorbance OD of the escherichia coli bacterial liquid 600 When the bacterial liquid is 0.4, 100 mu L of bacterial liquid is inoculated on the NGM culture medium for overnight, and the bacterial liquid can be used after being dried. When the number of nematodes needs to be amplified, a piece of culture medium containing the nematodes can be cut by using a toothpick, and the mixture is reversely and lightly stuck to a new NGM culture plate, and the plate is turned once, generally for 3-5 days.
Synchronization of nematodes
The synchronization treatment is to make caenorhabditis elegans in the same growth period so as to perform experiments on nematodes in the same development stage to ensure the accuracy of the experiments. Before the experiment, an NGM plate containing a large number of adult gestation period is prepared in advance, 2mLM9 buffer solution is taken to wash the nematodes down, the nematodes are transferred into a centrifuge tube, the nematodes are centrifuged for 1min at a rotating speed of 3000rpm, the supernatant is discarded, and the escherichia coli bacterial liquid and other impurities are repeatedly washed out for 3 times. After washing, 1mL of 5% NaOCl and 2.5MNaOH mixed lysate is added, the insect body is broken by intense shaking, after the insect eggs are dissolved out, the method is carried out rapidly at 3000rpm for 30 seconds, the supernatant is immediately discarded, then 1mLM9 buffer solution is added for washing, the residual lysate is repeatedly washed for 3 times, 100 mu L of liquid is reserved after the last washing, after uniform mixing, NGM culture medium is poured, and age-synchronous eggs can be obtained, which is nematode synchronization.
Nematode oil red O staining
Oil red O is an oil-soluble dye for staining neutral fat, i.e. staining triglycerides in nematodes, and the use of oil red O staining is a method for evaluating fat accumulation in nematodes (Xu Linli, gao Xuejuan, qu Changqing. J. Chem. Cell chemistry, 2010,19 (06): 615-616.) the method of fat staining in caenorhabditis elegans has a good correlation with triglyceride content.
The oil phase in the emulsion was stained with nile red solution and the protein with FITC (fluorescein isothiocyanate) solution. Nile red solution was added to the grease so that the concentration of Nile red was 0.5g/mol of oil and acetone was removed by rotary evaporation. FITC solution was added to the protein solution, stirred for 2h, left to stand at room temperature in the dark for 24h, and then the dyed emulsion was dialyzed with 0.01mol/L phosphate buffer solution in the dark until fluorescence was not detected. Mixing the FITC-labeled protein solution with nile red-dyed grease to prepare fluorescence-labeled perilla seed oil microemulsion, and observing the emulsion dyeing condition by using a fluorescence inversion microscope. The fluorescent-labeled perilla seed oil microemulsion is fed to caenorhabditis elegans, the nematodes are washed three times with deionized water, then fixed for 10min by using 4% paraformaldehyde, centrifuged for 1min at 5000rpm, and the supernatant is discarded. Washing with deionized water for three times, and observing nematode staining condition with a fluorescent inverted microscope. The fluorescence intensity is measured by an M5 enzyme-labeled instrument, the excitation wavelength is 520nm, and the emission wavelength is 600nm.
Oil red O staining of caenorhabditis elegans. Fixing the nematodes by using 4% paraformaldehyde, washing the nematodes with deionized water for three times, adding an oil red O solution for dyeing for 10min, washing the nematodes with deionized water for three times, and observing the nematode dyeing condition by using a fluorescent inverted microscope. Quantitative analysis was performed using imagej software.
Determination of triglyceride content in nematodes
To evaluate the effect of perilla seed oil nanoemulsion on reduction of nematode fat accumulation, fat accumulation was comprehensively evaluated by a triglyceride TG content assay method in addition to oil red O staining (Huimin Peng, zhaohan Wei, hujie Luo, yiting Yang, zhengxing Wu, yang Lu, xiangliang Gan, inhibition of Fat Accumulation by Hesperidin in Caenorhabditis elegans, journal of Agricultural and Food Chemistry,2016,64 (25): 5207-5214.).
A batch of 25L 4 larval stage nematodes, 1mL M9 buffer and hydrogel (blank, unencapsulated emulsion hydrogel, encapsulated emulsion hydrogel) was added to the tube. Incubate at 20℃for 24h. The nematodes of each group were eluted in PBS centrifuge tubes, centrifuged at 1500r/min for 2min, and the supernatants were taken. After 3 replicates, the nematodes were aspirated into the corresponding 10ml grind tubes, PBS was added to give a liquid volume of 300. Mu.L in each grind tube, appropriate amount of grind beads were added, run at 1500rpm for 60s,10s batch time, 3 grind cycles, and finally observed under an inverted microscope for nematode breakage (grinding and pre-chilling of grind beads on ice). After sufficient grinding, each set of grind tubes was centrifuged at 5000rpm for 10min at 4℃and the supernatant was transferred to a 1.5mL centrifuge tube. The assay was performed according to the triglyceride assay kit and BCA protein assay kit instructions.
Quantitative determination of nematode ZXW mutant lipid droplets.
Lipid droplets are a class of eukaryotic organelles used to store neutral fats, such as triglycerides and cholesterol esters. Lipid droplets play an important role in energy utilization and storage in mammals and caenorhabditis elegans (Chunxiu Lin, zuanxin Su, jia Luo, lin Jiang, shaodan Shen, wanyang Zheng, wenxin Gu, yong Cao, yonjiao Chen. Polysaccharide extracted from the leaves of Cyclocarya paliurus (Batal.) Iljinskaja enhanced stress resistance in Caenorhabditis elegans via skn-1and hsf-1[ J ]. International Journal ofBiological Macromolecules,2020, 143:243-254). The short-chain dehydrogenase DHS-3 was almost completely localized at the physiological level in the gut of C.elegans, in mutant nematodes ZXW, the lipid droplets were labeled with GFP and thus were green fluorescent labeled, which was clearly visible in a laser confocal microscope, and could be used to detect lipid droplets states (Zhang Peng, na Huimin, liu Zhenglong, zhang Shuyan, xue Peng, chen Yong, pu Jing, peng Gong, huang Xun, yang Fuquan, xie Zhensheng, xu Tao, xu Pingyong, ou Guangshuo, zhang ShaobO, liu Pingseng, proteomic Study and Marker Protein Identification of Caenorhabditis elegans Lipid Droplets, molecular & Cellular Proteomics;2012,11 (8): 317-328.
Nematodes were grown according to the above method and after incubation for 24h at 20℃lipid titration was performed on ZXW618 vermicular lipid drops. All worms were anesthetized in 100mM sodium azide drops and mounted on 2% agar slides prior to imaging. The fluorescence image adopts a confocal laser scanning microscope. Worms were observed using 488nm excitation filters and 525nm emission filters. More than 20 worms were detected under each condition and the images were analyzed using ImageJ.
The results are shown in fig. 4, 5 and 6.
In FIG. 4, OP50 is a blank control, PSO-Casein is a CK mixture, PSO-Casein-EGCG is a perilla seed oil microemulsion, and Casein-EGCG is an EGCG aqueous solution (equal amount of water is used for replacing perilla seed oil).
As can be seen from FIG. 4, the lipid lowering effect is PSO-Casein-EGCG emulsion > PSO-Casein > Casein-EGCG > OP50. The EGCG and the perilla seed oil have lipid-lowering effects. The fat accumulation in the nematode body is measured by using oil red O staining, and the staining intensity of the oil red O of the nematode in the PSO-Casein-EGCG emulsion is obviously reduced. In addition, quantification of staining intensity showed a significant decrease in nematode fat accumulation on media containing perilla seed oil compared to normal feeding nematodes. Compared to the OP50 group, the PSO-Casein treated nematodes and the PSO-Casein-EGCG emulsion treated nematodes showed a reduction in fat accumulation of 39.32% and 47.72% (p < 0.05), respectively, with a significantly higher reduction than the samples without perilla seed oil.
Experiments show that the perilla seed oil has remarkable effect of reducing fat accumulation in the nematode, and the PSO-Casein-EGCG emulsion has better effect of reducing the fat accumulation in the nematode than a mixture without EGCG. Therefore, the prepared perilla seed oil microemulsion has positive effect on reducing fat accumulation in the nematode body. Therefore, the perilla seed oil microemulsion formed by the perilla seed oil and the EGCG-CA compound improves the lipid-lowering effect of the perilla seed oil, and the perilla seed oil plays a leading role in reducing fat accumulation in a nematode body.
As can be seen from FIG. 5, the lipid lowering effect is PSO-Casein-EGCG emulsion > PSO-Casein > Casein-EGCG > OP50. The ability to reduce triglyceride levels ranked consistent with the oil red O staining ranking, indicating that the two experiments had good correlation. By measuring the triglyceride content, the accumulated fat amount of the nematodes fed with PSO-Casein and PSO-Casein-EGCG emulsion was reduced by 50.00% and 77.50%, respectively. The fat reduction values were large compared to the oil red O staining experiments, probably because of some subtle differences between the methods of determining triglyceride and oil red O staining. These data indicate that PSO-Casein-EGCG emulsion has a significant effect in reducing normal nematode fat accumulation.
Lipid droplets are a class of eukaryotic organelles used to store neutral fats, such as triglycerides and cholesterol esters. Lipid droplets play an important role in the energy utilization and storage of mammals and caenorhabditis elegans. The short-chain dehydrogenase DHS-3 is located almost completely in the intestinal tract of caenorhabditis elegans at physiological level, and in mutant nematode ZXW618, GFP is used for marking lipid droplets, so that the lipid droplets have green fluorescent marks, can be clearly seen by a laser confocal microscope, and can be used for detecting the lipid droplet state. The results are shown in FIG. 6, and the average particle size results of the mutants show that the particle size is PSO-Casein-EGCG emulsion < PSO-Casein < Casein-EGCG < OP50 from small to large. The lipid droplet sizes of nematodes treated with PSO-Casein and PSO-Casein-EGCG emulsions were reduced by 33.53% and 41.23%, respectively. The results show that the perilla seed oil nanoemulsion can reduce the size of the nematode lipid droplets, thereby reducing fat accumulation. Therefore, the PSO-Casein-EGCG emulsion has better lipid-lowering effect.
Experimental example 3
Hydrogels with different gellan gum contents (1% GG, 2% GG, 3% GG) were prepared according to the preparation method described in example 1, and then the hydrogels were adsorbed with perilla seed oil microemulsions to obtain the volumes of the perilla seed oil microemulsions-hydrogels with different GG concentrations, and the hydrogels were stored at 37℃for 1 to 5 days, and the MDA values were measured to evaluate the oxidative stability of the hydrogels by the MDA values.
As shown in FIG. 7, the MDA content of all samples increased significantly over time during storage for 1-5 days, indicating lipid oxidation of the lipids in the samples. Under the same storage conditions, the magnitudes of the MDA values are, in order: PSO > 3% GG-PSO complex > PSO-Casein > 1% GG-PSO complex > PSO-Casein-EGCG emulsion > 2% GG-PSO complex. After the purple Perilla Seed Oil (PSO) MDA value without any treatment is increased from 0.094mg/mL to 0.259mg/mL to prepare PSO-Casein-EGCG emulsion (purple perilla seed oil microemulsion) and the MDA value is increased from 0.040mg/mL to 0.116mg/mL, and after the PSO-Casein-EGCG emulsion is prepared into a hydrogel system, the MDA value of 2% GG-PSO compound (purple perilla seed oil microemulsion-hydrogel system) is increased from 0.028mg/mL to 0.093mg/mL; it can be seen that 2% GG-PSO complex (perilla seed oil microemulsion-hydrogel system) has better oxidation stability; this is probably due to the fact that after the hydrogel encapsulates the emulsion, the entry of oxygen is further isolated, so that the perilla seed oil is in an anaerobic environment, and the stability of the PSO-Casein-EGCG emulsion and the perilla seed oil is better improved.
Apparent and microstructure (SEM) characterization analysis of the perilla seed oil microemulsion-hydrogel system
As a result, as shown in fig. 8, the color gradually changed from colorless transparent to white opaque as the gellan gum content increased, the GG-PSO emulsion complex exhibited a white opaque state and the texture was softer than the hydrogel without the PSO emulsion.
Microscopic structural observation is carried out on the perilla seed oil microemulsion-hydrogel system with different gellan gum concentrations by using a Scanning Electron Microscope (SEM), so that the hydrogel and GG-PSO emulsion composite are subjected to freeze drying, a large number of gaps appear at the breaking positions of the 1% GG hydrogel and GG-PSO emulsion composite, the gaps are larger, and the breaking positions of the 2% GG hydrogel and the 3% GG hydrogel are smoother. The surface of the hydrogel of 1% GG and GG-PSO emulsion complex is rough and wrinkled and is full of cracks; the hydrogel surface of 2% GG was smoother, but had protrusions and wrinkles; the hydrogel surface with 3% GG is the smoother, flatter and denser.
Rheology determination
Rheology is a discipline in studying material flow and deformation. Rheology is mainly studied as a response of materials to mechanics in case of tension or external pressure. The storage modulus G' [ Pa ], also known as the elastic portion, refers to the amount of energy stored by a material as it deforms due to elastic deformation (reversibility). It reflects the strength of the material mechanics. The greater the storage modulus G' of the material, the better the mechanical properties of the material. The loss modulus G' [ Pa ], also known as the viscous fraction, refers to the amount of energy lost by the viscous deformation (irreversible) of a material as it is deformed. It reflects the strength of the material viscosity.
The rheology experiment was performed by means of a rheometer. The hydrogel was about 1.0 mm thick with a small spatula, removed from the preparation dish and carefully placed on the lower plate of the rheometer. The upper plate is then lowered until it reaches the hydrogel surface. The frequency sweep experiment was performed on a hydrogel in the linear viscoelastic region at 25, 37 ℃ in the range of 0.01-10 Hz.
Stress sweep experiments (25, 37 ℃ and 0.1 Hz) were performed with a pressure range of 0.01-30Pa.
The measurement of the yield stress (25 and 37 ℃) was carried out by applying a linearly increasing stress (0.1-4,000 pa in 4 minutes) on the hydrogel and the resulting deformation γ=f (σ) was recorded. Yield stress is calculated as the intersection point (flow point) of lines extrapolated from the linear part of the experimental curve. All experiments were performed in triplicate and the calculated values were recorded as arithmetic mean.
From fig. 9, it can be seen that the storage modulus of the 2% gg (hydrogel with 2% gellan gum content) and 2% gg-PSO emulsion complex (perilla seed oil microemulsion-hydrogel system after adsorption of PSO-Casein-EGCG emulsion by hydrogel with 2% gellan gum content) are not substantially changed with the change of angular frequency during the test, indicating that the mechanical properties are relatively stable. Adsorption PSO-Casein-EGCG emulsionAfter that, the gel strength of the hydrogel was lowered. As GG content increases, gel strength increases, possibly Mg 2+ And electrostatic action of GG results in an increase in gel strength of the hydrogel.
Lipid-lowering effect of perilla seed oil microemulsion-hydrogel system
Specific test methods are described in experimental example 2.
As shown in FIG. 10, the lipid-lowering effect was 2% GG-PSO complex > PSO-Casein-EGCG emulsion > 2% GG > OP50. This indicates that the 2% GG-PSO complex prepared by hydrogel has lipid-lowering effect. Fat accumulation in nematodes was measured by oil red O staining, and the staining intensity of nematode oil red O in 2% GG-PSO complex was significantly reduced. Furthermore, quantification of staining intensity showed a significant decrease in nematode fat accumulation in the medium containing 2% GG-PSO complex compared to normal feeding nematodes. Fat accumulation was reduced by 47.72% and 52.84% (p < 0.05) for PSO-Casein-EGCG emulsion treated nematodes and 2% GG-PSO complex treated nematodes, respectively, compared to OP50 group. It is possible that oxidation of the emulsion by oxygen and the like is further isolated by encapsulation of the hydrogel, thereby improving the stability of the emulsion such that nematode fat accumulation is reduced.
Experiments show that the PSO-Casein-EGCG emulsion is encapsulated in hydrogel to prepare 2% GG-PSO compound, which can effectively reduce nematode fat accumulation.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (11)
1. The preparation method of the purple perilla seed oil microemulsion-hydrogel system is characterized by comprising the following steps:
1) Mixing casein with water to obtain a water phase, mixing EGCG with perilla seed oil to obtain an oil phase, mixing the water phase with the oil phase, and homogenizing to obtain a perilla seed oil microemulsion;
2) Gellan gumSolution drop-added to chitosan-MgCl 2 Repeatedly freezing and thawing for 3-5 times in the solution to obtain hydrogel;
3) Mixing the hydrogel after freeze-drying with the perilla seed oil microemulsion for 10-14 hours to obtain a perilla seed oil microemulsion-hydrogel system;
there is no time sequence limitation between step 1) and step 2).
2. The preparation method of claim 1, wherein the mass percentage of casein in the perilla seed oil microemulsion is 0.4% -1.5%, the mass percentage of EGCG in the perilla seed oil microemulsion is 0.008% -0.05%, and the mass percentage of perilla seed oil in the perilla seed oil microemulsion is 5% -15%.
3. The production method according to claim 1 or 2, wherein the homogenizing comprises sequentially performing first homogenizing and second homogenizing; the rotation speed of the first homogenization is 8000-12000 rpm, the repetition number of the first homogenization is 2-4, and the time of each time is 0.5-1.5 min; the second homogenization is high-pressure homogenization, the pressure of the second homogenization is 450-550 mpa, the repetition number of the second homogenization is 4-6, and the time of each time is 20-40 s.
4. The method according to claim 1, wherein the method for preparing the gellan gum solution in step 2) comprises mixing gellan gum with water, and heating at 85-95 ℃ for 15-25 min.
5. The preparation method according to claim 1 or 4, wherein the chitosan-MgCl 2 The pH value of the solution is 3.4-3.6, and the chitosan-MgCl 2 MgCl in solution 2 The mass percentage of the chitosan-MgCl is 0.10% -0.15%, and the chitosan-MgCl is the same as the chitosan-MgCl 2 The mass percentage of chitosan in the solution is 2% -4%.
6. The preparation method of claim 1, wherein the mass percentage of gellan gum in the hydrogel obtained in the step 2) is 1% -3%; the mass ratio of the freeze-dried hydrogel to the perilla seed oil microemulsion in the step 3) is (1-2), namely (1-2).
7. The preparation method according to claim 1 or 6, wherein the repeated freezing and thawing is carried out at a freezing temperature of-3 to-6 ℃ for 7 to 9 hours; and the melting temperature of repeated freezing and thawing is 10-30 ℃.
8. The perilla seed oil microemulsion-hydrogel system prepared by the preparation method of any one of claims 1 to 7.
9. Use of the perilla seed oil microemulsion-hydrogel system of claim 8 for preparing a medicament with lipid-lowering efficacy.
10. Use of the perilla seed oil microemulsion-hydrogel system of claim 8 for preparing a functional food with auxiliary lipid-lowering effect.
11. The use according to claim 9 or 10, wherein the lipid lowering comprises lowering the content of triglycerides and/or the particle size of lipid droplets.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210302211.5A CN114698843B (en) | 2022-03-25 | 2022-03-25 | Perilla seed oil microemulsion-hydrogel system and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210302211.5A CN114698843B (en) | 2022-03-25 | 2022-03-25 | Perilla seed oil microemulsion-hydrogel system and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114698843A CN114698843A (en) | 2022-07-05 |
| CN114698843B true CN114698843B (en) | 2023-05-12 |
Family
ID=82171184
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210302211.5A Active CN114698843B (en) | 2022-03-25 | 2022-03-25 | Perilla seed oil microemulsion-hydrogel system and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114698843B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115812827A (en) * | 2022-11-07 | 2023-03-21 | 嘉必优生物技术（武汉）股份有限公司 | Hydrogel system suitable for preparing gel soft sweets and preparation method thereof |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103467755A (en) * | 2013-09-18 | 2013-12-25 | 薛巍 | Medicinal sustained-release hydrogel as well as preparation method and application thereof |
| CN103483604A (en) * | 2012-06-12 | 2014-01-01 | 江南大学 | Controlled-release chitosan-gellan gum calcium gel bead and preparation method thereof |
| CN107950684A (en) * | 2017-12-22 | 2018-04-24 | 暨南大学 | A kind of oleogel rich in unrighted acid and preparation method and application |
| CN109172517A (en) * | 2018-10-25 | 2019-01-11 | 山东师范大学 | A kind of preparation method of the microemulsion hydrogel as apiolin carrier with pH response |
| CN109288065A (en) * | 2018-11-01 | 2019-02-01 | 广州白云山汉方现代药业有限公司 | A kind of suspension emulsion gel and preparation method thereof loading liposoluble vitamin |
| CN110339393A (en) * | 2019-07-19 | 2019-10-18 | 吉林大学 | It is a kind of based on hydrogel-core-shell particles wound dressing and preparation method thereof |
| CN111034813A (en) * | 2019-11-19 | 2020-04-21 | 华南农业大学 | Preparation method of novel structured emulsion for improving oxidation stability of grease |
| CN113243529A (en) * | 2021-04-29 | 2021-08-13 | 东北农业大学 | Preparation method of carrier beta-carotene gel microspheres |
| CN113683811A (en) * | 2021-06-30 | 2021-11-23 | 青岛科技大学 | Efficient flame-retardant phosphorylated chitosan aerogel and preparation method thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070104778A1 (en) * | 2005-11-07 | 2007-05-10 | Hongxia Zeng | Controlled-release emulsion compositions |
-
2022
- 2022-03-25 CN CN202210302211.5A patent/CN114698843B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103483604A (en) * | 2012-06-12 | 2014-01-01 | 江南大学 | Controlled-release chitosan-gellan gum calcium gel bead and preparation method thereof |
| CN103467755A (en) * | 2013-09-18 | 2013-12-25 | 薛巍 | Medicinal sustained-release hydrogel as well as preparation method and application thereof |
| CN107950684A (en) * | 2017-12-22 | 2018-04-24 | 暨南大学 | A kind of oleogel rich in unrighted acid and preparation method and application |
| CN109172517A (en) * | 2018-10-25 | 2019-01-11 | 山东师范大学 | A kind of preparation method of the microemulsion hydrogel as apiolin carrier with pH response |
| CN109288065A (en) * | 2018-11-01 | 2019-02-01 | 广州白云山汉方现代药业有限公司 | A kind of suspension emulsion gel and preparation method thereof loading liposoluble vitamin |
| CN110339393A (en) * | 2019-07-19 | 2019-10-18 | 吉林大学 | It is a kind of based on hydrogel-core-shell particles wound dressing and preparation method thereof |
| CN111034813A (en) * | 2019-11-19 | 2020-04-21 | 华南农业大学 | Preparation method of novel structured emulsion for improving oxidation stability of grease |
| CN113243529A (en) * | 2021-04-29 | 2021-08-13 | 东北农业大学 | Preparation method of carrier beta-carotene gel microspheres |
| CN113683811A (en) * | 2021-06-30 | 2021-11-23 | 青岛科技大学 | Efficient flame-retardant phosphorylated chitosan aerogel and preparation method thereof |
Non-Patent Citations (6)
| Title |
|---|
| Tea polyphenols association to caseinate-stabilized oilewater interfaces;Somayeh Sabouri 等;Food Hydrocolloids;第51卷;95-100 * |
| 冻融法介导的多糖凝胶化及其在食品领域中的研究进展;李安琪 等;食品与发酵工业;第46卷(第23期);301-308 * |
| 壳聚糖-海藻酸钠凝胶微粒稳定的AKD乳液;陈学帅 等;中国造纸;第35卷(第12期);7-10 * |
| 紫苏油的药理活性及其缓释技术研究进展;杜艳;殷丽君;朱巧梅;刘海杰;;河南工业大学学报(自然科学版);第37卷(第4期);114-117，123 * |
| 紫苏籽油乳状液的制备及其稳定性评价;韩丽丽 等;食品与发酵工业;第42卷(第4期);144-149 * |
| 镁离子对PVA/GG-Mg~(2+)复合水凝胶的结构和物理化学性质的影响;王飞 等;化工学报;第71卷(第1期);376-387 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114698843A (en) | 2022-07-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Mohammadi et al. | Development of novel active packaging films based on whey protein isolate incorporated with chitosan nanofiber and nano-formulated cinnamon oil | |
| Hosseini et al. | Potential Pickering emulsion stabilized with chitosan-stearic acid nanogels incorporating clove essential oil to produce fish-oil-enriched mayonnaise | |
| Jiménez et al. | Influence of nanoliposomes incorporation on properties of film forming dispersions and films based on corn starch and sodium caseinate | |
| Liu et al. | Characterization of lipid emulsions during in vitro digestion in the presence of three types of nanocellulose | |
| Zhang et al. | Natural oil bodies from typical oilseeds: Structural characterization and their potentials as natural delivery system for curcumin | |
| Zhang et al. | Development of soy protein isolate/κ-carrageenan composite hydrogels as a delivery system for hydrophilic compounds: Monascus yellow | |
| Cao et al. | Enhancing the physicochemical performance of myofibrillar gels using Pickering emulsion fillers: Rheology, microstructure and stability | |
| Dong et al. | Oleogel-based Pickering emulsions stabilized by ovotransferrin–carboxymethyl chitosan nanoparticles for delivery of curcumin | |
| Chen et al. | Development of anti-photo and anti-thermal high internal phase emulsions stabilized by biomass lignin as a nutraceutical delivery system | |
| Xie et al. | Encapsulation of astaxanthin-enriched camelina oil extract in ovalbumin/gum arabic stabilized emulsion with/without crosslinking by tannic acid | |
| CN114698843B (en) | Perilla seed oil microemulsion-hydrogel system and preparation method and application thereof | |
| Najafi et al. | Pullulan films loading saffron extract encapsulated in nanoliposomes; preparation and characterization | |
| Lan et al. | Physicochemical stability of safflower oil body emulsions during food processing | |
| Zhou et al. | Effects of carboxymethyl cellulose on the emulsifying, gel and digestive properties of myofibrillar protein-soybean oil emulsion | |
| Hao et al. | The improvement of the physicochemical properties and bioaccessibility of lutein microparticles by electrostatic complexation | |
| Ribeiro et al. | In vitro digestion and bioaccessibility studies of vitamin E-loaded nanohydroxyapatite Pickering emulsions and derived fortified foods | |
| Jiang et al. | Improving probiotic survival using water-in-oil-in-water (W1/O/W2) emulsions: Role of fish oil in inner phase and sodium alginate in outer phase | |
| Yue et al. | Fabrication and stabilization mechanisms of Pickering emulsions based on gliadin/arabinoxylan complexes | |
| Wellala et al. | Juice related water-soluble pectin characteristics and bioaccessibility of bioactive compounds in oil and emulsion incorporated mixed juice processed by high pressure homogenization | |
| Liu et al. | Pickering emulsions stabilized with a spirulina protein–chitosan complex for astaxanthin delivery | |
| Wei et al. | Caffeic acid phenethyl ester loaded in a targeted delivery system based on a solid-in-oil-in-water multilayer emulsion: Characterization, stability, and fate of the emulsion during in vivo digestion | |
| Tang et al. | Pseudosciaena crocea roe protein‐stabilized emulsions for oral delivery systems: In vitro digestion and in situ intestinal perfusion study | |
| Akonjuen et al. | Development of protein isolate-alginate-based delivery system to improve oxidative stability of njangsa (Ricinodendron heudelotii) seed oil | |
| Wang et al. | Effect of simulated saliva components on the in vitro digestion of peanut oil body emulsion | |
| Yan et al. | Construction and characterization of egg white protein-gallic acid-xanthan gum-based emulsion and oleogel |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |