CN108938690B - Moringa oleifera leaf ethyl acetate extract and extraction method and application thereof - Google Patents
Moringa oleifera leaf ethyl acetate extract and extraction method and application thereof Download PDFInfo
- Publication number
- CN108938690B CN108938690B CN201811132481.6A CN201811132481A CN108938690B CN 108938690 B CN108938690 B CN 108938690B CN 201811132481 A CN201811132481 A CN 201811132481A CN 108938690 B CN108938690 B CN 108938690B
- Authority
- CN
- China
- Prior art keywords
- ethyl acetate
- acetate extract
- moringa
- moringa oleifera
- extracting
- 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
- 235000011347 Moringa oleifera Nutrition 0.000 title claims abstract description 57
- 244000179886 Moringa oleifera Species 0.000 title claims abstract description 39
- 239000002024 ethyl acetate extract Substances 0.000 title claims abstract description 38
- 238000000605 extraction Methods 0.000 title claims abstract description 11
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims abstract description 34
- 241000220215 Moringa Species 0.000 claims abstract description 18
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims abstract description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 11
- 239000000284 extract Substances 0.000 claims abstract description 8
- 238000000034 method Methods 0.000 claims abstract description 8
- 239000000706 filtrate Substances 0.000 claims abstract description 7
- 239000003208 petroleum Substances 0.000 claims abstract description 7
- 230000009758 senescence Effects 0.000 claims abstract description 6
- 239000003814 drug Substances 0.000 claims abstract description 5
- 210000001035 gastrointestinal tract Anatomy 0.000 claims abstract description 4
- 238000002360 preparation method Methods 0.000 claims abstract description 4
- 238000001914 filtration Methods 0.000 claims abstract description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 16
- IYRMWMYZSQPJKC-UHFFFAOYSA-N kaempferol Chemical compound C1=CC(O)=CC=C1C1=C(O)C(=O)C2=C(O)C=C(O)C=C2O1 IYRMWMYZSQPJKC-UHFFFAOYSA-N 0.000 claims description 8
- 239000007864 aqueous solution Substances 0.000 claims description 6
- 238000004519 manufacturing process Methods 0.000 claims description 5
- GYFFKZTYYAFCTR-JUHZACGLSA-N 4-O-trans-caffeoylquinic acid Chemical compound O[C@@H]1C[C@](O)(C(O)=O)C[C@@H](O)[C@H]1OC(=O)\C=C\C1=CC=C(O)C(O)=C1 GYFFKZTYYAFCTR-JUHZACGLSA-N 0.000 claims description 4
- DSHJQVWTBAAJDN-SMKXDYDZSA-N 4-caffeoylquinic acid Natural products CO[C@@]1(C[C@@H](O)[C@H](OC(=O)C=Cc2ccc(O)c(O)c2)[C@H](O)C1)C(=O)O DSHJQVWTBAAJDN-SMKXDYDZSA-N 0.000 claims description 4
- CWVRJTMFETXNAD-GMZLATJGSA-N 5-Caffeoyl quinic acid Natural products O[C@H]1C[C@](O)(C[C@H](OC(=O)C=Cc2ccc(O)c(O)c2)[C@@H]1O)C(=O)O CWVRJTMFETXNAD-GMZLATJGSA-N 0.000 claims description 4
- GYFFKZTYYAFCTR-UHFFFAOYSA-N 5-O-(6'-O-galloyl)-beta-D-glucopyranosylgentisic acid Natural products OC1CC(O)(C(O)=O)CC(O)C1OC(=O)C=CC1=CC=C(O)C(O)=C1 GYFFKZTYYAFCTR-UHFFFAOYSA-N 0.000 claims description 4
- JMGZEFIQIZZSBH-UHFFFAOYSA-N Bioquercetin Natural products CC1OC(OCC(O)C2OC(OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5)C(O)C2O)C(O)C(O)C1O JMGZEFIQIZZSBH-UHFFFAOYSA-N 0.000 claims description 4
- GYFFKZTYYAFCTR-ZNEHSRBWSA-N Cryptochlorogensaeure Natural products O[C@@H]1C[C@@](O)(C[C@@H](O)[C@@H]1OC(=O)C=Cc2ccc(O)c(O)c2)C(=O)O GYFFKZTYYAFCTR-ZNEHSRBWSA-N 0.000 claims description 4
- UBSCDKPKWHYZNX-UHFFFAOYSA-N Demethoxycapillarisin Natural products C1=CC(O)=CC=C1OC1=CC(=O)C2=C(O)C=C(O)C=C2O1 UBSCDKPKWHYZNX-UHFFFAOYSA-N 0.000 claims description 4
- CWVRJTMFETXNAD-JUHZACGLSA-N chlorogenic acid Chemical compound O[C@@H]1[C@H](O)C[C@@](O)(C(O)=O)C[C@H]1OC(=O)\C=C\C1=CC=C(O)C(O)=C1 CWVRJTMFETXNAD-JUHZACGLSA-N 0.000 claims description 4
- GYFFKZTYYAFCTR-LMRQPLJMSA-N cryptochlorogenic acid Natural products O[C@H]1C[C@@](O)(C[C@H](O)[C@H]1OC(=O)C=Cc2ccc(O)c(O)c2)C(=O)O GYFFKZTYYAFCTR-LMRQPLJMSA-N 0.000 claims description 4
- IVTMALDHFAHOGL-UHFFFAOYSA-N eriodictyol 7-O-rutinoside Natural products OC1C(O)C(O)C(C)OC1OCC1C(O)C(O)C(O)C(OC=2C=C3C(C(C(O)=C(O3)C=3C=C(O)C(O)=CC=3)=O)=C(O)C=2)O1 IVTMALDHFAHOGL-UHFFFAOYSA-N 0.000 claims description 4
- 235000008777 kaempferol Nutrition 0.000 claims description 4
- MWDZOUNAPSSOEL-UHFFFAOYSA-N kaempferol Natural products OC1=C(C(=O)c2cc(O)cc(O)c2O1)c3ccc(O)cc3 MWDZOUNAPSSOEL-UHFFFAOYSA-N 0.000 claims description 4
- UXOUKMQIEVGVLY-UHFFFAOYSA-N morin Natural products OC1=CC(O)=CC(C2=C(C(=O)C3=C(O)C=C(O)C=C3O2)O)=C1 UXOUKMQIEVGVLY-UHFFFAOYSA-N 0.000 claims description 4
- FDRQPMVGJOQVTL-UHFFFAOYSA-N quercetin rutinoside Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 FDRQPMVGJOQVTL-UHFFFAOYSA-N 0.000 claims description 4
- 235000005493 rutin Nutrition 0.000 claims description 4
- ALABRVAAKCSLSC-UHFFFAOYSA-N rutin Natural products CC1OC(OCC2OC(O)C(O)C(O)C2O)C(O)C(O)C1OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5 ALABRVAAKCSLSC-UHFFFAOYSA-N 0.000 claims description 4
- IKGXIBQEEMLURG-BKUODXTLSA-N rutin Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@@H]1OC[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 IKGXIBQEEMLURG-BKUODXTLSA-N 0.000 claims description 4
- 229960004555 rutoside Drugs 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract description 35
- 210000002490 intestinal epithelial cell Anatomy 0.000 abstract description 7
- -1 phenolic acid compounds Chemical class 0.000 abstract description 7
- 230000032683 aging Effects 0.000 abstract description 6
- 235000013305 food Nutrition 0.000 abstract description 5
- 239000000047 product Substances 0.000 abstract description 4
- 238000002474 experimental method Methods 0.000 abstract description 3
- 229930003935 flavonoid Natural products 0.000 abstract description 3
- 235000017173 flavonoids Nutrition 0.000 abstract description 3
- 230000036541 health Effects 0.000 abstract description 3
- 229940079593 drug Drugs 0.000 abstract description 2
- 230000001681 protective effect Effects 0.000 abstract description 2
- WQZGKKKJIJFFOK-SVZMEOIVSA-N (+)-Galactose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-SVZMEOIVSA-N 0.000 description 14
- 238000010186 staining Methods 0.000 description 10
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 9
- 238000004458 analytical method Methods 0.000 description 9
- 229940118019 malondialdehyde Drugs 0.000 description 9
- 102000004169 proteins and genes Human genes 0.000 description 9
- 108090000623 proteins and genes Proteins 0.000 description 9
- 102100025064 Cellular tumor antigen p53 Human genes 0.000 description 8
- 102000005936 beta-Galactosidase Human genes 0.000 description 8
- 108010005774 beta-Galactosidase Proteins 0.000 description 8
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 6
- 101000997832 Homo sapiens Tyrosine-protein kinase JAK2 Proteins 0.000 description 6
- 102000001712 STAT5 Transcription Factor Human genes 0.000 description 6
- 108010029477 STAT5 Transcription Factor Proteins 0.000 description 6
- 102100033444 Tyrosine-protein kinase JAK2 Human genes 0.000 description 6
- 239000012071 phase Substances 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 230000035897 transcription Effects 0.000 description 6
- 238000013518 transcription Methods 0.000 description 6
- 102100021569 Apoptosis regulator Bcl-2 Human genes 0.000 description 5
- 101000971171 Homo sapiens Apoptosis regulator Bcl-2 Proteins 0.000 description 5
- 102000052116 epidermal growth factor receptor activity proteins Human genes 0.000 description 5
- 108700015053 epidermal growth factor receptor activity proteins Proteins 0.000 description 5
- YOHYSYJDKVYCJI-UHFFFAOYSA-N n-[3-[[6-[3-(trifluoromethyl)anilino]pyrimidin-4-yl]amino]phenyl]cyclopropanecarboxamide Chemical compound FC(F)(F)C1=CC=CC(NC=2N=CN=C(NC=3C=C(NC(=O)C4CC4)C=CC=3)C=2)=C1 YOHYSYJDKVYCJI-UHFFFAOYSA-N 0.000 description 5
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 210000004379 membrane Anatomy 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 102000007469 Actins Human genes 0.000 description 3
- 108010085238 Actins Proteins 0.000 description 3
- 101100243447 Arabidopsis thaliana PER53 gene Proteins 0.000 description 3
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 3
- 101150080074 TP53 gene Proteins 0.000 description 3
- 229930003268 Vitamin C Natural products 0.000 description 3
- 210000005027 intestinal barrier Anatomy 0.000 description 3
- 230000007358 intestinal barrier function Effects 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 239000012192 staining solution Substances 0.000 description 3
- 235000019154 vitamin C Nutrition 0.000 description 3
- 239000011718 vitamin C Substances 0.000 description 3
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- 108091092584 GDNA Proteins 0.000 description 2
- 101710088172 HTH-type transcriptional regulator RipA Proteins 0.000 description 2
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 description 2
- 239000011575 calcium Substances 0.000 description 2
- 229910052791 calcium Inorganic materials 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 210000003855 cell nucleus Anatomy 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000000834 fixative Substances 0.000 description 2
- 235000019253 formic acid Nutrition 0.000 description 2
- 238000003119 immunoblot Methods 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- 230000002934 lysing effect Effects 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 230000036542 oxidative stress Effects 0.000 description 2
- 238000004445 quantitative analysis Methods 0.000 description 2
- 238000010814 radioimmunoprecipitation assay Methods 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 238000007619 statistical method Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 239000012224 working solution Substances 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- OPIFSICVWOWJMJ-AEOCFKNESA-N 5-bromo-4-chloro-3-indolyl beta-D-galactoside Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1OC1=CNC2=CC=C(Br)C(Cl)=C12 OPIFSICVWOWJMJ-AEOCFKNESA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 208000022559 Inflammatory bowel disease Diseases 0.000 description 1
- 206010051606 Necrotising colitis Diseases 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000013566 allergen Substances 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 238000003149 assay kit Methods 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000032677 cell aging Effects 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 229940116108 lactase Drugs 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 208000004995 necrotizing enterocolitis Diseases 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 230000030648 nucleus localization Effects 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000012261 overproduction Methods 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 201000006195 perinatal necrotizing enterocolitis Diseases 0.000 description 1
- 239000002985 plastic film Substances 0.000 description 1
- 229920006255 plastic film Polymers 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 238000005173 quadrupole mass spectroscopy Methods 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 238000001269 time-of-flight mass spectrometry Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 108700012359 toxins Proteins 0.000 description 1
- 238000001195 ultra high performance liquid chromatography Methods 0.000 description 1
- 238000003828 vacuum filtration Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
Images
Classifications
-
- 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)
-
- 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
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/35—Extraction with lipophilic solvents, e.g. Hexane or petrol ether
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Mycology (AREA)
- Public Health (AREA)
- Botany (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Alternative & Traditional Medicine (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Medical Informatics (AREA)
- General Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention discloses an ethyl acetate extract of moringa leaves and an extraction method, wherein the method comprises the following steps: s1: extracting Moringa oleifera leaves with an extractant, and filtering; s2: extracting the filtrate obtained in the step S1 by using petroleum ether to leave a water phase; s3: extracting the residual water phase in the step S2 by using ethyl acetate; s4: the extract in step S3 was concentrated and dried to obtain an ethyl acetate extract. The invention also discloses application of the moringa oleifera leaf ethyl acetate extract in reducing the aging condition of IEC-6 cells and reducing the generation of MDA in an IEC-6 cell model, and application of the moringa oleifera leaf ethyl acetate extract in preparing food, health care products or medicines for protecting intestinal tracts. The preparation method is simple and environment-friendly, the extracted moringa oleifera leaf ethyl acetate extract contains various phenolic acid compounds and flavonoid compounds, and experiments prove that the ethyl acetate extract has a protective effect on an intestinal epithelial cell line IEC-6 cell senescence model.
Description
Technical Field
The invention relates to the field of extraction and application of moringa leaves, in particular to a moringa leaf ethyl acetate extract and an extraction method and application thereof.
Background
Intestinal epithelial cells are directly exposed to food and the external environment, and under the action of harmful microorganisms, toxins and allergens, there is oxidative stress and aging. Evidence suggests that intestinal barrier failure in necrotizing enterocolitis is due, in part, to the overproduction of nitric oxide and other toxic oxidant species leading to intestinal epithelial cell death and intestinal barrier failure. The EGF signaling pathway plays an important role in reducing the mortality and necrosis counts of intestinal epithelial cells. The failure of damaged cells to repair in time will have an impact on the breakdown of intestinal barrier and increase the probability of inflammatory bowel disease.
Moringa leaves are approved as new resource food in 2012, the whole plant of Moringa can be utilized, roots and barks are traditional medicine raw materials, and tender leaves and fruit pods are nutritious vegetables. The moringa oleifera is rich in protein, vitamins and amino acids, wherein the moringa oleifera leaves contain a large amount of VA, VB and VC, calcium, iron, protein and the like, so the moringa oleifera is widely eaten. The edible moringa product can improve the immunity of human bodies, wherein 25g of moringa leaf powder contains 270% of VA, 42% of protein, 125% of calcium, 70% of iron and 22% of Vc required by infants every day.
However, the research on the moringa leaves is limited at present, and the extraction method, the components and the effects of the moringa leaves are yet to be further developed.
Disclosure of Invention
In view of the above, the present invention aims to provide a method for extracting moringa oleifera leaves with ethyl acetate.
The technical scheme is as follows.
A method for extracting moringa oleifera leaves with ethyl acetate comprises the following steps:
s1: extracting Moringa oleifera leaves with an extractant, and filtering;
s2: extracting the filtrate obtained in the step S1 by using petroleum ether to leave a water phase;
s3: extracting the residual water phase in the step S2 by using ethyl acetate;
s4: the extract in step S3 was concentrated and dried to obtain an ethyl acetate extract.
Further, in step S1, the extracting agent is anhydrous acetone or an aqueous acetone solution.
Further, in the step S1, the extracting agent is 75% acetone aqueous solution.
Further, in the step S1, the mass-to-volume ratio of the moringa leaves to the acetone aqueous solution is 1 (5-10). The applicant extracts moringa leaves in a ratio of 1:5, 1:6, 1:8 and 1:10 respectively, and can extract corresponding effective components from the moringa leaves.
Further, in the step S2, the volume ratio of the petroleum ether to the filtrate in the step S1 is 1:1, and the number of times of extraction is three.
Further, in the step S3, the volume ratio of ethyl acetate to the remaining aqueous phase was 1:1, and the number of extractions was three.
The invention also aims to provide the moringa oleifera leaf ethyl acetate extract.
Further, the components of the extract include 3-caffeoylquinic acid, 4-caffeoylquinic acid, rutin, kaempferol and 6 "-O-malonylgylcin. Corresponding molecular structures are described in the detailed description.
The invention also aims to provide the application of the moringa oleifera leaf ethyl acetate extract in reducing the aging condition of IEC-6 cells and reducing the generation of MDA in an IEC-6 cell model.
The fourth purpose of the invention is to provide the application of the moringa oleifera leaf ethyl acetate extract in preparing food, health care products or medicines for protecting intestinal tracts.
Compared with the prior art, the preparation method is simple and environment-friendly, and the extracted moringa oleifera leaf ethyl acetate extract contains various phenolic acid compounds and flavonoid compounds including 3-caffeoylquinic acid, 4-caffeoylquinic acid, rutin, kaempferol and 6' -O-malonylglycol.
Experiments prove that the ethyl acetate extract has a protective effect on an intestinal epithelial cell line IEC-6 cell aging model, and the specific expression is as follows: the aging condition of IEC-6 cells can be reduced, the production of MDA in an IEC-6 cell model can be reduced, the reduction of D-galactose, EGFR, JAK2 and STAT5 can be recovered to a certain degree, the reduction of Bcl-2 transcription level caused by D-galactose can be recovered to a certain degree, and the content of P53 in cell nucleus, the transcription level of P53 and the mRNA transcription amount of Bax can be reduced to a certain degree.
Drawings
FIG. 1 is a chemical formula diagram of each component in the moringa oleifera leaf ethyl acetate extract;
FIG. 2 is a graph showing the result of staining the senescence kit of the present invention;
FIG. 3 is a statistical plot of the levels of cellular MDA of the present invention;
FIG. 4 is a fluorescent map of EGFR, JAK2, STAT5, P53, Bcl-2, C-Jun, and β -actin expression in IEC-6 total cell protein of the present invention;
FIG. 5 is a graph showing the relative expression amounts of STAT5mRNA, P53mRNA and Bcl-2mRNA of cells.
Detailed Description
In order to facilitate understanding of the technical solutions of the present invention, the following detailed description is made with reference to the accompanying drawings and specific embodiments.
Example 1
The moringa oleifera leaf ethyl acetate extract is prepared by adopting the following method:
soaking 10kg of moringa leaves which are air-dried and ground into powder at room temperature, extracting the soaked moringa leaves with 75% acetone aqueous solution for three times, wherein the mass volume ratio of the moringa leaves to the acetone aqueous solution is 1:8, and then carrying out vacuum filtration at 50 ℃ to obtain filtrate. Diluting the filtrate with water, and extracting with petroleum ether at a volume ratio of 1:1 for three times. Extracting the residual water phase after extracting with petroleum ether with ethyl acetate three times, wherein the volume ratio of ethyl acetate to the water phase solution is 1: 1. After extraction, 200g of moringa oleifera leaf ethyl acetate extract (MoEF) is obtained by adopting a reduced pressure distillation mode, and the yield reaches 2.0%.
Firstly, component analysis:
and (3) performing component analysis on the moringa oleifera leaf ethyl acetate extract by adopting ultra-high performance liquid chromatography and quadrupole/time-of-flight mass spectrometry. Qualitative analysis was carried out at 30 ℃ using an ACQUITYUPLCCTMBEHC 18 (2.1X 100mm,1.7 μm) column, mobile phase 0.1% aqueous formic acid and acetonitrile. The elution procedure was: the ratio of 0.1% formic acid solution in water in mobile phase is increased linearly from 15% to 100% for 40min, and after 5min, the ratio is decreased linearly to 15% for 5 min. The flow rate is 0.8mL/min, and the eluent is detected in the 200-600nm wave band.
The analysis result of the components shows that the moringa oleifera leaf ethyl acetate extract contains a plurality of phenolic acid compounds and flavonoid compounds including 3-caffeoylquinic acid (A), 4-caffeoylquinic acid (B), rutin (C), kaempferol (D) and 6' -O-malonylgenin (E), the chemical formula of each component is shown in figure 1, and the content of each component is A: 0.75 percent; b: 0.75 percent; c: 2.60 percent; d: 1.47%; e: 0.91 percent.
Secondly, the function of the moringa oleifera leaf ethyl acetate extract in intestinal protection:
(1) cell culture (cells are IEC-6 intestinal epithelial cells): cells in logarithmic growth phase were inoculated into 24-well plates and cultured for 24 h. Adding different concentrations (5, 25, 50 μ g/mL respectively) of Moringa oleifera leaf ethyl acetate extract, and culturing for 4 hr. Then 50mmol/L D-galactose was added and the cultivation was continued for 24 h. Cells of a normal control group (corresponding to normal in the figure of the specification) were cultured for 24 hours with changing the medium every 12 hours. The model group (corresponding to the control in the attached drawings of the specification) is processed in the following way: directly treating the cells with 50mmol/L D-galactose for 24 h.
(2) And (3) performing in-situ staining experiments on beta-galactosidase, wherein the cells are IEC-6 intestinal epithelial cells: according to the instructions of the beta-galactosidase staining kit, the cells cultured according to (1) were inoculated into a 96-well plate, the cell culture solution was aspirated, washed once with PBS, and 200. mu.l of beta-galactosidase staining fixative was added and fixed at room temperature for 15 min. Cell fixative was aspirated and cells were washed 3 times with PBS for three minutes each. PBS was then aspirated and 200. mu.l of β -galactosidase staining working solution was added to each well. The 96-well plate was sealed with plastic film and incubated overnight at 37 ℃. Thereafter, observation was performed under a normal optical microscope.
The preparation method of the beta-galactosidase staining working solution comprises the following steps: staining solution A with beta-galactosidase: 10 μ l, β -galactosidase staining solution B: 10 μ l, β -galactosidase staining solution C: 930. mu.l of X-Gal solution 50. mu.l were mixed well.
(3) Determination of the level of oxidative stress
Cells were homogenized with RIPA tissue lysing agent and MDA levels in cells were tested using the relevant kits described in the literature.
(4) Immunoblotting
The expression of EGFR, JAK2, STAT5, P53, Bcl-2, C-Jun and beta-actin in the total protein of IEC-6 cells is detected by immunoblotting. The specific operation is as follows: the stored cells were first lysed into a homogenate using RIPA tissue lysing agent. Protein samples were quantitated using the BCA protein concentration assay kit. Total protein equivalents were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to PVDF membrane at 200mA for 1 h. The membrane was placed in blocking buffer (5% bovine serum albumin V) and the blot was stacked for 1h at room temperature. The membrane was incubated with the appropriate primary antibody overnight at 4 ℃ and then treated with the secondary anti-rabbitantibody (1:5000) at room temperature for 1 h. The membrane was then treated with a chemiluminescent indicator and detected with a fluorescent device.
(5) Real-time fluorescent quantitative Polymerase Chain Reaction (PCR)
RNA was extracted from cells using TRIzol and converted to cDNA using the primesttmRT kit and the gDNA eraser (preferably a real-time gDNA eraser) kit. Real-time instrument using ABISTEPOnePlus and
PremixExTaqTMII dye and online Taqman expression for quantitative RT-PCR detection.
(6) Statistical analysis means: quantitative analysis of the image of the half-lactase staining was performed using image-proplus6.0 software. Statistical analysis was performed using graphpadprism 5.0.
(7) And (3) analyzing a detection result:
FIG. 2 is a result of staining with a senescence kit, and the darker the color, the more severe senescence was shown. Wherein, the A-E pictures in figure 2 respectively correspond to the staining patterns of the normal control group, the model group and the experimental groups with the moringa oleifera leaf ethyl acetate extract concentration of 5, 25 and 50 mug/mL, and the F picture in figure 2 is a quantitative analysis statistical table of the staining results of the five experimental groups. As can be seen from FIG. 2, the ethyl acetate extract of Moringa oleifera leaves can reduce the senescence of IEC-6 cells.
FIG. 3 is a statistical table of cellular MDA levels of normal control group, model group, experimental group with Moringa oleifera leaf ethyl acetate extract concentration of 5, 25, 50. mu.g/mL, and cells treated with vitamin C. As can be seen in FIG. 3, the ethyl acetate extract of Moringa oleifera leaves reduced MDA production in the IEC-6 cell model.
FIG. 4 is a fluorescent spectrum showing the expression of EGFR, JAK2, STAT5, P53, Bcl-2, C-Jun and β -actin in the IEC-6 total cell protein of normal control group, model group, Moringa oleifera leaf ethyl acetate extract concentration of 50. mu.g/mL, and cells treated with vitamin C.
FIGS. 5A-D are graphs showing the relative expression amounts of STAT5mRNA, P53mRNA and Bcl-2mRNA of normal control group, model group, experimental group with Moringa oleifera leaf ethyl acetate extract concentration of 5, 25 and 50. mu.g/mL, and cells treated with vitamin C, respectively.
The aging model means that after the cells are treated by the D-galactose for 48 hours, the cells are damaged by free radicals, the content of Malondialdehyde (MDA) in the cells is obviously increased, the expression level of receptors related to growth signals and downstream signal proteins JAK2 and STAT5 in the cells is obviously reduced, and the nuclear localization of P53 related to apoptosis is obviously increased. From the analysis of the results in FIG. 2, it can be seen that the ethyl acetate extract of Moringa oleifera leaves can reduce the aging of IEC-6 cells; from the analysis in FIG. 3, it can be seen that the ethyl acetate extract of Moringa oleifera leaves can reduce the production of MDA in the IEC-6 cell model; from the analysis in fig. 4B, it is known that the ethyl acetate extract of moringa oleifera leaves can restore the decrease of EGFR, JAK2 and STAT5 caused by D-galactose to some extent; from the analysis in FIG. 5, the ethanol-acetate extract of Moringa oleifera leaves can restore the Bcl-2 transcription level reduction caused by D-galactose to some extent; from the analysis in fig. 4A, the moringa oleifera leaf ethyl acetate extract can reduce the P53 content in the cell nucleus to some extent; from the analysis in FIG. 5, it is found that the Moringa oleifera leaf ethyl acetate extract can reduce the transcription level of P53 and the mRNA transcription amount of Bax in the nucleus to some extent.
The moringa oleifera leaf ethyl acetate extract obtained in each embodiment is used as the only active ingredient, and is added with food, health care products or pharmaceutically acceptable auxiliary materials to prepare dosage forms suitable for people to take, such as tablets, capsules, powder and the like, so as to protect intestinal tracts.
The above is only a preferred embodiment of the present invention, and the scope of the present invention is defined by the appended claims, and several modifications and amendments made by those skilled in the art without departing from the spirit and scope of the present invention should be construed as the scope of the present invention.
Claims (4)
1. The method for extracting the moringa oleifera leaves by using the ethyl acetate is characterized by comprising the following steps: the method comprises the following steps:
s1: extracting moringa leaves with 75% acetone aqueous solution, and filtering, wherein the mass volume ratio of the moringa leaves to the acetone aqueous solution is 1 (5-10);
s2: extracting the filtrate in the step S1 by using petroleum ether, and then leaving a water phase, wherein the volume ratio of the petroleum ether to the filtrate in the step S1 is 1:1, and the extraction times are three times;
s3: extracting the residual water phase in the step S2 by using ethyl acetate, wherein the volume ratio of the ethyl acetate to the residual water phase is 1:1, and the extraction times are three times;
s4: concentrating and drying the extract obtained in the step S3 to obtain an ethyl acetate extract, wherein the ethyl acetate extract comprises 3-caffeoylquinic acid, 4-caffeoylquinic acid, rutin, kaempferol and 6' -O-malonylglobin.
2. An ethyl acetate extract of moringa oleifera leaf prepared by the method of claim 1.
3. Use of the ethyl acetate moringa leaf extract according to claim 2 for the manufacture of a medicament for reducing the IEC-6 cell senescence and reducing the production of MDA in IEC-6 cell models.
4. Use of the Moringa oleifera leaf ethyl acetate extract according to claim 2 for the preparation of a medicament for protecting the intestinal tract.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811132481.6A CN108938690B (en) | 2018-09-27 | 2018-09-27 | Moringa oleifera leaf ethyl acetate extract and extraction method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811132481.6A CN108938690B (en) | 2018-09-27 | 2018-09-27 | Moringa oleifera leaf ethyl acetate extract and extraction method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108938690A CN108938690A (en) | 2018-12-07 |
| CN108938690B true CN108938690B (en) | 2021-06-18 |
Family
ID=64471984
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811132481.6A Active CN108938690B (en) | 2018-09-27 | 2018-09-27 | Moringa oleifera leaf ethyl acetate extract and extraction method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108938690B (en) |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101732393A (en) * | 2008-11-11 | 2010-06-16 | 温州医学院 | Cuscuta chinessis Lam. ethyl acetate extract with anti-diarrhea effect and preparation method thereof |
| CN105616462B (en) * | 2016-03-11 | 2019-12-31 | 云南农业大学 | Moringa oleifera extract for promoting intestinal growth and preparation method and application thereof |
| CN105796615A (en) * | 2016-03-11 | 2016-07-27 | 田洋 | Moringa oleifera effective ingredient capable of relieving intestinal injury and preparation method and application thereof |
| CN106309528B (en) * | 2016-10-24 | 2021-12-24 | 贵州省中国科学院天然产物化学重点实验室 | Radix cynanchi wilfordii extract, preparation method thereof and application thereof in reducing blood sugar |
| CN107184621A (en) * | 2017-05-16 | 2017-09-22 | 云南省林业科学院 | A kind of extracting method of leaf of Moringa active ingredient and its application |
-
2018
- 2018-09-27 CN CN201811132481.6A patent/CN108938690B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN108938690A (en) | 2018-12-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Sharifi-Rad et al. | Prosopis plant chemical composition and pharmacological attributes: Targeting clinical studies from preclinical evidence | |
| Gracz-Bernaciak et al. | Functional studies of plant latex as a rich source of bioactive compounds: focus on proteins and alkaloids | |
| Singburaudom | Hydroxychavicol from Piper betel leave is an antifungal activity against plant pathogenic fungi | |
| Karatas et al. | Determination of in vitro free radical scavenging activities of various extracts from in vitro propagated Ceratophyllum demersum L | |
| Nazir et al. | High efficiency in vitro wound healing of Dictyophora indusiata extracts via anti-Inflammatory and collagen stimulating (MMP-2 inhibition) mechanisms | |
| Hama Hasan et al. | Effect of Equisetum Arvense Phenolic Extract in Treatment of Entamoeba Histolytica Infection. | |
| Castro et al. | Potent schistosomicidal constituents from Garcinia brasiliensis | |
| Kim et al. | Anti-inflammatory effect of the extract from fermented Asterina pectinifera with Cordyceps militaris mycelia in LPS-induced RAW264. 7 macrophages | |
| KR102232572B1 (en) | Manufacturing method for cosmetic pomposition comprising centella asiatica extract | |
| Freitas et al. | UPLC-QTOF-MS/MS analysis and antibacterial activity of the Manilkara zapota (L.) P. Royen against Escherichia coli and other MDR bacteria | |
| Lee et al. | Protective role of fermented mulberry leave extract in LPS‑induced inflammation and autophagy of RAW264. 7 macrophage cells | |
| Lai et al. | Antioxidant and antiedema properties of solid-state cultured honey mushroom, Armillaria mellea (higher Basidiomycetes), extracts and their polysaccharide and polyphenol contents | |
| CN108938690B (en) | Moringa oleifera leaf ethyl acetate extract and extraction method and application thereof | |
| de Souza et al. | Modulatory effect of Senecio brasiliensis (Spreng) Less. in a murine model of inflammation induced by carrageenan into the pleural cavity | |
| Okpala et al. | Effects of n-butanol fraction of Gongronema latifolium leave extract on some biochemical parameters in CCl4-induced oxidative damage in Wistar albino rats | |
| Chen et al. | Moutai Distiller's grains Polyphenol extracts and rutin alleviate DSS-induced colitis in mice: Modulation of gut microbiota and intestinal barrier function (R2) | |
| Tavares et al. | Toxicity and Anti-Inflammatory Activity of Phenolic-Rich Extract from Nopalea cochenillifera (Cactaceae): A Preclinical Study on the Prevention of Inflammatory Bowel Diseases | |
| Gandhi et al. | Antifungal and haemolytic activities of organic extracts of Tecoma stans (Bignoniaceae) | |
| Setyawati et al. | Characterization of Fraction of Carica papaya L. Leaves Ethyl Acetate Extract to African Catfish Clarias gariepinus Leucocytes Using UV-Vis, FTIR and GC-MS Methods | |
| Aniq et al. | Chemical composition of isoflavones compounds and antioxidants from Feijoa sellowiana leaves | |
| CN114588184A (en) | Potentilla anserine extract and preparation method and application thereof | |
| Eddine et al. | Antioxidant and antimicrobial activity of flavonoids fraction extract from Arnebia decumbens (Vent) growing in South East Algeria | |
| GADE et al. | Phytochemical investigation and thin layer chromatography of Aegle marmelos leaves Methanolic extract | |
| CN105106476B (en) | Application of saffron crocus in relieving fruit fly intestinal inflammation injury | |
| CN112174848B (en) | Oleoylethanolamide compound with antibacterial activity in parasitic loranthus, preparation method and application thereof |
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 |