CN113264828A - 马齿苋中一种苯甲酸类化合物及其提取分离方法 - Google Patents
马齿苋中一种苯甲酸类化合物及其提取分离方法 Download PDFInfo
- Publication number
- CN113264828A CN113264828A CN202110642677.5A CN202110642677A CN113264828A CN 113264828 A CN113264828 A CN 113264828A CN 202110642677 A CN202110642677 A CN 202110642677A CN 113264828 A CN113264828 A CN 113264828A
- Authority
- CN
- China
- Prior art keywords
- benzoic acid
- extraction
- methanol
- ethanol
- elution
- 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.)
- Granted
Links
- WPYMKLBDIGXBTP-UHFFFAOYSA-N Benzoic acid Natural products OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 title claims abstract description 58
- 239000005711 Benzoic acid Substances 0.000 title claims abstract description 43
- 235000010233 benzoic acid Nutrition 0.000 title claims abstract description 43
- -1 Benzoic acid compound Chemical class 0.000 title claims abstract description 32
- 238000000926 separation method Methods 0.000 title claims abstract description 30
- 235000001855 Portulaca oleracea Nutrition 0.000 title claims abstract description 23
- 241000219304 Portulacaceae Species 0.000 title claims abstract description 23
- 238000000605 extraction Methods 0.000 title claims abstract description 23
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 53
- 150000001875 compounds Chemical class 0.000 claims abstract description 32
- 238000000034 method Methods 0.000 claims abstract description 19
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 claims abstract description 11
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims abstract description 9
- 239000004952 Polyamide Substances 0.000 claims abstract description 8
- 229920002647 polyamide Polymers 0.000 claims abstract description 8
- 238000010992 reflux Methods 0.000 claims abstract description 8
- 238000004440 column chromatography Methods 0.000 claims abstract description 7
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 6
- 238000010898 silica gel chromatography Methods 0.000 claims abstract description 5
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 78
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 45
- 239000003814 drug Substances 0.000 claims description 19
- 238000010828 elution Methods 0.000 claims description 18
- 238000010829 isocratic elution Methods 0.000 claims description 14
- 239000000463 material Substances 0.000 claims description 13
- 239000007788 liquid Substances 0.000 claims description 10
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 9
- 239000000741 silica gel Substances 0.000 claims description 9
- 229910002027 silica gel Inorganic materials 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 239000000126 substance Substances 0.000 claims description 8
- 238000004809 thin layer chromatography Methods 0.000 claims description 8
- 230000003110 anti-inflammatory effect Effects 0.000 claims description 6
- 238000013375 chromatographic separation Methods 0.000 claims description 6
- 239000012141 concentrate Substances 0.000 claims description 6
- 239000002024 ethyl acetate extract Substances 0.000 claims description 6
- 238000001704 evaporation Methods 0.000 claims description 6
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 6
- 230000003078 antioxidant effect Effects 0.000 claims description 5
- 239000000499 gel Substances 0.000 claims description 5
- 238000001816 cooling Methods 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 3
- 239000003963 antioxidant agent Substances 0.000 claims description 3
- 239000000469 ethanolic extract Substances 0.000 claims description 3
- 239000000284 extract Substances 0.000 claims description 3
- 239000000706 filtrate Substances 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 235000019253 formic acid Nutrition 0.000 claims description 3
- 230000036541 health Effects 0.000 claims description 3
- 230000008569 process Effects 0.000 claims description 3
- HQVFCQRVQFYGRJ-UHFFFAOYSA-N formic acid;hydrate Chemical compound O.OC=O HQVFCQRVQFYGRJ-UHFFFAOYSA-N 0.000 claims description 2
- 238000011068 loading method Methods 0.000 claims description 2
- 239000000047 product Substances 0.000 claims description 2
- 238000002791 soaking Methods 0.000 claims description 2
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 abstract description 5
- 238000005160 1H NMR spectroscopy Methods 0.000 abstract description 5
- 230000000144 pharmacologic effect Effects 0.000 abstract description 5
- 238000011160 research Methods 0.000 abstract description 5
- 206010061218 Inflammation Diseases 0.000 abstract description 3
- 230000003064 anti-oxidating effect Effects 0.000 abstract description 3
- 239000002547 new drug Substances 0.000 abstract description 3
- 238000000746 purification Methods 0.000 abstract description 3
- 229940126680 traditional chinese medicines Drugs 0.000 abstract description 3
- 238000009509 drug development Methods 0.000 abstract description 2
- 239000002994 raw material Substances 0.000 abstract description 2
- 230000003334 potential effect Effects 0.000 abstract 1
- 238000010183 spectrum analysis Methods 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- OVYUSNXZFBGMAL-UHFFFAOYSA-N CC(C(C=C1)=CC=C1O)C(C(O)=C1)=CC(O)=C1C(O)=O Chemical compound CC(C(C=C1)=CC=C1O)C(C(O)=C1)=CC(O)=C1C(O)=O OVYUSNXZFBGMAL-UHFFFAOYSA-N 0.000 description 8
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 7
- 238000001228 spectrum Methods 0.000 description 7
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 108090001005 Interleukin-6 Proteins 0.000 description 6
- 210000002540 macrophage Anatomy 0.000 description 6
- 229930013930 alkaloid Natural products 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- 238000002835 absorbance Methods 0.000 description 4
- 150000001558 benzoic acid derivatives Chemical class 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 230000002757 inflammatory effect Effects 0.000 description 4
- 230000003595 spectral effect Effects 0.000 description 4
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 4
- XILIYVSXLSWUAI-UHFFFAOYSA-N 2-(diethylamino)ethyl n'-phenylcarbamimidothioate;dihydrobromide Chemical compound Br.Br.CCN(CC)CCSC(N)=NC1=CC=CC=C1 XILIYVSXLSWUAI-UHFFFAOYSA-N 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 238000003919 heteronuclear multiple bond coherence Methods 0.000 description 3
- 150000003254 radicals Chemical class 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 229930182555 Penicillin Natural products 0.000 description 2
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 2
- 102100040247 Tumor necrosis factor Human genes 0.000 description 2
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 2
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 description 2
- 150000003797 alkaloid derivatives Chemical class 0.000 description 2
- POJWUDADGALRAB-UHFFFAOYSA-N allantoin Chemical compound NC(=O)NC1NC(=O)NC1=O POJWUDADGALRAB-UHFFFAOYSA-N 0.000 description 2
- 230000003698 anagen phase Effects 0.000 description 2
- 235000006708 antioxidants Nutrition 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 description 2
- 208000001848 dysentery Diseases 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- 229940049954 penicillin Drugs 0.000 description 2
- 125000001997 phenyl group Chemical class [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 2
- 229960005322 streptomycin Drugs 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 description 2
- SFLSHLFXELFNJZ-QMMMGPOBSA-N (-)-norepinephrine Chemical compound NC[C@H](O)C1=CC=C(O)C(O)=C1 SFLSHLFXELFNJZ-QMMMGPOBSA-N 0.000 description 1
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 1
- BVCOHOSEBKQIQD-UHFFFAOYSA-N 2-tert-butyl-6-methoxyphenol Chemical compound COC1=CC=CC(C(C)(C)C)=C1O BVCOHOSEBKQIQD-UHFFFAOYSA-N 0.000 description 1
- 229930024421 Adenine Natural products 0.000 description 1
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 1
- POJWUDADGALRAB-PVQJCKRUSA-N Allantoin Natural products NC(=O)N[C@@H]1NC(=O)NC1=O POJWUDADGALRAB-PVQJCKRUSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 206010007247 Carbuncle Diseases 0.000 description 1
- 101800005151 Cholecystokinin-8 Proteins 0.000 description 1
- 102400000888 Cholecystokinin-8 Human genes 0.000 description 1
- 201000004624 Dermatitis Diseases 0.000 description 1
- 108010016626 Dipeptides Proteins 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 201000000297 Erysipelas Diseases 0.000 description 1
- XWDDIZKKSZLMEB-UHFFFAOYSA-N Feruloyl tyramine Natural products COc1cc(C=CC(=O)Oc2ccc(CCN)cc2)ccc1O XWDDIZKKSZLMEB-UHFFFAOYSA-N 0.000 description 1
- 206010017553 Furuncle Diseases 0.000 description 1
- 208000012671 Gastrointestinal haemorrhages Diseases 0.000 description 1
- 206010054787 Haemorrhoidal haemorrhage Diseases 0.000 description 1
- 208000032843 Hemorrhage Diseases 0.000 description 1
- 208000006877 Insect Bites and Stings Diseases 0.000 description 1
- 206010027514 Metrorrhagia Diseases 0.000 description 1
- NPNNKDMSXVRADT-WEVVVXLNSA-N N-feruloyltyramine Chemical compound C1=C(O)C(OC)=CC(\C=C\C(=O)NCCC=2C=CC(O)=CC=2)=C1 NPNNKDMSXVRADT-WEVVVXLNSA-N 0.000 description 1
- AVBCARAQLFOQID-UHFFFAOYSA-N N-trans-feruloyltyramine Natural products COc1cc(C=CC(=O)CNCc2ccc(O)cc2)ccc1O AVBCARAQLFOQID-UHFFFAOYSA-N 0.000 description 1
- MIBATSHDJRIUJK-ROJLCIKYSA-N Neferine Chemical compound C1=CC(OC)=CC=C1C[C@@H]1C2=CC(OC=3C(=CC=C(C[C@@H]4C5=CC(OC)=C(OC)C=C5CCN4C)C=3)O)=C(OC)C=C2CCN1C MIBATSHDJRIUJK-ROJLCIKYSA-N 0.000 description 1
- MIBATSHDJRIUJK-UHFFFAOYSA-N Neferine Natural products C1=CC(OC)=CC=C1CC1C2=CC(OC=3C(=CC=C(CC4C5=CC(OC)=C(OC)C=C5CCN4C)C=3)O)=C(OC)C=C2CCN1C MIBATSHDJRIUJK-UHFFFAOYSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 241000219295 Portulaca Species 0.000 description 1
- 230000002292 Radical scavenging effect Effects 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- 241000270295 Serpentes Species 0.000 description 1
- 108010087230 Sincalide Proteins 0.000 description 1
- 208000004078 Snake Bites Diseases 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 229960000643 adenine Drugs 0.000 description 1
- 229960005305 adenosine Drugs 0.000 description 1
- 150000001336 alkenes Chemical class 0.000 description 1
- 229960000458 allantoin Drugs 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 208000010668 atopic eczema Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- ZYGHJZDHTFUPRJ-UHFFFAOYSA-N benzo-alpha-pyrone Natural products C1=CC=C2OC(=O)C=CC2=C1 ZYGHJZDHTFUPRJ-UHFFFAOYSA-N 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- NPNNKDMSXVRADT-UHFFFAOYSA-N cis-N-feruloyl tyramine Natural products C1=C(O)C(OC)=CC(C=CC(=O)NCCC=2C=CC(O)=CC=2)=C1 NPNNKDMSXVRADT-UHFFFAOYSA-N 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 238000005100 correlation spectroscopy Methods 0.000 description 1
- 235000001671 coumarin Nutrition 0.000 description 1
- 150000004775 coumarins Chemical class 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000009795 derivation Methods 0.000 description 1
- VILAVOFMIJHSJA-UHFFFAOYSA-N dicarbon monoxide Chemical compound [C]=C=O VILAVOFMIJHSJA-UHFFFAOYSA-N 0.000 description 1
- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 1
- 229960003638 dopamine Drugs 0.000 description 1
- 238000001425 electrospray ionisation time-of-flight mass spectrometry Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 150000002213 flavones Chemical class 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- 239000006481 glucose medium Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 208000035861 hematochezia Diseases 0.000 description 1
- 235000008216 herbs Nutrition 0.000 description 1
- 238000001052 heteronuclear multiple bond coherence spectrum Methods 0.000 description 1
- 238000000990 heteronuclear single quantum coherence spectrum Methods 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 125000000896 monocarboxylic acid group Chemical group 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- SFLSHLFXELFNJZ-UHFFFAOYSA-N norepinephrine Natural products NCC(O)C1=CC=C(O)C(O)=C1 SFLSHLFXELFNJZ-UHFFFAOYSA-N 0.000 description 1
- 229960002748 norepinephrine Drugs 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- JRZJOMJEPLMPRA-UHFFFAOYSA-N olefin Natural products CCCCCCCC=C JRZJOMJEPLMPRA-UHFFFAOYSA-N 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 1
- 230000002040 relaxant effect Effects 0.000 description 1
- 238000001896 rotating frame Overhauser effect spectroscopy Methods 0.000 description 1
- 210000002460 smooth muscle Anatomy 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- 239000012085 test solution Substances 0.000 description 1
- 229940113082 thymine Drugs 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 229940035893 uracil Drugs 0.000 description 1
- 239000000341 volatile oil Substances 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C65/00—Compounds having carboxyl groups bound to carbon atoms of six—membered aromatic rings and containing any of the groups OH, O—metal, —CHO, keto, ether, groups, groups, or groups
- C07C65/01—Compounds having carboxyl groups bound to carbon atoms of six—membered aromatic rings and containing any of the groups OH, O—metal, —CHO, keto, ether, groups, groups, or groups containing hydroxy or O-metal groups
- C07C65/105—Compounds having carboxyl groups bound to carbon atoms of six—membered aromatic rings and containing any of the groups OH, O—metal, —CHO, keto, ether, groups, groups, or groups containing hydroxy or O-metal groups polycyclic
-
- 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
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
- C07C51/42—Separation; Purification; Stabilisation; Use of additives
- C07C51/47—Separation; Purification; Stabilisation; Use of additives by solid-liquid treatment; by chemisorption
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- General Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Mycology (AREA)
- Toxicology (AREA)
- Biochemistry (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Nutrition Science (AREA)
- Botany (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Pain & Pain Management (AREA)
- Rheumatology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
本发明涉及中药提取、分离领域,尤其涉及从马齿苋中提取、分离和鉴别出的苯甲酸类化合物及其提取分离方法。所述的苯甲酸类化合物,分子式为C15H14O5,命名为2,5‑dihydroxy‑4‑(1‑(4‑hydroxyphenyl)ethyl)benzoicacid,还提供上述苯甲酸类化合物的提取分离方法,依次采用乙醇回流提取、硅胶柱层析、聚酰胺柱层析、Sephadex LH‑20纯化及HPLC分离制备。其结构采用1H‑NMR、13C‑NMR及二维核磁波谱解析的方法确定为一种新的苯甲酸类化合物。该化合物具有潜在的抗炎和抗氧化等活性,可作为新药开发和药理活性研究的原料,为开发新药和开发新成分提供先导物和理论依据。
Description
技术领域
本发明涉及中药提取、分离领域,尤其涉及从马齿苋药材中提取、分离和鉴别出的一种苯甲酸类化合物及其提取分离方法。
背景技术
马齿苋(Portulaca oleracea L.)别称寿菜、五行草、五方草、马蜂菜等,全草供药用,广泛分布于世界各地。马齿苋是一年生肉质草本植物,全株无毛,为田间常见杂草,是国家卫生部划定的70余种药材之一。马齿苋收载于2020版《中华人民共和国药典》,具有清热解毒、凉血止血、止痢等功效,用于热毒血痢、痈肿疔疮、湿疹、丹毒、蛇虫咬伤、便血、痔血、崩漏下血等。
现代药理学研究表明,马齿苋具有降血脂、降血糖、抗炎、抗肿瘤、抗菌、松弛或兴奋平滑肌及增强免疫力等功效。马齿苋中含有多种化学成分,主要包括:黄酮类、生物碱类、萜类、香豆素类、有机酸类、挥发油、多糖、氨基酸、各种色素类和矿物质类等,为其具有丰富的药理作用提供了科学依据。其中生物碱是马齿苋中的一大类活性成分,目前已报道的生物碱类成分有去甲肾上腺素、尿囊素、多巴胺、金莲花碱、胸腺嘧啶、尿嘧啶、腺嘌呤腺苷、N,N-二环己基脲、N-反式-阿魏酰基酪胺;还有环二肽生物碱和酰胺类生物碱。
目前从马齿苋中分离出的化学成分大多数是已知的,且结构新颖性较低,因此,对马齿苋中新化合物的开发和分离是亟待需要的。
发明内容
针对上述问题,本发明提供从马齿苋中提取的一种苯甲酸类化合物,经研究发现本发明的一种苯甲酸类化合物具有抗炎、抗氧化的作用,同时提供一种针对本发明一种苯甲酸类化合物的简便、快速、环保、纯度高的提取分离方法。
为了实现上述目的,本发明提供了如下技术方案。
本发明提供一种从马齿苋药材中分离出的苯甲酸类化合物,其特征在于,所述化合物的分子式为:C15H14O5,并且根据结构命名为2,5-dihydroxy-4-(1-(4-hydroxyphenyl)ethyl)benzoicacid,其化学结构式如下:
本发明还提供一种从马齿苋药材中分离出的苯甲酸类化合物的提取分离方法,其特征在于,所述提取分离方法的具体步骤包括:
步骤1、取马齿苋干燥药材,采用乙醇回流提取,醇提液滤过,合并滤液减压浓缩,放凉至室温,得药液备用;
步骤2、将步骤1中药液蒸干后上硅胶柱,用乙酸乙酯洗脱,减压回收乙酸乙酯至浸膏,得到乙酸乙酯提取物;
步骤3、将步骤2中乙酸乙酯提取物经聚酰胺柱分离,采用乙醇∶水梯度洗脱,将95%乙醇洗脱部分合并蒸干,后经硅胶柱层析分离,用乙酸乙酯∶甲醇梯度洗脱,经薄层色谱进行检测,显色,合并显色部位,经减压浓缩至干,备用;
步骤4、将步骤3中所得浓缩物经预处理的Sephadex LH-20层析分离,以甲醇等度洗脱,经薄层色谱进行检测,显色,将显色的洗脱部位分别减压浓缩至干,得浓缩物备用;
步骤5、将步骤4中所得显色部位再经预处理的Sephadex LH-20柱层析,以甲醇等度洗脱,经薄层色谱进行检测,显色,将显色部位合并,减压浓缩至干,备用;
步骤6、将步骤5中所得浓缩物通过HPLC分离制备,以甲醇∶0.1%甲酸水为流动相进行等度洗脱,最终得到本发明所述的化合物。
进一步地,所述步骤1中乙醇回流提取两次,每次煎煮2小时,乙醇用量为药材的10倍。
进一步地,所述步骤4和步骤5中Sephadex LH-20凝胶的预处理过程为甲醇浸泡过24小时,上柱,以初始流动相平衡。
进一步地,所述步骤2中所用流动相洗脱程序为等度洗脱。
进一步地,所述步骤3中聚酰胺柱分离依次用乙醇,体积比为30∶70、50∶50、70∶30、95∶5、100∶0的乙醇∶水梯度洗脱,硅胶层析分离依次用乙酸乙酯,体积比为5:1、2:1的乙酸乙酯∶甲醇梯度洗脱。
进一步地,所述步骤4和步骤5中所用流动相洗脱程序为等度洗脱。
进一步地,所述步骤6中所用甲醇∶0.1%甲酸等度洗脱中甲醇和水的体积比为50∶50。
本发明还提供了一种如上所述的苯甲酸类化合物的用途,其特征在于,所述用途可用于制备抗炎和抗氧化的药物或保健品。
与现有技术相比本发明的有益效果。
本发明中所述马齿苋中一种苯甲酸类化合物的分离和药理活性研究未被发现有论文期刊所报道;本发明提供来源于马齿苋的一种苯甲酸类化合物及一种针对本发明化合物的提取分离方法,依次采用乙醇回流提取、硅胶柱层析、聚酰胺柱分离、Sephadex LH-20及HPLC进行分离纯化与制备,成功提取分离出一种新的苯甲酸类化合物,该方法操作步骤仅为六步,操作方法简便及快速,提取分离过程主要采用水提取,工艺方法环保,且经该方法分离得到的化合物纯度较高均大于90%,此外经研究表明以上化合物具有抗炎和抗氧化作用,因此本发明一种苯甲酸化合物及其盐和衍生物可以作为其他化合物合成先导物,以及新药开发和药理活性研究的原料,亦可用于制备抗炎和抗氧化的药物。
附图说明
图1为本发明苯甲酸类类化合物2,5-dihydroxy-4-(1-(4-hydroxyphenyl)ethyl)benzoicacid的1H-NMR光谱图。
图2为本发明苯甲酸类化合物2,5-dihydroxy-4-(1-(4-hydroxyphenyl)ethyl)benzoicacid的13C-NMR光谱图。
图3为本发明苯甲酸类化合物2,5-dihydroxy-4-(1-(4-hydroxyphenyl)ethyl)benzoicacid的DEPT 135光谱图。
图4为本发明苯甲酸类化合物2,5-dihydroxy-4-(1-(4-hydroxyphenyl)ethyl)benzoicacid的1H-1HCOSY光谱图。
图5为本发明苯甲酸类化合物2,5-dihydroxy-4-(1-(4-hydroxyphenyl)ethyl)benzoicacid的HSQC光谱图。
图6为本发明苯甲酸类化合物2,5-dihydroxy-4-(1-(4-hydroxyphenyl)ethyl)benzoicacid的HMBC光谱图。
图7为本发明苯甲酸类化合物2,5-dihydroxy-4-(1-(4-hydroxyphenyl)ethyl)benzoicacid的ROESY光谱图。
图8为本发明苯甲酸类化合物2,5-dihydroxy-4-(1-(4-hydroxyphenyl)ethyl)benzoicacid的高分辨质谱图。
具体实施方式
下面结合具体实施例对本发明做详细的说明。
实施例1。
本发明提供一种苯甲酸类化合物,分子式为C15H14O5,命名为2,5-dihydroxy-4-(1-(4-hydroxyphenyl)ethyl)benzoicacid,化学结构式为:
所述一种苯甲酸类化合物根据结构命名为2,5-dihydroxy-4-(1-(4-hydroxyphenyl)ethyl)benzoicacid,表1为该一种苯甲酸类化合物的核磁数据:1H-NMR与13C-NMR在MeOD-d 4中。
表1:本发明苯甲酸类化合物
2,5-dihydroxy-4-(1-(4-hydroxyphenyl)ethyl)benzoicacid的核磁数据
本发明一种苯甲酸类化合物2,5-dihydroxy-4-(1-(4-hydroxyphenyl)ethyl)benzoicacid的结构鉴定与推导。
2,5-dihydroxy-4-(1-(4-hydroxyphenyl)ethyl)benzoicacid:淡黄色粉末状物,易溶于甲醇,不溶、微溶于水。点样于硅胶薄层板后,喷三氯化铁试液斑点显青色,提示该化合物含有酚羟基类成分。UHPLC-ESI-TOF-MS给出m/z:273.0769 [M-H]-的准分子离子峰,分子量为274.0841。结合1H-NMR、13C-NMR以及DEPT 135数据,推测该化合物可能的分子式为C15H14O5,不饱和度为9。13C-NMR谱和DEPT 135谱显示15个碳信号,包括1个CH3(δC23.39),7个CH(δC38.74、116.98、118.83、129.29、116.59、116.59、129.29),7个季碳(1个羰基碳δC182.04;6个烯烃碳δC128.72、150.7、136.90、145.97、135.30、157.24)。
1H-NMR信号δH6.91(d,J=8.58HZ,H-2'',H-6''),δH6.70(d,J=8.58HZ,H-3'',H-5'')以及13C-NMR谱信号δC129.29(C-2'',C-6'',重叠),δC116.59(C-3'',C-5'',重叠)显示一个AA'BB'系统,HMBC谱相关性显示H-2'',H-6''与C-4''相关,H-3'',H-5''与C-1'',C-4''相关,又根据化学位移提示,推断羟基与C-4''位相连。此外,H-6(δH7.50,s)与C-2,C-4,C-5相关,H-3(δH6.54,s)与C-1,C-5相关,提示存在一个四取代苯环。1H-NMR谱提示存在两个芳香族单峰信号δH6.54,δH7.50,提示C-2,C-5位存在对位取代羟基,又根据HMBC谱相关性,H-6与COOH相关,又根据化学位移提示,推断羰基与C-1相连。再根据HMBC谱相关性,H-1'(δH4.10,m)与C-5,C-1',C-2'',C-6''相关,推断两个苯环均与H-1'相连,且1H-1H COSY谱相关信息显示,CH3与H-1'互相相关,推断甲基与C-1'连接。根据以上信息,可确定此化合物为上述结构。
本发明还提供上述此苯甲酸类化合物的提取分离方法,具体步骤为:
步骤1:称取马齿苋干燥药材150kg,采用50%乙醇提取,50%乙醇用量为药材的10倍,回流提取两次,每次煎煮2h,醇提液滤过,合并滤液,减压浓缩至150L,放凉至室温,得药液备用;
步骤2:将步骤1中所得药液蒸干后经硅胶柱层析分离,用乙酸乙酯(115L)等度洗脱,其中硅胶为100目~200目,40℃以下减压回收乙酸乙酯至浸膏,得到乙酸乙酯提取物;
步骤3:将步骤2中乙酸乙酯提取物经聚酰胺柱分离,采用乙醇∶水(0∶100、30∶70、50∶50、70∶30、95∶5、100∶0,v/v)梯度洗脱,将水和95%(体积百分数)乙醇部位的显色部分合并蒸干后经硅胶柱层析分离,其中硅胶为200目~300目,依次用乙酸乙酯,乙酸乙酯∶甲醇(5∶1、2∶1,v/v)梯度洗脱,将乙酸乙酯洗脱得到部位合并后显色,并于室温以上,40℃以下减压浓缩至干;
步骤4:将步骤3中所得显色部位经预处理的Sephadex LH-20柱层析,以甲醇等度洗脱,得到10个部位(即梯度洗脱得10个瓶,每瓶20mL),经薄层色谱进行检测,显色,留下显色的第4部位,50℃以下减压浓缩至干,备用;
步骤5:将步骤4中所得显色部位再经预处理的Sephadex LH-20柱层析,以甲醇等度洗脱,得到8个部位(即梯度洗脱得8个瓶,每瓶10mL),经薄层色谱进行检测,显色,将显色的4~6部位合并,50℃以下减压浓缩至干,备用;
步骤6:将步骤5中所得显色部位经HPLC分离制备,以甲醇和0.1%甲酸(50∶50,v/v)作为流动相,检测波长为210nm和280nm,分离制备得到本发明苯甲酸类化合物,归一法测定纯度均为90%~99%。
所述步骤4和步骤5中的Sephadex LH-20凝胶的预处理过甲醇浸泡过24h,上柱,初始流动相平衡。
实施例2 本发明苯甲酸类化合物的抗炎作用。
1主要材料。
1.1、药品和试剂:实验所用化合物由上述方法制备,纯度为90%~99%,精密称取,用DMSO稀释至下述各剂量组所需溶液。DMEM高糖培养基、胎牛血清(美国Hyclone公司);青霉素、链霉素(杭州四季青公司);LPS(美国Sigma公司);IL-6、TNF-α的ELISA试剂盒(美国Cayman公司);细胞裂解液。
1.2 细胞株:RAW264.7巨噬细胞(美国ATCC细胞库)。
1.3 分组:对照组、LPS组和实验组,各一组。
2 实验方法。
2.1 细胞培养,DMEM高糖培养基,加入10%的胎牛血清,l%抗菌素(100U/mL青霉素和100μg/mL链霉素),置于37℃,5%CO2培养箱中培养。
2.2 CCK8法测定细胞活力,上述三组分别取对数生长期RAW264.7巨噬细胞接种于96孔培养板中,细胞密度为1×104个/mL,每孔100μL,温度37℃,5%CO2条件下培养过夜后,实验组加入不同浓度的本发明苯甲酸类化合物2,5-dihydroxy-4-(1-(4-hydroxyphenyl)ethyl)benzoicacid(1μM~20μM),孵育1h后向LPS组和实验组分别加入浓度为1μg/mL的LPS,另设调零组(含DMSO溶媒的培养液),每组设3个复孔,考察加入药物后对细胞的影响。上述各组细胞培养24h后,在各孔细胞中加入10μL的CCK-8,温度37℃,5%CO2条件下继续孵育4h后,酶标仪450nm波长处测定各孔吸光值。
2.3 ELISA法测定炎症因子IL-6和TNF-α:将对数生长期RAW264.7巨噬细胞接种于24孔培养板中,细胞密度为1×105个/mL,每孔1mL,温度37℃,5%CO2条件下培养过夜,实验组加入本发明苯甲酸类化合物2,5-dihydroxy-4-(1-(4-hydroxyphenyl)ethyl)benzoicacid(1μM~20μM),培育1h后,在每孔加入LPS(终浓度为1μg/mL),共孵育24h,每组处理重复3孔。ELISA法测定马齿苋来源新化合物处理后的RAW264.7巨噬细胞分泌的IL-6和TNF-α的含量。
3实验结果。
实验结果表明本发明苯甲酸化合物对LPS诱导的巨噬细胞RAW264.7的增殖无影响,安全无毒;并可有效抑制LPS诱导的巨噬细胞RAW264.7所产生过量炎症细胞因子IL-6和TNF-α炎症介质,且呈浓度依赖。
细胞相对存活率实验结果如表2所示。
表2:本发明对RAW264.7巨噬细胞相对存活率的影响
注:*P<0.05与对照组比较(高浓度组有显著性差异)。
ELISA法测定炎症因子IL-6和TNF-α炎症介质结果如表3所示。
表3:本发明对LPS诱导的RAW264.7细胞分泌的IL-6和TNF-α含量的影响
注:*P<0.05与对照组比较,#P<0.05与LPS组比较,均数±SD,n=3。
实施例3 本发明苯甲酸类化合物的抗氧化作用。
1 主要材料。
1.1 药品和试剂:实验所用化合物由上述方法制备,纯度为90%~99%,精密称取,用甲醇稀释至下述各剂量组所需溶液。DPPH(1,1-二苯基-2-苦基肼自由基,Sigma-Fluka公司);BHA(叔丁基羟茴香醚,上海祥瑞科技有限公司);甲醇(色谱纯,昌泰兴业有限公司)。
1.2分组:对照组,实验组,空白组。
2 实验方法。
比色法测定消除DPPH自由基的能力,样品组取1.5mL的DPPH溶液(80μM)加入到4mL比色皿中,再加入1.5mL不同浓度的样品溶液(2.5μM、5μM、10μM、20μM和40μM);对照组取1.5mL甲醇溶液加入到4mL比色皿中,再加入1.5mL不同浓度的样品溶液;空白组取1.5mL的DPPH溶液加入到4mL比色皿中,再加入1.5mL甲醇溶液。三组均充分混匀,室温避光静置10min,517nm下测定吸光值,静置30min后,按同样方法操作。每个样品平均测定三次取平均值,阳性对照为不同浓度的BHA溶液。根据以下公式计算样品对DPPH自由基的清除率,并进一步计算其自由基清除率IC50值。
DPPH清除率(%)=1-(A1-A2)/A0×100%。
其中,A1为样品组的吸光度值;A2为对照组的吸光度值;A0为空白组的吸光度值。
3 实验结果。
实验结果表明本发明化合物对DPPH自由基具有清除作用,且随药物浓度增大,清除率也明显升高。本发明化合物对DPPH自由基IC50值见表4。
表4:本发明化合物对DPPH自由基的清除作用
综上所述,本发明提供特殊化合物及其提取分离方法,依次采用乙醇回流提取、硅胶柱层析、聚酰胺柱层析、葡聚糖凝胶柱层析及HPLC分离纯化,成功的分离得到一种新化合物,该方法简便,快速,环保,且经该方法分离得到的化合物纯度较高,由于所得化合物化学结构独特,从常用中药马齿苋中提取出来,其具有抗炎和抗氧化作用,因此本发明特殊化合物及其盐和衍生物可以作为天然产物开发中药新药,具有广阔的前景。
Claims (9)
2.一种如权利要求1所述的苯甲酸类化合物的提取分离方法,其特征在于,所述提取分离方法的具体步骤包括:
步骤1、取马齿苋干燥药材,采用乙醇回流提取,醇提液滤过,合并滤液减压浓缩,放凉至室温,得药液备用;
步骤2、将步骤1中药液蒸干后上硅胶柱,用乙酸乙酯洗脱,减压回收乙酸乙酯至浸膏,得到乙酸乙酯提取物;
步骤3、将步骤2中乙酸乙酯提取物经聚酰胺柱分离,采用乙醇∶水梯度洗脱,将95%乙醇洗脱部分合并蒸干,后经硅胶柱层析分离,用乙酸乙酯∶甲醇梯度洗脱,经薄层色谱进行检测,显色,合并显色部位,经减压浓缩至干,备用;
步骤4、将步骤3中所得浓缩物经预处理的Sephadex LH-20层析分离,以甲醇等度洗脱,经薄层色谱进行检测,显色,将显色的洗脱部位分别减压浓缩至干,得浓缩物备用;
步骤5、将步骤4中所得显色部位再经预处理的Sephadex LH-20柱层析,以甲醇等度洗脱,经薄层色谱进行检测,显色,将显色部位合并,减压浓缩至干,备用;
步骤6、将步骤5中所得浓缩物通过HPLC分离制备,以甲醇∶0.1%甲酸水为流动相进行等度洗脱,最终得到本发明所述的化合物。
3.如权利要求2所述的提取分离方法,其特征在于,所述步骤1中乙醇回流提取两次,每次2小时,乙醇用量为药材体积的10倍。
4.如权利要求2所述的提取分离方法,其特征在于,所述步骤4和步骤5中的SephadexLH-20凝胶的预处理过程为甲醇浸泡过24小时,上柱,以初始流动相平衡。
5.如权利要求2所述的提取分离方法,其特征在于,所述步骤2中所用流动相洗脱程序为等度洗脱。
6.如权利要求2所述的提取分离方法,其特征在于,所述步骤3中聚酰胺柱分离依次用乙醇,体积比为30∶70、50∶50、70∶30、95∶5、100∶0的乙醇∶水梯度洗脱,硅胶层析分离依次用乙酸乙酯,体积比为5:1、2:1乙酸乙酯∶甲醇梯度洗脱。
7.如权利要求2所述的提取分离方法,其特征在于,所述步骤4和步骤5中所用流动相洗脱程序为等度洗脱。
8.如权利要求2所述的提取分离方法,其特征在于,所述步骤6中所用甲醇∶0.1%甲酸等度洗脱中甲醇和水的体积比为50∶50。
9.一种如权利要求1所述的苯甲酸类化合物的用途,其特征在于,所述用途可用于制备抗炎和抗氧化的药物或保健品。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110642677.5A CN113264828B (zh) | 2021-06-09 | 2021-06-09 | 马齿苋中一种苯甲酸类化合物及其提取分离方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110642677.5A CN113264828B (zh) | 2021-06-09 | 2021-06-09 | 马齿苋中一种苯甲酸类化合物及其提取分离方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113264828A true CN113264828A (zh) | 2021-08-17 |
CN113264828B CN113264828B (zh) | 2022-05-17 |
Family
ID=77234707
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110642677.5A Active CN113264828B (zh) | 2021-06-09 | 2021-06-09 | 马齿苋中一种苯甲酸类化合物及其提取分离方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN113264828B (zh) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113968785A (zh) * | 2021-11-23 | 2022-01-25 | 辽宁中医药大学 | 马齿苋中一种含氧苯甲酸及其提取分离方法与应用 |
CN115521245A (zh) * | 2022-10-19 | 2022-12-27 | 辽宁中医药大学 | 马齿苋中一种生物碱类化合物及其提取分离方法与应用 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4845264A (en) * | 1987-03-05 | 1989-07-04 | Teijin Limited | Phenoxycarboxylic acid and herbicide comprising it as active ingredient |
CN1505603A (zh) * | 2001-09-17 | 2004-06-16 | ��һ��������ҩ��ʽ���� | 具有nf-kb抑制作用的取代苯甲酸衍生物 |
CN107827726A (zh) * | 2017-11-28 | 2018-03-23 | 辽宁中医药大学 | 马齿苋中化合物Oleracone E及其提取分离方法 |
CN110272342A (zh) * | 2019-07-16 | 2019-09-24 | 辽宁中医药大学 | 马齿苋中一种萘酸化合物及其提取分离方法与用途 |
-
2021
- 2021-06-09 CN CN202110642677.5A patent/CN113264828B/zh active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4845264A (en) * | 1987-03-05 | 1989-07-04 | Teijin Limited | Phenoxycarboxylic acid and herbicide comprising it as active ingredient |
CN1505603A (zh) * | 2001-09-17 | 2004-06-16 | ��һ��������ҩ��ʽ���� | 具有nf-kb抑制作用的取代苯甲酸衍生物 |
CN107827726A (zh) * | 2017-11-28 | 2018-03-23 | 辽宁中医药大学 | 马齿苋中化合物Oleracone E及其提取分离方法 |
CN110272342A (zh) * | 2019-07-16 | 2019-09-24 | 辽宁中医药大学 | 马齿苋中一种萘酸化合物及其提取分离方法与用途 |
Non-Patent Citations (1)
Title |
---|
SHOU-FANG WU, ET AL.: "Anti-inflammatory and Cytotoxic Neoflavonoids and Benzofurans", 《JOURNAL OF NATURAL PRODUCTS》 * |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113968785A (zh) * | 2021-11-23 | 2022-01-25 | 辽宁中医药大学 | 马齿苋中一种含氧苯甲酸及其提取分离方法与应用 |
CN113968785B (zh) * | 2021-11-23 | 2022-09-30 | 辽宁中医药大学 | 马齿苋中一种含氧苯甲酸及其提取分离方法与应用 |
CN115521245A (zh) * | 2022-10-19 | 2022-12-27 | 辽宁中医药大学 | 马齿苋中一种生物碱类化合物及其提取分离方法与应用 |
CN115521245B (zh) * | 2022-10-19 | 2024-01-19 | 辽宁中医药大学 | 马齿苋中一种生物碱类化合物及其提取分离方法与应用 |
Also Published As
Publication number | Publication date |
---|---|
CN113264828B (zh) | 2022-05-17 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN109897077B (zh) | 马齿苋中化合物Oleraciamide E及其提取分离方法与应用 | |
CN110272369B (zh) | 马齿苋中一种吡咯二羧酸类化合物及其提取分离方法与用途 | |
CN108558809B (zh) | 马齿苋中化合物Oleracone F及其提取分离方法 | |
CN113264828B (zh) | 马齿苋中一种苯甲酸类化合物及其提取分离方法 | |
CN108084060B (zh) | 马齿苋中生物碱oleraurea及其提取分离方法 | |
CN115716790B (zh) | 马齿苋中一种酰胺酯类生物碱的提取分离方法及其应用 | |
CN107827726B (zh) | 马齿苋中化合物Oleracone E及其提取分离方法 | |
CN113264886B (zh) | 马齿苋中一种哒嗪类化合物的提取分离方法及其应用 | |
CN114213473A (zh) | 马齿苋中三种生物碱类化合物及其提取分离方法 | |
CN115724812B (zh) | 马齿苋中一种呋喃酯类生物碱的提取分离方法及其应用 | |
CN115521245B (zh) | 马齿苋中一种生物碱类化合物及其提取分离方法与应用 | |
CN113698446B (zh) | 马齿苋中一种生物碱类化合物及其提取分离方法 | |
CN114989084B (zh) | 马齿苋中一种四氢异喹啉类生物碱的提取分离方法及其应用 | |
CN113968862B (zh) | 马齿苋中两种新生物碱及其提取分离方法 | |
CN114989064B (zh) | 马齿苋中一种新型吡咯生物碱类化合物及其提取分离方法 | |
CN113264829B (zh) | 马齿苋中四种木脂素及其提取分离方法 | |
CN114369076B (zh) | 马齿苋中两种茚类化合物及其提取分离方法 | |
CN114369022B (zh) | 马齿苋中一种有机酸类化合物及其提取分离方法 | |
CN110305094B (zh) | 马齿苋中两种黄酮类化合物及其提取分离方法与用途 | |
CN116606286B (zh) | 马齿苋中呋喃类生物碱及其提取分离方法 | |
CN114989083B (zh) | 马齿苋中一种新型异喹啉类生物碱及其提取分离方法 | |
CN113912657B (zh) | 马齿苋中三种吲哚类生物碱及其提取分离方法与用途 | |
CN116284005B (zh) | 马齿苋中一种新生物碱及其提取分离方法 | |
CN113968817B (zh) | 马齿苋中两种四氢异喹啉的提取分离方法及其应用 | |
CN115385884B (zh) | 马齿苋中一种新色酮醇类的提取分离方法及其应用 |
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 |