CN104431995A - Preparation method of fresh tricholoma matsutake active material - Google Patents
Preparation method of fresh tricholoma matsutake active material Download PDFInfo
- Publication number
- CN104431995A CN104431995A CN201510018428.3A CN201510018428A CN104431995A CN 104431995 A CN104431995 A CN 104431995A CN 201510018428 A CN201510018428 A CN 201510018428A CN 104431995 A CN104431995 A CN 104431995A
- Authority
- CN
- China
- Prior art keywords
- matsutake
- solution
- active material
- fresh
- preparation
- 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
- 241000121220 Tricholoma matsutake Species 0.000 title claims abstract description 187
- 239000011149 active material Substances 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 239000000243 solution Substances 0.000 claims abstract description 67
- 238000000034 method Methods 0.000 claims abstract description 32
- 238000000605 extraction Methods 0.000 claims abstract description 28
- 239000002994 raw material Substances 0.000 claims abstract description 16
- 230000001476 alcoholic effect Effects 0.000 claims abstract description 15
- 238000003809 water extraction Methods 0.000 claims abstract description 15
- 238000004821 distillation Methods 0.000 claims abstract description 10
- 238000001914 filtration Methods 0.000 claims abstract description 6
- 239000000284 extract Substances 0.000 claims description 84
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 24
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 21
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 20
- 239000012266 salt solution Substances 0.000 claims description 20
- 230000008569 process Effects 0.000 claims description 18
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 14
- VSMOENVRRABVKN-UHFFFAOYSA-N oct-1-en-3-ol Chemical compound CCCCCC(O)C=C VSMOENVRRABVKN-UHFFFAOYSA-N 0.000 claims description 14
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 12
- 239000003518 caustics Substances 0.000 claims description 10
- 238000006243 chemical reaction Methods 0.000 claims description 10
- 239000011780 sodium chloride Substances 0.000 claims description 10
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 8
- 230000006837 decompression Effects 0.000 claims description 8
- 239000011259 mixed solution Substances 0.000 claims description 8
- 238000009210 therapy by ultrasound Methods 0.000 claims description 8
- DUXYWXYOBMKGIN-UHFFFAOYSA-N trimyristin Chemical compound CCCCCCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCCCCCC)COC(=O)CCCCCCCCCCCCC DUXYWXYOBMKGIN-UHFFFAOYSA-N 0.000 claims description 8
- 239000012153 distilled water Substances 0.000 claims description 7
- 108010059892 Cellulase Proteins 0.000 claims description 6
- 108090000526 Papain Proteins 0.000 claims description 6
- 239000004365 Protease Substances 0.000 claims description 6
- 229940106157 cellulase Drugs 0.000 claims description 6
- 238000001976 enzyme digestion Methods 0.000 claims description 6
- 229940055729 papain Drugs 0.000 claims description 6
- 235000019834 papain Nutrition 0.000 claims description 6
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 6
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 5
- 238000002137 ultrasound extraction Methods 0.000 claims description 5
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 4
- 241000238631 Hexapoda Species 0.000 claims description 4
- 229920002302 Nylon 6,6 Polymers 0.000 claims description 4
- 241000607479 Yersinia pestis Species 0.000 claims description 4
- CEYULKASIQJZGP-UHFFFAOYSA-L disodium;2-(carboxymethyl)-2-hydroxybutanedioate Chemical compound [Na+].[Na+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O CEYULKASIQJZGP-UHFFFAOYSA-L 0.000 claims description 4
- 238000001704 evaporation Methods 0.000 claims description 4
- 239000000835 fiber Substances 0.000 claims description 4
- 238000007598 dipping method Methods 0.000 claims description 3
- 239000000758 substrate Substances 0.000 claims description 3
- 239000007788 liquid Substances 0.000 abstract description 85
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 abstract description 26
- 230000000694 effects Effects 0.000 abstract description 22
- 102000004169 proteins and genes Human genes 0.000 abstract description 19
- 108090000623 proteins and genes Proteins 0.000 abstract description 19
- 229930003944 flavone Natural products 0.000 abstract description 16
- 235000011949 flavones Nutrition 0.000 abstract description 16
- 150000004676 glycans Chemical class 0.000 abstract description 16
- 229920001282 polysaccharide Polymers 0.000 abstract description 16
- 239000005017 polysaccharide Substances 0.000 abstract description 16
- 150000008442 polyphenolic compounds Chemical class 0.000 abstract description 15
- 235000013824 polyphenols Nutrition 0.000 abstract description 15
- 150000003648 triterpenes Chemical class 0.000 abstract description 14
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 abstract description 12
- 229930003268 Vitamin C Natural products 0.000 abstract description 12
- 235000019154 vitamin C Nutrition 0.000 abstract description 12
- 239000011718 vitamin C Substances 0.000 abstract description 12
- 230000001965 increasing effect Effects 0.000 abstract description 6
- 230000003647 oxidation Effects 0.000 abstract description 5
- 238000007254 oxidation reaction Methods 0.000 abstract description 5
- 238000010521 absorption reaction Methods 0.000 abstract description 4
- 230000008901 benefit Effects 0.000 abstract description 4
- 230000002708 enhancing effect Effects 0.000 abstract description 2
- 239000004480 active ingredient Substances 0.000 abstract 2
- 239000012267 brine Substances 0.000 abstract 2
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 abstract 2
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 abstract 1
- 150000002212 flavone derivatives Chemical class 0.000 abstract 1
- 239000004615 ingredient Substances 0.000 abstract 1
- 238000002390 rotary evaporation Methods 0.000 abstract 1
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 abstract 1
- 238000012360 testing method Methods 0.000 description 55
- 241000699666 Mus <mouse, genus> Species 0.000 description 49
- 210000002966 serum Anatomy 0.000 description 44
- 238000003304 gavage Methods 0.000 description 37
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 28
- 241000699670 Mus sp. Species 0.000 description 22
- 238000011088 calibration curve Methods 0.000 description 21
- 230000031700 light absorption Effects 0.000 description 18
- 235000018102 proteins Nutrition 0.000 description 18
- 210000000952 spleen Anatomy 0.000 description 18
- 230000009182 swimming Effects 0.000 description 18
- 210000001541 thymus gland Anatomy 0.000 description 18
- 239000002585 base Substances 0.000 description 17
- 150000002213 flavones Chemical class 0.000 description 15
- 239000004310 lactic acid Substances 0.000 description 14
- 235000014655 lactic acid Nutrition 0.000 description 14
- 210000002540 macrophage Anatomy 0.000 description 11
- 230000000242 pagocytic effect Effects 0.000 description 11
- PNNCWTXUWKENPE-UHFFFAOYSA-N [N].NC(N)=O Chemical compound [N].NC(N)=O PNNCWTXUWKENPE-UHFFFAOYSA-N 0.000 description 10
- 210000004369 blood Anatomy 0.000 description 10
- 239000008280 blood Substances 0.000 description 10
- 239000000203 mixture Substances 0.000 description 10
- 235000001014 amino acid Nutrition 0.000 description 9
- 229940024606 amino acid Drugs 0.000 description 9
- 150000001413 amino acids Chemical class 0.000 description 9
- 230000006870 function Effects 0.000 description 9
- 235000016709 nutrition Nutrition 0.000 description 9
- 235000019441 ethanol Nutrition 0.000 description 7
- 235000015097 nutrients Nutrition 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 6
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 6
- 230000002929 anti-fatigue Effects 0.000 description 6
- 239000003513 alkali Substances 0.000 description 5
- 230000003064 anti-oxidating effect Effects 0.000 description 5
- 230000037396 body weight Effects 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- 230000012010 growth Effects 0.000 description 5
- 230000007365 immunoregulation Effects 0.000 description 5
- 238000002372 labelling Methods 0.000 description 5
- 210000001519 tissue Anatomy 0.000 description 5
- 206010015719 Exsanguination Diseases 0.000 description 4
- 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 4
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 230000003078 antioxidant effect Effects 0.000 description 4
- 210000005252 bulbus oculi Anatomy 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000011156 evaluation Methods 0.000 description 4
- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical compound OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 230000036541 health Effects 0.000 description 4
- 239000000976 ink Substances 0.000 description 4
- 238000007689 inspection Methods 0.000 description 4
- 230000035764 nutrition Effects 0.000 description 4
- 230000000050 nutritive effect Effects 0.000 description 4
- -1 pectase Proteins 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 230000001953 sensory effect Effects 0.000 description 4
- 210000000697 sensory organ Anatomy 0.000 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 description 3
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 102000019197 Superoxide Dismutase Human genes 0.000 description 3
- 108010012715 Superoxide dismutase Proteins 0.000 description 3
- 239000003963 antioxidant agent Substances 0.000 description 3
- 235000006708 antioxidants Nutrition 0.000 description 3
- 239000004202 carbamide Substances 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 230000001186 cumulative effect Effects 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 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 description 3
- 230000003203 everyday effect Effects 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 150000002632 lipids Chemical class 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 238000004321 preservation Methods 0.000 description 3
- 238000013441 quality evaluation Methods 0.000 description 3
- 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 description 3
- 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 description 3
- 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 description 3
- 235000005493 rutin Nutrition 0.000 description 3
- 229960004555 rutoside Drugs 0.000 description 3
- 229910000029 sodium carbonate Inorganic materials 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 3
- 210000003462 vein Anatomy 0.000 description 3
- 239000002699 waste material Substances 0.000 description 3
- WCGUUGGRBIKTOS-GPOJBZKASA-N (3beta)-3-hydroxyurs-12-en-28-oic acid Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CC[C@@H](C)[C@H](C)[C@H]5C4=CC[C@@H]3[C@]21C WCGUUGGRBIKTOS-GPOJBZKASA-N 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000006538 anaerobic glycolysis Effects 0.000 description 2
- 238000013475 authorization Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 229940098773 bovine serum albumin Drugs 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 235000021557 concentrated beverage Nutrition 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 238000000354 decomposition reaction Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 239000003205 fragrance Substances 0.000 description 2
- 229940074391 gallic acid Drugs 0.000 description 2
- 235000004515 gallic acid Nutrition 0.000 description 2
- 239000012362 glacial acetic acid Substances 0.000 description 2
- 230000036737 immune function Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 230000033001 locomotion Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 239000012460 protein solution Substances 0.000 description 2
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 2
- 150000003254 radicals Chemical class 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 230000000630 rising effect Effects 0.000 description 2
- 229930182490 saponin Natural products 0.000 description 2
- 150000007949 saponins Chemical class 0.000 description 2
- 230000002000 scavenging effect Effects 0.000 description 2
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 229940096998 ursolic acid Drugs 0.000 description 2
- PLSAJKYPRJGMHO-UHFFFAOYSA-N ursolic acid Natural products CC1CCC2(CCC3(C)C(C=CC4C5(C)CCC(O)C(C)(C)C5CCC34C)C2C1C)C(=O)O PLSAJKYPRJGMHO-UHFFFAOYSA-N 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- BNGXYYYYKUGPPF-UHFFFAOYSA-M (3-methylphenyl)methyl-triphenylphosphanium;chloride Chemical compound [Cl-].CC1=CC=CC(C[P+](C=2C=CC=CC=2)(C=2C=CC=CC=2)C=2C=CC=CC=2)=C1 BNGXYYYYKUGPPF-UHFFFAOYSA-M 0.000 description 1
- 235000001674 Agaricus brunnescens Nutrition 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 102100026189 Beta-galactosidase Human genes 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 229940123457 Free radical scavenger Drugs 0.000 description 1
- 241000222336 Ganoderma Species 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 229920002527 Glycogen Polymers 0.000 description 1
- 206010021143 Hypoxia Diseases 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- 241000581650 Ivesia Species 0.000 description 1
- 238000012449 Kunming mouse Methods 0.000 description 1
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 1
- LEVWYRKDKASIDU-IMJSIDKUSA-N L-cystine Chemical compound [O-]C(=O)[C@@H]([NH3+])CSSC[C@H]([NH3+])C([O-])=O LEVWYRKDKASIDU-IMJSIDKUSA-N 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- UCUNFLYVYCGDHP-BYPYZUCNSA-N L-methionine sulfone Chemical compound CS(=O)(=O)CC[C@H](N)C(O)=O UCUNFLYVYCGDHP-BYPYZUCNSA-N 0.000 description 1
- 108010059881 Lactase Proteins 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- UEZVMMHDMIWARA-UHFFFAOYSA-N Metaphosphoric acid Chemical compound OP(=O)=O UEZVMMHDMIWARA-UHFFFAOYSA-N 0.000 description 1
- 241000699660 Mus musculus Species 0.000 description 1
- 238000012356 Product development Methods 0.000 description 1
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 1
- 230000024932 T cell mediated immunity Effects 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 108010005774 beta-Galactosidase Proteins 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 238000010241 blood sampling Methods 0.000 description 1
- 229910021538 borax Inorganic materials 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 238000005352 clarification Methods 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 229960003067 cystine Drugs 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 230000035622 drinking Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 229940096919 glycogen Drugs 0.000 description 1
- 230000034659 glycolysis Effects 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 210000003128 head Anatomy 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 239000008236 heating water Substances 0.000 description 1
- 230000004727 humoral immunity Effects 0.000 description 1
- 230000007954 hypoxia Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 229960000310 isoleucine Drugs 0.000 description 1
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 1
- 229940116108 lactase Drugs 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000002932 luster Substances 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 230000000527 lymphocytic effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000000116 mitigating effect Effects 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- 210000000865 mononuclear phagocyte system Anatomy 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000003387 muscular Effects 0.000 description 1
- 235000006408 oxalic acid Nutrition 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000008557 oxygen metabolism Effects 0.000 description 1
- 238000005502 peroxidation Methods 0.000 description 1
- 210000001539 phagocyte Anatomy 0.000 description 1
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 239000002516 radical scavenger Substances 0.000 description 1
- 230000000384 rearing effect Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 229910052711 selenium Inorganic materials 0.000 description 1
- 239000011669 selenium Substances 0.000 description 1
- 235000010288 sodium nitrite Nutrition 0.000 description 1
- 239000004328 sodium tetraborate Substances 0.000 description 1
- 235000010339 sodium tetraborate Nutrition 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 230000001550 time effect Effects 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
Classifications
-
- 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
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
Landscapes
- Cosmetics (AREA)
Abstract
The invention provides a preparation method of a fresh tricholoma matsutake active material. The method includes the steps of firstly, raw material pretreatment, secondly, ultrasonic-assisted water extraction, thirdly, weak-base brine solution extraction, fourthly, alcoholic solution extraction, and fifthly, combination of an ultrasonic extracting solution, a weak-base brine solution extracting solution and an alcoholic solution extracting solution and obtainment of the fresh tricholoma matsutake active material after filtration, rotary evaporation and distillation. By the adoption of the preparation method, the limitation that the utilization rate of a tricholoma matsutake active material is low is broken through, comprehensive extraction is achieved in multiple extraction modes, the yield of tricholoma matsutake active ingredients is greatly increased, the effective ingredient content of a tricholoma matsutake concentrated solution is increased, the extracted active ingredients such as water-soluble polysaccharide, vitamin C, flavone, alkaline protein, polyphenol and triterpene have the effects of enhancing immune adjustment and resisting to oxidation and fatigue, human body absorption is facilitated, the fresh tricholoma matsutake active material has the advantages of being easy and convenient to use and convenient to preserve in combination with oral liquid, and the utilization value and economic benefits of tricholoma matsutake are greatly increased.
Description
Technical field
The present invention relates to a kind of preparation method of matsutake active material.
Background technology
Matsutake is as the rare edible and medicinal fungi of a kind of preciousness, and it contains abundant nutrient substance, has important health care and medical value, and its application is paid close attention to widely.The commercial value of matsutake, the always dark favor by national consumers such as Japan, US and Europeans.Due to matsutake growth, gather restriction by the factors such as season, region, weather, be difficult to fresh-keeping and storing, cause matsutake expensive, float frequent, only have only a few people to eat both at home and abroad, most people is difficult to taste matsutake goods, enjoys delicious food and the health-care effect of matsutake goods.The domestic and international research to matsutake is fewer at present, and cultivation aspect is also only limitted to manually urge high yield technology, and processing aspect is also simple preservation technique, and on market, matsutake converted products is less, there is no both at home and abroad and extracts and product development matsutake functionality active component.
Carrying out the functional concentrated beverage exploitation of matsutake, can, realizing under the resource continuous efficiency utilization prerequisite of matsutake, be the thinking that pine mushroom health product market development is new.In addition, as prospective basic research, also can be and develop the functional concentrated beverage of the edible fungi such as Agricus blazei, glossy ganoderma from now on successively foundation is provided.
But by the restriction of the factors such as season, region, weather, it is difficult to fresh-keeping and storing, causes a large amount of active material to run off, nutritive value reduces.Matsutake is fresh-keeping had been once global problem.How saving matsutake nutritional labeling to the full extent from damage can long term storage be also problem demanding prompt solution, is also the inexorable trend of future studies.The more important thing is that matsutake off standard and leftover pieces can not be used by current method, the waste of resource can not be reduced, turn waste into wealth, increase income and economize on spending, creating new way for comprehensively effectively utilizing matsutake resource.
Summary of the invention
The object of the invention is to solve above-mentioned Problems existing, and adopt that ultrasonic assistant water extraction is got, weak base salt solution extracts, alcoholic solution extracts, extract matsutake Middle nutrition and functional components, obtain a kind of preparation method of fresh matsutake active material.
The preparation method of a kind of fresh matsutake active material of the present invention, it carries out according to following steps:
One, select fresh, without matsutake that is invalid, free from insect pests, in clear water clean, draining, Tricholoma matsutake (lto et lmai) Singer sporophore is carried out slicing treatment, add its 20 times of volume sterile distilled waters, being immersed in containing mass percentage by the matsutake decompression after section is the citric acid of 0.03%, mass percentage is 0.02% n-butanol and mass percentage is in the mixed solution of 0.02% myristin, after decompression dipping 10min, to add mass percentage be 0.02%EDTA and mass percentage is 0.2% natrium citricum, then matsutake tissue is smashed to pieces, again by volume for 1:1:(1 ~ 2) ratio add the cellulase of 6 ~ 7g/L, the pectase of 6 ~ 7g/L and the papain of 6 ~ 7g/L carry out enzyme digestion reaction and obtain matsutake process raw material, wherein, reaction conditions is pH=5, temperature is 50 DEG C, enzymolysis time is 1.5h, enzyme-to-substrate concentration ratio is 1:3.3,
Two, ultrasonic assistant water extraction is got: the water that matsutake process raw material step one obtained is placed in 20 times of matsutake process raw material volume carries out ultrasonic extraction, collects the ultrasonic extract of matsutake and residue; Wherein, ultrasonic treatment conditions are: acoustic power 5 ~ 150w, extraction time 5 ~ 25min, ultrasonic wave working time 1 ~ 5s, interval time 1 ~ 5s;
Three, weak base salt solution extracts: the weak caustic solution adding 20 times of residue volumes in the residue that step 2 is collected extracts, wherein, weak caustic solution contain mass percentage be 0.1 ~ 0.5% sodium bicarbonate solution and mass percentage be 0.5 ~ 2.5% sodium chloride solution, collect matsutake weak base salt solution extract and residue; Wherein, extraction time is 30 ~ 150min, and Extracting temperature is 50 ~ 90 DEG C;
Four, alcoholic solution extracts: step 3 being collected the volumn concentration that residue is placed in 20 times of residue volumes is the ethanol of 50 ~ 90%, is then extract 30 ~ 150min under the condition of 50 ~ 90 DEG C at bath temperature, obtains matsutakealcohol solution extract;
Five, matsutake weak base salt solution extract that the ultrasonic extract of matsutake, step 3 that step 2 obtains obtain and the matsutakealcohol solution extract that step 4 obtains is collected, after mixing, filter with 100 object nylon66 fiber filtering layers, utilize rotary evaporating device with 50 DEG C, the condition of 60r/min carries out distillation and concentration to above-mentioned mixed solution, distillation and concentration, to 1/10 of former mixed liquor volume, namely completes described matsutake active material and extracts.
The present invention comprises following beneficial effect:
The present invention has broken the low limitation of matsutake active material utilization, by changing extracting mode, add the recovery rate of matsutake concentrate, and integrated oral liquid use is simple, convenient, be convenient to the advantages such as preservation, substantially increase value and the economic benefit of matsutake.
At present, the research major part of matsutake is also confined to extraction to matsutake single component and utilization, and have ignored the interaction between other nutritional labelings and each composition.Common Hot water extraction, water boiling and precipitation with ethanol method can only extract a small amount of nutriment, and it is extremely difficult for all nutritional labelings being extracted, and this causes waste to the utilization of matsutake.Utilization rate and the matsutake overall nutritive value of the present invention in order to improve matsutake, step by step arithmetic method is adopted to extract comprehensively and effectively the composition in matsutake, the various nutritional labelings in matsutake can be extracted to greatest extent, nutritive value can be kept again not suffer a loss, be also conducive to the application of matsutake.
The optimised process that ultrasonic assistant water extraction of the present invention is got is: ultrasonic power 125w, extraction time 10min, ultrasonic wave working time 4s, interval time 1s; The optimised process that weak base salt solution extracts is: sodium acid carbonate concentration 0.3%, sodium chloride concentration 0.5%, extraction time 120min, Extracting temperature 80 DEG C; The optimised process that alcoholic solution extracts is: concentration of alcohol 80%, extraction time 30min, Extracting temperature 70 DEG C.The ultrasonic assistant water extraction recorded under optimum process condition get in the yield of matsutake water-soluble polysaccharide be 0.52g/100g, ascorbic yield is 3.0mg/100g, and the yield of flavones is 7.1mg/100g; The yield of the alkaline protein that weak base salt solution extracts is 1.16g/100g; The yield of the polyphenol that alcoholic solution extracts and triterpene is respectively 1.16mg/100g and 0.23g/100g.
The quality of the present invention to matsutake concentrated oral liquid is evaluated, and detects its sense organ, relative density, pH value, Hygienic Index and nutrient composition content.Result shows, matsutake concentrated oral liquid is clear and bright yellowish-brown liquid, and without muddy and precipitation, taste is sweet, relative density is 1.10 ± 0.05g/mL, pH value is 7.50 ± 0.05, and often propping up loading amount is 10mL, and brown bottle is packed, Hygienic determination meets standard, containing polysaccharide 86.7 ± 0.2mg, vitamin C 0.50 ± 0.04mg, flavones 1.18 ± 0.04mg, protein 193.3 ± 0.3mg, polyphenol 0.19 ± 0.02mg, triterpene 38.3 ± 0.1mg in 100mL, detect 17 seed amino acids, its quality is good.
Using the immunoregulation capability of mouse, non-oxidizability and anti-fatigue ability as Testing index, matsutake concentrated oral liquid prepared by the inventive method is detected, with gavage physiological saline mouse in contrast, the situation of change of the indices of mouse after gavage matsutake concentrated oral liquid is examined.Result shows: after gavage matsutake concentrated oral liquid 21d, in immunoregulation capability, thymus index improves 26%, index and spleen index improves 9.1%, serum IgG content improves 11%, macrophages phagocytic capacity improves 51%, significant difference (P < 0.05) compared with control group; In oxidation resistance detects, after test group gavage 21d, MDA content reduces by 57%, SOD content and improves 11%, significant difference (P < 0.05) compared with control group; In anti-fatigue ability detects, after gavage 21d, swimming time improves 39%, there was no significant difference; Serum lactic acid content and serum urea nitrogen reduce 29% and 49% respectively, significant difference (P < 0.05) compared with control group.
Accompanying drawing explanation
The glucose standard curve that Fig. 1 is;
Fig. 2 is vitamin C calibration curve;
Fig. 3 is flavones calibration curve;
Fig. 4 is protein standard curve;
Fig. 5 is polyphenol calibration curve;
Fig. 6 is triterpene calibration curve.
Detailed description of the invention
Detailed description of the invention one: the preparation method of a kind of fresh matsutake active material of present embodiment, it carries out according to following steps:
One, select fresh, without matsutake that is invalid, free from insect pests, in clear water clean, draining, Tricholoma matsutake (lto et lmai) Singer sporophore is carried out slicing treatment, add its 20 times of volume sterile distilled waters, being immersed in containing mass percentage by the matsutake decompression after section is the citric acid of 0.03%, mass percentage is 0.02% n-butanol and mass percentage is in the mixed solution of 0.02% myristin, after decompression dipping 10min, to add mass percentage be 0.02%EDTA and mass percentage is 0.2% natrium citricum, then matsutake tissue is smashed to pieces, again by volume for 1:1:(1 ~ 2) ratio add the cellulase of 6 ~ 7g/L, the pectase of 6 ~ 7g/L and the papain of 6 ~ 7g/L carry out enzyme digestion reaction and obtain matsutake process raw material, wherein, reaction conditions is pH=5, temperature is 50 DEG C, enzymolysis time is 1.5h, enzyme-to-substrate concentration ratio is 1:3.3,
Two, ultrasonic assistant water extraction is got: the water that matsutake process raw material step one obtained is placed in 20 times of matsutake process raw material volume carries out ultrasonic extraction, collects the ultrasonic extract of matsutake and residue; Wherein, ultrasonic treatment conditions are: acoustic power 5 ~ 150w, extraction time 5 ~ 25min, ultrasonic wave working time 1 ~ 5s, interval time 1 ~ 5s;
Three, weak base salt solution extracts: the weak caustic solution adding 20 times of residue volumes in the residue that step 2 is collected extracts, wherein, weak caustic solution contain mass percentage be 0.1 ~ 0.5% sodium bicarbonate solution and mass percentage be 0.5 ~ 2.5% sodium chloride solution, collect matsutake weak base salt solution extract and residue; Wherein, extraction time is 30 ~ 150min, and Extracting temperature is 50 ~ 90 DEG C;
Four, alcoholic solution extracts: step 3 being collected the volumn concentration that residue is placed in 20 times of residue volumes is the ethanol of 50 ~ 90%, is then extract 30 ~ 150min under the condition of 50 ~ 90 DEG C at bath temperature, obtains matsutakealcohol solution extract;
Five, matsutake weak base salt solution extract that the ultrasonic extract of matsutake, step 3 that step 2 obtains obtain and the matsutakealcohol solution extract that step 4 obtains is collected, after mixing, filter with 100 object nylon66 fiber filtering layers, utilize rotary evaporating device with 50 DEG C, the condition of 60r/min carries out distillation and concentration to above-mentioned mixed solution, distillation and concentration, to 1/10 of former mixed liquor volume, namely completes described matsutake active material and extracts.
The yield that present embodiment ultrasonic assistant water extraction gets middle matsutake water-soluble polysaccharide is 0.52g/100g: ascorbic yield is 3.0mg/100g: the yield of flavones is 7.1mg/100g;
The yield of the alkaline protein that weak base salt solution extracts is 1.16g/100g;
The yield of the polyphenol that alcoholic solution extracts and triterpene is respectively 1.16mg/100g and 0.23g/100g.
Detailed description of the invention two: present embodiment and detailed description of the invention one unlike: the papain adding the cellulase of 6.8g/L, the pectase of 6.8g/L and 6.8g/L for the ratio of 1:1:1.4 by volume in step one carries out enzyme digestion reaction.Other is identical with detailed description of the invention one.
Detailed description of the invention three: present embodiment and detailed description of the invention one unlike: in step 2, ultrasonic treatment conditions are: acoustic power 100 ~ 150w, extraction time 5 ~ 10min, ultrasonic wave working time 2 ~ 4s, interval time 1 ~ 3s.Other is identical with detailed description of the invention one.
Detailed description of the invention four: present embodiment and detailed description of the invention one unlike: in step 2, ultrasonic treatment conditions are: acoustic power 125w, extraction time 10min, ultrasonic wave working time 4s, interval time 1s.Other is identical with detailed description of the invention one.
Detailed description of the invention five: present embodiment and detailed description of the invention one unlike: in step 3 weak caustic solution contain mass percentage be 0.3% sodium bicarbonate solution and mass percentage be 0.5% sodium chloride solution.Other is identical with detailed description of the invention one.
Detailed description of the invention six: present embodiment and detailed description of the invention one unlike: in step 3, extraction time is 90 ~ 150min, and Extracting temperature is 70 ~ 90 DEG C.Other is identical with detailed description of the invention one.
Detailed description of the invention seven: present embodiment and detailed description of the invention one unlike: in step 3, extraction time is 120min, and Extracting temperature is 80 DEG C.Other is identical with detailed description of the invention one.
Detailed description of the invention eight: present embodiment and detailed description of the invention one unlike: in step 4, the volumn concentration of ethanol is 80%.Other is identical with detailed description of the invention one.
Detailed description of the invention nine: present embodiment and detailed description of the invention one unlike: be extract 30 ~ 90min under the condition of 60 ~ 80 DEG C at bath temperature in step 4.Other is identical with detailed description of the invention one.
Detailed description of the invention ten: present embodiment and detailed description of the invention one unlike: be extract 30min under the condition of 70 DEG C at bath temperature in step 4.Other is identical with detailed description of the invention one.
Content of the present invention is not limited only to the content of the respective embodiments described above, and the combination of one of them or several detailed description of the invention equally also can realize the object of inventing.
Beneficial effect of the present invention is verified by following examples:
Embodiment one
Containing a large amount of nutrition and functional components in matsutake, particularly fresh matsutake is useful to health, but fresh matsutake exists the shortcomings such as not easily preservation, utilization rate of active components are low.
For the problems referred to above, the present embodiment is in order to extract nutriment in matsutake and functional components to greatest extent, according to its character, adopt that ultrasonic assistant water extraction is got, weak base salt solution extracts, alcoholic solution extracts, extract matsutake Middle nutrition and functional components, consider the complexity of extraction process, stability and production cost simultaneously, obtain the preparation technology of the dense active material of fresh matsutake.
The fresh matsutake of Mudanjiang City of Heilongjiang Province, matsutake source DongNing needed for following examples.
1, concrete grammar
2, pre-treatment of raw material
Select fresh, without matsutake that is invalid, free from insect pests, in clear water clean, draining.Tricholoma matsutake (lto et lmai) Singer sporophore is carried out slicing treatment; Use electronic balance weigh, add 20 times of volume sterile distilled waters, the matsutake decompression after section is immersed in containing 0.03% citric acid, 0.02% n-butanol and 0.02% myristin mixed solution in, 10min is flooded in decompression.Add 0.02%EDTA, 0.2% natrium citricum, with tissue mincer by disorganization.Add cellulase, pectase, papain, add carry out enzyme digestion reaction by 6.8g/L, the ratio of three kinds of enzymes is 1:1:1.4.
3, to pretreated raw material, carry out ultrasonic assistant water extraction with this and get → weak base salt solution extraction → alcoholic solution extraction:
One, ultrasonic assistant water extraction is got: matsutake process raw material step one obtained is placed in water and carries out ultrasonic extraction (the usage ratio relation of matsutake raw material and water is 1:30), collects the ultrasonic extract of matsutake and residue; Wherein, ultrasonic treatment conditions are: acoustic power 125w, extraction time 10min, ultrasonic wave working time 4s, interval time 1s;
Two, weak base salt solution extracts: the weak caustic solution adding 20 times of residue volumes in the residue that step 2 is collected extracts, wherein, weak caustic solution contain mass percentage be 0.3% sodium bicarbonate solution and mass percentage be 0.5% sodium chloride solution, collect matsutake weak base salt solution extract and residue; Wherein, extraction time is 120min, and Extracting temperature is 80 DEG C;
Three, alcoholic solution extracts: step 3 being collected the volumn concentration that residue is placed in 20 times of residue volumes is the ethanol of 80%, extracts 30min, obtain matsutakealcohol solution extract under the temperature of alcoholic solution is the condition of 70 DEG C.
To matsutake through ultrasonic assistant water extraction get → weak base salt solution extraction → alcoholic solution leaching process after, the yield that the ultrasonic assistant water extraction recorded gets middle matsutake water-soluble polysaccharide is 0.52g/100g, ascorbic yield is 3.0mg/100g, and the yield of flavones is 7.1mg/100g; The yield of the alkaline protein that weak base salt solution extracts is 1.16g/100g; The yield of the polyphenol that alcoholic solution extracts and triterpene is respectively 1.16mg/100g and 0.23g/100g.
Embodiment two
1, matsutake concentrated oral liquid is prepared
(1) fresh matsutake is carried out pretreatment of raw material (pretreatment of raw material with part 2);
(2) ultrasonic assistant water extraction is got: upper matsutake after pretreatment is carried out ultrasonic extraction in ultrasonic cell disruption instrument, regulate ultrasonic power 125w, ultrasonic time 10min, working time 4s, interval time 1s, be separated 10min with the centrifuge of 4000r/min, obtain extract 1 and residue 1;
(3) weak base salt solution extracts: residue 1 is immersed in matsutake quality 20 times of volumes and contains in 0.3% sodium acid carbonate, 0.5% sodium chloride solution, and 80 DEG C of constant temperature extract 120min.Then be separated 10min with 4000r/min centrifugation, obtain extract 2 and residue 2;
(4) ethanolic solution extracts: residue 2 is immersed in matsutake quality 20 times of volumes and contains in 80% ethanolic solution and extract 30min in 70 DEG C of thermostat water baths, obtain extract 3;
(5) concentrated distillation: by No. three extract mixing, then filter, filter with the nylon66 fiber filtering layer of 100 meshes.Utilize rotary evaporating device with 50 DEG C, 60r/min carries out distillation and concentration to above-mentioned mixed solution, distillation and concentration to 1/10 of cumulative volume, sterilization filling.Obtain yellowish-brown clear oral liquid.
2, obtained matsutake concentrated oral liquid carries out quality evaluation analysis
2.1 quality evaluation indexs
(1) sensory evaluation
Get 3 batches of matsutake concentrated oral liquids, by 10 professional persons, its color and luster, fragrance and structural state are marked, average and evaluate.
(2) relative density inspection
Get 3 batches of matsutake concentrated oral liquid sample densimeters respectively and measure its relative density, every batch sample replication 3 times, averages.
(3) pH value inspection
Get 3 batches of matsutake concentrated oral liquid samples respectively, with reference to 2010 editions NF annex VII G pH value determination methods, measure with acidometer, correct with borax standard buffer solution before measuring, every batch sample replication 3 times, averages.
(4) health examination
With reference to the microbial limit pertinent regulations of " Chinese Pharmacopoeia " (2010 editions one) annex IG mixture, according to step described in attached Ⅻ C microbial limit detection method, respectively bacterial population, mould and EHEC number in matsutake concentrated oral liquid are detected.
(5) nutrient composition content measures
1) polysaccharide
(1) yield determination of water-soluble polysaccharide
1. the drafting of glucose standard curve
Phenol sulfuric acid procedure is adopted to measure blazei polysaccharide content.Accurately take glucose standard items 100mg, distilled water is settled to 100mL, obtains the standard liquid that concentration is 1.0mg/mL.Accurate absorption glucose standard 0.1mL, 0.2mL, 0.4mL, 0.6mL, 0.8mL, 1.0mL, moisturizing is to 2.0mL.Add the phenol solution 1.0mL of 50g/L, concentrated sulfuric acid 5.0mL, mixing, boiling water bath 2min.Light absorption value is surveyed at 495nm place, with polysaccharide concentration (mg/mL) for abscissa, with light absorption value (OD after cooling
495) be ordinate, as shown in Figure 1.
2. the calculating of water-soluble polysaccharide yield
Get 1mL extract, measure light absorption value according to above-mentioned calibration curve method, obtain the concentration of polysaccharide in extract according to calibration curve, can calculate:
Water-soluble polysaccharide quality/matsutake quality × 100% in water-soluble polysaccharide yield=extract.
2) vitamin C
Ascorbic yield
1. the drafting of vitamin C calibration curve
Accurately take oxalic acid 6.30g, EDTA0.0584g, 100mL is settled to after abundant dissolving, accurately take dry vitamin C 5mg, 500mL is settled to oxalic acid-EDTA solution, the standard ascorbic acid solution drawing 0.8mL, 1.2mL, 1.6mL, 2.0mL, 2.4mL, 2.8mL, 3.2mL respectively, in the volumetric flask of 50mL, then adds people's oxalic acid-EDTA solution, makes cumulative volume reach 10mL.Add metaphosphoric acid one acetum of 1.0mL and the sulfuric acid 2.0mL of 5% again, shake up the ammonium aluminate adding people 4.0mL, under 705nm, measure light absorption value, with vitamin C concentration (μ g/mL) for abscissa, with light absorption value (OD
705) be ordinate, as shown in Figure 2.
2. the calculating of vitamin C yield
Get 1mL extract, measure light absorption value according to above-mentioned calibration curve method, obtain ascorbic concentration in extract according to calibration curve, can calculate:
Vitamin C quality/matsutake quality × 100% in vitamin C yield=extract.
3. flavones
3) yield of flavones
1. the drafting of flavones calibration curve
Accurately take 20mg rutin standard items, be settled to 100mL with 50% ethanolic solution, obtain 0.2mg/mL rutin standard liquid, measure standard liquid 1.0mL, 3.0mL, 5.0mL, 7.0mL, 13.0mL, 17.0mL, 23.0mL.Be placed in 100mL volumetric flask respectively, 5% sodium nitrite solution 1.0mL is added in each volumetric flask, after placing 6min, add 10% aluminum nitrate solution 1.0mL more respectively, add 4% sodium hydroxide solution 10mL respectively after leaving standstill 6min, use 50% ethanolic solution constant volume, after leaving standstill 10min, light absorption value is surveyed, with rutin concentration (μ g/mL) for abscissa, with light absorption value (OD in 508nm place
508) be ordinate, as shown in Figure 3.
2. the calculating of flavones yield
Get 1mL extract, measure light absorption value according to above-mentioned calibration curve method, obtain the concentration of flavones in extract according to calibration curve, can calculate:
Flavones quality/matsutake quality × 100% in flavones yield=extract.
4) protein
The yield of alkali solubility protein
1. the drafting of protein standard curve
First the sodium chloride solution of bovine serum albumin(BSA) 0.15mol/mL is made into the standard protein solution of 1.0mg/mL.Accurately take 100mg Coomassie brilliant G-250 again, after being dissolved in the ethanol of 50mL95%, add the phosphoric acid of 120mL 85%, with distilled water diluting to 1L.Get standard protein solution 0.1mL, 0.2mL, 0.3mL, 0.4mL, 0.5mL, 0.6mL that 6 test tubes add 1.0mg/mL respectively, then add to 1mL with 0.15mol/mL sodium chloride solution, a last test tube adds 5.0mL Coomassie brilliant G-250 reagent respectively.Often add a pipe, mix on vortex mixer immediately.After adding reagent 2-5min, survey light absorption value in 595nm place.With bovine serum albumin(BSA) concentration (mg/mL) for abscissa, with light absorption value (OD
595) be ordinate, as shown in Figure 4.
2. the calculating of alkali solubility protein yield
Get 1mL extract, measure light absorption value according to above-mentioned calibration curve method, obtain the concentration of alkali solubility protein in extract according to calibration curve, can calculate:
Alkali solubility albumen quality/matsutake quality × 100% in alkali solubility protein yield=extract.
5) polyphenol
The yield of polyphenol
1. the drafting of polyphenol calibration curve
Accurately take gallic acid standard items 25mg, be settled to 250mL by water-soluble solution, obtain the standard liquid of 0.1mg/mL.Accurate standard liquid 0.1mL, 0.2mL, 0.3mL, 0.4mL, 0.5mL, 0.6mL of drawing is in 100mL volumetric flask, add 1mL forint phenol developer, 2mL15% sodium carbonate liquor is added again after shaking up, be settled to 100mL, light absorption value is surveyed in 760nm place after reacting 2h under room temperature, with gallic acid concentration (μ g/mL) for abscissa, with light absorption value (OD
760) be ordinate, as shown in Figure 5.
2. the calculating of polyphenol yield
Get 1mL extract, measure light absorption value according to above-mentioned calibration curve method, obtain the concentration of polyphenol in extract according to calibration curve, can calculate
Polyphenol quality/matsutake quality × 100% in polyphenol yield=extract.
6) triterpene
The calculating of triterpene yield
1. the drafting of triterpene calibration curve
Accurately take ursolic acid standard items 1.150mg, be dissolved in 10mL absolute ethyl alcohol, obtain 0.115mg/mL standard liquid.Accurate absorption 0.1mL, 0.2mL, 0.4mL, 0.6mL, 0.8mL, 1.0mL, 1.2mL standard liquid in 10mL test tube, water bath method solvent.5% vanillic aldehyde-glacial acetic acid 0.5mL, perchloric acid 1mL is added, sealing respectively to often propping up in test tube.60 DEG C of heating water bath 15min, taking-up is placed in frozen water and cools.Adding 5mL glacial acetic acid solution to often propping up in test tube again, shaking up.Light absorption value is surveyed, with ursolic acid quality (μ g) for abscissa, with light absorption value (OD in 548nm place
548) be ordinate, as shown in Figure 6.
2. the calculating of triterpene yield
Get 1mL extract, measure light absorption value according to above-mentioned calibration curve method, obtain the concentration of polysaccharide in extract according to calibration curve, can calculate
Triterpene quality/matsutake quality × 100% in triterpene yield=extract.
7. amino acid
Amino-acid analyzer is utilized to measure.
3, results and analysis
3.1 sensory evaluation
The sense organ of oral liquid is that consumer is familiar with for first of oral liquid and evaluates, so the sense organ of oral liquid directly affects the quality of its quality, this test is evaluated the sense organ of matsutake concentrated oral liquid through professional training personage by 10, and result is as shown in table 1.
The sensory evaluation of table 1 matsutake concentrated oral liquid proterties
As shown in Table 1, the proterties often criticizing matsutake concentrated oral liquid is good, and sensory evaluation score is all more than 90 points, and obtained matsutake concentrated oral liquid is yellowish-brown, gives off a strong fragrance, clarification.
3.2 the detection of relative density
The relative density of matsutake concentrated oral liquid 3 batch sample is 1.10 ± 0.05g/mL.
The detection of 3.3pH value
Have detected the pH value of matsutake concentrated oral liquid 3 batch sample respectively, the pH value preparing matsutake concentrated oral liquid is 7.50 ± 0.05.
3.4 Hygienic determination
In matsutake concentrated oral liquid, bacterium, mould and EHEC number meet national standard after testing, do not detect pathogenic bacteria.
The detection of 3.5 nutrient composition contents
The content of nutritional labeling determines the quality of oral liquid, and this test detects each nutrient composition content in obtained matsutake concentrated oral liquid, and result is as table 2 and table 3.
The each component content of table 2 matsutake concentrated oral liquid
As shown in Table 2, during 3 batches of oral liquids often prop up, each nutrient composition content difference is little, and wherein polyoses content is 86.7 ± 0.2mg/100mL, vitamin C is 0.50 ± 0.04mg/100mL, flavones is 1.18 ± 0.04mg/100mL, protein is 193.3 ± 0.3mg/100mL, polyphenol content is 0.19 ± 0.02mg/100mL, triterpene content is 38.3 ± 0.1mg/100mL.Compared with Tricholoma matsutake (lto et lmai) Singer sporophore, matsutake concentrated oral liquid more comprehensively remains its nutriment and functional component.
Table 3 matsutake concentrated oral liquid amino acid content
Can be obtained by table 3, except cystine, methionine sulfone, cysteine do not detect in matsutake concentrated oral liquid, detect 17 seed amino acids.Compared with the literature, the amino acid classes detected and document basically identical, the content of glutamic acid, methionine, isoleucine and lysine is slightly less than document, and other 13 seed amino acid content are all higher than document mean value.
The present embodiment detects the appearance characteristics of Matsutske oral liquid, pH value, relative density, hygiene and nutrient composition content etc., establishes the Preliminary Standards of quality evaluation.After testing, matsutake concentrated oral liquid is clear and bright yellowish-brown liquid, without muddy and precipitation, taste is sweet, relative density is 1.10 ± 0.05g/mL, pH value is 7.50 ± 0.05, often propping up loading amount is 10mL, brown bottle is packed, Hygienic determination meets standard, containing polysaccharide 86.7 ± 0.2mg, vitamin C 0.50 ± 0.04mg, flavones 1.18 ± 0.04mg, protein 193.3 ± 0.3mg, polyphenol 0.19 ± 0.02mg, triterpene 38.3 ± 0.1mg in 100mL, amino acid classes is consistent with document, and 13 seed amino acid content are greater than the mean value of document.
Embodiment three
Matsutake concentrated oral liquid functional authorization
Matsutake, as the rare famous and precious mushroom of one, has very high nutritive value and medical value.Research shows, matsutake has the multiple bioactive functions such as antitumor, develop immunitypty, anti-oxidant, anti-sudden change.The present embodiment is verified the enhancing immunocompetence of matsutake concentrated oral liquid, non-oxidizability and fatigue resistance.
1, experimental animal
Kunming mice (Mus musculus) is female, SPF level, 6-8 week age, body weight 18 ± 2g, be purchased from Harbin Veterinary Medicine Inst., China Academy of Agriculture SPF level animal center, Mouse feeder is in cleaning grade Animal House.Mouse feeder is (humidity 50% ± 10% under standard conditions; 12h daytime/12h night alternately), feed through the mouse grain of sterilizing and distilled water.
2, test method
2.1 test mice grouping and process
Mouse, after 3 days adaptability is raised, is divided into four groups at random, often organizes 12.With stainless steel hamper and plastics drinking bottle sub-cage rearing, every day respectively adds feed once sooner or later.On-test, at the end of respectively weigh once, all mouse can contact free to feed and water.Wherein two groups of mouse stomach SPSSs, other two groups of gavage matsutake concentrated oral liquids, every mouse gavage every day once, each 0.2mL/10g, continuous 21d.
The mensuration of 2.2 immunoregulation capability indexs
2.2.1 thymus index, spleen index measure
7d, 14d and 21d respectively at test pluck eyeball sacrificed by exsanguination mouse, peel off thymus gland (spleen), and surrounding tissue are peeled off totally, weigh.
Be calculated as follows immune organ index: thymus gland (spleen) index=thymus gland (spleen) weight (mg)/body weight (g).
2.2.2 serum IgG content
Specific IgG antibody in Immunoglobulin IgG in serum and reality, forms antigen antibody complex and produces turbidity, and its turbidity height becomes to compare with IgG in serum when a certain amount of antibody exists.By measuring the absorbance of specific wavelength, the content of IgG in serum can be calculated with reference to calibration curve.
7d, 14d and 21d respectively at test pluck eyeball sacrificed by exsanguination mouse, operate according to IgG kit description.
2.2.3 macrophage phagocytic function
(1) india ink is pressed 1:3 dilution proportion, calculate by the every 10g body weight 0.1mL of small white mouse in 7d, 14d and 21d of test, be injected in vivo through tail vein, treat the complete timing immediately of Ink injection.
(2), when injecting 2min, 10min after prepared Chinese ink, getting blood 20 μ L from retroorbital venous clump respectively, shake up, survey OD value with ultraviolet specrophotometer under 600nm wavelength in 2mL sodium carbonate liquor, take sodium carbonate as blank.
Index (Clearance index ofcarbon particles, CICP) is cleaned up and index (Adjusted clearance index ofcarbon particles, ACICP) is cleaned up in correction according to formulae discovery carbon granule.
OD in formula
1, OD
2be respectively 2min (t
1), 10min (t
2) time the blood sampling absorbance that records; t
1, t
2for the time of taking a blood sample after injection india ink.
The mensuration of 2.3 antioxidation activity indexs
2.3.1 mda content measures
MDA in lipid peroxide catabolite can with thiobarbituricacidα-condensation, formed red product, have maximum absorption band at 532nm place.
7d, 14d and 21d respectively at test pluck eyeball sacrificed by exsanguination mouse, operate according to MDA kit description.
Calculate Content of MDA (nmol/mL) as follows:
2.3.2 Determination of erythrocyte superoxide dismutase activity
7d, 14d and 21d respectively at test pluck eyeball sacrificed by exsanguination mouse, operate according to SOD kit description.
(1) by the inhibiting rate of formulae discovery SOD:
(2) by the vigor of formulae discovery SOD:
Extension rate before SOD vigor=SOD inhibiting rate ÷ 50% × reflection system extension rate × test sample.
The mensuration of 2.4 anti-fatigue effect indexs
2.4.1 swimming time measures
The walking weight load of mouse is measured according to " health food inspection and assessment technical specification " (2003 editions).Every day, gavage 1 time, distinguished continuous gavage 7d, 14d and 21d.After last gavage 30min, be placed in swimming trunk went swimming at mouse tail root load 5% body weight galvanized wire.The depth of water is not less than 30cm, water temperature 25 ± 1.0 DEG C.Exhaust criterion (i.e. mouse head submerged can't return the water surface through 10s and be considered as power and exhaust) with reference to power, record from swimming beginning to the time that power exhausts, as swimming time.Data with mean and standard deviation (
) represent.
2.4.2 Serum lactic acid content measures
Respectively at 7d, 14d and 21d after gavage, the swimming trunk went swimming 10min of 30 DEG C put into by galvanized wire test group and control group mice afterbody being born 5% body weight; At once get mouse angular vein blood 20 μ L after swimming, operate according to Serum lactic acid content kit description.
2.4.3 serum urea nitrogen analysis
Respectively at 7d, 14d and 21d after gavage, test group and control group mice are put into the swimming trunk went swimming 60min of 30 DEG C simultaneously, at once get mouse vena ophthalmica clump blood, centrifugal, get serum, operate according to serum urea nitrogen content kit description.
Computing formula:
2.4.4 statistical method
The data obtained with
represent, application T-Test inspection process, P < 0.05 represents significant difference, and P < 0.01 represents that difference is extremely remarkable.
3, results and analysis
3.1, matsutake concentrated oral liquid is on the impact of mouse immune regulating power
3.1.1 on the impact of thymus index
Containing abundant lymphocyte in thymus gland, it is absolutely necessary to the development of the lymphocytic maturation of T and cell-mediated immunity.Tentatively can think that matsutake concentrated oral liquid can promote the growth of mouse thymus by test, result is as table 4.
Table 4 matsutake concentrated oral liquid is to mouse thymus exponential effect
Note: * represents the significant difference of P < 0.05 level.
As can be seen from Table 4, along with the increase of time, each group thymus index is in rising trend, and wherein test group thymus index is all higher than control group.After gavage matsutake concentrated oral liquid 7d, 14d, test group thymus index, higher than control group, improve 12% and 16% respectively, but difference is not remarkable.After 21d, the thymus index of test group, apparently higher than control group, improves 26%, and significant difference (P < 0.05).
3.1.2 on the impact of index and spleen index
Spleen contains a large amount of phagocyte, is the chief component of mononuclear phagocyte system.The main immunologic function of spleen is microorganism by damming in blood and produces with it immune response to carry out filtering blood.Test finds that matsutake concentrated oral liquid can promote the growth of mouse spleen.Result is as table 5.
Table 5 matsutake concentrated oral liquid is to mouse spleen exponential effect
Note: * represents the significant difference of P < 0.05 level.
Mouse spleen index increases gradually as can be seen from Table 5, and test group is all higher than control group, 7d, 14d and 21d index and spleen index improves 6.4%, 7.3% and 9.1% respectively, during 21d, test group significant difference (P < 0.05) compared with control group, has statistical significance.After illustrating that mouse is subject to the effect of matsutake concentrated oral liquid, facilitate the growth of spleen.
3.1.3 on the impact of serum IgG content
In serum, IgG content is the humoral immunity molecule that the humoral activity cell in Immune Organs of Body tissue produces, and is the important indicator of immunity system functional status.Test shows that the impact of matsutake concentrated oral liquid on mice serum IgG content has the trend of increase compared with control group, and matsutake concentrated oral liquid has the effect increasing IgG content in serum, as table 6.
Table 6 matsutake concentrated oral liquid affects mice serum IgG content
Note: * * represents the significant difference of P < 0.01 level.
Be not difficult to find out from table 6, after gavage oral liquid 7d, test group IgG content and control group change not quite, there is no significant difference, and after gavage 14d and 21d, mice serum IgG content improves 10% and 11% respectively, difference extremely significantly (P < 0.01).
3.1.4 on the impact of macrophage phagocytic function
After vein injects the inertia carbon granules of specific size, it can be engulfed rapidly by macrophage, and cleans up from blood flow.Therefore the ability of macrophage phagocytic foreign matter can be reflected by the disappearance speed measuring carbon granules in serum.Find after overtesting, matsutake concentrated oral liquid is improved the effect of the phagocytic function of macrophage.Result is as table 7.
Table 7 matsutake concentrated oral liquid affects mouse macrophage phagocytic index
Note: * represents the significant difference of P < 0.05 level.
Can find from table 7, after gavage matsutake concentrated oral liquid 7d and 14d, test group is compared with control group, and phagocytic activity improves 28%, 32% respectively, but does not have significant difference.After gavage 21d, test group, far above control group, improves 51%, and otherness significantly (P < 0.05).
3.2, matsutake concentrated oral liquid is on the impact of mouse antioxidation activity
3.2.1, on the impact of mda content
MDA is one of end-product of polynary unsaturated lipids oxidation in animal body, and the height of its concentration can the extent of peroxidation of unsaturated lipids in antimer, indirectly reflects the degree of injury of cell membrane.Matsutake concentrated oral liquid on the impact of MDA content in Mice Body in table 8.
Table 8 matsutake concentrated oral liquid affects mouse MDA content
Note: * * represents the significant difference of P < 0.01 level.
As shown in Table 8, test group MDA content is all lower than blank group, and when 7d and 14d, MDA content have dropped 10% and 16% respectively, and after gavage 21d, control group is compared with test group, reduces 57%, and otherness extremely significantly (P ﹤ 0.01).Illustrate that matsutake concentrated oral liquid can to reduce in Mice Body MDA content in serum, thus alleviate membrane lipid peroxidatio process in Mice Body.
3.2.2, on the impact of superoxide dismutase activity
SOD is a kind of important enzyme free radical scavenger in organism, has and maintains active oxygen metabolism balance, the function of diaphragm structure.SOD can O in purged body
2 --, thus important use has been balanced to the oxidative and anti-oxidative of body.The height of SOD vigor can reflect the power of body Scavenging ability indirectly.SOD vigor in this test determination serum, concrete vigor the results are shown in Table 9.
Table 9 matsutake concentrated oral liquid is to SOD in Mice effect of vigor
Note: * represents the significant difference of P < 0.05 level.
As can be seen from Table 9, the SOD vigor of test group small mouse serum all higher than blank group, but when gavage 7d and 14d and there was no significant difference, improves 7% and 6% respectively.After gavage 21d, the SOD vigor in test group mice serum, then apparently higher than control group, improves 11%, significant difference (P < 0.05).The power that in Mice Body, the vigor height of anti-oxidant ferment reflects body Scavenging ability illustrates that the matsutake concentrated oral liquid of test group can remove ultra-oxygen anion free radical excessive in Mice Body, improves the oxidation resistance of serum in Mice Body.
3.3, matsutake concentrated oral liquid is on the impact of mouse anti-reflecting fatigue function
3.3.1, on the impact of mouse swimming time
Swim to the time that power exhausts be the common counter of reflection locomitivity, and the raising of locomitivity to be body resisting fatigue ability strengthen the strongest macroscopic view embodies.Long strenuous exercise will cause body relatively hypoxia and glycolysis to be accelerated, and then produce a large amount of lactic acid, and lactic acid accumulation too much can affect the normal metabolic processes in the relatively stable of vivo environment and body.
Table 10 matsutake concentrated oral liquid is to mice burden swimming time effects
From table 10, after gavage matsutake concentrated oral liquid 7d, 14d and 21d, test group compares with blank group, swimming time extends to some extent, improve 43%, 41% and 39% respectively, but there was no significant difference, tentatively can think that matsutake concentrated oral liquid can extend the mice burden swimming time, but machine-processed its also needs further verification experimental verification.
3.3.2, on the impact of Mouse Blood lactic acid content
Lactic acid is the product of anaerobic glycolysis, and after short time large intensity strenuous exercise, anaerobic glycolysis, makes lactic acid production increase sharply.The amplitude that after strenuous exercise, blood lactase acid reduces in short-term is one of important indicator judging micro fatigue crack speed, body recovery degree.
Table 11 matsutake concentrated oral liquid affects Serum lactic acid content
Note: * represents the significant difference of P < 0.05 level.
Can find from table 11, after gavage matsutake concentrated oral liquid 7d, test group and control group are without significant change.After 14d and 21d, in test group mice serum, Serum lactic acid content is starkly lower than control group group, reduces 21% and 29% respectively, has significant difference (P < 0.05), along with the increase of gavage time, in serum, Serum lactic acid content is obvious downward trend.
3.3.3 on the impact of mice serum urea nitrogen content
The change of plasma wrea content can illustrate nitrogen substance catabolism situation in body, plasma wrea content can be made to increase after long period motion, vigorous exercise and strong physical work.When energy balance is destroyed, the catabolism of protein and nitrogen-containing compound is strengthened, and the formation along with urea increases, body urea nitrogen content with work or exercise load increase and increase, body is poorer to load performance, and blood urea nitrogen increases more obvious.Plasma wrea content is a sensitive index for the reaction of body when load, therefore by reducing the decomposition of protein, reducing serum urea nitrogen level, can improve the adaptive capacity to exercise load.
Table 12 matsutake concentrated oral liquid affects serum urea nitrogen content
Note: * * represents the significant difference of P < 0.01 level.
Can find from table 12, after gavage matsutake concentrated oral liquid 7d, control group and test group urea in serum nitrogen content substantially constant, after 14d and 21d, control group serum urea nitrogen content compared with test group all obviously declines, have dropped 38% and 49% respectively, difference extremely significantly (P < 0.01).
The present embodiment has investigated the impact of matsutake concentrated oral liquid on mouse immune regulating action, antioxidation activity and anti-fatigue effect.Result shows, mouse swimming time increases, but difference is not obvious; Thymus index, index and spleen index, macrophage phagocytic function, MDA and superoxide dismutase content have conspicuousness to improve or reduce, significant difference (P < 0.05) after gavage 21d; Extremely significant difference (P < 0.01) is just there is in serum IgG content and urea in serum nitrogen content after 14d after gavage.Thus tentatively can think that matsutake concentrated oral liquid has certain humidification to the immunoregulation capability of mouse, antioxidation activity and anti-fatigue effect.
4, matsutake concentrated oral liquid functional authorization interpretation of result
4.1, immunologic function checking
This test with the thymus index of mouse, index and spleen index, serum IgG content and macrophages phagocytic capacity for Testing index.Result of study shows, gavage matsutake concentrated oral liquid group compares with physiological saline group mouse by mouse, Thymus and Spleen index is in rising trend, and test group mouse thymus index and index and spleen index all significantly rise (P < 0.05) as gavage 21d, show that matsutake concentrated oral liquid effectively can promote the growth of mouse thymus and spleen.Find when IgG content and macrophages phagocytic capacity in Study Mouse serum, test group IgG content is all higher than control group, and when gavage 14d and 21d, difference extremely significantly (P < 0.01).Macrophages phagocytic capacity is 21d generation significant difference (P < 0.05) after gavage then.Can tentatively illustrate, matsutake concentrated oral liquid can significantly improve the immunoregulation capability of mouse, consistent with other results of study.
4.2, anti-oxidation function checking
Found by this experimental study, control group is compared with test group, in test group mice serum, MDA content is all lower than control group, and along with the increase of gavage time, MDA content significantly declines, control group and test group exists and significant difference (P < 0.01) after gavage 21d.This may be because the nutriment in matsutake participates in the approach of quench free radicals in vivo directly, and by participating in the endogenous antioxidant transferring or activate body, its quantity is increased and increased activity to the level of body requirement, thus avoid or alleviate the damage of radical pair body.And although the SOD vigor of test group small mouse serum is higher than control group, but when gavage 7d and 14d and no significant difference, when after gavage 21d, just apparently higher than control group, there is notable difference (P < 0.05) in the SOD vigor in test group mice serum.Although this may be because matsutake concentrated oral liquid can improve SOD vigor in serum, the ability improving SOD vigor can not complete fast with reparation damaged cell, and needs the cumulative function of long period.In summary, by long period gavage matsutake concentrated oral liquid, the oxidation resistance of mouse obtains the raising of conspicuousness, consistent with other results of study.
4.3, anti-fatigue effect checking
Find after exhausting swimming by mouse power, control group is compared with test group, although the time that mouse power exhausts swimming is improved, not obvious difference, this may be due to test mice negligible amounts, and the gavage time is short, and the difference between individuality does not cause very much.And Serum lactic acid content and BUN concentration decline all to some extent in mice serum, after gavage matsutake concentrated oral liquid 14d, just start to have significantly (P < 0.05) and and the difference of remarkable (P < 0.01).Containing various trace elements such as saponin, polysaccharide, flavones, multiple amino and acid, selenium, zinc, copper in matsutake, wherein saponin(e can increase muscle glycogen storage to provide the energy needed for motion, also can increase LDH activity, accelerate lactic acid metabolism, prevent lactic acid from assembling in vivo; Carbohydrate is main energy matter, breaks down proteins energy supply can be passed through when it is under-supply, and then produce a large amount of urea nitrogen and be discharged in blood, the BUN content criterion that whether sufficient carbohydrate energy supply is just, BUN elimination efficiency is negative correlation with the muscular fatigue effect produced of moving; Flavonoid substances, can reduce the decomposition rate of protein, strengthens the synthesis capability of protein, and it is relevant to impel insulin level to raise.Just because of matsutake concentrated oral liquid Middle nutrition material abundance, so matsutake concentrated oral liquid can strengthen mouse anti-reflecting fatigue and have certain mitigation to mouse fatigue, consistent with other results of study.
Claims (10)
1. a preparation method for fresh matsutake active material, is characterized in that it carries out according to following steps:
One, select fresh, without matsutake that is invalid, free from insect pests, in clear water clean, draining, Tricholoma matsutake (lto et lmai) Singer sporophore is carried out slicing treatment, add its 20 times of volume sterile distilled waters, being immersed in containing mass percentage by the matsutake decompression after section is the citric acid of 0.03%, mass percentage is 0.02% n-butanol and mass percentage is in the mixed solution of 0.02% myristin, after decompression dipping 10min, to add mass percentage be 0.02%EDTA and mass percentage is 0.2% natrium citricum, then matsutake tissue is smashed to pieces, again by volume for 1:1:(1 ~ 2) ratio add the cellulase of 6 ~ 7g/L, the pectase of 6 ~ 7g/L and the papain of 6 ~ 7g/L carry out enzyme digestion reaction and obtain matsutake process raw material, wherein, reaction conditions is pH=5, temperature is 50 DEG C, enzymolysis time is 1.5h, enzyme-to-substrate concentration ratio is 1:3.3,
Two, ultrasonic assistant water extraction is got: the water that matsutake process raw material step one obtained is placed in 20 times of matsutake process raw material volume carries out ultrasonic extraction, collects the ultrasonic extract of matsutake and residue; Wherein, ultrasonic treatment conditions are: acoustic power 5 ~ 150w, extraction time 5 ~ 25min, ultrasonic wave working time 1 ~ 5s, interval time 1 ~ 5s;
Three, weak base salt solution extracts: the weak caustic solution adding 20 times of residue volumes in the residue that step 2 is collected extracts, wherein, weak caustic solution contain mass percentage be 0.1 ~ 0.5% sodium bicarbonate solution and mass percentage be 0.5 ~ 2.5% sodium chloride solution, collect matsutake weak base salt solution extract and residue; Wherein, extraction time is 30 ~ 150min, and Extracting temperature is 50 ~ 90 DEG C;
Four, alcoholic solution extracts: step 3 being collected the volumn concentration that residue is placed in 20 times of residue volumes is the ethanol of 50 ~ 90%, is then extract 30 ~ 150min under the condition of 50 ~ 90 DEG C at bath temperature, obtains matsutakealcohol solution extract;
Five, matsutake weak base salt solution extract that the ultrasonic extract of matsutake, step 3 that step 2 obtains obtain and the matsutakealcohol solution extract that step 4 obtains is collected, after mixing, filter with 100 object nylon66 fiber filtering layers, utilize rotary evaporating device with 50 DEG C, the condition of 60r/min carries out distillation and concentration to above-mentioned mixed solution, distillation and concentration, to 1/10 of former mixed liquor volume, namely completes described matsutake active material and extracts.
2. the preparation method of a kind of fresh matsutake active material according to claim 1, is characterized in that the papain adding the cellulase of 6.8g/L, the pectase of 6.8g/L and 6.8g/L for the ratio of 1:1:1.4 by volume in step one carries out enzyme digestion reaction.
3. the preparation method of a kind of fresh matsutake active material according to claim 1, is characterized in that in step 2, ultrasonic treatment conditions are: acoustic power 100 ~ 150w, extraction time 5 ~ 10min, ultrasonic wave working time 2 ~ 4s, interval time 1 ~ 3s.
4. the preparation method of a kind of fresh matsutake active material according to claim 3, is characterized in that in step 2, ultrasonic treatment conditions are: acoustic power 125w, extraction time 10min, ultrasonic wave working time 4s, interval time 1s.
5. the preparation method of a kind of fresh matsutake active material according to claim 1, it is characterized in that weak caustic solution in step 3 contain mass percentage be 0.3% sodium bicarbonate solution and mass percentage be 0.5% sodium chloride solution.
6. the preparation method of a kind of fresh matsutake active material according to claim 1, it is characterized in that in step 3, extraction time is 90 ~ 150min, Extracting temperature is 70 ~ 90 DEG C.
7. the preparation method of a kind of fresh matsutake active material according to claim 6, it is characterized in that in step 3, extraction time is 120min, Extracting temperature is 80 DEG C.
8. the preparation method of a kind of fresh matsutake active material according to claim 1, is characterized in that the volumn concentration of ethanol in step 4 is 80%.
9. the preparation method of a kind of fresh matsutake active material according to claim 1, is characterized in that under bath temperature is the condition of 60 ~ 80 DEG C, extracting 30 ~ 90min in step 4.
10. the preparation method of a kind of fresh matsutake active material according to claim 1, is characterized in that under bath temperature is the condition of 70 DEG C, extracting 30min in step 4.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510018428.3A CN104431995B (en) | 2015-01-14 | 2015-01-14 | A kind of preparation method of fresh Tricholoma matsutake (lto et lmai) Singer active substance |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510018428.3A CN104431995B (en) | 2015-01-14 | 2015-01-14 | A kind of preparation method of fresh Tricholoma matsutake (lto et lmai) Singer active substance |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104431995A true CN104431995A (en) | 2015-03-25 |
| CN104431995B CN104431995B (en) | 2016-07-06 |
Family
ID=52879342
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510018428.3A Active CN104431995B (en) | 2015-01-14 | 2015-01-14 | A kind of preparation method of fresh Tricholoma matsutake (lto et lmai) Singer active substance |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104431995B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104906149A (en) * | 2015-05-28 | 2015-09-16 | 吉林大学 | Medical application of tricholoma matsutake high-yield strain (T35) in resisting fatigue |
| CN104906148A (en) * | 2015-05-28 | 2015-09-16 | 吉林大学 | Medical application of tricholoma matsutake high-yield strain (T35) in antioxidant drugs |
| CN106977592A (en) * | 2016-01-18 | 2017-07-25 | 临沂格瑞食品有限公司 | A kind of method for isolating and purifying Agricus blazei fructification activated protein |
| CN113087762A (en) * | 2021-05-17 | 2021-07-09 | 吉林农业大学 | Preparation method of protein-derived antioxidant hydrolyzed peptide of tricholoma matsutake fruiting body |
| CN115005379A (en) * | 2022-05-27 | 2022-09-06 | 黑龙江农业工程职业学院 | Tonic tricholoma matsutake powder electuary and preparation method thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101768614A (en) * | 2010-03-05 | 2010-07-07 | 浙江省农业科学院 | Method for efficiently extracting polysaccharide and polyphenol from agaricus blazei |
| CN101933610A (en) * | 2010-07-23 | 2011-01-05 | 江苏安惠生物科技有限公司 | Edible and medical fungus cell component comprehensive extraction method and use of extract |
| CN103070388A (en) * | 2013-02-04 | 2013-05-01 | 云南茂昽生物科技有限公司 | Delicate tricholoma matsutake active extract and preparation method and application thereof |
| CN103131195A (en) * | 2013-02-04 | 2013-06-05 | 云南茂昽生物科技有限公司 | Matsutake active extractives and preparation method and application thereof |
| CN103550261A (en) * | 2013-11-01 | 2014-02-05 | 王颖 | Method for extracting matsutake essences |
| CN103694365A (en) * | 2013-12-12 | 2014-04-02 | 舒梅 | Ultrahigh-pressure extraction process of tricholoma matsutake polysaccharide |
-
2015
- 2015-01-14 CN CN201510018428.3A patent/CN104431995B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101768614A (en) * | 2010-03-05 | 2010-07-07 | 浙江省农业科学院 | Method for efficiently extracting polysaccharide and polyphenol from agaricus blazei |
| CN101933610A (en) * | 2010-07-23 | 2011-01-05 | 江苏安惠生物科技有限公司 | Edible and medical fungus cell component comprehensive extraction method and use of extract |
| CN103070388A (en) * | 2013-02-04 | 2013-05-01 | 云南茂昽生物科技有限公司 | Delicate tricholoma matsutake active extract and preparation method and application thereof |
| CN103131195A (en) * | 2013-02-04 | 2013-06-05 | 云南茂昽生物科技有限公司 | Matsutake active extractives and preparation method and application thereof |
| CN103550261A (en) * | 2013-11-01 | 2014-02-05 | 王颖 | Method for extracting matsutake essences |
| CN103694365A (en) * | 2013-12-12 | 2014-04-02 | 舒梅 | Ultrahigh-pressure extraction process of tricholoma matsutake polysaccharide |
Non-Patent Citations (3)
| Title |
|---|
| 尹秀莲等: "松茸不同溶剂提取物及其抗氧化活性", 《食品与发酵工业》, vol. 37, no. 5, 31 December 2011 (2011-12-31), pages 194 - 198 * |
| 张娅等: "松茸多糖超声提取物抗氧化活性研究", 《化学与生物工程》, vol. 28, no. 9, 31 December 2011 (2011-12-31) * |
| 胡尚勤等: "松茸发酵物中活性成分的提取与作用的研究", 《安徽农业大学学报》, vol. 33, no. 4, 31 December 2006 (2006-12-31), pages 499 - 501 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104906149A (en) * | 2015-05-28 | 2015-09-16 | 吉林大学 | Medical application of tricholoma matsutake high-yield strain (T35) in resisting fatigue |
| CN104906148A (en) * | 2015-05-28 | 2015-09-16 | 吉林大学 | Medical application of tricholoma matsutake high-yield strain (T35) in antioxidant drugs |
| CN104906149B (en) * | 2015-05-28 | 2018-08-03 | 吉林大学 | Matsutake superior strain（T35）Medical application in antifatigue |
| CN104906148B (en) * | 2015-05-28 | 2018-08-03 | 吉林大学 | Matsutake superior strain（T35）Medical usage in anti-oxidation medicine |
| CN106977592A (en) * | 2016-01-18 | 2017-07-25 | 临沂格瑞食品有限公司 | A kind of method for isolating and purifying Agricus blazei fructification activated protein |
| CN113087762A (en) * | 2021-05-17 | 2021-07-09 | 吉林农业大学 | Preparation method of protein-derived antioxidant hydrolyzed peptide of tricholoma matsutake fruiting body |
| CN113087762B (en) * | 2021-05-17 | 2024-05-31 | 吉林农业大学 | Preparation method of tricholoma matsutake fruiting body protein source antioxidant hydrolysis peptide |
| CN115005379A (en) * | 2022-05-27 | 2022-09-06 | 黑龙江农业工程职业学院 | Tonic tricholoma matsutake powder electuary and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104431995B (en) | 2016-07-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104431995B (en) | A kind of preparation method of fresh Tricholoma matsutake (lto et lmai) Singer active substance | |
| CN108887673B (en) | Preparation method of stauntonvine enzyme rich in superoxide dismutase (SOD) | |
| CN101940344B (en) | Processing method of instant hot and sour laver | |
| CN104450488B (en) | A kind of ginseng medlar vinegar and preparation method thereof | |
| CN103710237B (en) | A kind of bee wine and preparation method thereof | |
| CN103284153B (en) | Cod liver oil soft capsule and preparation method thereof | |
| CN114246281A (en) | Oyster peptide and sea cucumber peptide sports flavor beverage and preparation method thereof | |
| CN109349467B (en) | Giant salamander beverage | |
| CN103099248A (en) | Method for preparing oyster active component | |
| CN110463992A (en) | A kind of fingered citron ferment powder and production method | |
| CN114097970A (en) | Marine active peptide energy beverage and preparation method thereof | |
| CN102433251A (en) | Russian olive health-care wine and preparation method thereof | |
| CN109527190A (en) | The preparation method and applications of the compound small molecule Gly-His-Lys of black fungus | |
| CN102318865B (en) | Functional beverage and preparation method | |
| CN103054096B (en) | Biological fermented dark chestnut and preparation method thereof | |
| CN105483160B (en) | A kind of Antrodia camphorata culture composition and preparation method thereof | |
| CN107308436B (en) | Biological product with physical fatigue relieving and antioxidant functions and preparation method thereof | |
| CN107281317A (en) | A kind of oral liquid for improving body immunity and preparation method thereof | |
| CN106489835A (en) | The hatching technique of embryo egg | |
| CN1252241C (en) | Health wine for strengthening immunity and its preparing method | |
| CN112075634A (en) | Anti-aging compound nutrient containing coenzyme and bioactive hydrogen and preparation method and application thereof | |
| CN106490507A (en) | Improve the embryo egg hatching technique of immunoglobulin content | |
| CN100391476C (en) | Medicine composition for delaying senility and raising immunity and its prepn | |
| CN105192823A (en) | Chlorella fermented beverage and preparation technology thereof | |
| CN105249465A (en) | Health food capable of improving immunity and preparation method of health food |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |