CN106421047A - Honey pomelo leaf extract, preparation method thereof and application - Google Patents
Honey pomelo leaf extract, preparation method thereof and application Download PDFInfo
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- CN106421047A CN106421047A CN201610971469.9A CN201610971469A CN106421047A CN 106421047 A CN106421047 A CN 106421047A CN 201610971469 A CN201610971469 A CN 201610971469A CN 106421047 A CN106421047 A CN 106421047A
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- citri grandis
- folium citri
- extract
- honey
- sweet
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- 239000000284 extract Substances 0.000 title claims abstract description 97
- 235000012907 honey Nutrition 0.000 title claims abstract description 42
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- 240000000560 Citrus x paradisi Species 0.000 title abstract 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 59
- 230000000694 effects Effects 0.000 claims abstract description 35
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 claims abstract description 25
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 claims abstract description 25
- 229940116269 uric acid Drugs 0.000 claims abstract description 25
- 239000000843 powder Substances 0.000 claims abstract description 15
- 238000000605 extraction Methods 0.000 claims abstract description 14
- 238000007873 sieving Methods 0.000 claims abstract description 10
- 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 claims abstract description 6
- 229930003944 flavone Natural products 0.000 claims abstract description 6
- 150000002212 flavone derivatives Chemical class 0.000 claims abstract description 6
- 235000011949 flavones Nutrition 0.000 claims abstract description 6
- 150000008442 polyphenolic compounds Chemical class 0.000 claims abstract description 6
- 235000013824 polyphenols Nutrition 0.000 claims abstract description 6
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 claims abstract description 6
- 235000009508 confectionery Nutrition 0.000 claims description 87
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 30
- 108010093894 Xanthine oxidase Proteins 0.000 claims description 20
- 102100033220 Xanthine oxidase Human genes 0.000 claims description 20
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 claims description 17
- 230000002000 scavenging effect Effects 0.000 claims description 16
- 239000000469 ethanolic extract Substances 0.000 claims description 14
- 239000007788 liquid Substances 0.000 claims description 12
- -1 DPPH free radical Chemical class 0.000 claims description 10
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 claims description 9
- 239000003814 drug Substances 0.000 claims description 9
- 239000000706 filtrate Substances 0.000 claims description 7
- 235000013402 health food Nutrition 0.000 claims description 7
- 230000001629 suppression Effects 0.000 claims description 7
- 201000005569 Gout Diseases 0.000 claims description 6
- 230000006378 damage Effects 0.000 claims description 5
- 208000027418 Wounds and injury Diseases 0.000 claims description 3
- 208000014674 injury Diseases 0.000 claims description 3
- 238000009098 adjuvant therapy Methods 0.000 claims description 2
- 238000004737 colorimetric analysis Methods 0.000 claims description 2
- 238000001514 detection method Methods 0.000 claims description 2
- 230000029142 excretion Effects 0.000 claims description 2
- 238000000227 grinding Methods 0.000 claims description 2
- 210000003734 kidney Anatomy 0.000 claims description 2
- 230000008439 repair process Effects 0.000 claims description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims 1
- 238000000338 in vitro Methods 0.000 claims 1
- 238000001035 drying Methods 0.000 abstract description 6
- 230000003647 oxidation Effects 0.000 abstract description 2
- 238000007254 oxidation reaction Methods 0.000 abstract description 2
- 235000019441 ethanol Nutrition 0.000 abstract 5
- 238000004108 freeze drying Methods 0.000 abstract 1
- LRFVTYWOQMYALW-UHFFFAOYSA-N 9H-xanthine Chemical compound O=C1NC(=O)NC2=C1NC=N2 LRFVTYWOQMYALW-UHFFFAOYSA-N 0.000 description 16
- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical compound OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 description 16
- 229940074391 gallic acid Drugs 0.000 description 8
- 235000004515 gallic acid Nutrition 0.000 description 8
- 229940075420 xanthine Drugs 0.000 description 8
- 239000012153 distilled water Substances 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 230000031700 light absorption Effects 0.000 description 7
- 239000008363 phosphate buffer Substances 0.000 description 7
- 239000008367 deionised water Substances 0.000 description 6
- 229910021641 deionized water Inorganic materials 0.000 description 6
- 230000002401 inhibitory effect Effects 0.000 description 6
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 5
- 239000003153 chemical reaction reagent Substances 0.000 description 5
- 239000006101 laboratory sample Substances 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical compound N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 description 4
- DGEZNRSVGBDHLK-UHFFFAOYSA-N [1,10]phenanthroline Chemical compound C1=CN=C2C3=NC=CC=C3C=CC2=C1 DGEZNRSVGBDHLK-UHFFFAOYSA-N 0.000 description 4
- 201000010099 disease Diseases 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 3
- 229910019142 PO4 Inorganic materials 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 229960003459 allopurinol Drugs 0.000 description 3
- OFCNXPDARWKPPY-UHFFFAOYSA-N allopurinol Chemical compound OC1=NC=NC2=C1C=NN2 OFCNXPDARWKPPY-UHFFFAOYSA-N 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- SURQXAFEQWPFPV-UHFFFAOYSA-L iron(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+2].[O-]S([O-])(=O)=O SURQXAFEQWPFPV-UHFFFAOYSA-L 0.000 description 3
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 3
- 239000010452 phosphate Substances 0.000 description 3
- 239000000523 sample Substances 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 229910002567 K2S2O8 Inorganic materials 0.000 description 2
- 230000003064 anti-oxidating effect Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 229960002163 hydrogen peroxide Drugs 0.000 description 2
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical compound O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 description 2
- 238000009413 insulation Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 150000003254 radicals Chemical class 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 201000001431 Hyperuricemia Diseases 0.000 description 1
- UGQMRVRMYYASKQ-UHFFFAOYSA-N Hypoxanthine nucleoside Natural products OC1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 UGQMRVRMYYASKQ-UHFFFAOYSA-N 0.000 description 1
- IGLKELDWPZFFKF-UHFFFAOYSA-N OC(C1=CC=CC=C1C(O)=O)=O.OC(C1=CC=CC=C1C(O)=O)=O.OC(C1=CC=CC=C1C(O)=O)=O.P.P Chemical compound OC(C1=CC=CC=C1C(O)=O)=O.OC(C1=CC=CC=C1C(O)=O)=O.OC(C1=CC=CC=C1C(O)=O)=O.P.P IGLKELDWPZFFKF-UHFFFAOYSA-N 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
- 230000036592 analgesia Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 229960002708 antigout preparations Drugs 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 239000011790 ferrous sulphate Substances 0.000 description 1
- 235000003891 ferrous sulphate Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 1
- 230000002439 hemostatic effect Effects 0.000 description 1
- 241000411851 herbal medicine Species 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 206010025482 malaise Diseases 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 230000004792 oxidative damage Effects 0.000 description 1
- USHAGKDGDHPEEY-UHFFFAOYSA-L potassium persulfate Chemical compound [K+].[K+].[O-]S(=O)(=O)OOS([O-])(=O)=O USHAGKDGDHPEEY-UHFFFAOYSA-L 0.000 description 1
- 235000019394 potassium persulphate Nutrition 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 150000005837 radical ions Chemical class 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000013558 reference substance Substances 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 230000001835 salubrious effect Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 239000000341 volatile oil Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/75—Rutaceae (Rue family)
- A61K36/752—Citrus, e.g. lime, orange or lemon
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines Containing Plant Substances (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Jellies, Jams, And Syrups (AREA)
Abstract
The invention discloses a honey pomelo leaf extract, a preparation method thereof and an application. The preparation method includes the steps: (1) drying honey pomelo leave, performing stemming for the dried honey pomelo leave, crushing, and sieving; (2) performing heat extraction for the sieved honey pomelo leaf powder by the aid of ethyl alcohol to obtain honey pomelo leaf ethyl alcohol extracting solution; (3) concentrating the honey pomelo leaf ethyl alcohol extracting solution, and freeze-drying the concentrated honey pomelo leaf ethyl alcohol extracting solution to obtain the honey pomelo leaf extract. Main active components in the honey pomelo leaf extract are flavone and polyphenol. The honey pomelo leaf extract is obtained by the aid of an ethyl alcohol heat extraction method. The honey pomelo leaf extract has obvious functions of resisting oxidation and reducing uric acid activity.
Description
Technical field
The present invention relates to the exploitation of leaf of Fructus Citri grandis, and in particular to a kind of honey Folium Citri grandis extract and preparation method thereof and should
With.
Background technology
China's Fructus Citri grandiss aboundresources, widely distributed, the ground such as Guangdong, Guangxi, Fujian, Yunnan are distributed mainly on, because which is salubrious
Mouthfeel, unique fragrance and extremely people's favor.At present, people focus primarily upon Fructus Citri grandiss leaf essential oil to the research of leaf of Fructus Citri grandis
Extract, the research to leaf of Fructus Citri grandis polyphenol, flavone is rarely reported.Polyphenol, flavone be widely present in plant peel, root, leaf, in fruit
A class natural organic-compound, with extensive biological activity, such as antioxidation, antitumor, antibacterial, adjust immunity, antiinflammatory and resist
Allergy, hemostatic analgesia etc., have been listed in a class functional factor of health food.
The oxidative damage that free radical causes has close relationship with numerous disease, safely and effectively antioxygen in plant
Agent is the focus that studies at this stage, but leaf of Fructus Citri grandis non-oxidizability is also rarely reported.Xanthine oxidase (Xanthine
Oxidase, XOD) it is important enzyme in nucleic acid in vivo metabolism, xanthine can be catalyzed and hypoxanthine oxidation generates uric acid, while
Peroxide radical is produced, uric acid concentration is too high to cause hyperuricemia, cause gout to be shown effect, therefore, suppress the work of XOD
Property be effectively reduced the generation of internal uric acid, prevent or delay the outbreak of gout, reduce body injury.
With the change of people life style, high purine food is ingested in a large number, causes gout sickness rate in rising year by year
Trend, it has also become China is inferior to the second largest metabolism class disease of diabetes.But its medicine is often cured the symptoms, not the disease at present, and
With untoward reaction, therefore natural safely and effectively anti-gout drugs are increasingly paid close attention to by people.In recent years, many research
The effect that concern Chinese herbal medicine extract suppresses to xanthine oxidase, but in terms of leaf of Fructus Citri grandis extract is to xanthine oxidase activity
Probe into and be rarely reported.The extraction process of leaf of Fructus Citri grandis active component polyphenol and flavone is mainly studied in this experiment, and research is carried first
Inhibitory activity of the thing to xanthine oxidase is taken, which is inquired into as the probability of uric acid resisting functional factor, while for leaf of Fructus Citri grandis
Comprehensive development and utilization provides certain reference frame.
Content of the invention
The primary and foremost purpose of the present invention be to provide a kind of sweet Folium Citri grandis extract with antioxidation and uric acid resisting effect and its
Preparation method, the sweet Folium Citri grandis extract can be used to develop the health product and medicine with prevention or treatment gout class diseases effect.
The present invention is achieved through the following technical solutions.
A kind of preparation method of honey Folium Citri grandis extract, comprises the steps：
(1) sweet Folium Citri grandis are dried destemming, grinding and sieving；
(2) the sweet Folium Citri grandis powder after sieving carries out hot extraction with ethanol, obtains sweet Folium Citri grandis ethanol extract；
(3) by lyophilizing after the concentration of sweet Folium Citri grandis ethanol extract, sweet Folium Citri grandis extract is obtained.
Further, in step (1), described sieve for cross 60～80 mesh sieves.
Further, in step (2), the ethanol is the ethanol of volume fraction 50～80%.
Further, in step (2), sweet Folium Citri grandis powder is 1 with the solid-liquid ratio of ethanol:15～1:30g/mL.
Further, in step (2), the heat is extracted and is：6～10h is extracted at a temperature of 40～80 DEG C, in triplicate,
Merging filtrate.
Further, in step (3), described concentrate be concentrated into respect to water density be 1.15～1.30.
The sweet Folium Citri grandis extract that preparation method described in any of the above-described is obtained.
Further, the main component of the honey Folium Citri grandis extract is flavone and polyphenol.
Further, the honey Folium Citri grandis extract has non-oxidizability and uric acid resisting activity.
Further, the non-oxidizability be presented as to DPPH free radical, hydroxy radical, superoxide anion and
ABTS+activity removing.
Further, the activity to DPPH free radical, hydroxy radical, superoxide anion and ABTS.+ is removed
IC50 value is respectively 1.24mg/mL, 1.04mg/mL, 1.49mg/mL and 3.31mg/mL.
Further, the uric acid resisting activity is presented as external uric acid resisting activity, and the detection of external uric acid resisting activity is adopted
Use colorimetry.
Further, the uric acid resisting activity is presented as the activity of suppression xanthine oxidase, to xanthine oxidase
The IC50 value of activity suppression is 4.8mg/mL.
The honey Fructus Citri grandiss extract is used as to include the adjuvant therapy medicaments of gout or for preparing to have with uric acid resisting is
The health food of functional factor and medicine.
The honey Folium Citri grandis extract is used for the medicine for preparing uric acid resisting, promoting uric acid renal excretion, repair injury of kidney；By sweet Fructus Citri grandiss
Leaf extract makes health food with pharmaceutically acceptable carrier.
Compared with prior art, the invention has the advantages that and beneficial effect；
(1) extracting method of the present invention is simple, convenient and swift；
(2) present invention obtains sweet Folium Citri grandis extract by way of ethanol heat is extracted, and the sweet Folium Citri grandis extract for obtaining has aobvious
The uric acid resisting activity of work, can be further used for screening and prepare and exploitation uric acid resisting medicine and health food.
Description of the drawings
Fig. 1 is sweet active scavenging action curve chart of the Folium Citri grandis extract to DPPH free radical in embodiment 1；
Fig. 2 is sweet active scavenging action curve chart of the Folium Citri grandis extract to hydroxy radical in embodiment 2；
Fig. 3 is sweet active scavenging action curve chart of the Folium Citri grandis extract to superoxide anion in embodiment 3；
Fig. 4 be in embodiment 4 sweet Folium Citri grandis extract to ABTS+active scavenging action curve chart；
The block diagram that Fig. 5 is the sweet Folium Citri grandis extract of embodiment 1～4 to the IC50 value that each free radical ion activity is removed；
Fig. 6 is sweet inhibitory activity curve chart of the Folium Citri grandis extract to xanthine oxidase in embodiment 5.
Specific embodiment
The present invention is further illustrated below by the mode of embodiment, but therefore do not limit the present invention to described reality
Apply among a scope, the experimental technique of unreceipted actual conditions in the following example, all conventionally and condition, or according to
Catalogue is selected, and the raw material in following examples is commercially available.
Embodiment 1
Scavenging action of the sweet Folium Citri grandis extract to DPPH free radical
1st, the extraction of sweet Folium Citri grandis extract
(1) will be destemming for the drying of sweet Folium Citri grandis, 60 mesh sieves are crossed after crushing；
(2) the sweet Folium Citri grandis powder volume fraction after sieving be 80% ethanol at 80 DEG C heat extract 6h, in triplicate,
Merging filtrate, obtains sweet Folium Citri grandis ethanol extract；Sweet Folium Citri grandis powder is 1 with the solid-liquid ratio of ethanol:30g/mL；
(3) by sweet Folium Citri grandis ethanol extract be concentrated into respect to water density be 1.15 after lyophilizing, obtain sweet Folium Citri grandis extraction
Thing.
2nd, prepared by laboratory sample and reagent
Sweet Folium Citri grandis extract solution：20mg honey Folium Citri grandis extract is accurately weighed, and 2.5mg/ml solution is made into distilled water, point
Be not diluted to 0.5,1.0,1.5, the sweet Folium Citri grandis extract solution of 2.0mg/ml, standby.
The DPPH solution of 0.2mmol/L：Accurately weigh DPPH 0.00788g and be dissolved in 95% ethanol of 100ml, lucifuge
Preserve.
3rd, experimentation
2mLDPPH solution (0.2mmol/L) is placed in test tube, 2mL honey Folium Citri grandis extract solution is added, concussion is equal
Even, room temperature determines light absorption value (Ai) after placing 30min at 517nm；
The zeroing of 2mL distilled water is added with 2mL95% ethanol；
Compare and be：2mL DPPH solution determines the light absorption value (Ac) under wavelength at 517nm plus 2mL95% ethanol,
The light absorption value (Aj) of wavelength is determined after 2mL honey Folium Citri grandis extract solution and the mixing of 2mL95% ethanol at 517nm.
Extract represents to the Scavenging activity suppression ratio R of DPPH：R=[1- (Ai-Aj)/Ac] × 100%.
With the Vc of variable concentrations and gallic acid solution as standard substance, and with same determination of experimental method variable concentrations
The activity removing of Vc and gallic acid solution to DPPH free radical.
4th, experimental result
The experimental result such as Fig. 1 for the scavenging action to DPPH free radical for obtaining, as shown in Figure 1, sweet Folium Citri grandis extract
The IC50 value removed by the activity of DPPH free radical is 1.24mg/mL.
Embodiment 2
Scavenging action of the sweet Folium Citri grandis extract to hydroxy radical
1st, the extraction of sweet Folium Citri grandis extract
(1) will be destemming for the drying of sweet Folium Citri grandis, 60 mesh sieves are crossed after crushing；
(2) the sweet Folium Citri grandis powder volume fraction after sieving be 70% ethanol at 80 DEG C heat extract 8h, in triplicate,
Merging filtrate, obtains sweet Folium Citri grandis ethanol extract；Sweet Folium Citri grandis powder is 1 with the solid-liquid ratio of ethanol:15g/mL；
(3) sweet Folium Citri grandis ethanol extract is concentrated into lyophilizing after the density 1.15 with respect to water, obtains sweet Folium Citri grandis extract.
2nd, prepared by laboratory sample and reagent
Sweet Folium Citri grandis extract solution：20mg honey Folium Citri grandis extract is accurately weighed, and 2.5mg/ml solution is made into distilled water, point
Be not diluted to 0.5,1.0,1.5, the sweet Folium Citri grandis extract solution of 2.0mg/ml, standby.
5mmol/L orthophenanthroline solution：0.0991g orthophenanthroline solid, adds 98ml after adding 2ml anhydrous alcohol solution
Deionized water.
7.4 phosphate buffer of pH：3.12g NaH2PO4·2H2O constant volume obtains solution A to 100ml；7.16g
Na2HPO4·12H2O constant volume obtains solution B to 100ml；Take 9.5ml solution A and 40.5ml solution B is mixed in beaker, adjust pH
To 7.4.
15mmol/L EDTA solution：1.0959g EDTA solid deionized water constant volume is in 250ml volumetric flask, standby.
5mmol/L copperas solution：0.1390g ferrous sulfate solid, adds 100ml deionized water (now with the current).
0.1wt% hydrogenperoxide steam generator：The 30%H2O2 of 0.335ml adds 100ml deionized water (now with the current).
3rd, experimentation
3 groups of sample cell (numbering is 1. 2. 3.)：Take 1.2mL orthophenanthroline solution (5mmol/L) and add 0.8mL phosphate-buffered
After liquid (0.2M, pH 7.4) is mixed, 1.2mL honey Folium Citri grandis extract solution and 1.2mLEDTA (15mmol/L) solution is added, again
Mix.
Damage pipe, do not damage pipe (numbering is 4. 5.)：Take 1.2mL orthophenanthroline solution (5mmol/L) and add 0.8mL phosphate
After buffer (0.2M, pH 7.4) is mixed, 1.2mL deionized water and 1.2mL EDTA (15mmol/L) is added.
Test tube 1.～be 5. separately added into 1.2ml 5mmlol/L copperas solution, after fully mixing, test tube is 1. 2. 3. 4.
1.6ml 0.1wt% hydrogenperoxide steam generator is separately added into, 5. test tube adds 1.6ml deionized water；Test tube 1.～be 5. placed in 37 DEG C
Thermostat water bath is incubated 1h, determines light absorption value at 536nm, record data.
With the Vc of variable concentrations and gallic acid solution as standard substance, and with same determination of experimental method variable concentrations
The activity removing of Vc and gallic acid solution to hydroxy radical ion.
4th, experimental result
The experimental result such as Fig. 2 of the sweet Folium Citri grandis extract for obtaining to the scavenging action of hydroxy radical, as shown in Figure 2, sweet Fructus Citri grandiss
The IC50 value that leaf extract is removed to the activity of hydroxy radical is 1.04mg/mL.
Embodiment 3
Scavenging capacity of the sweet Folium Citri grandis extract to superoxide anion
1st, the extraction of sweet Folium Citri grandis extract
(1) will be destemming for the drying of sweet Folium Citri grandis, 70 mesh sieves are crossed after crushing；
(2) the sweet Folium Citri grandis powder volume fraction after sieving be 60% ethanol at 40 DEG C heat extract 6h, in triplicate,
Merging filtrate, obtains sweet Folium Citri grandis ethanol extract；Sweet Folium Citri grandis powder is 1 with the solid-liquid ratio of ethanol:25g/mL；
(3) sweet Folium Citri grandis ethanol extract is concentrated into lyophilizing after the density 1.30 with respect to water, obtains sweet Folium Citri grandis extract.
2nd, prepared by laboratory sample and reagent
Sweet Folium Citri grandis extract solution：20mg extract is accurately weighed, and 2.5mg/ml solution is made into distilled water, is diluted respectively
Become 0.5,1.0,1.5, the sweet Folium Citri grandis extract solution of 2.0mg/ml, standby.
3rd, experimentation
Xanthine oxidase can be catalyzed xanthine and be converted into uric acid under conditions of aerobic, at the same produce super oxygen cloudy from
Sub- free radical (O2-), O2- combined with NBT rear in blueness；Sweet Folium Citri grandis extract Scavenging activity is bigger, is combined with NBT
O2- fewer, solution colour will be more shallow.
Assay method is：20 μ L 3mmol/L xanthine add 180 μ L 0.6mmol/L NBT and 20 μ L honey Folium Citri grandis to extract
Thing solution, reacts 5min in 37 DEG C；The xanthine oxidase of 20 μ L 0.1u/mLs in 96 orifice plates in 37 DEG C insulations are added afterwards
After 30min, light absorption value is determined at 560nm, be designated as Asample.
Replace sweet Folium Citri grandis extract solution right as blank with phosphate buffer (pH=7.5, concentration is 0.2mol/L)
According at 560nm, measure light absorption value, is designated as Acontrol.
Clearance rate (%)=(Acontrol-Asample)/Acontrol × 100
With the Vc of variable concentrations and gallic acid solution as standard substance, and with same determination of experimental method variable concentrations
The activity removing of Vc and gallic acid solution to ultra-oxygen anion free radical.
4th, experimental result
Experimental result is as shown in figure 3, from the figure 3, it may be seen that IC50 of the sweet Folium Citri grandis extract to the activity removing of superoxide anion
It is worth for 1.49mg/mL.
Embodiment 4
Sweet Fructus Citri grandiss extract to ABTS+scavenging capacity
1st, the extraction of sweet Folium Citri grandis extract
(1) will be destemming for the drying of sweet Folium Citri grandis, 80 mesh sieves are crossed after crushing；
(2) the sweet Folium Citri grandis powder volume fraction after sieving is 80% ethanol heat extraction 10h, repetition three at 60 DEG C
Secondary, merging filtrate, obtain sweet Folium Citri grandis ethanol extract；Sweet Folium Citri grandis powder is 1 with the solid-liquid ratio of ethanol:30g/mL；
(3) sweet Folium Citri grandis ethanol extract is concentrated into lyophilizing after the density 1.20 with respect to water, obtains sweet Folium Citri grandis extract.
2nd, prepared by laboratory sample and reagent
ABTS storing solution (7.4mmol/L, 0.4mL)：Take ABTS 3mg, plus distilled water 0.735mL.(MW=548.7)
K2S2O8Storing solution (2.6mmol/L, 1.43mL)：Take K2S2O81mg, plus distilled water 1.43mL.(MW=270.32)
Sweet Folium Citri grandis extract solution：20mg extract is accurately weighed, and 2.5mg/ml solution is made into distilled water, is diluted respectively
Become 0.5,1.0,1.5, the sweet Folium Citri grandis extract solution of 2.0mg/ml, standby.
3rd, experimentation
Prepare ABTS+ storing solution：ABTS deposit with the potassium peroxydisulfate storing solution dissolving 7.4mmol/L of 2.6mmol/L
Liquid, is made into the ABTS+ storing solution of 7mmol/L, stands 12h under the conditions of room temperature, lucifuge.
Prepare ABTS+ and determine liquid：ABTS+ storing solution is diluted with phosphate buffer (10mmol/L, pH7.4), is made
Its absorbance reaches 0.700 ± 0.020 at 734nm wavelength.
Determine：Take 0.8mLABTS+ and liquid is determined, 0.2ml honey Folium Citri grandis extract solution is added, accurately vibrates 10s, standing 6
Minute, determine the absorbance A at 734nm wavelength.
Blank：Take 0.8mLABTS+ and liquid is determined, 0.2ml95% ethanol is added, accurately 10s is vibrated, 6 minutes are stood, survey
Determine the absorbance A at 734nm wavelength1.
Clearance rate=(A1-A)/A1× 100%
With the Vc of variable concentrations and gallic acid solution as standard substance, and with same determination of experimental method variable concentrations
The activity removing of Vc and gallic acid solution to ABTS+.
4th, measurement result
Sweet Fructus Citri grandiss extract to ABTS+scavenging capacity result as shown in figure 4, as shown in Figure 4, sweet Folium Citri grandis extract pair
The IC50 value that the activity of ABTS+ is removed is 3.31mg/mL.
Fig. 5 is block diagram of the sweet Folium Citri grandis extract to the IC50 value of each radical ion scavenging capacity in embodiment 1～4；
As shown in Figure 5, sweet Folium Citri grandis extract to DPPH free radical, hydroxy radical, superoxide anion and ABTS+activity remove
IC50 value is respectively 1.24mg/mL, 1.04mg/mL, 1.49mg/mL and 3.31mg/mL.
Embodiment 5
Inhibitory activity of the sweet Folium Citri grandis extract to xanthine oxidase
1st, the extraction of sweet Folium Citri grandis extract
(1) will be destemming for the drying of sweet Folium Citri grandis, 75 mesh sieves are crossed after crushing；
(2) the sweet Folium Citri grandis powder volume fraction after sieving be 75% ethanol at 50 DEG C heat extract 8h, in triplicate,
Merging filtrate, obtains sweet Folium Citri grandis ethanol extract；Sweet Folium Citri grandis powder is 1 with the solid-liquid ratio of ethanol:20g/mL；
(3) sweet Folium Citri grandis ethanol extract is concentrated into lyophilizing after the density 1.18 with respect to water, obtains sweet Folium Citri grandis extract.
2nd, prepared by laboratory sample and reagent
Sweet Folium Citri grandis extract solution：Sweet Folium Citri grandis extract is configured to 2,4,6,8,10mg/mL honey Folium Citri grandis extract solution.
Positive reference substance solution：Allopurinol is first configured to 50 μ g/mL solution with the NaOH dissolving of 1mol/L, then uses
0.2mol/L phosphate buffer (pH=7.5) is diluted to 2.5,7.5,14,20,27.5 μ g/mL allopurinol solution, standby.
3rd, experimentation
By 50 μ L concentration be the xanthine oxidase solution of 0.1U/mL, 50 μ L concentration for 0.2mol/L (PH=7.5) phosphorus
Phthalate buffer, sweet Folium Citri grandis extract solution are added sequentially in 96 micropore versions, and after shaking 30s, 37 DEG C of insulation 15min are (during balance
Between result is had an impact, this time ensure operation possibility), using multichannel pipettor quickly to 96 microwell plates add 150 μ L dense
The xanthine for 0.2mM is spent, record reaction 30min in microplate reader；50 μ L concentration are added afterwards for the HCl terminating reaction of 1M, in
Light absorption value is determined at 290nm.
4th, according to formula, inhibitory activity of the extract sample to XOD is calculated
Suppression ratio (%)=[(A-B)-(C-D)]/(A-B) × 100%
A:+ 150 μ L of phosphate buffer of+50 μ L 0.2mol/L (PH=7.5) of 50 μ L 0.1U/mL xanthine oxidase
The xanthine of 0.2mM
B:The phosphate of+50 μ L 0.2mol/L (PH=7.5) of phosphate buffer of 50 μ L0.2mol/L (PH=7.5)
The xanthine of+150 μ L0.2mM of buffer
C:The xanthine of+150 μ L 0.2mM of+50 μ L of 50 μ L 0.1U/mL xanthine oxidase honey Folium Citri grandis extract solution
D:+ 150 μ L0.2mM of+50 μ L of phosphate buffer honey Folium Citri grandis extract solution of 50 μ L0.2mol/L (PH=7.5)
Xanthine
With the allopurinol solution of variable concentrations as standard substance, and with the not fast of same determination of experimental method variable concentrations
Purine alcoholic solution suppresses to xanthine oxidase activity.
5th, measurement result
Inhibitory activity result to xanthine oxidase is as shown in fig. 6, it will be appreciated from fig. 6 that sweet Folium Citri grandis extract has preferably
Xanthine oxidase inhibitory activity, to xanthine oxidase activity suppression IC50 value be 4.8mg/mL.
Claims (10)
1. a kind of honey Folium Citri grandis extract preparation method, it is characterised in that comprise the steps：
（1）Sweet Folium Citri grandis are dried destemming, grinding and sieving；
（2）Sweet Folium Citri grandis powder after sieving carries out hot extraction with ethanol, obtains sweet Folium Citri grandis ethanol extract；
（3）Lyophilizing after sweet Folium Citri grandis ethanol extract is concentrated, obtains sweet Folium Citri grandis extract.
2. according to claim 1 a kind of honey Folium Citri grandis extract preparation method, it is characterised in that step（1）In, described
Sieve as crossing 60 ~ 80 mesh sieves.
3. according to claim 1 a kind of honey Folium Citri grandis extract preparation method, it is characterised in that step（2）In, described
Ethanol is the ethanol of volume fraction 50 ~ 80%, and sweet Folium Citri grandis powder is 1 with the solid-liquid ratio of ethanol:15~1:30g/mL, the heat is extracted
It is：6 ~ 10h, in triplicate, merging filtrate are extracted at a temperature of 40 ~ 80 DEG C；Step（3）In, described concentration is to be concentrated into relatively
It is 1.15 ~ 1.30 in the density of water.
4. the sweet Folium Citri grandis extract that preparation method described in any one of claim 1 ~ 3 is obtained, it is characterised in that the honey Folium Citri grandis
The main component of extract is flavone and polyphenol；The honey Folium Citri grandis extract has non-oxidizability and uric acid resisting activity.
5. according to claim 4 a kind of honey Folium Citri grandis extract, it is characterised in that it is right that the non-oxidizability is presented as
DPPH free radical, hydroxy radical, superoxide anion and ABTS·+ scavenging capacity.
6. according to claim 5 a kind of honey Folium Citri grandis extract, it is characterised in that described to DPPH free radical, hydroxyl freedom
Base, superoxide anion and ABTS·+ scavenging capacity IC50 value be respectively 1.24 mg/mL, 1.04 mg/mL, 1.49 mg/mL
With 3.31 mg/mL.
7. according to claim 4 a kind of honey Folium Citri grandis extract, it is characterised in that the uric acid resisting activity is presented as in vitro
Uric acid resisting activity, the detection of external uric acid resisting activity adopts colorimetry.
8. according to claim 4 a kind of honey Folium Citri grandis extract, it is characterised in that the uric acid resisting activity is presented as suppression
The activity of xanthine oxidase, the IC50 value to xanthine oxidase activity suppression is 4.8 mg/mL.
9. the application of a kind of honey Folium Citri grandis extract described in claim 4, it is characterised in that the honey Folium Citri grandis extract is used for wrapping
In the preparation of the adjuvant therapy medicaments for including gout, or the health food for having with uric acid resisting as functional factor or medicine
Preparation in.
10. the application of a kind of honey Folium Citri grandis extract described in claim 4, it is characterised in that the honey Folium Citri grandis extract is used for
Uric acid resisting, promote uric acid renal excretion, repair injury of kidney medicine preparation in；Or in the preparation of health food, i.e., with pharmacy
Upper acceptable carrier makes health food.
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| TWI713829B (en) * | 2018-03-23 | 2020-12-21 | 田錦衡 | Plant-based black hair dye and hair dyeing method |
| CN115477977A (en) * | 2021-11-08 | 2022-12-16 | 木由子(上海)生物科技有限公司 | Extraction process of shaddock leaf essential oil for essence and spice and shaddock leaf essential oil |
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| CN103599123A (en) * | 2013-12-03 | 2014-02-26 | 广东世信药业有限公司 | Drug for treating purine metabolic disorder disease |
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| CN103599123A (en) * | 2013-12-03 | 2014-02-26 | 广东世信药业有限公司 | Drug for treating purine metabolic disorder disease |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI713829B (en) * | 2018-03-23 | 2020-12-21 | 田錦衡 | Plant-based black hair dye and hair dyeing method |
| CN115477977A (en) * | 2021-11-08 | 2022-12-16 | 木由子(上海)生物科技有限公司 | Extraction process of shaddock leaf essential oil for essence and spice and shaddock leaf essential oil |
| CN115477977B (en) * | 2021-11-08 | 2024-04-12 | 木由子(上海)生物科技有限公司 | Extraction process of shaddock leaf essential oil for essence and spice and shaddock leaf essential oil |
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