CN104435071A - Active part of mallotus oblongifolius as well as extraction method and application thereof - Google Patents

Active part of mallotus oblongifolius as well as extraction method and application thereof Download PDF

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Publication number
CN104435071A
CN104435071A CN201410618288.9A CN201410618288A CN104435071A CN 104435071 A CN104435071 A CN 104435071A CN 201410618288 A CN201410618288 A CN 201410618288A CN 104435071 A CN104435071 A CN 104435071A
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China
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extract
mountain
folium ligustri
ligustri pubescentis
petroleum ether
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CN201410618288.9A
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CN104435071B (en
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康文艺
朱晓娣
尹震花
王培卿
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Henan University
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Henan University
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    • 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)
    • A61K36/47—Euphorbiaceae (Spurge family), e.g. Ricinus (castorbean)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/35—Extraction with lipophilic solvents, e.g. Hexane or petrol ether
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps

Abstract

The invention belongs to the technical field of plant extraction, and particularly relates to an active part of mallotus oblongifolius as well as an extraction method and an application thereof. The active part of mallotus oblongifolius is a petroleum ether part and/or an ethyl acetate part and/or an n-butanol part of mallotus oblongifolius stems or leaves. The effective part alpha-glucosidase has an inhibiting effect.

Description

Mountain Folium Ligustri pubescentis effective site and extracting method thereof, application
 
technical field
The invention belongs to technical field of plant extraction, be specifically related to a kind of mountain Folium Ligustri pubescentis effective site and extracting method thereof, application.
background technology
Mountain Folium Ligustri pubescentis ( mallotus oblongifolius), be Euphorbiaceae Mallotus plant, the Hainan Island of main distribution China, the middle print peninsula and Sumatra.Its fragrance is pleasant, is rich in several mineral materials, and essential amino acid, has heat-clearing and toxic substances removing, effect such as heat syndrome cough, diuresis of eliminating the phlegm.People from Hainan custom mountain Folium Ligustri pubescentis leaf beverage brewed of drying.Therefore mountain Folium Ligustri pubescentis be among the people a kind of in Hainan have strong local characteristic and national characters for tea beverage plant and important medicinal plants.In recent years, the research of closing Yushan Hill Folium Ligustri pubescentis increases, and finds that mountain Folium Ligustri pubescentis has function of gallbladder promoting, analgesia and antibacterial effect, but for can Inhibiting α-glucosidase activity wait blood sugar reducing function to have no report.
Alpha-glucosidase extensively distributes at nature, of a great variety, distinct, is almost present in all organisms.It has important physiological function in the carbohydate metabolism of the glycogen degradation of the mankind and animal, plant, microorganism.
Alpha-glucosidase (α-Glucosidase, EC 3.2.1.20) be again alpha-D-glucose glycosides hydrolytic enzyme.Mainly comprise the enzymes such as maltase, saccharase, isomeric maltose enzyme, Lactose enzyme, it is mainly distributed in small intestine epithelium chorion brush along upper, has important function to catabolism of carbohydrate.Main mechanism is: the polysaccharide in food, as starch direct oral cavity saliva, pancreatic amylase are digested to the oligosaccharide containing minority glucose molecule, alpha-glucosidase cuts α-1 at the non-reducing end of these oligosaccharide, 4 glycosidic bonds, discharge glucose, the latter enters blood after being absorbed by small intestine epithelium, just becomes blood glucose.
Alpha-glucosidase inhibitor is the activity of the alpha-glucosidase by reversible inhibition or competitive inhibition intestinal brush border, thus postpone polysaccharide, disaccharidase is converted into absorbable glucose, slows down the rising of post-prandial glycemia.This type of medicine is called as third generation oral hypoglycemic, clinical practice have acarbose, Voglibose tablet, miglitol etc.From Chinese herbal medicine, more particularly can find natural, that side effect is little alpha-glucosidase inhibitor there is prospect widely in the use plant of safety.
summary of the invention
The object of the present invention is to provide a kind of mountain Folium Ligustri pubescentis effective site with Inhibiting α-glucosidase active function, and furthermore present the extracting method of mountain Folium Ligustri pubescentis effective site.
Based on above object, the present invention adopts following technical scheme:
Mountain Folium Ligustri pubescentis effective site is the petroleum ether part of mountain Folium Ligustri pubescentis stem or leaf and/or ethyl acetate extract and/or n-butanol portion.
The petroleum ether part of mountain Folium Ligustri pubescentis stem or leaf, ethyl acetate extract and n-butanol portion obtain by the following method: leaf and the stem of getting mountain Folium Ligustri pubescentis respectively, and pulverize, add ethanol, heating and refluxing extraction, the liquid concentration of extraction obtains total extractum; Be scattered in water by total extractum, extract with petroleum ether, ethyl acetate, n-butyl alcohol, namely the petroleum ether extract obtained, acetic acid ethyl ester extract and n-butyl alcohol extract are respectively petroleum ether part, ethyl acetate extract and n-butanol portion.
The concentration of ethanol is 65-80V%; Reflux 2-3 time, each 0.5-2h.
The extracting method of mountain Folium Ligustri pubescentis effective site, the first drying of the stem of mountain Folium Ligustri pubescentis or leaf, pulverizing, then add alcohol heating reflux and extract 2-3 time, each 0.5-2h, merge extractive liquid, filtration, concentratedly to obtain always extractum; Be scattered in water by total extractum, extract, volatilize solvent with petroleum ether, ethyl acetate, n-butyl alcohol, namely the petroleum ether extract obtained, acetic acid ethyl ester extract and n-butyl alcohol extract are respectively petroleum ether part, ethyl acetate extract and n-butanol portion.
Mountain Folium Ligustri pubescentis effective site is in preparation αapplication in-glucosidase inhibitor class medicine.
The present invention for positive control drug carries out in vitro tests, observes the petroleum ether part of mountain Folium Ligustri pubescentis and mountain Folium Ligustri pubescentis stem, ethyl acetate extract and n-butanol portion pair with acarbose (Acarbose) respectively αthe inhibitory action of-glucosidase activity.Result of the test shows, except the inhibitory action of mountain Folium Ligustri pubescentis n-butanol portion is more weak, and other mountain Folium Ligustri pubescentis effective sites pair α-glucosidase all has obvious inhibitory action, and in dose-effect relationship within the scope of mass concentration, rising suppression ratio with concentration also increases, but when but concentration declines higher than suppression ratio during 750 μ g/mL, when 750 μ g/mL, suppression ratio reaches maximum, and now the suppression ratio of these 5 effective sites all reaches more than 95%, the results are shown in Figure 1 and Fig. 2.Positive control drug acarbose suppression ratio when concentration is 1500 μ g/mL is 62.62%, is 37.03% when when 750 μ g/mL, suppression ratio drops to; 5 effective sites except the Folium Ligustri pubescentis n-butanol portion of mountain, compared with acarbose, are all better than acarbose.
accompanying drawing explanation
Fig. 1 is each effective site of mountain Folium Ligustri pubescentis stem α-Glucosidase inhibitor rate is with the variation diagram of drug level;
Fig. 2 is each effective site of mountain Folium Ligustri pubescentis α-Glucosidase inhibitor rate is with the variation diagram of drug level.
detailed description of the invention
The embodiment extracted mountain Folium Ligustri pubescentis and mountain Folium Ligustri pubescentis stem is respectively as follows:
Embodiment 1, each effective site of mountain Folium Ligustri pubescentis
Get dry mountain Folium Ligustri pubescentis 145.3g, by heating and refluxing extraction under the ethanol water slight boiling condition of 70V% 2 times, each 2h, each extracting solution filters, and merges 2 filtrates and is condensed into extractum; Extractum is scattered in certain water and forms dispersion liquid, use petroleum ether (about 1 times of volume of dispersion liquid), ethyl acetate (about 1 times of volume of dispersion liquid), n-butyl alcohol (about 1 times of volume of dispersion liquid) to extract extractum respectively, each extraction three times; Volatilize solvent, obtain petroleum ether (PE) position of mountain Folium Ligustri pubescentis, ethyl acetate (EA) position and n-butyl alcohol (BU) position.Yield corresponding to different solvents is in Table 1.
Embodiment 2
As different from Example 1, concentration of alcohol is 80V%, reflux 3 times, each 0.5h.
Embodiment 3
As different from Example 1, concentration of alcohol is 65V%, reflux 3 times, each 1h.
Embodiment 4, each effective site of mountain Folium Ligustri pubescentis
Get dry mountain Folium Ligustri pubescentis stem 80.3g, by heating and refluxing extraction under the ethanol water slight boiling condition of 70V% 2 times, each 2h, each extracting solution filters, and merges 2 filtrates and is condensed into extractum; Extractum is scattered in certain water, uses petroleum ether (about 1 times of volume), ethyl acetate (about 1 times of volume), n-butyl alcohol (about 1 times of volume) to extract extractum respectively, each extraction three times; Volatilize solvent, obtain petroleum ether (PE) position of mountain Folium Ligustri pubescentis stem, ethyl acetate (EA) position and n-butyl alcohol (BU) position.Yield corresponding to different solvents is in Table 1.
Embodiment 5
As different from Example 4, concentration of alcohol is 80V%, reflux 3 times, each 0.5h.
Embodiment 6
As different from Example 4, concentration of alcohol is 65V%, reflux 3 times, each 1h.
Effect experimental: mountain Folium Ligustri pubescentis (leaf) is tested with the external Inhibiting enzyme activity of mountain Folium Ligustri pubescentis (stem) effective site
1.1 test methods: Microdilution plate method
1.1.1 principle: α-D-Glucose glycosides enzyme can catalyzing hydrolysis 4-Nitrobenzol- α-D-pyranglucoside (PNP-G), produce nitrophenol (PNP), it is yellow substance, has maximum absorption band at about 400nm.And α-glycosidase inhibitor can suppress αthe combination of-glucoside enzyme-to-substrate, thus the burst size reducing PNP.The inhibition of enzyme activity rate of effective site is calculated with the changes of contents of PNP in reaction system in certain hour.
instrument:rotary Evaporators (German Heidolph company); Electronic balance (Mettler-Toledo company of the U.S.); CS-HI type blender; DELTA 320 type PH counts (Mettler-Toledo); Multiskan MK3 microplate reader (Thermo Electron company of the U.S.); LRH-150 constant incubator (the permanent Science and Technology Ltd. in Shanghai one); Pipettor (German BRAND company); Rifle head; Centrifuge tube etc.
reagent: α-glucosidase (Japan and the meeting of light pure pharmaceutical worker's industry strain formula, Lot:22003201), p-nitrobenzophenone- α- d-pyranglucoside (PNPG, Calbiochem company, Lot:D00129651), kaliumphosphate buffer (PH 6.8), acarbose (German SERVE, Lot:100434), other reagent are analytical pure.
the preparation of sample solution
Each effective site: the effective site taking certain mass, adds the DMSO of certain volume, and being made into initial concentration is 30mg/mL solution.
Acarbose preparation is with each effective site.
detection method and date processing
Experimental group arranges as follows:
Blank group: 8 μ L DMSO+152 μ L kaliumphosphate buffers, surveys OD value under 405 nm wavelength;
Blank group: 8 μ L DMSO+112 μ L kaliumphosphate buffer+20 μ L α-glucosidase+20 μ LPNPG, surveys OD value under 405 nm wavelength;
Sample controls group: 8 μ L sample solution+152 μ L kaliumphosphate buffers, surveys OD value under 405 nm wavelength;
Sample sets: 8 μ L sample solution+20 μ L α-glucosidase+112 μ L kaliumphosphate buffer+20 μ LPNPG, surveys OD value after reaction terminates under 405 nm wavelength; (detailed process: 112 μ L kaliumphosphate buffers (pH 6.8), adding concentration is 0.2 U/mL αthe sample solution of-glucosidase 20 μ L, 8 μ L respective concentration, 37 DEG C of constant temperature 15 min, add 20 μ L 2.5 mmol/L PNPG, 37 DEG C of isothermal reaction 15 min.Add 80 μ L again, terminator Na that concentration is 0.2 mol/L 2cO 3solution, surveys OD value under 405 nm wavelength.）
Do simultaneously the blank group under same system, blank group, sample controls group, sample sets, calculate suppression ratio by computing formula below, and obtain corresponding IC with Origin 6.0 software 50value.
Said method is adopted to determine suppression ratio and the IC of mountain Folium Ligustri pubescentis (leaf) and the sample such as mountain Folium Ligustri pubescentis (stem) each effective site (petroleum ether part, ethyl acetate extract and n-butanol portion) and positive control acarbose in embodiment 1 and embodiment 4 respectively 50value.
experimental result
Shown in table 1 is various sample α-glucosidase inhibitory active, Fig. 1 is mountain Folium Ligustri pubescentis (stem) each sample α-Glucosidase inhibitor rate is with the variation diagram of drug level.Fig. 2 is mountain Folium Ligustri pubescentis (leaf) each sample α-Glucosidase inhibitor rate is with the variation diagram of drug level.
As can be seen from Table 1, except the inhibitory action at mountain Folium Ligustri pubescentis (leaf) BU position is more weak, the IC of other mountain Folium Ligustri pubescentis (leaf) and mountain Folium Ligustri pubescentis (stem) effective site 50value is all much smaller than positive control drug acarbose, and this shows that the 5 groups of active sites except mountain Folium Ligustri pubescentis (leaf) BU position extracted from mountain Folium Ligustri pubescentis (stem) and mountain Folium Ligustri pubescentis (leaf) all have well α-glucosidase activity inhibitory action.Wherein, the IC of mountain Folium Ligustri pubescentis (leaf) ethyl acetate extract 50being worth minimum, showing its inhibition best, is secondly mountain Folium Ligustri pubescentis (stem) n-butanol portion, and the IC of mountain Folium Ligustri pubescentis (leaf) petroleum ether part 50be worth maximum.
Fig. 1 and Fig. 2 all shows, mountain of the present invention Folium Ligustri pubescentis (stem) and mountain Folium Ligustri pubescentis (leaf) blood sugar lowering effective site pair αthe inhibit activities of-glucosidase is all in dose dependent.Petroleum ether part suppression ratio when reaching finite concentration of 3 positions of the mountain Folium Ligustri pubescentis (stem) of Fig. 1 and the mountain Folium Ligustri pubescentis (leaf) of Fig. 2 levels off to stable, but be all when concentration is higher than 750 μ g/mL, suppression ratio declines on the contrary to some extent, namely suppression ratio when concentration is 1500 μ g/mL on the contrary lower than concentration during 750 μ g/mL, the particularly petroleum ether part of mountain Folium Ligustri pubescentis (leaf).Wherein the concentration of the petroleum ether part of mountain Folium Ligustri pubescentis (stem), ethyl acetate extract and n-butanol portion respectively when 93.75 μ g/mL, 46.86 μ g/mL, 23.44 μ g/mL suppression ratio level off to stable, and the ethyl acetate extract of mountain Folium Ligustri pubescentis (leaf) dense be eventually that 11.72 μ g/mL suppression ratio level off to stable.For the reason declined higher than the suppression ratio after 750 μ g/mL may be: when concentration reaches 1500 μ g/mL, excessive concentration is right on the contrary αthe suppression of-glucosidase activity weakens to some extent.
To sum up, several groups of positions of several groups of mountain Folium Ligustri pubescentiss (stem) that the present invention provides and mountain Folium Ligustri pubescentis (leaf), except mountain Folium Ligustri pubescentis (leaf) n-butanol portion, other 5 effective sites pair α-glucosidase all has very strong inhibitory action, wherein obvious with the ethyl acetate extract effect of the n-butanol portion of mountain Folium Ligustri pubescentis (stem) and mountain Folium Ligustri pubescentis (leaf), is good α-glucosidase inhibitor, may be used for preparing hypoglycemic drug.

Claims (5)

1. mountain Folium Ligustri pubescentis effective site, is characterized in that: be the petroleum ether part of mountain Folium Ligustri pubescentis stem or leaf and/or ethyl acetate extract and/or n-butanol portion.
2. mountain as claimed in claim 1 Folium Ligustri pubescentis effective site, it is characterized in that, the petroleum ether part of mountain Folium Ligustri pubescentis stem or leaf, ethyl acetate extract and n-butanol portion obtain by the following method: leaf or the stem of getting mountain Folium Ligustri pubescentis, pulverize, add ethanol, heating and refluxing extraction, the liquid concentration of extraction obtains total extractum; Be scattered in water by total extractum, extract respectively with petroleum ether, ethyl acetate, n-butyl alcohol, namely the petroleum ether extract obtained, acetic acid ethyl ester extract and n-butyl alcohol extract are respectively petroleum ether part, ethyl acetate extract and n-butanol portion.
3. mountain as claimed in claim 2 Folium Ligustri pubescentis effective site, it is characterized in that, the concentration of ethanol is 65-80V%; Reflux 2-3 time, each 0.5-2h.
4. the extracting method of mountain Folium Ligustri pubescentis effective site described in claim 1, is characterized in that, the first drying of the stem of mountain Folium Ligustri pubescentis or leaf, pulverizing, then add alcohol heating reflux and extract 2-3 time, each 0.5-2h, merge extractive liquid, filtration, concentratedly to obtain always extractum; Total extractum is scattered in water, extract respectively with petroleum ether, ethyl acetate, n-butyl alcohol, volatilize solvent, namely the petroleum ether extract obtained, acetic acid ethyl ester extract and n-butyl alcohol extract are respectively petroleum ether part, ethyl acetate extract and n-butanol portion.
5. mountain according to claim 1 Folium Ligustri pubescentis effective site is in preparation αapplication in-glucosidase inhibitor class medicine.
CN201410618288.9A 2014-11-06 2014-11-06 Mountain herb mixtures tea active component and its extracting method, application Expired - Fee Related CN104435071B (en)

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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103860611A (en) * 2014-01-16 2014-06-18 河南大学 Crape myrtle extract as well as extraction method and application thereof

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103860611A (en) * 2014-01-16 2014-06-18 河南大学 Crape myrtle extract as well as extraction method and application thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
唐春红: "《天然防腐剂与抗氧化剂》", 31 May 2010 *
郭玲，等: "海南山苦茶叶低极性油状物成分的分析", 《中国药学杂志》 *

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