CN103893306A - Blackberry seed active ingredient and extraction method and application thereof - Google Patents
Blackberry seed active ingredient and extraction method and application thereof Download PDFInfo
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- CN103893306A CN103893306A CN201410081276.7A CN201410081276A CN103893306A CN 103893306 A CN103893306 A CN 103893306A CN 201410081276 A CN201410081276 A CN 201410081276A CN 103893306 A CN103893306 A CN 103893306A
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- petroleum ether
- blackberry seed
- blackberry
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- ethyl acetate
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Abstract
The invention belongs to the technical field of plant extraction, and particularly relates to a blackberry seed active ingredient and an extraction method and application thereof. The method comprises the following steps: performing cold soaking on blackberry seeds by using petroleum ether for degreasing; performing cold soaking on degreased blackberry seed residues by using ethanol; concentrating an ethanol extracting solution to obtain an ethanol total extract; dispersing the ethanol total extract into water; and extracting by using petroleum ether, ethyl acetate and n-butyl alcohol to obtain an petroleum ether extract, an ethyl acetate extraction and an n-butyl alcohol extract, namely, the petroleum ether fraction, ethyl acetate fraction and n-butyl alcohol fraction of blackberry seeds respectively. The petroleum ether fraction, the ethyl acetate fraction, the n-butyl alcohol fraction and blackberry seed oil are extracted from blackberry seeds by adopting the novel extraction method, and these active ingredients have certain functions of increasing the content of liver glycogen as proved by researches.
Description
technical field
The invention belongs to technical field of plant extraction, be specifically related to a kind of blackberry seed effective ingredient and extracting method thereof, application.
background technology
Blackberry (
rubusspp. Blackberry) belong to Rosaceae (Rasaceae) rubus polymerization fruit plant, its fruit has very high nutritive value and medical value, there is the human body of improvement metabolism, remove the functions such as free radical, slow down aging, enhancing immunity, have especially and reduce cholesterol level, control heart disease, anticancer (reducing the toxic and side effects that chemotherapy causes) and antithrombotic effect.
Blackberry seed is the by-product of the converted productss such as blackberry fruit wine, blackberry juice, contains abundant polyphenol compound, is abandoned in a large number for many years, causes a large amount of wastes of resource.In recent years, technical staff had extracted blackberry seed oil from blackberry seed.
Research shows, blackberry seed oil contains higher unsaturated fatty acid, as Palmic acid, stearic acid, oleic acid, linoleic acid plus linolenic acid etc., can offer the basic fatty acid of human body, and these fatty acids can not be synthetic by any known chemistry route, must from food, obtain.In addition, seed oil has effective removing free radical, effectively suppresses H
2o
2mouse red blood cell oxidative hemolysis and the VC-FeSO of induction
4the murine liver tissue malonaldehyde of system induction generates and VC-FeSO
4the effect of the Mouse Liver mitochondrial swelling of system induction, cardiovascular lipid oxidation anti-with it and active anticancer are closely bound up; In addition, also find that the main effective ingredient of blackberry seed after defat is procyanidin, it is the polyphenol compound being formed by connecting with different polymerization degree by monomer flavan-3-alcohol, there is the ability of efficient removing free radical, there is the extensive uses such as anti-swollen, the mutation for the treatment of arteriosclerosis, antiinflammatory and prophylaxis of cancer.
summary of the invention
The present invention is in order to expand the application of blackberry seed, for reasonable development with utilize blackberry seed Resource Supply scientific basis, provides a kind of blackberry seed effective ingredient and extracting method thereof, application.
The present invention is by the following technical solutions:
A kind of extracting method of blackberry seed effective ingredient, effective ingredient is the petroleum ether part of blackberry seed, ethyl acetate extract, n-butanol portion and/or blackberry seed oil, when extraction, comprise the following steps: the defat of blackberry seed petroleum ether merceration, blackberry seed after defat is ethanol (65-80V%) merceration for residue, ethanol extract concentrates to obtain total ethanol extractum, total ethanol extractum is scattered in water, use successively petroleum ether, ethyl acetate, n-butyl alcohol extracts, the petroleum ether extract obtaining, acetic acid ethyl ester extract and n-butyl alcohol extract, be respectively the petroleum ether part of blackberry seed, ethyl acetate extract and n-butanol portion.
In order to improve the extraction effect of blackberry seed effective ingredient, can before petroleum ether merceration, blackberry seed be pulverized.
Extracting solution after defat reclaims petroleum ether and obtains blackberry seed oil.
When the defat of petroleum ether merceration, repeat 4-5 time each 3-4d, merge extractive liquid.
The consumption of each petroleum ether is 8-10 times of blackberry seed quality, and merceration temperature is room temperature.
When ethanol merceration, repeat 2-3 time, each 3-4d, merges ethanol extract.
The consumption of each ethanol is 8-10 times of blackberry seed residue quality, and merceration temperature is room temperature.
The blackberry seed effective ingredient that adopts above extracting method to obtain, is characterized in that, is petroleum ether part, ethyl acetate extract, n-butanol portion and/or the blackberry seed oil of blackberry seed.
The application of blackberry seed effective ingredient aspect preparation rising liver glycogen medicine.
The present invention adopts new extracting method from blackberry seed, to extract petroleum ether part, ethyl acetate extract, n-butanol portion, blackberry seed oil, and adopt tail vein injection alloxan to induce diabetic mice model, study first the impact of blackberry seed on diabetic mice liver glycogen, studies confirm that these effective ingredient all have the effect of certain rising liver glycogen content, can infer that it may have the effect of synthesizing, reduce the decomposition of liver glycogen that promotes liver glycogen.
detailed description of the invention
Embodiment 1
The extracting method of blackberry seed effective ingredient, step is as follows: 500g blackberry seed is pulverized, and uses the defat of 4000mL petroleum ether merceration at every turn, repeats 4 times, and each 4d, filters, merge extractive liquid,, extracting solution reclaims petroleum ether and obtains blackberry seed oil 145.04g;
2800mL ethanol (70V%) merceration for blackberry seed residue 341g after defat, repeat 3 times, each 3d, merge ethanol extract, ethanol extract concentrates to obtain total ethanol extractum, and total ethanol extractum is scattered in water, extracts successively with petroleum ether, ethyl acetate, n-butyl alcohol, petroleum ether extract 2.34g, the acetic acid ethyl ester extract 5.46g and the n-butyl alcohol extract 13.42g that obtain, be respectively petroleum ether part, ethyl acetate extract and the n-butanol portion of blackberry seed.
Embodiment 2
The extracting method of blackberry seed effective ingredient, step is as follows: 500g blackberry seed is pulverized, and uses the defat of 5000mL petroleum ether merceration at every turn, repeats 5 times, and each 3d, filters, merge extractive liquid,, extracting solution reclaims petroleum ether and obtains blackberry seed oil 162.50g;
3000mL ethanol (65V%) merceration for blackberry seed residue 332g after defat, repeat 2 times, each 4d, merge ethanol extract, ethanol extract concentrates to obtain total ethanol extractum, and total ethanol extractum is scattered in water, extracts successively with petroleum ether, ethyl acetate, n-butyl alcohol, petroleum ether extract 2.02g, the acetic acid ethyl ester extract 5.10g and the n-butyl alcohol extract 14.28g that obtain, be respectively petroleum ether part, ethyl acetate extract and the n-butanol portion of blackberry seed.
Embodiment 3
The extracting method of blackberry seed effective ingredient, step is as follows: 500g blackberry seed is pulverized, and uses the defat of 4500mL petroleum ether merceration at every turn, repeats 4 times, each 4d, merge extractive liquid,, extracting solution reclaims petroleum ether and obtains blackberry seed oil 162.26g;
3300mL ethanol (80V%) merceration for blackberry seed residue 335g after defat, repeat 3 times, each 4d, merge ethanol extract, ethanol extract concentrates to obtain total ethanol extractum, and total ethanol extractum is scattered in water, extracts successively with petroleum ether, ethyl acetate, n-butyl alcohol, petroleum ether extract 2.26g, the acetic acid ethyl ester extract 5.54g and the n-butyl alcohol extract 12.94g that obtain, be respectively petroleum ether part, ethyl acetate extract and the n-butanol portion of blackberry seed.
The impact of the liver glycogen of the diabetic mice on alloxan induction in effect experiment blackberry seed effective ingredient body
Petroleum ether part, ethyl acetate extract, n-butanol portion, blackberry seed oil that experimental raw: embodiment 1 makes.
Laboratory animal: Kunming mouse, male, body weight 22 ± 2 g, SPF level, is provided production licence number: SCXD Shandong 20080002 by Lukang Medical Co., Ltd., Shandong's Product Quality Verification Centers Animal Lab..
Experiment reagent: glucose assays test kit (ShangHai RongSheng Biology Pharmacy Co., Ltd, 20131006147); Liver/inose unit measures test kit (Bioengineering Research Institute is built up in Nanjing, 20131216); Acarbose (Acarbose, 130523, Hua Cheng pharmaceutical Co. Ltd of Hangzhou Sino-U.S.); Alloxan (Alloxan monohydrate 98%, Lot:10150059, Alfa Aesar).
Experimental apparatus: LRH-150 constant incubator; DELTA 320 type pH meters; Multiskan MK3 microplate reader; Electronic balance; Rotary Evaporators; UV-2000 type ultraviolet-uisible spectrophotometer; CS-H1 type blender; Micropipettor and the rifle of each specification are first-class.
Experimental technique and result
1, experimental technique
Animal grouping and modeling method: 150 of healthy kunming mices, be divided at random two groups, 10 of Normal groups, remaining is diabetes modeling group.Diabetes modeling group be can't help after water 12 h in fasting, tail vein injection 80mg/kg alloxan induction diabetic mice.Normal nursing after 72 h, water 12 h are can't help in fasting, eye socket is got blood, surveys fasting blood glucose level, and according to blood glucose homeostatic principle, by blood sugar level, the mice more than 11.1 mmol/L is divided into 14 groups: the high, medium and low dosage group of blackberry seed petroleum ether part, the high, medium and low dosage group of blackberry seed ethyl acetate extract, the high, medium and low dosage group of blackberry seed n-butanol portion, the high, medium and low dosage group of blackberry seed oil, acarbose positive controls, and diabetic model group, Normal group is as blank group.Except diabetic model group and blank group, other groups corresponding medicine of gavages respectively, 1 time/d, successive administration 7 d, and can't help water 12 h in fasting on that night, the 8th d wins eyeball and gets blood execution animal, get liver and weigh, preparing liver glycogen and measure liquid, for the mensuration of liver glycogen content.
2, experimental result is as follows
The impact of the different effective ingredient of table 1 on blood glucose in diabetic mice (
`
x±
s ,
n=10)
Note: the positive contrast of acarbose;
With the comparison of blank group:
△ △ △ p< 0.001,
△ △ p< 0.01,
△ p< 0.05;
With model group comparison:
* * p< 0.001,
* p< 0.01,
* p< 0.05;
With positive controls comparison:
### p< 0.001,
## p< 0.01,
# p< 0.05.
The impact of the different effective ingredient of table 2 on diabetic mice liver glycogen (
`
x±
s ,
n=10)
Note: the positive contrast of acarbose;
With the comparison of blank group:
△ △ △ p< 0.001,
△ △ p< 0.01,
△ p< 0.05;
With model group comparison:
* * p< 0.001,
* p< 0.01,
* p< 0.05.
(1) as can be seen from Table 1, before administration, compared with blank group, diabetic groups (petroleum ether part, ethyl acetate extract, n-butanol portion, blackberry seed oil, positive controls and model group) fasting glucose be all significance raise (
p<0.001), alloxan induction diabetic mice modeling success is described, and there was no significant difference between each administration group (
p>0.001).
(2) as can be seen from Table 2, compared with blank group, the reduction of model group Mouse Liver glycogen content significance (
p<0.01); After drug treatment 7 d, compared with model group, the content of the equal energy of low dose group, ethyl acetate extract senior middle school dosage group, blackberry seed oil low dose group and positive control significance ground rising liver glycogen in petroleum ether part (
p<0.05), though the content there was no significant difference of other each dosage group rising liver glycogens, liver glycogen content is all higher than model group.Visible, petroleum ether part, ethyl acetate extract, n-butanol portion and blackberry seed oil all have the effect of certain rising liver glycogen content, infer and have the decomposition of synthesizing, reduce liver glycogen that promotes liver glycogen.
Claims (6)
1. the extracting method of a blackberry seed effective ingredient, it is characterized in that, effective ingredient is the petroleum ether part of blackberry seed, ethyl acetate extract, n-butanol portion and/or blackberry seed oil, when extraction, comprise the following steps: the defat of blackberry seed petroleum ether merceration, blackberry seed residue ethanol merceration after defat, ethanol extract concentrates to obtain total ethanol extractum, total ethanol extractum is scattered in water, with petroleum ether, ethyl acetate, n-butyl alcohol extracts, the petroleum ether extract obtaining, acetic acid ethyl ester extract and n-butyl alcohol extract are respectively the petroleum ether part of blackberry seed, ethyl acetate extract and n-butanol portion.
2. the extracting method of blackberry seed effective ingredient as claimed in claim 1, is characterized in that, the extracting solution after defat reclaims petroleum ether and obtains blackberry seed oil.
3. the extracting method of blackberry seed effective ingredient as claimed in claim 1, is characterized in that, when the defat of petroleum ether merceration, repeats 4-5 time, each 3-4d, merge extractive liquid.
4. the extracting method of blackberry seed effective ingredient as claimed in claim 1, is characterized in that, when ethanol merceration, repeats 2-3 time, and each 3-4d, merges ethanol extract.
5. the blackberry seed effective ingredient that adopts the extracting method of claim 1 or 2 to obtain, is characterized in that, is petroleum ether part, ethyl acetate extract, n-butanol portion and/or the blackberry seed oil of blackberry seed.
6. the application of blackberry seed effective ingredient claimed in claim 5 aspect preparation rising liver glycogen medicine.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104478714A (en) * | 2014-12-24 | 2015-04-01 | 淮海工学院 | Method for extracting ethyl linolenate from blackberry seed oil |
| CN104783271A (en) * | 2015-02-25 | 2015-07-22 | 南京舜安食品保鲜有限公司 | Low-temperature hot-filling method for producing blackberry raw solution |
| CN105017375A (en) * | 2015-08-14 | 2015-11-04 | 河南大学 | Anticoagulation blackberry seed effective component and extraction and separation method and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1788755A (en) * | 2004-12-14 | 2006-06-21 | 北京市农林科学院 | Framberry leaf extract effective part and its composition and uses |
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2014
- 2014-03-07 CN CN201410081276.7A patent/CN103893306B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1788755A (en) * | 2004-12-14 | 2006-06-21 | 北京市农林科学院 | Framberry leaf extract effective part and its composition and uses |
Non-Patent Citations (5)
| Title |
|---|
| 张东升等: "黑莓籽油脂肪酸成分及抗氧化活性研究", 《中国粮油学报》 * |
| 张东升等: "黑莓籽油脂肪酸成分及抗氧化活性研究", 《中国粮油学报》, vol. 26, no. 11, 30 November 2011 (2011-11-30), pages 55 - 58 * |
| 李思睿: "黑莓籽油的提取、精炼及氧化稳定性的研究", 《中国优秀硕士学位论文全文数据库医药卫生科技辑》 * |
| 李立: "黑莓果渣提取物抗氧化活性评价及提取工艺研究", 《中国优秀硕士学位论文全文数据库医药卫生科技辑》 * |
| 李立: "黑莓果渣提取物抗氧化活性评价及提取工艺研究", 《中国优秀硕士学位论文全文数据库医药卫生科技辑》, no. 10, 15 October 2011 (2011-10-15) * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104478714A (en) * | 2014-12-24 | 2015-04-01 | 淮海工学院 | Method for extracting ethyl linolenate from blackberry seed oil |
| CN104783271A (en) * | 2015-02-25 | 2015-07-22 | 南京舜安食品保鲜有限公司 | Low-temperature hot-filling method for producing blackberry raw solution |
| CN105017375A (en) * | 2015-08-14 | 2015-11-04 | 河南大学 | Anticoagulation blackberry seed effective component and extraction and separation method and application thereof |
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