CN109954043A - A method of the extraction purification saponin(e from cherry tomato - Google Patents
A method of the extraction purification saponin(e from cherry tomato Download PDFInfo
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- CN109954043A CN109954043A CN201910358945.3A CN201910358945A CN109954043A CN 109954043 A CN109954043 A CN 109954043A CN 201910358945 A CN201910358945 A CN 201910358945A CN 109954043 A CN109954043 A CN 109954043A
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/81—Solanaceae (Potato family), e.g. tobacco, nightshade, tomato, belladonna, capsicum or jimsonweed
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- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
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- A61K2236/30—Extraction of the material
- A61K2236/31—Extraction of the material involving untreated material, e.g. fruit juice or sap obtained from fresh plants
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- A61K2236/30—Extraction of the material
- A61K2236/37—Extraction at elevated pressure or temperature, e.g. pressurized solvent extraction [PSE], supercritical carbon dioxide extraction or subcritical water extraction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- 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
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- 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
- A61K2236/55—Liquid-liquid separation; Phase separation
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Abstract
The invention belongs to active constituent extractive technique field, a kind of method for specifically disclosing extraction purification saponin(e from cherry tomato.Method for extraction and purification of the present invention is the following steps are included: (1) is beaten: cherry tomato is beaten and adjusts pH；(2) vacuum enzymolysis processing: constantly reducing vacuum degree while being continuously increased enzyme concentration and temperature to carry out enzymolysis processing to holy girl's pulp；(3) further extraction process supercritical carbon dioxide treatment: is carried out using supercritical carbon dioxide enzymatic hydrolysis slurry；(4) it is centrifuged；(5) it purifies: being carried out after separation elution using large pore resin absorption column with the further abstraction purification of n-butanol；(6) dry.Method of pectinase is combined to be vacuum-treated and be extracted with supercritical carbon dioxide extracting to cherry tomato saponin(e by the present invention, it is combined again using adsorption resin column and n-butanol and cherry tomato saponin(e is purified, not only increase the diversity of cherry tomato saponin extraction method, and extraction effect is good, saponin(e yield and purity is high.
Description
[technical field]
The present invention relates to active constituent extractive technique fields, and in particular to a kind of side of the extraction purification saponin(e from cherry tomato
Method.
[background technique]
Cherry tomato is Solanaceae cherry tomato platymiscium, and fruit type is small, moisture is more, and vitamin content is higher than common cherry tomato,
Substance also containing the cancer-resistings such as glutathione and cherry tomato red pigment, the deep favor by the majority of consumers, by the agriculture of joint League of Nations
Tissue is classified as one of the fruits and vegetables preferentially promoted.Main nutritional ingredient is exactly cherry tomato red pigment in cherry tomato, is carrotene
It is a kind of.Cherry tomato red pigment is a kind of fat-soluble unsaturated hydrocarbon, in cherry tomato other than fat-soluble compound still
There is the not low water-soluble soap glycoside compound of content.Cherry tomato saponin(e has strong reducing blood lipid, norcholesterol, anti-artery hard
Change, is anti-oxidant, inhibiting the physiological activity such as cancer cell multiplication.
Currently, generalling use pectin enzymatic isolation method to extract cherry tomato saponin(e, extracting method is single and recovery rate is lower.
[summary of the invention]
Goal of the invention of the invention is: in view of the above problems, providing a kind of extracting method of cherry tomato saponin(e.
Method of pectinase is combined to be vacuum-treated and be extracted with supercritical carbon dioxide extracting to cherry tomato saponin(e by the present invention, then using suction
Attached resin column and n-butanol, which combine, purifies cherry tomato saponin(e, not only increases the multiplicity of cherry tomato saponin extraction method
Property, and extraction effect is good, cherry tomato saponin(e yield and purity is high.
To achieve the goals above, The technical solution adopted by the invention is as follows:
A kind of extracting method of cherry tomato saponin(e, comprising the following steps:
(1) it is beaten: after fresh clear cherry tomato wash clean, being put into beater and be beaten, and adjusting pH is 4.0-4.5,
Obtain holy girl's pulp；
(2) vacuum enzymolysis processing: it is holy girl's pulp quality 0.03-0.05% that quality is added into holy girl's pulp
Pectase mix to after uniformly, digest 10-15min at a temperature of the vacuum degree of 0.06-0.08MPa and 48-50 DEG C, obtain the
One enzymatic hydrolysis slurry；It is mixed that the pectase that quality is the first enzymatic hydrolysis slurry quality 0.02-0.03% is added into the first enzymatic hydrolysis slurry
After being bonded to uniformly, 15-20min is digested at a temperature of the vacuum degree of 0.2-0.5MPa and 43-46 DEG C, obtains the second enzymatic hydrolysis slurry；It is past
It is described second enzymatic hydrolysis slurry in be added quality be it is described second enzymatic hydrolysis slurry quality 0.01-0.02% pectase mix to uniformly after,
15-20min is digested at a temperature of the vacuum degree of 0.7-0.9MPa and 40-43 DEG C, obtains third enzymatic hydrolysis slurry；
(3) supercritical carbon dioxide treatment: above-mentioned third enzymatic hydrolysis slurry is put into supercritical carbon dioxide extracting device, in 40
DEG C temperature and 10-12MPa pressure under extract 20-35min, slurry must be extracted；
(4) it is centrifuged: mixing toward extractions slurry addition water to after uniform, put into a centrifuge centrifugation, filter to obtain supernatant；
((5) purifying: above-mentioned supernatant being adsorbed up to D101 large pore resin absorption column and is enriched with, first with 6-8 times of column volume
Deionized water elutes impurity, then the ethanol elution for being 80-90% with volume fraction, collects eluent, adds into the eluent
It is colourless to n-butanol layer to enter to be saturated extracting n-butyl alcohol, collects n-butanol phase；
(6) dry: the n-butanol is mutually dried after being concentrated under reduced pressure to get the cherry tomato saponin(e.
Further, in step (3), the third enzymatic hydrolysis slurry is put into before supercritical carbon dioxide extracting device, first by pH
It is adjusted to 3.5.
Further, in step (4), the additional amount of the water is the 1/2 of the extraction slurry product.
Further, in step (4), the revolving speed of the centrifuge is 2500-3000r/min.
Further, in step (5), the absorption amount of resin of the large pore resin absorption column is 130-140mL.
Further, in step (5), the dosage of the ethyl alcohol is 3-4 times of column volume.
In conclusion by adopting the above-described technical solution, the beneficial effects of the present invention are:
The present invention mentions cherry tomato saponin(e using vacuum aided pectinase enzymatic hydrolysis combination supercritical carbon dioxide treatment
It takes, is extracted compared to only with pectinase treatment, effectively improve the recovery rate of cherry tomato saponin(e, extraction effect is good, mentions
It takes efficiency and DNA purity high, a kind of new, efficient method is provided for cherry tomato saponin(e, conducive to promotion cherry tomato
Saponin(e further researchs and develops.Wherein, vacuum aided pectase is digested, entire enzymolysis process only time-consuming 40-55min,
35min or more is shortened relative to enzymatic hydrolysis time-consuming is carried out only with pectase, it is high-efficient, meanwhile, vacuum-assisted processing facilitates
The cell wall for destroying cherry tomato cell, promotes the dissolution of the intracellular active principle of cherry tomato, and then improve cherry tomato saponin(e
Yield further in enzymolysis process, constantly reduce vacuum degree and temperature while being continuously increased enzyme concentration, enzyme concentration,
The variation of vacuum degree and temperature combines, and further promotes the dissolution of the intracellular active principle of cherry tomato, cherry tomato saponin(e yield
It is improved than being all made of the cherry tomato saponin(e yield that constant enzyme concentration, vacuum degree and temperature carry out enzymolysis processing to holy girl's pulp
7.13-8.11%, meanwhile, than only constant enzyme concentration, only constant vacuum or only steady temperature handles holy girl's pulp
Cherry tomato saponin(e yield improve 4.29-5.54%, 4.38-5.77% and 2.15-2.68% respectively；Supercritical carbon dioxide
Suitable for the extraction of liposoluble substance, and cherry tomato saponin(e is water-soluble substances, if carrying out only with supercritical carbon dioxide holy
Female's fruit saponin extraction extracts difficulty greatly and extract impurity is more, and cherry tomato saponin(e yield is lower than 0.1%, purity lower than 93%, and
The application extracts in conjunction with supercritical carbon dioxide after vacuum aided enzymatic hydrolysis, has effectively facilitated the molten of active principle instead
Out, the destruction of effective component is effectively prevented, and avoids the dissolution mixing of impurity, to further improve cherry tomato soap
The yield and purity of glycosides.Finally, successively being purified using macroporous absorbent resin and n-butanol to supernatant, effectively removes and mentioned
The impurity in object is taken, the purity of saponin(e is improved, compared to being purified by flash only with macroporous absorbent resin or only with just
Butanol carries out the saponin(e that abstraction purification obtains, and the saponin(e purity that the application purifies improves 2.1-3.3%.
[specific embodiment]
Below in conjunction with specific embodiment, the present invention is further illustrated.
Embodiment 1
A method of the extraction purification saponin(e from cherry tomato, comprising the following steps:
(1) it is beaten: after fresh clear cherry tomato wash clean, being put into beater and be beaten, and adjusting pH is 4.0, is obtained holy
Female's pulp；
(2) fruit that quality is holy girl's pulp quality 0.03% vacuum enzymolysis processing: is added into holy girl's pulp
Glue enzyme mix to uniformly after, digest 10min at a temperature of the vacuum degree of 0.06MPa and 48 DEG C, obtain the first enzymatic hydrolysis slurry；Described in
First enzymatic hydrolysis slurry in be added quality be it is described first enzymatic hydrolysis slurry quality 0.02% pectase mix to uniformly after, in 0.2MPa's
15min is digested at a temperature of vacuum degree and 43 DEG C, obtains the second enzymatic hydrolysis slurry；It is described that quality, which is added, into the second enzymatic hydrolysis slurry
Two enzymatic hydrolysis slurry quality 0.01% pectases mix to uniformly after, digested at a temperature of the vacuum degree of 0.7MPa and 40 DEG C
15min obtains third enzymatic hydrolysis slurry；
(3) supercritical carbon dioxide treatment: after the pH of above-mentioned third enzymatic hydrolysis slurry is adjusted to 3.5, it is put into overcritical titanium dioxide
In carbon extractor, 20min is extracted at 40 DEG C of temperature and the pressure of 10MPa, slurry must be extracted；
(4) be centrifuged: 2:1 mixes extractions slurry and water to after uniform by volume, puts into a centrifuge in 2500r/
It is centrifuged under the revolving speed of min, filters to obtain supernatant；
(5) it purifies: above-mentioned supernatant is adsorbed into richness up to the D101 large pore resin absorption column that absorption amount of resin is 130mL
Collection first elutes impurity, then the ethanol elution for being 80% with 3 times of column volumes and volume fraction with the deionized water of 6 times of column volumes, receives
Collect eluent, addition saturation extracting n-butyl alcohol is colourless to n-butanol layer into the eluent, collects n-butanol phase；
(6) dry: the n-butanol is mutually dried after being concentrated under reduced pressure to get the cherry tomato saponin(e.
Embodiment 2
A method of the extraction purification saponin(e from cherry tomato, comprising the following steps:
(1) it is beaten: after fresh clear cherry tomato wash clean, being put into beater and be beaten, and adjusting pH is 4.2, is obtained holy
Female's pulp；
(2) fruit that quality is holy girl's pulp quality 0.04% vacuum enzymolysis processing: is added into holy girl's pulp
Glue enzyme mix to uniformly after, digest 12min at a temperature of the vacuum degree of 0.07MPa and 49 DEG C, obtain the first enzymatic hydrolysis slurry；Described in
First enzymatic hydrolysis slurry in be added quality be it is described first enzymatic hydrolysis slurry quality 0.025% pectase mix to uniformly after, in 0.4MPa
Vacuum degree and 45 DEG C at a temperature of digest 17min, obtain the second enzymatic hydrolysis slurry；It is described that quality, which is added, into the second enzymatic hydrolysis slurry
Second enzymatic hydrolysis slurry quality 0.015% pectase mix to uniformly after, digested at a temperature of the vacuum degree of 0.8MPa and 42 DEG C
18min obtains third enzymatic hydrolysis slurry；
(3) supercritical carbon dioxide treatment: after the pH of above-mentioned third enzymatic hydrolysis slurry is adjusted to 3.5, it is put into overcritical titanium dioxide
In carbon extractor, 30min is extracted at 40 DEG C of temperature and the pressure of 11MPa, slurry must be extracted；
(4) be centrifuged: 2:1 mixes extractions slurry and water to after uniform by volume, puts into a centrifuge in 2700r/
It is centrifuged under the revolving speed of min, filters to obtain supernatant；
(5) it purifies: above-mentioned supernatant is adsorbed into richness up to the D101 large pore resin absorption column that absorption amount of resin is 135mL
Collection first elutes impurity, then the ethanol elution for being 85% with 3.5 times of column volumes and volume fraction with the deionized water of 7 times of column volumes,
Eluent is collected, addition saturation extracting n-butyl alcohol is colourless to n-butanol layer into the eluent, collects n-butanol phase；
(6) dry: the n-butanol is mutually dried after being concentrated under reduced pressure to get the cherry tomato saponin(e.
Embodiment 3
A method of the extraction purification saponin(e from cherry tomato, comprising the following steps:
(1) it is beaten: after fresh clear cherry tomato wash clean, being put into beater and be beaten, and adjusting pH is 4.5, is obtained holy
Female's pulp；
(2) fruit that quality is holy girl's pulp quality 0.05% vacuum enzymolysis processing: is added into holy girl's pulp
Glue enzyme mix to uniformly after, digest 15min at a temperature of the vacuum degree of 0.08MPa and 50 DEG C, obtain the first enzymatic hydrolysis slurry；Described in
First enzymatic hydrolysis slurry in be added quality be it is described first enzymatic hydrolysis slurry quality 0.03% pectase mix to uniformly after, in 0.5MPa's
20min is digested at a temperature of vacuum degree and 46 DEG C, obtains the second enzymatic hydrolysis slurry；It is described that quality, which is added, into the second enzymatic hydrolysis slurry
Two enzymatic hydrolysis slurry quality 0.02% pectases mix to uniformly after, digested at a temperature of the vacuum degree of 0.9MPa and 43 DEG C
20min obtains third enzymatic hydrolysis slurry；
(3) supercritical carbon dioxide treatment: after the pH of above-mentioned third enzymatic hydrolysis slurry is adjusted to 3.5, it is put into overcritical titanium dioxide
In carbon extractor, 35min is extracted at 40 DEG C of temperature and the pressure of 12MPa, slurry must be extracted；
(4) be centrifuged: 2:1 mixes extractions slurry and water to after uniform by volume, puts into a centrifuge in 3000r/
It is centrifuged under the revolving speed of min, filters to obtain supernatant；
(5) it purifies: above-mentioned supernatant is adsorbed into richness up to the D101 large pore resin absorption column that absorption amount of resin is 135mL
Collection first elutes impurity, then the ethanol elution for being 90% with 4 times of column volumes and volume fraction with the deionized water of 8 times of column volumes, receives
Collect eluent, addition saturation extracting n-butyl alcohol is colourless to n-butanol layer into the eluent, collects n-butanol phase；
(6) dry: the n-butanol is mutually dried after being concentrated under reduced pressure to get the cherry tomato saponin(e.
Compliance test result:
Using the cherry tomato saponin extraction purification process of 1-3 of the embodiment of the present invention as experimental group 1-3；
After being digested only with pectase, centrifugation immediately, purifying, dry cherry tomato saponin extraction method are the most right
According to group 1；
With cherry tomato saponin extraction purification process substantially the same manner as Example 1 as a control group 2, the difference is that:
Enzymolysis processing is not only carried out using vacuum aided；
With cherry tomato saponin extraction purification process substantially the same manner as Example 1 as a control group 3, the difference is that:
Only do not handled using supercritical carbon dioxide.
In the case where remaining is consistent, holy girl's fruit total saponin that above-mentioned 4 groups of method for extraction and purification extract is calculated
Yield and purity (related assays method is the prior art, then this is not just repeated), the results are shown in Table 1:
The cherry tomato total saponin and purity (%) of 1 each group extracting method of table
| Group | Experimental group 1 | Experimental group 2 | Experimental group 3 | Control group 1 | Control group 2 | Control group 3 |
| Total saponin | 4.45 | 4.59 | 4.49 | 0.24 | 0.35 | 0.29 |
| Saponin(e purity | 98.3 | 99.2 | 98.7 | 94.6 | 88.5 | 91.4 |
As shown in Table 1, cherry tomato total saponin: experimental group 1-3 > 3 > control group of control group, 2 > control group 1, purity: real
A group 1-3 > 1 > control group of control group, 3 > control group 2 is tested, illustrates the application by vacuum-assisted processing and supercritical carbon dioxide treatment
It is combined with pectinase enzymatic hydrolysis processing, can produce synergistic effect, can effectively promote the yield and purity of holy girl's fruit total saponin, mention
Take effect good.
Above description is the detailed description for the present invention preferably possible embodiments, but embodiment is not limited to this hair
Bright patent claim, it is all the present invention suggested by technical spirit under completed same changes or modifications change, should all belong to
In the covered the scope of the patents of the present invention.
Claims (6)
1. a kind of method of the extraction purification saponin(e from cherry tomato, which comprises the following steps:
(1) it is beaten: after fresh clear cherry tomato wash clean, being put into beater and be beaten, and adjusting pH is 4.0-4.5, is obtained holy
Female's pulp；
(2) fruit that quality is holy girl's pulp quality 0.03-0.05% vacuum enzymolysis processing: is added into holy girl's pulp
Glue enzyme mix to uniformly after, digest 10-15min at a temperature of the vacuum degree of 0.06-0.08MPa and 48-50 DEG C, obtain the first enzyme
Solution slurry；Toward it is described first enzymatic hydrolysis slurry in be added quality be it is described first enzymatic hydrolysis slurry quality 0.02-0.03% pectase mix to
After uniformly, 15-20min is digested at a temperature of the vacuum degree of 0.2-0.5MPa and 43-46 DEG C, obtains the second enzymatic hydrolysis slurry；Described in
Second enzymatic hydrolysis slurry in be added quality be it is described second enzymatic hydrolysis slurry quality 0.01-0.02% pectase mix to uniformly after, in
15-20min is digested at a temperature of the vacuum degree of 0.7-0.9MPa and 40-43 DEG C, obtains third enzymatic hydrolysis slurry；
(3) supercritical carbon dioxide treatment: above-mentioned third enzymatic hydrolysis slurry is put into supercritical carbon dioxide extracting device, in 40 DEG C
20-35min is extracted under temperature and the pressure of 10-12MPa, slurry must be extracted；
(4) it is centrifuged: mixing toward extractions slurry addition water to after uniform, put into a centrifuge centrifugation, filter to obtain supernatant；
(5) purify: above-mentioned supernatant being adsorbed up to D101 large pore resin absorption column and is enriched with, first with 6-8 times of column volume go from
Sub- water elution impurity, then the ethanol elution for being 80-90% with volume fraction collect eluent, are added into the eluent full
It is colourless to n-butanol layer with extracting n-butyl alcohol, collect n-butanol phase；
(6) dry: the n-butanol is mutually dried after being concentrated under reduced pressure to get the cherry tomato saponin(e.
2. a kind of method of the extraction purification saponin(e from cherry tomato according to claim 1, which is characterized in that in step (3),
The third enzymatic hydrolysis slurry is put into before supercritical carbon dioxide extracting device, and pH is first adjusted to 3.5.
3. a kind of method of the extraction purification saponin(e from cherry tomato according to claim 1, which is characterized in that in step (4),
The additional amount of the water is the 1/2 of the extraction slurry product.
4. a kind of method of the extraction purification saponin(e from cherry tomato according to claim 1, which is characterized in that in step (4),
The revolving speed of the centrifuge is 2500-3000r/min.
5. a kind of method of the extraction purification saponin(e from cherry tomato according to claim 1, which is characterized in that in step (5),
The absorption amount of resin of the large pore resin absorption column is 130-140mL.
6. a kind of method of the extraction purification saponin(e from cherry tomato according to claim 1, which is characterized in that in step (5),
The dosage of the ethyl alcohol is 3-4 times of column volume.
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Citations (3)
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|---|---|---|---|---|
| CN101804122A (en) * | 2010-04-09 | 2010-08-18 | 广西壮族自治区中国科学院广西植物研究所 | Tomato water-soluble saponin extract and preparation method and application thereof |
| CN106963844A (en) * | 2017-03-28 | 2017-07-21 | 广西壮族自治区中国科学院广西植物研究所 | Application of the tomato water-soluble saponin extract in treatment antihyperuricemic disease drug or health products are prepared |
| JP2017203012A (en) * | 2016-05-13 | 2017-11-16 | 株式会社Ｎｉｌ | Hair remedy |
-
2019
- 2019-04-30 CN CN201910358945.3A patent/CN109954043A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101804122A (en) * | 2010-04-09 | 2010-08-18 | 广西壮族自治区中国科学院广西植物研究所 | Tomato water-soluble saponin extract and preparation method and application thereof |
| JP2017203012A (en) * | 2016-05-13 | 2017-11-16 | 株式会社Ｎｉｌ | Hair remedy |
| CN106963844A (en) * | 2017-03-28 | 2017-07-21 | 广西壮族自治区中国科学院广西植物研究所 | Application of the tomato water-soluble saponin extract in treatment antihyperuricemic disease drug or health products are prepared |
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| 林顺权: "《园艺植物生物技术》", 31 August 2007, 中国农业出版社 * |
| 梁艺英: "《保健食品研发与审评》", 30 November 2012, 中国医药科技出版社 * |
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| 赵陆华: "《中药高效液相色谱法应用》", 31 December 2004, 中国医药科技出版社 * |
| 韩丽: "《实用中药制剂新技术》", 31 October 2002, 化学工业出版社 * |
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