CN102086220A - Extraction and purification method of phlorizin in litchi peels - Google Patents
Extraction and purification method of phlorizin in litchi peels Download PDFInfo
- Publication number
- CN102086220A CN102086220A CN 201010617724 CN201010617724A CN102086220A CN 102086220 A CN102086220 A CN 102086220A CN 201010617724 CN201010617724 CN 201010617724 CN 201010617724 A CN201010617724 A CN 201010617724A CN 102086220 A CN102086220 A CN 102086220A
- Authority
- CN
- China
- Prior art keywords
- phlorizin
- litchi
- high performance
- behind
- performance liquid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention discloses an extraction and purification method of phlorizin in litchi peels, belonging to the field of cosmetic raw material development in natural plants. The extraction and purification method comprises the steps of smashing, extracting crude extract liquid, enriching litchi rind crude extract liquid, carrying out solid-phase extraction and purifying. Radically found in the litchi by the extraction and purification method, the phlorizin is purified out from the litchi peels and then is applied to cosmetic to serve as a natural raw material with skin whitening and anti-aging activities. The litchi peels has wide sources in Lingnan area of China, and wastes are turned into valuable things.
Description
Technical field
The present invention relates to a kind of extraction and purification process of phlorizin, particularly relate to the extraction and the purification process of phlorizin in a kind of Fructus Litchi, belong to cosmetic material development field in the natural phant.
Background technology
Phloretin and glucosides phlorizin thereof belong to the dihydrochalcone in the flavonoid, are a kind of novel skin whitening agent and the antioxidants of researching and developing recently both at home and abroad.Dihydrochalcone is less in distributed in nature, and Phloretin and phlorizin are present among some plant rhizome and the Gen Pi, and content is more in apple according to the literature, and how exists with the form of phlorizin
[1-2]Lichee (
Litchi chinensis Sonn.) be the characteristic fruit that area, the Chinese south of the Five Ridges abounds with, the research of Fructus Litchi is inferred that tentatively it contains Flavonoid substances
[3]But before this subject study, still from lichee, do not find the bibliographical information of phlorizin both at home and abroad.At present, the method for extracting phlorizin both at home and abroad in natural phant mainly is organic solvent extraction method, ultrasonic wave extraction and microwave assisting method, and the method for separation and purification mainly is high performance liquid chromatography, silica gel column chromatography and thin layer chromatography.
Summary of the invention
Lichee is the bigger seasonal characteristic fruit of Chinese south of the Five Ridges area output, is loved by the people.If can from Fructus Litchi, extract phlorizin and be applied in the makeup as the natural matter of a kind of Pear Power and activity of fighting against senium, wide material sources not only, and turn waste into wealth, both can bring huge economic benefit, can promote the development of local makeup industry core technology again.
The object of the present invention is to provide the extraction and the purification process of phlorizin in a kind of Fructus Litchi.
Purpose of the present invention is achieved by the following technical programs.
The extraction of phlorizin and purification process in a kind of Fructus Litchi, it may further comprise the steps:
1, pulverizes: after fresh lichee pericarp (connecting clothing) dries in the shade, pulverized 40 mesh sieves;
2, extract crude extract: accurately take by weighing Fructus Litchi powder 10.0 g, add 70% ethanol 150mL, supersound process 30 minutes, power 100%; Repeat after the filtration to extract once merging filtrate; Filtrate is evaporated to 70 mL through Rotary Evaporators, add 180 mL dehydrated alcohols and leave standstill, behind the 16h with filter paper filtering; Filtrate decompression is concentrated into ethanol and all volatilizees, and adds distilled water 150mL then, and with ultrasonic assist in dissolving, obtains crude extract after filtering at last;
3, enrichment litchi rind crude extract: with sample on the above-mentioned crude extract to macroporous resin column 16 g, last sample speed 1 mL/min, then with 450 mL distilled water drip washing to colourless, again with 70% ethanol 50mL desorption, flow velocity 1mL/min obtains the litchi rind extracting solution, and gives 4 ℃ and keep in Dark Place;
4, Solid-Phase Extraction: get the solid phase extraction column after above-mentioned litchi rind extracting solution 5mL passes through to activate with the 1mL/min flow velocity, preceding 3mL effluent liquid discards, and all the other effluent liquid are collected; The activation of described solid phase extraction column needs to pass through solid phase extraction column with 10mL methyl alcohol earlier, and with 10mL distilled water flush away methyl alcohol, speed is 1mL/min again;
5, purifying: described purifying adopts high performance liquid chromatography to carry out, and it may further comprise the steps:
(1) high performance liquid chromatography detects the phlorizin standard substance: precision takes by weighing phlorizin standard substance 0.0050 g, be dissolved in the methyl alcohol, be mixed with concentration 0.002 mg/mL standard solution, behind 0.45 μ m filtering with microporous membrane, make high performance liquid chromatography, obtain phlorizin standard substance retention time 21.444min, see Fig. 1;
(2) the collection liquid after the high performance liquid chromatography detection Solid-Phase Extraction: precision takes by weighing collection liquid 0.0050 g after the Solid-Phase Extraction, be dissolved in the methyl alcohol, be mixed with concentration 0.002 mg/mL solution, behind 0.45 μ m filtering with microporous membrane, make high performance liquid chromatography, obtain this collection liquid retention time 21.518min, see Fig. 2.Collect the 21.518min effluent liquid, because the retention time of this retention time and standard substance is roughly the same, so this crest is the target substance phlorizin probably;
(3) effluent liquid that step (2) is collected is concentrated into 120 μ L with Nitrogen evaporator, makes high performance liquid chromatography behind 0.45 μ m filtering with microporous membrane, and the demonstration retention time is that the single crest of 21.758min is monomeric compound---the phlorizin behind the purifying;
Structure is identified: precision takes by weighing phlorizin standard substance 0.0050 g, is dissolved in the methyl alcohol, is mixed with concentration 0.005 mg/mL standard solution.Monomeric compound behind this standard solution and the purifying is made mass spectrometric detection respectively.The main mass spectra peak (see figure 4) of phlorizin standard substance is consistent with the main mass spectra peak (see figure 5) of monomeric compound behind the Fructus Litchi extracting solution purifying.
High performance liquid chromatography is on the liquid column chromatography basis of classics, adopts high-pressure pump, efficient stationary phase and high sensitivity detector, introduces the chromatographic technique that grows up after the gas-chromatography theory.Its principle is to utilize the retention time of every kind of material in stationary phase different and reach isolating purpose.High performance liquid chromatography has following characteristics: (1) chromatographic column can be used repeatedly, and the moving phase range of choice is wide, flows out component and collects easily; (2) separation efficiency height, analysis speed is fast, and is highly sensitive, operation automation.Behind the extraction liquid sample introduction of plant, the different crests that occur can seen soon on the detector on different retention time points, each crest is a kind of compound, area is the content of this compound under the crest, therefore, this is a kind of method of not only can purifying substance but also can carry out quantitative analysis.In addition, high performance liquid chromatography not only can the separate complex sample, and aspect quantitative analysis, has superiority, and Agilent 1100 high performance liquid chromatographs that use in this experiment have photodiode array (PDAD) ultraviolet photometric detector, the scanning of ultraviolet wave spectrum can be provided, and preliminary evaluation flows out the chemical structure of component.
Even with the same material and same chromatographic condition, at every turn also can not be in full accord by the retention time behind the high performance liquid chromatograph, allow certain error.
The discovery that method of the present invention is breakthrough have phlorizin in the lichee, and its natural matter that comes out as a kind of Pear Power and activity of fighting against senium of purifying from Fructus Litchi is applied in the makeup, Fructus Litchi is wide material sources not only in area, the Chinese south of the Five Ridges, and turn waste into wealth, both huge economic benefit can be brought, the development of local makeup industry core technology can be promoted again.
Description of drawings
Fig. 1 is phlorizin standard substance (concentration 0.002 mg/ml) HPLC collection of illustrative plates, retention time=21.444 min;
Fig. 2 is a Fructus Litchi extracting solution HPLC collection of illustrative plates, retention time=visible color spectrum peak, 21.518min place;
Fig. 3 is the monomeric compound HPLC collection of illustrative plates of Fructus Litchi extracting solution after purified, retention time=21.758min;
Fig. 4 is phlorizin standard substance (concentration 0.005 mg/mL) mass spectrums;
Fig. 5 is the monomeric compound mass spectrum behind the Fructus Litchi extracting solution purifying.
Embodiment
The present invention is further illustrated below in conjunction with specific embodiment.
One, prepares key instrument, reagent and material
Agilent 1100 high performance liquid chromatographs (photodiode array detector), Ultimate C18 post (250 * 4.6mm, 5 μ m), and Dikma C18 solid phase extraction column (500 mg, 3mL).
API 4000 QTrap liquid chromatography-triple quadrupole bar/linear ion GC-MS (u.s.a. applied biosystem company), data auto-associating-enhanser ion scan, lock 435.4 m/z, collision energy-15V ,-30V ,-45V three's stack, sweep limit 50-500 m/z.
KQ5200DA type numerical control supersonic cleanser, power 100W.
HGC-12 Nitrogen evaporator (the permanent company difficult to understand in Tianjin), 50 ℃ of temperature.
D101 macroporous resin (Tianjin Nankai with become company).
The phlorizin standard substance are available from U.S. Sigma company, purity 〉=99.0%.
The used methyl alcohol of moving phase is chromatographically pure (Dikma company), and experimental water is a deionized water.All the other reagent are analytical pure.
The lichee kind is the osmanthus flavor, picks up from Chuan Keng orchard, Zengcheng, Guangzhou.
Two, high-efficient liquid phase chromatogram condition
Mobile phase A is a methyl alcohol, Mobile phase B be 0.1% acetate (v:v, pH3.20), sample size 20 μ L, 35 ℃ of column temperatures detect wavelength 286 nm.Eluent gradient and flow velocity see Table 1.
Table 1 moving phase ratio and flow velocity
Three, the extraction and the purification process of phlorizin in a kind of Fructus Litchi of present embodiment, it may further comprise the steps:
The extraction of phlorizin and purification process in a kind of Fructus Litchi, it may further comprise the steps:
1, pulverizes: after fresh lichee pericarp (connecting clothing) dries in the shade, pulverized 40 mesh sieves;
2, extract crude extract: accurately take by weighing Fructus Litchi powder 10.0 g, add 70% ethanol 150mL, supersound process 30 minutes, power 100%; Repeat after the filtration to extract once merging filtrate; Filtrate is evaporated to 70 mL through Rotary Evaporators, add 180 mL dehydrated alcohols and leave standstill, behind the 16h with filter paper filtering; Filtrate decompression is concentrated into ethanol and all volatilizees, and adds distilled water 150mL then, and with ultrasonic assist in dissolving, obtains crude extract after filtering at last;
3, enrichment litchi rind crude extract: with sample on the above-mentioned crude extract to macroporous resin column 16 g, last sample speed 1 mL/min, then with 450 mL distilled water drip washing to colourless, again with 70% ethanol 50mL desorption, flow velocity 1mL/min obtains the litchi rind extracting solution, and gives 4 ℃ and keep in Dark Place;
4, Solid-Phase Extraction: get the solid phase extraction column after above-mentioned litchi rind extracting solution 5mL passes through to activate with the 1mL/min flow velocity, preceding 3mL effluent liquid discards, and all the other effluent liquid are collected; The activation of described solid phase extraction column needs to pass through solid phase extraction column with 10mL methyl alcohol earlier, and with 10mL distilled water flush away methyl alcohol, speed is 1mL/min again;
5, purifying: described purifying adopts high performance liquid chromatography to carry out, and it may further comprise the steps:
A) high performance liquid chromatography detects the phlorizin standard substance: precision takes by weighing phlorizin standard substance 0.0050 g, be dissolved in the methyl alcohol, be mixed with concentration 0.002 mg/mL standard solution, behind 0.45 μ m filtering with microporous membrane, make high performance liquid chromatography, obtain phlorizin standard substance retention time 21.444min, see Fig. 1;
B) the collection liquid after the high performance liquid chromatography detection Solid-Phase Extraction: precision takes by weighing collection liquid 0.0050 g after the Solid-Phase Extraction, be dissolved in the methyl alcohol, be mixed with concentration 0.002 mg/mL solution, behind 0.45 μ m filtering with microporous membrane, make high performance liquid chromatography, obtain this collection liquid retention time 21.518min, see Fig. 2.Collect the 21.518min effluent liquid, because the retention time of this retention time and standard substance is roughly the same, so this crest is the target substance phlorizin probably;
C) effluent liquid that step b) is collected is concentrated into 120 μ L with Nitrogen evaporator, makes high performance liquid chromatography behind 0.45 μ m filtering with microporous membrane, and the demonstration retention time is that the single crest of 21.758min is monomeric compound---the phlorizin behind the purifying;
6, structure is identified: precision takes by weighing phlorizin standard substance 0.0050 g, is dissolved in the methyl alcohol, is mixed with concentration 0.005 mg/mL standard solution.Monomeric compound behind this standard solution and the purifying is made mass spectrometric detection respectively.The main mass spectra peak (see figure 4) of phlorizin standard substance is consistent with the main mass spectra peak (see figure 5) of monomeric compound behind the Fructus Litchi extracting solution purifying, confirmed with method of the present invention extract and purifying after material be exactly phlorizin.
Claims (1)
1. the extraction and the purification process of phlorizin in the Fructus Litchi, it is characterized in that: it may further comprise the steps:
Pulverize: after fresh lichee pericarp (connecting clothing) dries in the shade, pulverized 40 mesh sieves;
Extract crude extract: accurately take by weighing Fructus Litchi powder 10.0 g, add 70% ethanol 150mL, supersound process 30 minutes, power 100%; Repeat after the filtration to extract once merging filtrate; Filtrate is evaporated to 70 mL through Rotary Evaporators, add 180 mL dehydrated alcohols and leave standstill, behind the 16h with filter paper filtering; Filtrate decompression is concentrated into ethanol and all volatilizees, and adds distilled water 150mL then, and with ultrasonic assist in dissolving, obtains crude extract after filtering at last;
Enrichment litchi rind crude extract: with sample on the above-mentioned crude extract to macroporous resin column 16 g, last sample speed 1 mL/min, then with 450 mL distilled water drip washing to colourless, again with 70% ethanol 50mL desorption, flow velocity 1mL/min obtains the litchi rind extracting solution, and gives 4 ℃ and keep in Dark Place;
Solid-Phase Extraction: get the solid phase extraction column after above-mentioned litchi rind extracting solution 5mL passes through to activate with the 1mL/min flow velocity, preceding 3mL effluent liquid discards, and all the other effluent liquid are collected; The activation of described solid phase extraction column needs to pass through solid phase extraction column with 10mL methyl alcohol earlier, and with 10mL distilled water flush away methyl alcohol, speed is 1mL/min again;
Purifying: described purifying adopts high performance liquid chromatography to carry out, and may further comprise the steps:
High performance liquid chromatography detects the phlorizin standard substance: precision takes by weighing phlorizin standard substance 0.0050 g, be dissolved in the methyl alcohol, be mixed with concentration 0.002 mg/mL standard solution, behind 0.45 μ m filtering with microporous membrane, make high performance liquid chromatography, obtain phlorizin standard substance retention time 21.444min;
High performance liquid chromatography detects the collection liquid after the Solid-Phase Extraction: precision takes by weighing collection liquid 0.0050 g after the Solid-Phase Extraction, be dissolved in the methyl alcohol, be mixed with concentration 0.002 mg/mL solution, behind 0.45 μ m filtering with microporous membrane, make high performance liquid chromatography, obtain this collection liquid retention time 21.518min, collect the 21.518min effluent liquid;
The effluent liquid that step b) is collected is concentrated into 120 μ L with Nitrogen evaporator, makes high performance liquid chromatography behind 0.45 μ m filtering with microporous membrane, and the demonstration retention time is that the single crest of 21.758min is monomeric compound---the phlorizin behind the purifying.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010617724 CN102086220A (en) | 2010-12-31 | 2010-12-31 | Extraction and purification method of phlorizin in litchi peels |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010617724 CN102086220A (en) | 2010-12-31 | 2010-12-31 | Extraction and purification method of phlorizin in litchi peels |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102086220A true CN102086220A (en) | 2011-06-08 |
Family
ID=44098242
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201010617724 Pending CN102086220A (en) | 2010-12-31 | 2010-12-31 | Extraction and purification method of phlorizin in litchi peels |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102086220A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102286037A (en) * | 2011-08-02 | 2011-12-21 | 桂林三宝药业有限公司 | Process for extracting phlorhizin from litchi rind |
| CN103239546A (en) * | 2013-04-28 | 2013-08-14 | 王惠莹 | Method for extracting general flavone from litchi shells |
| CN103271304A (en) * | 2013-04-09 | 2013-09-04 | 广东药学院 | Composition for improving litchi rind extract stability and method for improving litchi rind extract stability |
| CN103864864A (en) * | 2014-03-27 | 2014-06-18 | 江苏斯威森生物医药工程研究中心有限公司 | Method for efficiently extracting phlorizin from plants |
| CN104262423A (en) * | 2014-09-30 | 2015-01-07 | 桂林三宝药业有限公司 | Method for extracting phlorhizin from litchi rind |
| CN105924482A (en) * | 2016-04-29 | 2016-09-07 | 中南林业科技大学 | Method for extracting phloretin from camellia oleifera leaves |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1683382A (en) * | 2005-03-11 | 2005-10-19 | 天津市尖峰天然产物研究开发有限公司 | Process for extracting phlorhizin from bark, root, branch and leaf of rosaceous plant and juice pressing waste material |
| CN101392008A (en) * | 2007-09-18 | 2009-03-25 | 兴化格林生物制品有限公司 | Extraction technique of high-purity phlorizin |
-
2010
- 2010-12-31 CN CN 201010617724 patent/CN102086220A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1683382A (en) * | 2005-03-11 | 2005-10-19 | 天津市尖峰天然产物研究开发有限公司 | Process for extracting phlorhizin from bark, root, branch and leaf of rosaceous plant and juice pressing waste material |
| CN101392008A (en) * | 2007-09-18 | 2009-03-25 | 兴化格林生物制品有限公司 | Extraction technique of high-purity phlorizin |
Non-Patent Citations (1)
| Title |
|---|
| 《中国美容医学》 20080731 谢阳等 荔枝果皮中的美白剂根皮苷的提取、纯化及鉴定 第17卷, 第07期 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102286037A (en) * | 2011-08-02 | 2011-12-21 | 桂林三宝药业有限公司 | Process for extracting phlorhizin from litchi rind |
| CN103271304A (en) * | 2013-04-09 | 2013-09-04 | 广东药学院 | Composition for improving litchi rind extract stability and method for improving litchi rind extract stability |
| CN103239546A (en) * | 2013-04-28 | 2013-08-14 | 王惠莹 | Method for extracting general flavone from litchi shells |
| CN103864864A (en) * | 2014-03-27 | 2014-06-18 | 江苏斯威森生物医药工程研究中心有限公司 | Method for efficiently extracting phlorizin from plants |
| CN104262423A (en) * | 2014-09-30 | 2015-01-07 | 桂林三宝药业有限公司 | Method for extracting phlorhizin from litchi rind |
| CN105924482A (en) * | 2016-04-29 | 2016-09-07 | 中南林业科技大学 | Method for extracting phloretin from camellia oleifera leaves |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102086220A (en) | Extraction and purification method of phlorizin in litchi peels | |
| CN1974527B (en) | Process of preparing high purity chlorogenic acid and flavonid with eucommia leaf | |
| CN107011308B (en) | The method of polymethoxyflavone class compound is isolated and purified from bowl mandarin orange fruit | |
| CN103421077B (en) | Method for separating and purifying limonin compounds from pomelo fruits | |
| CN103044503A (en) | Method for rapidly and efficiently extracting paeoniflorin and albiflorin | |
| CN104892687B (en) | The method that high speed adverse current chromatogram isolates and purifies monomeric compound in Chinese mahonia leaf | |
| CN101921277B (en) | Method for simultaneously preparing vasicine and vasicinone from peganum harmala | |
| CN101337876B (en) | Process for extracting xanthohumol from lupulus | |
| CN102229638B (en) | Method for extracting oleanolic acid from chaenomeles fruit and preparing oleanolic acid standard | |
| CN101525328B (en) | Method for extracting alpha-mangostin from mangosteen fruit peel | |
| CN103788167A (en) | Preparation method for glycyrrhetinic acid monoglucuronide (GAMG) | |
| CN104370895B (en) | A kind of preparation method of orientin and Lutonaretin | |
| CN106632546A (en) | Method for preparing two chemical reference substances of Rhoifolin and naringin simultaneously | |
| CN106496292A (en) | A kind of method for preparing 6 iridoid glycoside constituents from Fructus Gardeniae simultaneously | |
| CN102532077B (en) | Method for preparing salvianolic acid B through separation by means of flash chromatography | |
| CN102040500B (en) | Method for extracting and separating xanthohumol and flavone compounds | |
| CN102040635B (en) | Method for efficiently separating and purifying forsythiaside B monomer and poliumoside monomer | |
| CN101658598B (en) | Method for extracting and enriching alisma total triterpenic ketone alcohol components from alisma | |
| CN104327026B (en) | A kind of method extracting separation costunolide and Decahydro-3,6,9-tris(methylene)azuleno[4,5-b | |
| CN103570647A (en) | Method for preparing high-purity paclitaxel from taxus chinensis cell culture fluid | |
| CN103145775A (en) | Preparation and quality control method for high purity cleidion brevipetiolatum glycoside A | |
| CN101045719A (en) | Method for high efficiency separating and purifying 1-deacetyl Baccatins III (10-DABIII) | |
| CN105385716A (en) | Method for separating sulfur-containing compounds in garlic by adopting counter-current chromatography | |
| CN100484931C (en) | Isolation preparing process for a plurality of isoflavones components in astragalus root | |
| CN101210039B (en) | Method for separating and preparing madecassoside chemical reference substance |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20110608 |