CN102660141A - Method for extracting anthocyanin from fruit epidermis - Google Patents
Method for extracting anthocyanin from fruit epidermis Download PDFInfo
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- CN102660141A CN102660141A CN2012101616990A CN201210161699A CN102660141A CN 102660141 A CN102660141 A CN 102660141A CN 2012101616990 A CN2012101616990 A CN 2012101616990A CN 201210161699 A CN201210161699 A CN 201210161699A CN 102660141 A CN102660141 A CN 102660141A
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Abstract
The invention belongs to the field of plant effective constituent extraction, and discloses a method for extracting anthocyanin from fruit epidermis, which comprises the following steps: under the dissolution assisting action of ultrasonic waves, taking fruit epidermis, grinding with liquid nitrogen to obtain powder, adding 1% hydrochloride methanol extracting solution, evenly mixing to obtain a pulp, leaching at low temperature in a dark place, and carrying out ultrasonic assisting extraction in an ultrasonic instrument. The method disclosed by the invention is simple, convenient and quick, can effectively extract anthocyanin from fruit, and is carried out at low temperature in the whole process, thereby being beneficial to maintaining the stability of anthocyanin. Therefore, the ultrasonic assisting dissolution can quickly and efficiently extract anthocyanin from fruit.
Description
Technical field
The invention belongs to effective ingredients in plant and extract the field, relate to a kind of method of from fruit surface, extracting anthocyanin.
Background technology
The color and luster of pericarp is the core index of fruit appearance quality, also is the important factor that influences its commodity value.Research shows that fruit colour is mainly determined by the content and the ratio thereof of chlorophyll, carrotenoid and phenols pigment.Fruit color is owing to chlorophyll degradation in the fruit cell, forms and manifest the result of carrotenoid or the synthesis of anthocyanins simultaneously.Anthocyanin is a main moity red, red-purple pericarp color and luster; Intensive polar solvents such as soluble in water, methyl alcohol, ethanol; Under acidic conditions, can keep original color and luster, pH is from low to high the time, and anthocyanin solution appears by redness to colourless variation until blueness; And under high temperature, the high light condition, anthocyanin is prone to degraded.Therefore, anthocyanin is many in the tissue extracts under lucifuge, low temperature, adopts acidified aqueous solution or acidifying organic solution to extract.
Under coldcondition, acidic aqueous solution dissolving anthocyanin ability is lower, and the pigment extraction effect is relatively poor in the fruit, along with temperature improves; Its extraction efficiency raises, but anthocyanin is better stable at coldcondition, along with temperature rises; The anthocyanin stability decreases, degradation speed is accelerated, and research shows that anthocyanin is under 100 ℃; 2h i.e. degraded fully, and under 5 ℃, the transformation period of anthocyanin is 79d.Therefore, when adopting the acidified aqueous solution water-bath to extract anthocyanin, there is a pair of contradiction that can't coordinate, all can is that the anthocyanin content that extracts does not meet the actual value in the fruit under any condition, thereby can't accurately measures.
When adopting acidifying organic solution to extract, its extraction efficiency is higher, but needs long lixiviate; Be in static condition for a long time, the raw material deposition hardens, and is unfavorable for pigment in the complete dissolving raw material of extracting solution; And need repetition repeatedly, united extraction liquid, volume was bigger when this will make last constant volume; And anthocyanin content was just less originally in the fruit, and last extracting liquid volume makes anthocyanin concentration step-down greatly, and instrument (like performance liquid chromatography) is when measuring its content; Minimum detectability is arranged, and concentration is crossed low can making can't measure its content.Therefore; When adopting in the acidifying organic solution extracted many times material anthocyanin; Need behind the united extraction liquid constant volume to concentrate, can accelerate the anthocyanin degraded and heat in the rotary evaporation process, and anthocyanin sticks on the rotary evaporation bottle after concentrating through Rotary Evaporators; Less water is wash-out thoroughly, makes experimental error increase.
Therefore,, press for a kind of new process for extracting, to satisfy requirement to anthocyanin content and composition Study in the fruit because there is weak point in the anthocyanin process for extracting at present.To the appeal method, the present invention utilizes the effect of UW hydrotropy, low-loss, has high efficiencyly extracted anthocyanin in the pear fruit, and this method is easy and simple to handle, and is consuming time less.
Summary of the invention
The objective of the invention is above-mentioned defective, a kind of method of from fruit surface, extracting anthocyanin is provided to prior art.
The object of the invention can be realized through following technical scheme:
A kind of method of from fruit surface, extracting anthocyanin comprises the steps:
1) get fruit surface, liquid nitrogen grinding is to Powdered, and the real epidermis of every gram fresh fruit adds 3～6mL, 1% hydrochloric acid methanol (V/V), and homogenate gets homogenate;
2) with homogenate in 0～4 ℃, lucifuge, lixiviate 4～5h;
3) vat liquor is put into the ultrasonic instrument supersound process;
4) after the ultrasonication, vat liquor is got supernatant and is promptly got anthocyanin at 4 ℃ of centrifugal 18～25min of following 12000g.
Described fruit surface is the fruit surface of red skin pears, apple, grape, peach, Lee, cherry; Preferred red skin pear fruit epidermis.
Described fruit surface thickness is 0.8～1.2cm.
The concentration of hydrochloric acid that adds when preparing described 1% hydrochloric acid methanol is 36～37%,
Described ultrasound condition is that described ultrasound condition is under 0～4 ℃, and ultrasonic power is 100W, ultrasonic frequency 40KHZ, ultrasonic 0.5～1.5h.
The described supernatant of step 4) is used for anthocyanin composition or content detection after constant volume, filtration.
Beneficial effect:
1, anthocyanin is a main moity red, red-purple fruit complexion pool; To anthocyanin component and Determination on content thereof in the fruit is the basis of its fruit coloration being carried out further investigation, and process for extracting is a precondition of accurately measuring anthocyanin content in the fruit the most efficiently.Present method has been taken all factors into consideration the characteristic distributions and the character of anthocyanin in the pericarp; On the basis of traditional lixiviate, add supersound process; Select extraction temperature, extraction time, the ultrasound condition of specified range, to solve loss height in the anthocyanin traditional extraction process, problems such as length consuming time; In leaching process, be in coldcondition, and step is simple always, weak point consuming time, extraction efficiency is high, effectively reduces the degraded and the loss of anthocyanin, lays a good foundation for accurately measuring fruit anthocyanin content and component.
2, the present invention has shortened anthocyanin extraction time, has improved extraction efficiency, and anthocyanin in the disposable extraction fruit, and it is higher that it extracts concentration, need not subsequent operations, thereby reduced the anthocyanin loss.
3, method of use of the present invention is simple, and is convenient, fast.Only need mortar during extraction, the plastics film box, ultrasonic instrument, whizzer, centrifuge tube, volumetric flask, it is conventional instrument, and simple to operate, and the individual can accomplish all experiment contents.
4, generalization of the present invention and applicability are very strong, can be used to extract the anthocyanin of pears, apple, grape, peach, Lee, the multiple fruit tree of cherry, and therefore, this method has boundless application prospect.
Embodiment
Embodiment 1
1. anthocyanin extracts
1) pear fruit of gathering is cleaned, dries, get 2g pear fruit epidermis, thickness is put into mortar and is added liquid nitrogen and be ground to Powderedly about 0.1cm, adds 8mL 1% hydrochloric acid methanol (V/V) homogenate and gets homogenate; Described 1% hydrochloric acid methanol compound method adds the ultrapure water constant volume to 100mL for getting 1mL36% hydrochloric acid.
2) from mortar, change homogenate over to the 10mL centrifuge tube, put into and biological ice bag ice chest lucifuge is housed, 0～4 ℃ of following lixiviate 4h;
3) centrifuge tube is put into the ultrasonic 1h of ultrasonic instrument, ultrasonic power is 100W, and ultrasonic frequency 40KHZ puts into biological ice bag in ultrasonic instrument, and temperature is remained under 0～4 ℃, prevents that temperature raises in ultrasonic procedure, makes anthocyanin degrade;
4) after the ultrasonication, 4 ℃ of centrifugal 20min of following 12000g get supernatant and are settled to 10mL and get the anthocyanin extracting solution, are used for the anthocyanin assay;
2. anthocyanin assay
1) gets 1mL anthocyanin extracting solution, use 0.4molL respectively
-1KCl-HCl damping fluid (pH 1.0) and 0.4molL
-1Hydrocerol A/Sodium phosphate, dibasic damping fluid (pH 5.0) is diluted to 5mL, compares with zero(ppm) water behind the mixing, under 510nm, measures OD value (D).
2) every gram fresh fruit ginned cotton pigment glycosides content (mgg
-1)=△ D * 5 * 10 * 445.2/29600 * 2, wherein △ D=D 510 (pH 1.0)-D510 (pH 5.0); 5 is extension rate; 10 are the lixiviate volume; 445.2 be Minor centaury galactoside relative molecular weight; 29600 is Minor centaury galactoside molar absorptivity (mol
-1Cm
-1); 2 is pericarp quality (g).
3. test-results
Ultrasonication assisted extraction anthocyanin and 0～4 ℃ of lixiviate 4h do not carry out supersound process and 0～4 ℃ of lixiviate 24h does not carry out supersound process (CK) absorbance and the contrast of anthocyanin content are as shown in table 1:
The anthocyanin assay of table 1 Different Extraction Method relatively
Can be known by table 1, carry out the ultrasonic-assisted extraction anthocyanin behind the leach at low temperature 4h, its extraction yield does not carry out ultrasonication after being significantly higher than leach at low temperature 4h, and not carry out ultrasonication extraction yield difference not remarkable with leach at low temperature 24h.Explanation thus, the ultrasonic-assisted extraction anthocyanin can significantly shorten extraction time, and extraction efficiency is high, is suitable for the pericarp anthocyanin and extracts and assay.
Claims (6)
1. a method of from fruit surface, extracting anthocyanin is characterized in that comprising the steps:
1) get fruit surface, liquid nitrogen grinding is to Powdered, and the real epidermis of every gram fresh fruit adds 3～6mL, 1% hydrochloric acid methanol (V/V), and homogenate gets homogenate;
2) with homogenate in 0～4 ℃, lucifuge, lixiviate 4～5h;
3) vat liquor is put into the ultrasonic instrument supersound process;
4) after the ultrasonication, vat liquor is got supernatant and is promptly got anthocyanin at 4 ℃ of centrifugal 18～25min of following 12000g.
2. the method for from fruit surface, extracting anthocyanin according to claim 1 is characterized in that described fruit surface is the fruit surface of red skin pears, apple, grape, peach, Lee, cherry; Preferred red skin pear fruit epidermis.
3. the method for from fruit surface, extracting anthocyanin according to claim 2 is characterized in that described fruit surface thickness is 0.8～1.2cm.
4. the method for from fruit surface, extracting anthocyanin according to claim 1, the concentration of hydrochloric acid that adds when it is characterized in that preparing described 1% hydrochloric acid methanol is 36～37%.
5. the method for from fruit surface, extracting anthocyanin according to claim 1 is characterized in that described ultrasound condition is that described ultrasound condition is under 0～4 ℃, and ultrasonic power is 100W, ultrasonic frequency 40KHZ, ultrasonic 0.5～1.5h.
6. the method for from fruit surface, extracting anthocyanin according to claim 1 is characterized in that the described supernatant of step 4) is used for anthocyanin composition or content detection after constant volume, filtration.
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|---|---|---|---|
| CN2012101616990A CN102660141B (en) | 2012-05-22 | 2012-05-22 | Method for extracting anthocyanin from fruit epidermis |
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| CN2012101616990A CN102660141B (en) | 2012-05-22 | 2012-05-22 | Method for extracting anthocyanin from fruit epidermis |
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| CN102660141B CN102660141B (en) | 2013-12-11 |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103183967A (en) * | 2013-04-28 | 2013-07-03 | 湖南农业大学 | Method for producing purpurin by utilizing fresh camphor tree pericarp |
| CN104181017A (en) * | 2013-09-27 | 2014-12-03 | 天津天隆种业科技有限公司 | New method for screening variety of anthocyanin-rich highly-stable black rice |
| CN107722071A (en) * | 2017-11-06 | 2018-02-23 | 河南省林业科学研究院 | The stability extracting method of anthocyanin in a kind of red autumnal leaves poplar blade |
| CN109620853A (en) * | 2017-10-09 | 2019-04-16 | 董玲 | A kind of extraction process of medicinal material |
| CN109646351A (en) * | 2019-01-08 | 2019-04-19 | 百草堂医药股份有限公司 | A kind of pure and fresh clearing damp shower cream of wormwood and preparation method thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103183967A (en) * | 2013-04-28 | 2013-07-03 | 湖南农业大学 | Method for producing purpurin by utilizing fresh camphor tree pericarp |
| CN104181017A (en) * | 2013-09-27 | 2014-12-03 | 天津天隆种业科技有限公司 | New method for screening variety of anthocyanin-rich highly-stable black rice |
| CN109620853A (en) * | 2017-10-09 | 2019-04-16 | 董玲 | A kind of extraction process of medicinal material |
| CN107722071A (en) * | 2017-11-06 | 2018-02-23 | 河南省林业科学研究院 | The stability extracting method of anthocyanin in a kind of red autumnal leaves poplar blade |
| CN109646351A (en) * | 2019-01-08 | 2019-04-19 | 百草堂医药股份有限公司 | A kind of pure and fresh clearing damp shower cream of wormwood and preparation method thereof |
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|---|---|
| CN102660141B (en) | 2013-12-11 |
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