CN102660141B - Method for extracting anthocyanin from fruit epidermis - Google Patents
Method for extracting anthocyanin from fruit epidermis Download PDFInfo
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- CN102660141B CN102660141B CN2012101616990A CN201210161699A CN102660141B CN 102660141 B CN102660141 B CN 102660141B CN 2012101616990 A CN2012101616990 A CN 2012101616990A CN 201210161699 A CN201210161699 A CN 201210161699A CN 102660141 B CN102660141 B CN 102660141B
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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 field, relate to a kind of method of extracting anthocyanin from fruit surface.
Background technology
The color and luster of pericarp is the core index of fruit appearance quality, is also the important factor that affects its commodity value.Research shows, fruit colour is mainly determined by content and the ratio thereof of chlorophyll, carotenoid and phenols pigment.Fruit color, be due to the fruit cell Chlorophyll Degradation, forms simultaneously and manifest the result of carotenoid or the synthesis of anthocyanins.Anthocyanin is chief component composition red, red-purple pericarp color and luster, the intensive polar solvents such as soluble in water, methyl alcohol, ethanol, can keep original color and luster under acidic conditions, pH is from low to high the time, anthocyanin solution presents by redness to colourless until blue variation, and, under high temperature, high light condition, anthocyanin is easily degraded.Therefore, in tissue, anthocyanin is many extracts under lucifuge, low temperature, adopts acidified aqueous solution or acidifying organic solution to extract.
Under cold condition, acidic aqueous solution dissolving anthocyanin ability is lower, and in fruit, the pigment extraction effect is poor, along with temperature improves, its extraction efficiency raises, but anthocyanin is at cold condition, stability is better, along with temperature rises, and the anthocyanin stability decreases, degradation speed is accelerated, research shows, anthocyanin is under 100 ℃, and 2h is degradable, and, under 5 ℃, the transformation period of anthocyanin is 79d.Therefore, while adopting the acidified aqueous solution water-bath to extract anthocyanin, there is a pair of contradiction that can't coordinate, can be all that the Anthocyanin content extracted does not meet the actual value in fruit under any condition, thereby can't accurately measure.
While adopting acidifying organic solution to extract, its extraction efficiency is higher, but need long lixiviate, for a long time in static condition, the raw material precipitation hardens, be unfavorable for that extracting solution dissolves pigment in raw material fully, and need repetition repeatedly, united extraction liquid, when this will make last constant volume, volume is larger, and anthocyanin content in fruits is originally just less, last extracting liquid volume makes greatly anthocyanin concentration step-down, instrument (as high performance liquid chromatography), when measuring its content, has minimum detectability, and the too low meeting of concentration makes can't measure its content.Therefore, while adopting in acidifying organic solution extracted many times material anthocyanin, after united extraction liquid constant volume, need to concentrate by Rotary Evaporators, and in the rotary evaporation process, heating can be accelerated the anthocyanin degraded, and after concentrated, anthocyanin sticks on the rotary evaporation bottle, a small amount of water is wash-out thoroughly, makes experimental error increase.
Therefore, due to current anthocyanin extracting method Shortcomings part, in the urgent need to a kind of new extracting method, to meet the requirement to anthocyanin content in fruits and composition Study.For the appeal method, the present invention utilizes the effect of ultrasonic wave hydrotropy, low-loss, has high efficiencyly extracted anthocyanin in pear fruit, and the method is easy and simple to handle, consuming time less.
Summary of the invention
The objective of the invention is the above-mentioned defect for prior art, a kind of method of extracting anthocyanin from fruit surface is provided.
Purpose of the present invention can be achieved through the following technical solutions:
A kind of method of extracting anthocyanin from fruit surface, comprise 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 obtains homogenate;
2) by homogenate in 0～4 ℃, lucifuge, lixiviate 4～5h;
3) vat liquor is put into to the ultrasonic instrument supersound process;
4), after ultrasonication, vat liquor, at 4 ℃ of centrifugal 18～25min of lower 12000g, is got supernatant liquor and is obtained anthocyanin.
The fruit surface that described fruit surface is 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 added while 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 liquor of step 4) through constant volume, filter after for Anthocyanin Compositions or content detection.
Beneficial effect:
1, anthocyanin is chief component composition red, red-purple fruit complexion pool, mensuration to anthocyanin component and content thereof in fruit is its fruit coloration to be carried out to the basis of further investigation, and the most efficient extracting method is the precondition of Accurate Measurement anthocyanin content in fruits.Present method has considered characteristic distributions and the character of anthocyanin in the pericarp, add supersound process on the basis of traditional lixiviate, select extraction temperature, extraction time, the ultrasound condition of specified range, high with loss in solution anthocyanin traditional extraction process, the problems such as length consuming time; In leaching process, always in cold condition, and step is simple, and consuming time short, extraction efficiency is high, effectively reduces degraded and the loss of anthocyanin, for Accurate Measurement fruit Anthocyanin content and component are laid a good foundation.
2, the present invention has shortened anthocyanin extraction time, has improved extraction efficiency, and anthocyanin in disposable extraction fruit, and its extracting concentration is higher, without subsequent operations, thereby has reduced the anthocyanin loss.
3, using method of the present invention is simple, 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 complete all experiment contents.
4, generalization of the present invention and applicability are very strong, can be for extracting the anthocyanin of pears, apple, grape, peach, Lee, the multiple fruit tree of cherry, and therefore, the method has boundless application prospect.
Embodiment
Embodiment 1
1. anthocyanin extracts
1) pear fruit of collection is cleaned, dries, get 2g pear fruit epidermis, thickness is in the 0.1cm left and right, puts into mortar and adds liquid nitrogen and be ground to Powderedly, adds 8mL 1% hydrochloric acid methanol (V/V) homogenate and obtains homogenate; Described 1% hydrochloric acid methanol compound method adds the ultrapure water constant volume to 100mL for getting 1mL36% hydrochloric acid.
2) homogenate is proceeded to from mortar to the 10mL centrifuge tube, put into biological ice bag ice chest lucifuge is housed, 0～4 ℃ of lower lixiviate 4h;
3) centrifuge tube is put into to 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 the anthocyanin degraded;
4), after ultrasonication, 4 ℃ of centrifugal 20min of lower 12000g, get supernatant liquor and be settled to 10mL and obtain the anthocyanin extracting solution, for determination of anthocyanin content;
2. determination of anthocyanin content
1) get 1mL anthocyanin extracting solution, use respectively 0.4molL
-1kCl-HCl damping fluid (pH 1.0) and 0.4molL
-1citric acid/Sodium phosphate dibasic damping fluid (pH 5.0), be diluted to 5mL, mixes with distilled water, to compare afterwards, measures optical density value (D) under 510nm.
2) every gram fresh fruit skin Anthocyanin 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 is 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 Anthocyanin content contrast are as shown in table 1:
The determination of anthocyanin content of table 1 Different Extraction Method relatively
As shown in Table 1, carry out the ultrasonic-assisted extraction anthocyanin after 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 (3)
1. a method of extracting anthocyanin from fruit surface, 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 ~ 6 mL 1% hydrochloric acid methanols (V/V), and homogenate obtains homogenate;
2) by homogenate in 0 ~ 4 ℃, lucifuge, lixiviate 4 ~ 5 h;
3) vat liquor is put into to the ultrasonic instrument supersound process, described ultrasound condition is under 0 ~ 4 ℃, and ultrasonic power is 100 W, ultrasonic frequency 40KHZ, ultrasonic 0.5 ~ 1.5 h;
4), after ultrasonication, centrifugal 18 ~ 25 min of vat liquor 12000 g under 4 ℃, get supernatant liquor and obtain anthocyanin;
The fruit surface that described fruit surface is red skin pears, apple, grape, peach, Lee, cherry.
2. the method for extracting anthocyanin from fruit surface according to claim 1, the concentration of hydrochloric acid added while it is characterized in that preparing described 1% hydrochloric acid methanol is 36 ~ 37%.
3. the method for extracting anthocyanin from fruit surface according to claim 1, it is characterized in that the described supernatant liquor of step 4) through constant volume, filter after for Anthocyanin Compositions or content detection.
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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 true CN102660141B (en) | 2013-12-11 |
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| 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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| DE3310340A1 (en) * | 1982-03-31 | 1983-10-06 | Gen Foods Corp | METHOD FOR PRODUCING AN ANTHOCYANIN DYE FROM GRAPE VINE |
| JPS59223756A (en) * | 1983-06-02 | 1984-12-15 | San Ei Chem Ind Ltd | Production of anthocyanin pigment |
| JPH0737251B2 (en) * | 1990-04-09 | 1995-04-26 | 東洋自動機株式会社 | Bag holding device for rotary table type automatic packaging machine |
| JPH0926024A (en) * | 1995-07-12 | 1997-01-28 | Toyota Motor Corp | Shift controller of automatic transmission |
| JPH09206024A (en) * | 1996-01-30 | 1997-08-12 | Sanei Gen F F I Inc | Coloration of cherry |
| KR100485032B1 (en) * | 2002-04-09 | 2005-04-22 | 김선호 | The method for manufacturing of natural dyes |
| CN102229633B (en) * | 2011-05-31 | 2014-01-08 | 江南大学 | Method for separating and preparing five high-purity anthocyanidin monomers from grape skins |
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