CN111995609A - Extraction method and application of passion fruit anthocyanin - Google Patents
Extraction method and application of passion fruit anthocyanin Download PDFInfo
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- CN111995609A CN111995609A CN202010669615.9A CN202010669615A CN111995609A CN 111995609 A CN111995609 A CN 111995609A CN 202010669615 A CN202010669615 A CN 202010669615A CN 111995609 A CN111995609 A CN 111995609A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
- C07D311/60—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2
- C07D311/62—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2 with oxygen atoms directly attached in position 3, e.g. anthocyanidins
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Abstract
The invention discloses an extraction method of passion fruit anthocyanin, which comprises the following steps: 1) removing endocarp of purple passion fruit to obtain purple peel and passion fruit pulp, mixing and crushing to obtain crushed substances; 2) adding the crushed material into glacial acetic acid in a dark environment at the temperature of 20-30 ℃ and under normal pressure, mixing for 1-2min, then sequentially adding tartaric acid with the concentration of 0.5-1mol/L and citric acid with the concentration of 0.5-1mol/L, mixing and stirring for 1-2h, then adding ethanol with the concentration of 75-85%, performing reflux extraction for 3-4h, filtering, performing rotary evaporation, and then placing in an organic solvent for extraction to obtain passion fruit anthocyanin; wherein the dosage proportion is the mass of the crushed material: volume of ethanol: volume of glacial acetic acid: volume of tartaric acid: the volume of the citric acid is 1g:15-20ml:1-2ml:3ml:3 ml. The modified passion fruit anthocyanin extracted by the invention can be applied to preparation of a chicken feed additive.
Description
Technical Field
The invention relates to the field of passion fruit anthocyanidin. More specifically, the invention relates to an extraction method and application of passion fruit anthocyanin.
Background
Anthocyanins (also called anthocyanidins) are water-soluble natural pigments widely present in plants in nature, and are colored aglycones obtained by hydrolyzing anthocyanins (anthocyanins). The main color-producing substances in fruits, vegetables and flowers are mostly related to the plants. Under the condition of different pH values of plant cell vacuoles, the anthocyanin enables the petals to present colorful colors. 20 kinds of anthocyanidins are determined, the anthocyanidins extracted from different plants are different in types, natural anthocyanidins exist in a glucoside form, are called anthocyanin, and few free anthocyanidins exist. Anthocyanins are mainly used for coloring food, and can also be used in dye, medicine, and cosmetic. The optimized extraction process of the peucedanum praeruptorum dunn and the like performs the extraction process and the stability research of anthocyanin in the exocarp of the passion fruit, the pH value of the optimized extraction process is 2, the material-liquid ratio is 1:26, the time is 3.8h, the temperature is 34 ℃, the ethanol concentration is 82%, and the process obtains better extraction rate. However, when the passion fruit anthocyanin extracted by the process is applied to the chicken feed additive, the inhibition effect on salmonella and escherichia coli of chicken is small, and the application of the passion fruit anthocyanin in the chicken feed additive is influenced.
Disclosure of Invention
An object of the present invention is to solve at least the above problems and/or disadvantages and to provide at least the advantages described hereinafter.
The invention also aims to provide an extraction method of passion fruit anthocyanin, which is characterized in that various procyanidins are obtained by using purple peel and pulp of passion fruit, a plurality of hydroxyl groups of the various procyanidins and carboxyl groups of glacial acetic acid, tartaric acid and citric acid are subjected to dehydration, condensation and esterification reaction, partial redundant hydroxyl groups are removed, various procyanidin molecules are chemically modified, and the extracted modified passion fruit anthocyanin can be applied to preparation of chicken feed additives and can have an inhibiting effect on chicken salmonella and escherichia coli.
To achieve these objects and other advantages of the present invention, there is provided a method for extracting passion fruit anthocyanidin, comprising the steps of:
removing endocarp of the purple passion fruit to obtain purple peel and passion fruit pulp, and crushing to obtain crushed substances;
step two, adding the crushed material into glacial acetic acid in a light-proof environment at the temperature of 20-30 ℃ and under normal pressure, mixing for 1-2min, then sequentially adding tartaric acid with the concentration of 0.5-1mol/L and citric acid with the concentration of 0.5-1mol/L, mixing and stirring for 1-2h, then adding ethanol with the concentration of 75-85%, performing reflux extraction for 3-4h, filtering, performing rotary evaporation, and then placing in an organic solvent for extraction to obtain passion fruit anthocyanin; wherein the dosage proportion is the mass of the crushed material: volume of ethanol: volume of glacial acetic acid: volume of tartaric acid: the volume of the citric acid is 1g:15-20ml:1-2ml:3ml:3 ml.
Preferably, the organic solvent is one or more of n-hexane, petroleum ether or diethyl ether.
Preferably, the method further comprises the following steps of pretreating the passion fruit purple peel: shredding purple peel in dark environment, soaking in the solution, placing in a reaction kettle at 20 deg.C and 0.4MPa, maintaining the temperature and pressure for 1 hr, taking out, air drying at room temperature, and pulverizing to 100 mesh; wherein the dissolution liquid is a mixed aqueous solution of 0.3mg/mL of muramidase and 0.4mg/mL of cellulase, and the mass-volume ratio of the purple skin to the dissolution liquid is 1g:1.2 mL.
Preferably, the method further comprises the following pretreatment of the passion fruit pulp: ultrasonically crushing the pulp in a dark environment, soaking the pulp in the soaking solution, placing the soaked pulp in a reaction kettle at the temperature of 20 ℃ and the pressure of 0.4MPa, repeatedly stirring, keeping the temperature and the pressure for 1 hour, taking out the pulp, and air-drying the pulp at room temperature; wherein the soaking solution is 0.4mg/mL lywallzyme water solution, and the mass volume ratio of the pulp to the soaking solution is 1g:1 mL.
The invention also provides application of the passion fruit anthocyanin in preparation of a chicken feed additive.
The passion fruit anthocyanin extracted by the invention is used for preparing a chicken feed additive for resisting salmonella and escherichia coli.
The invention at least comprises the following beneficial effects:
1. the method obtains various procyanidins by using the purple peel and the pulp of the passion fruit, multiple hydroxyl groups of the various procyanidins and carboxyl groups of glacial acetic acid, tartaric acid and citric acid are subjected to dehydration condensation esterification reaction, partial redundant hydroxyl groups are removed, various procyanidin molecules are chemically modified, and the extracted modified passion fruit anthocyanin can be applied to preparation of a chicken feed additive and has an inhibiting effect on chicken salmonella resistance and escherichia coli resistance.
2. According to the method, the cell wall structure of purple peel is degraded by soaking in a mixed aqueous solution of muralytic enzyme and cellulase, the degraded cell wall is mechanically crushed and torn, the cell wall is further crushed, the purple peel cell wall is degraded, the procyanidine is favorably exposed and is beneficial to dissolution, a plurality of hydroxyl groups of the exposed procyanidine and carboxyl groups of glacial acetic acid, tartaric acid and citric acid are subjected to dehydration condensation esterification reaction, part of redundant hydroxyl groups are removed, and procyanidine molecules are chemically modified to obtain the modified passion fruit anthocyanin. Adding ethanol for extraction and organic solvent for extraction in the acidic environment of glacial acetic acid, tartaric acid and citric acid, and separating to obtain the modified passion fruit anthocyanin.
3. According to the method, the flesh cell wall structure cracked by ultrasonic vibration is degraded by soaking in the lywallzyme aqueous solution, so that the procyanidine is exposed and dissolved out easily, a plurality of hydroxyl groups of the exposed procyanidine and carboxyl groups of glacial acetic acid, tartaric acid and citric acid are subjected to dehydration condensation esterification reaction, part of redundant hydroxyl groups are removed, and procyanidine molecules are chemically modified to obtain the modified passion fruit anthocyanin. Adding ethanol for extraction and organic solvent for extraction in the acidic environment of glacial acetic acid, tartaric acid and citric acid, and separating to obtain the modified passion fruit anthocyanin.
Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention.
Detailed Description
The present invention is described in further detail below to enable those skilled in the art to practice the invention with reference to the description.
It will be understood that terms such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.
Example 1
A method for extracting passion fruit anthocyanin comprises the following steps:
removing endocarp of purple passion fruit to obtain purple peel and passion fruit pulp, mixing and crushing to obtain crushed substances;
step two, adding 10g of the crushed material into glacial acetic acid to mix for 1min in a light-proof environment at 30 ℃ and under normal pressure, then sequentially adding tartaric acid with the concentration of 0.5mol/L and citric acid with the concentration of 0.5mol/L, mixing and stirring for 1h, then adding ethanol with the concentration of 82%, refluxing and extracting for 4h, filtering, performing rotary evaporation, and then sequentially placing into n-hexane and petroleum ether to extract to obtain 39.12mg of passion fruit anthocyanin; wherein the dosage proportion is the mass of the crushed material: volume of ethanol: volume of glacial acetic acid: volume of tartaric acid: the volume of citric acid was 1g:20ml:1ml:3ml:3 ml.
Example 2
A method for extracting passion fruit anthocyanin comprises the following steps:
removing endocarp of the purple passion fruit to obtain purple peel and passion fruit pulp;
step two, pretreating purple peel of passion fruits: shredding purple peel in dark environment, soaking in the solution, placing in a reaction kettle at 20 deg.C and 0.4MPa, maintaining the temperature and pressure for 1 hr, taking out, air drying at room temperature, and pulverizing to 100 mesh; wherein the dissolution liquid is a mixed aqueous solution of 0.3mg/mL of muramidase and 0.4mg/mL of cellulase, and the mass-volume ratio of the purple skin to the dissolution liquid is 1g:1.2 mL;
step three, mixing and crushing the crushed purple peel and the passion fruit pulp to obtain a crushed material;
step four, adding 10g of the crushed material into glacial acetic acid to mix for 1min under normal pressure in a light-proof environment at the temperature of 30 ℃, then sequentially adding tartaric acid with the concentration of 0.5mol/L and citric acid with the concentration of 0.5mol/L, mixing and stirring for 1h, then adding ethanol with the concentration of 82%, refluxing and extracting for 4h, filtering, performing rotary evaporation, and then sequentially placing into n-hexane and petroleum ether to extract to obtain 42.91mg of passion fruit anthocyanin; wherein the dosage proportion is the mass of the crushed material: volume of ethanol: volume of glacial acetic acid: volume of tartaric acid: the volume of citric acid was 1g:20ml:1ml:3ml:3 ml.
Example 3
A method for extracting passion fruit anthocyanin comprises the following steps:
removing endocarp of the purple passion fruit to obtain purple peel and passion fruit pulp;
step two, pretreating purple peel of passion fruits: shredding purple peel in dark environment, soaking in the solution, placing in a reaction kettle at 20 deg.C and 0.4MPa, maintaining the temperature and pressure for 1 hr, taking out, air drying at room temperature, and pulverizing to 100 mesh; wherein the dissolution liquid is a mixed aqueous solution of 0.3mg/mL of muramidase and 0.4mg/mL of cellulase, and the mass-volume ratio of the purple skin to the dissolution liquid is 1g:1.2 mL;
step three, pretreating passion fruit pulp: ultrasonically crushing the pulp in a dark environment, soaking the pulp in the soaking solution, placing the soaked pulp in a reaction kettle at the temperature of 20 ℃ and the pressure of 0.4MPa, repeatedly stirring, keeping the temperature and the pressure for 1 hour, taking out the pulp, and air-drying the pulp at room temperature; wherein the soaking solution is 0.4mg/mL lywallzyme aqueous solution, and the mass-volume ratio of the pulp to the soaking solution is 1g:1 mL;
step four, mixing and crushing the crushed purple peel and the crushed passion fruit pulp to obtain a crushed material;
step five, adding 10g of the crushed material into glacial acetic acid to mix for 1min under normal pressure in a light-proof environment at the temperature of 30 ℃, then sequentially adding tartaric acid with the concentration of 0.5mol/L and citric acid with the concentration of 0.5mol/L, mixing and stirring for 1h, then adding ethanol with the concentration of 82%, refluxing and extracting for 4h, filtering, performing rotary evaporation, and then sequentially placing into n-hexane and petroleum ether to extract to obtain 46.85mg of passion fruit anthocyanin; wherein the dosage proportion is the mass of the crushed material: volume of ethanol: volume of glacial acetic acid: volume of tartaric acid: the volume of citric acid was 1g:20ml:1ml:3ml:3 ml.
Comparative example 1
The method for extracting the passion fruit anthocyanin is basically the same as that in the embodiment 1, and the only difference is that the passion fruit anthocyanin is extracted by using purple peel, and the method specifically comprises the following steps:
removing endocarp of purple passion fruit, and crushing purple peel of the remained epicarp to obtain a crushed material;
step two, adding 10g of the crushed material into glacial acetic acid to mix for 1min in a light-proof environment at 30 ℃ and under normal pressure, then sequentially adding tartaric acid with the concentration of 0.5mol/L and citric acid with the concentration of 0.5mol/L, mixing and stirring for 1h, then adding ethanol with the concentration of 82%, refluxing and extracting for 4h, filtering, performing rotary evaporation, and then sequentially placing into n-hexane and petroleum ether to extract to obtain 34.05mg of passion fruit anthocyanin; wherein the dosage proportion is the mass of the crushed material: volume of ethanol: volume of glacial acetic acid: volume of tartaric acid: the volume of citric acid was 1g:20ml:1ml:3ml:3 ml.
Comparative example 2
The extraction method of passion fruit anthocyanin is basically the same as that in the embodiment 2, and the only difference is that no glacial acetic acid is added for mixing, and the method specifically comprises the following steps:
removing endocarp of the purple passion fruit to obtain purple peel and passion fruit pulp;
step two, pretreating purple peel of passion fruits: shredding purple peel in dark environment, soaking in the solution, placing in a reaction kettle at 20 deg.C and 0.4MPa, maintaining the temperature and pressure for 1 hr, taking out, air drying at room temperature, and pulverizing to 100 mesh; wherein the dissolution liquid is a mixed aqueous solution of 0.3mg/mL of muramidase and 0.4mg/mL of cellulase, and the mass-volume ratio of the purple skin to the dissolution liquid is 1g:1.2 mL;
step three, mixing and crushing the crushed purple peel and the passion fruit pulp to obtain a crushed material;
step four, adding 10g of the crushed material into 0.5mol/L tartaric acid for mixing in a dark environment at the temperature of 30 ℃ and under normal pressure, adding 0.5mol/L citric acid, mixing and stirring for 1h, then adding 82% ethanol, carrying out reflux extraction for 4h, filtering, carrying out rotary evaporation, and then sequentially placing into n-hexane and petroleum ether for extraction to obtain 39.04mg of passion fruit anthocyanin; wherein the dosage proportion is the mass of the crushed material: volume of ethanol: volume of glacial acetic acid: volume of tartaric acid: the volume of citric acid was 1g:20ml:1ml:3ml:3 ml.
Comparative example 3
The method for extracting the passion fruit anthocyanin is basically the same as that in the embodiment 3, and the only difference is that tartaric acid and citric acid are not added, and the method specifically comprises the following steps:
removing endocarp of the purple passion fruit to obtain purple peel and passion fruit pulp;
step two, pretreating purple peel of passion fruits: shredding purple peel in dark environment, soaking in the solution, placing in a reaction kettle at 20 deg.C and 0.4MPa, maintaining the temperature and pressure for 1 hr, taking out, air drying at room temperature, and pulverizing to 100 mesh; wherein the dissolution liquid is a mixed aqueous solution of 0.3mg/mL of muramidase and 0.4mg/mL of cellulase, and the mass-volume ratio of the purple skin to the dissolution liquid is 1g:1.2 mL;
step three, pretreating passion fruit pulp: ultrasonically crushing the pulp in a dark environment, soaking the pulp in the soaking solution, placing the soaked pulp in a reaction kettle at the temperature of 20 ℃ and the pressure of 0.4MPa, repeatedly stirring, keeping the temperature and the pressure for 1 hour, taking out the pulp, and air-drying the pulp at room temperature; wherein the soaking solution is 0.4mg/mL lywallzyme aqueous solution, and the mass-volume ratio of the pulp to the soaking solution is 1g:1 mL;
step four, mixing and crushing the crushed purple peel and the crushed passion fruit pulp to obtain a crushed material;
step five, adding 10g of crushed materials into glacial acetic acid in a light-proof environment at the temperature of 30 ℃, mixing and stirring for 1h under normal pressure, then adding 82% ethanol, performing reflux extraction for 4h, filtering, performing rotary evaporation, and then sequentially placing the mixture into n-hexane and petroleum ether for extraction to obtain 44.06mg of passion fruit anthocyanin; wherein the dosage proportion is the mass of the crushed material: volume of ethanol: volume of glacial acetic acid: volume of tartaric acid: the volume of citric acid was 1g:20ml:1ml:3ml:3 ml.
Test No.)
Salmonella inhibition and Escherichia coli inhibition assay
1.1 strain: salmonella and Escherichia coli from the microbiological laboratory of Hunan agriculture university
1.2 preparation of bacterial liquid
Inoculating Salmonella and Escherichia coli respectively in meat soup culture solution, culturing at 37 deg.C for 18 hr, diluting the solution to 104～105Bacteria/ml.
1.3 Minimum Inhibitory Concentration (MIC) determination
Respectively collecting the above two strains at a concentration of 2.5 × 104Inoculating the bacterial solution/ml into 96-well plate, adding 0.1ml bacterial solution and 0.5ml broth culture solution into each well, placing 96-well plate at 37 deg.C and 5% CO2The culture medium of (1) is cultured in a saturated humidity incubator, and after 12 hours, 0mg, 1.5mg, 3.0mg, 4.5mg, 6.0mg, 7.5mg, 9.0mg, 10.5mg of passion fruit anthocyanin extracted in example 1 is added into the holes according to groups0.1ml of mg and 12.0mg aqueous solution is used as a positive control group, a test group S1-1, a test group S1-2, a test group S1-3, a test group S1-4, a test group S1-5, a test group S1-6, a test group S1-7 and a test group S1-8, wherein the water of the aqueous solution is sterile water; adding meat soup culture solution 0.5ml and water solution 0.1ml containing passion fruit anthocyanidin 0.30mg per ml as negative control group into the hole without bacteria solution, and making the above groups at 37 deg.C and 5% CO2After culturing for 48 hours in an incubator with saturated humidity, the strain was taken out, observed and judged for the Minimum Inhibitory Concentration (MIC) of the aqueous solution containing passion fruit anthocyanin against the strain, as shown in table 1. According to the same test procedure as described above, the test data of the passion fruit anthocyanins extracted in test example 2, example 3, comparative example 1, comparative example 2 and comparative example 3 for inhibiting salmonella and escherichia coli are shown in tables 2, 3, 4, 5 and 6, respectively.
Table 1: experimental data for the inhibition of Salmonella and Escherichia coli with Passion fruit anthocyanins extracted in example 1
Table 2: experimental data for the inhibition of Salmonella and Escherichia coli with Passion fruit anthocyanins extracted in example 2
| Salmonella | Escherichia coli | |
| Test groupS2-1(1.5mg/ml) | ++ | +++ |
| Test group S2-2(3.0mg/ml) | ++ | ++ |
| Test group S2-3(4.5mg/ml) | + | + |
| Test group S2-4(6.0mg/ml) | + | + |
| Test group S2-5(7.5mg/ml) | — | — |
| Test group S2-6(9.0mg/ml) | — | — |
| Test group S2-7(10.5mg/ml) | — | — |
| Test group S2-8(12.0mg/ml) | — | — |
| Positive control group (0mg/ml) | +++ | +++ |
| Negative control group (3.0mg/ml) | — | — |
| Minimum inhibitory concentration (mg/ml) | 7.5mg/ml | 7.5mg/ml |
Table 3: experimental data for the inhibition of Salmonella and Escherichia coli with Passion fruit anthocyanins extracted in example 3
| Salmonella | Escherichia coli | |
| Test group S3-1(1.5mg/ml) | ++ | +++ |
| Test group S3-2(3.0mg/ml) | + | ++ |
| Test group S3-3(4.5mg/ml) | + | + |
| Test group S3-4(6.0mg/ml) | — | + |
| Test group S3-5(7.5mg/ml) | — | — |
| Test group S3-6(9.0mg/ml) | — | — |
| Test group S3-7(10.5mg/ml) | — | — |
| Test group S3-8(12.0mg/ml) | — | — |
| Positive control group (0mg/ml) | +++ | +++ |
| Negative control group (3.0mg/ml) | — | — |
| Minimum inhibitory concentration (mg/ml) | 6.0mg/ml | 7.5mg/ml |
Table 4: experimental data for inhibiting salmonella and escherichia coli by using passion fruit anthocyanidin extracted in comparative example 1
| Salmonella | Escherichia coli | |
| Test group D1-1(1.5mg/ml) | +++ | +++ |
| Test group D1-2(3.0mg/ml) | +++ | ++ |
| Test group D1-3(4.5mg/ml) | +++ | ++ |
| Test group D1-4(6.0mg/ml) | ++ | ++ |
| Test group D1-5(7.5mg/ml) | + | + |
| Test group D1-6(9.0mg/ml) | + | — |
| Test group D1-7(10.5mg/ml) | — | — |
| Test group D1-8(12.0mg/ml) | — | — |
| Positive control group (0mg/ml) | +++ | +++ |
| Negative control group (3.0mg/ml) | — | — |
| Minimum inhibitory concentration (mg/ml) | 10.5mg/ml | 9.0mg/ml |
Table 5: experimental data for inhibiting salmonella and escherichia coli by using passion fruit anthocyanin extracted in comparative example 2
| Salmonella | Escherichia coli | |
| Test group D2-1(1.5mg/ml) | +++ | +++ |
| Test group D2-2(3.0mg/ml) | +++ | ++ |
| Test group D2-3(4.5mg/ml) | ++ | ++ |
| Test group D2-4(6.0mg/ml) | ++ | ++ |
| Test group D2-5(7.5mg/ml) | ++ | + |
| Test group D2-6(9.0mg/ml) | + | + |
| Test group D2-7(10.5mg/ml) | — | — |
| Test group D2-8(12.0mg/ml) | — | — |
| Positive control group (0mg/ml) | +++ | +++ |
| Negative control group (3.0mg/ml) | — | — |
| Minimum inhibitory concentration (mg/ml) | 10.5mg/ml | 10.5mg/ml |
Table 6: experimental data for inhibiting salmonella and escherichia coli by using passion fruit anthocyanin extracted in comparative example 3
| Salmonella | Escherichia coli | |
| Test group D3-1(1.5mg/ml) | +++ | +++ |
| Test group D3-2(3.0mg/ml) | +++ | ++ |
| Test group D3-3(4.5mg/ml) | ++ | ++ |
| Test group D3-4(6.0mg/ml) | + | ++ |
| Test group D3-5(7.5mg/ml) | + | + |
| Test group D3-6(9.0mg/ml) | — | + |
| Test group D3-7(10.5mg/ml) | — | — |
| Test group D3-8(12.0mg/ml) | — | — |
| Positive control group (0mg/ml) | +++ | +++ |
| Negative control group (3.0mg/ml) | — | — |
| Minimum inhibitory concentration (mg/ml) | 9.0mg/ml | 10.5mg/ml |
Note: "-" indicates no bacterial growth, "+" indicates bacterial growth, "+ + +" indicates strong bacterial growth, and "+ + + +" indicates strong growth.
As can be seen from the examples, the comparative examples and the test data in tables 1 to 6, the passion fruit anthocyanin extracted by the method has good inhibitory effect on salmonella and escherichia coli, and the passion fruit anthocyanin extracted by only using purple peel has poorer inhibitory effect on salmonella and escherichia coli than the passion fruit anthocyanin extracted by using purple peel and pulp together; the participation of glacial acetic acid is avoided, the forward movement of the esterification reaction is influenced, part of redundant hydroxyl groups are difficult to remove, and the effect of inhibiting salmonella and escherichia coli and the extraction rate are influenced; the tartaric acid and the citric acid are not involved, the modified passion fruit anthocyanin is insufficient in variety, and the effect and the extraction rate of inhibiting salmonella and escherichia coli are also influenced.
Test No. two
2 feeding experiment
90 sick chickens infected with salmonella and escherichia coli are randomly divided into two groups, the first group of 10 chickens serves as a control group, the daily feed serves as the daily ration, the first group of chickens is raised for 3 days, then the daily feed plus 0.5% of the passion fruit anthocyanin extracted in example 3 serves as the daily ration, the second group of chickens 80 serves as an experimental group, the daily feed plus 0.5% of the passion fruit anthocyanin extracted in example 3 serves as the daily ration, the chickens are fed and managed at normal time, observation is carried out for 1 week, and statistical observation data are shown in table 7.
Table 7:
from the experimental data in table 7, it can be known that the passion fruit anthocyanin extracted by the invention has an inhibitory effect on salmonella and escherichia coli of chickens, and can be applied to preparation of chicken feed additives for inhibiting salmonella and escherichia coli.
While embodiments of the invention have been disclosed above, it is not intended to be limited to the uses set forth in the specification and examples. It can be applied to all kinds of fields suitable for the present invention. Additional modifications will readily occur to those skilled in the art. Therefore, the invention is not to be limited to the specific details and embodiments shown and described herein, without departing from the general concept defined by the appended claims and their equivalents.
Claims (6)
1. The extraction method of passion fruit anthocyanin is characterized by comprising the following steps:
removing endocarp of purple passion fruit to obtain purple peel and passion fruit pulp, mixing and crushing to obtain crushed substances;
step two, adding the crushed material into glacial acetic acid in a light-proof environment at the temperature of 20-30 ℃ and under normal pressure, mixing for 1-2min, then sequentially adding tartaric acid with the concentration of 0.5-1mol/L and citric acid with the concentration of 0.5-1mol/L, mixing and stirring for 1-2h, then adding ethanol with the concentration of 75-85%, performing reflux extraction for 3-4h, filtering, performing rotary evaporation, and then placing in an organic solvent for extraction to obtain passion fruit anthocyanin; wherein the dosage proportion is the mass of the crushed material: volume of ethanol: volume of glacial acetic acid: volume of tartaric acid: the volume of the citric acid is 1g:15-20ml:1-2ml:3ml:3 ml.
2. The extraction method of passion fruit anthocyanidin according to claim 1, wherein the organic solvent is one or more of n-hexane, petroleum ether or diethyl ether.
3. The extraction method of passion fruit anthocyanidin according to claim 2, further comprising pretreatment of passion fruit purple peel: shredding purple peel in dark environment, soaking in the solution, placing in a reaction kettle at 20 deg.C and 0.4MPa, maintaining the temperature and pressure for 1 hr, taking out, air drying at room temperature, and pulverizing to 100 mesh; wherein the dissolution liquid is a mixed aqueous solution of 0.3mg/mL of muramidase and 0.4mg/mL of cellulase, and the mass-volume ratio of the purple skin to the dissolution liquid is 1g:1.2 mL.
4. The extraction method of passion fruit anthocyanidin according to claim 3, further comprising the following steps of preprocessing passion fruit pulp: ultrasonically crushing the pulp in a dark environment, soaking the pulp in the soaking solution, placing the soaked pulp in a reaction kettle at the temperature of 20 ℃ and the pressure of 0.4MPa, repeatedly stirring, keeping the temperature and the pressure for 1 hour, taking out the pulp, and air-drying the pulp at room temperature; wherein the soaking solution is 0.4mg/mL lywallzyme water solution, and the mass volume ratio of the pulp to the soaking solution is 1g:1 mL.
5. Use of passion fruit anthocyanidin for the preparation of a chicken feed additive according to any one of claims 1-4.
6. The use of passion fruit anthocyanidin according to claim 5, wherein the passion fruit anthocyanidin is used for preparing a chicken feed additive for inhibiting salmonella and escherichia coli.
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| CN112608356A (en) * | 2021-01-21 | 2021-04-06 | 广西师范大学 | Separation and purification method of passion fruit spiro ketoside monomer and application thereof |
| CN114044837A (en) * | 2021-12-03 | 2022-02-15 | 湖南科技学院 | Method for jointly extracting passion fruit anthocyanin and cellulose biomass |
| CN116283875A (en) * | 2023-02-03 | 2023-06-23 | 陕西省生物农业研究所 | Corncob anthocyanin extraction method and extraction composition thereof |
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| CN112608356A (en) * | 2021-01-21 | 2021-04-06 | 广西师范大学 | Separation and purification method of passion fruit spiro ketoside monomer and application thereof |
| CN112608356B (en) * | 2021-01-21 | 2022-04-29 | 广西师范大学 | Separation and purification method of passion fruit spiro ketoside monomer and application thereof |
| CN114044837A (en) * | 2021-12-03 | 2022-02-15 | 湖南科技学院 | Method for jointly extracting passion fruit anthocyanin and cellulose biomass |
| CN116283875A (en) * | 2023-02-03 | 2023-06-23 | 陕西省生物农业研究所 | Corncob anthocyanin extraction method and extraction composition thereof |
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