CN111481962A - Method for extracting salmonella-inhibiting substance from olive pomace and application - Google Patents
Method for extracting salmonella-inhibiting substance from olive pomace and application Download PDFInfo
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- CN111481962A CN111481962A CN202010326300.4A CN202010326300A CN111481962A CN 111481962 A CN111481962 A CN 111481962A CN 202010326300 A CN202010326300 A CN 202010326300A CN 111481962 A CN111481962 A CN 111481962A
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- salmonella
- ethyl acetate
- olive pomace
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/04—Solvent extraction of solutions which are liquid
- B01D11/0492—Applications, solvents used
Abstract
The invention provides a method for extracting a salmonella inhibiting substance from olive pomace and application thereof, belonging to the technical field of deep processing of food byproducts and comprising the following steps: the method comprises the following steps: 1) mixing olive pomace with an acid solution, adjusting the pH value to be neutral, standing, centrifuging the obtained standing matter, and removing an acid layer to obtain an acid-removing solution; 2) mixing the deacidification solution obtained in the step 1) with ethyl acetate, whirling, filtering, and removing the ethyl acetate to obtain the antibacterial substance. By adopting the method provided by the invention, about 3ml of salmonella inhibiting substances can be extracted from 250g of olive pomace, the total phenol content is 351mg/kg, salmonella can be inhibited, the additional value of the olive is increased, and environmental pollution is avoided.
Description
Technical Field
The invention belongs to the technical field of deep processing of food byproducts, and particularly relates to a method for extracting a salmonella inhibiting substance from olive pomace and application of the salmonella inhibiting substance.
Background
The fructus Canarii albi is a arbor plant of Canarium of Burseraceae, and has oval to spindle shape, yellow green color at maturity and thick peel. The olive pomace is the solid matter left after squeezing olive fruits and extracting juice or oil, and comprises pericarp, pulp, seeds, stalks and the like, and the pomace is usually discarded. The effective utilization of olive pomace and the environmental protection are inevitable trends, and if the olive pomace cannot be effectively utilized, the waste of resources is caused on one hand, and the environmental pollution is caused on the other hand.
Disclosure of Invention
In view of the above, the invention aims to provide a method for extracting a salmonella inhibiting substance from olive pomace and application thereof, and the method provided by the invention can be used for extracting the salmonella inhibiting substance from the olive pomace, has an inhibiting effect on salmonella, increases the additional value of olives and reduces environmental pollution.
In order to achieve the above purpose, the invention provides the following technical scheme:
the invention provides a method for extracting a salmonella inhibiting substance from olive pomace, which comprises the following steps:
1) mixing olive pomace with an acid solution, adjusting the pH value to be neutral, standing, centrifuging the obtained standing matter, and removing an acid layer to obtain an acid-removing solution;
2) mixing the deacidification solution obtained in the step 1) with ethyl acetate, whirling, filtering, and removing the ethyl acetate to obtain the antibacterial substance.
Preferably, the centrifugation conditions of step 1) include: the rotating speed of the centrifugation is 3000-5000 rpm, and the time of the centrifugation is 10-30 min.
Preferably, the step 1) acid solution comprises a hydrochloric acid solution.
Preferably, the volume ratio of the mass of the olive pomace in the step 1) to the volume of the acid solution is 200-300 g: 200-300 ml.
Preferably, the vortex time in the step 2) is 30-600 s.
Preferably, the standing time in the step 2) is 1.5-2.5 h.
Preferably, the mass ratio of the volume of the ethyl acetate in the step 2) to the olive pomace is 450-550 ml to 200-300 g.
Preferably, the method for removing the ethyl acetate in the step 2) comprises distillation, and the distillation temperature is 50-60 ℃.
The invention also provides application of the salmonella-inhibiting substance obtained by the method in the technical scheme in inhibiting salmonella.
The invention provides a method for extracting a salmonella inhibiting substance from olive pomace, which comprises the following steps: mixing olive pomace with an acid solution, adjusting the pH value to be neutral, standing, centrifuging the obtained standing matter, and removing an acid layer to obtain an acid-removing solution; 2) mixing the deacidification solution obtained in the step 1) with ethyl acetate, whirling, filtering, and removing the ethyl acetate to obtain the antibacterial substance. The method provided by the invention can be used for extracting and obtaining the salmonella inhibition bacteria from the olive pomace, inhibiting the salmonella, increasing the additional value of the olive and causing no environmental pollution.
Drawings
FIG. 1 shows the results of the zone of inhibition of Salmonella typhimurium by the stock solution of the extract of example 3, the diameter of the zone of inhibition being 14.2 mm.
Detailed Description
The invention provides a method for extracting a salmonella inhibiting substance from olive pomace, which comprises the following steps:
1) mixing olive pomace with an acid solution, adjusting the pH value to be neutral, standing, centrifuging the obtained standing matter, and removing an acid layer to obtain an acid-removing solution;
2) mixing the deacidification solution obtained in the step 1) with ethyl acetate, whirling, filtering, and removing the ethyl acetate to obtain the antibacterial substance.
Preferably, the olive pomace is filtered and then mixed with an acid solution.
In the present invention, the olive pomace is the remainder of cold pressing of olive fruits.
Preferably, the olive pomace is mixed with an acid solution, the pH value is adjusted to be neutral, then the mixture is kept stand, the obtained standing material is centrifuged, and an acid layer is discarded to obtain the deacidification solution.
In the invention, the acid solution preferably comprises a hydrochloric acid solution, and the volume ratio of the mass of the olive pomace to the volume of the acid solution is preferably 200-300 g: 200-300 ml, and more preferably 250g:250 ml. In the invention, the rotation speed of the centrifugation is preferably 3000-5000 rpm, and the time of the centrifugation is preferably 10-30 min. In the invention, the standing time is preferably 1.5-2.5 h, and more preferably 2 h.
The obtained deacidification solution is mixed with ethyl acetate and swirled, and the ethyl acetate is removed after filtration to obtain the antibacterial substance.
In the present invention, the mixing method preferably includes a vortex, and the time of the vortex is preferably 30 to 600 seconds.
In the invention, the mass ratio of the volume of the ethyl acetate to the olive pomace is preferably 450-550 ml to 200-300 g, and more preferably 500ml to 250 g.
In the invention, the method for removing the ethyl acetate preferably comprises distillation, and the distillation temperature is preferably 50-60 ℃, and more preferably 55 ℃. In the invention, the bacteriostatic substances are tyrosol and hydroxytyrosol, and the contents of the bacteriostatic substances are 11mg/kg and 28mg/kg respectively.
The invention also provides application of the salmonella-inhibiting substance obtained by the method in the technical scheme in inhibiting salmonella.
In the present invention, the Salmonella is preferably a standard strain of Salmonella typhimurium (Salmonella typhimurium) ATCC 14028.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Filtering olive pomace (a three-phase system product) by using gauze, taking 250g of filtered olive pomace, adding 250m of L hydrochloric acid for hydrolysis, stirring, adjusting the pH value to be neutral, standing for 2 hours, centrifuging at 3000-5000 rpm for 15min, removing a hydrochloric acid layer, adding 500m of L ethyl acetate (pomace/ethyl acetate, v/v and 1/2), swirling for 30-600 s, filtering, removing the pomace, removing the ethyl acetate by using a rotary evaporator through vacuum evaporation, and setting the temperature to be between room temperature and 55 ℃ until an extract with antibacterial activity is obtained, wherein the results are shown in table 1.
Example 2
Filtering olive pomace (a 'two-phase' system product) by using gauze, taking 250g of filtered olive pomace, adding 250m of L hydrochloric acid for hydrolysis, stirring, adjusting the pH to be neutral, standing for 2 hours, centrifuging at 3000-5000 rpm for 15min, removing a hydrochloric acid layer, adding 500m of L ethyl acetate (pomace/ethyl acetate, v/v and 1/2), swirling for 30-600 s, filtering, removing pomace, removing ethyl acetate by using a rotary evaporator through vacuum evaporation, and setting the temperature to be between room temperature and 55 ℃ until an extract with antibacterial activity is obtained, wherein the results are shown in table 1.
Example 3
Preparing extract solution with mass concentration of 0.00 (sterile water), 0.00078125, 0.0015625, 0.03125, 0.0625, 0.25 and 0.5 (stock solution) mg/m L by adopting a Minimum Inhibitory Concentration (MIC) method and adopting an antibacterial ring method, wherein the sample adding volume is 60 mu L, and 0.1m L-1 m L salmonella typhimurium standard strain suspension (10)7CFU/m L) was added to 100m L nutrient agar cooled to 45-50 deg.C, mixed well, poured onto a plate, left horizontally to stand for coagulation, the extract was injected into the well (phi 6mm), incubated at 37 deg.C for 24h, and the zone of inhibition was measured, the results are shown in Table 2.
Table 1 extraction results of examples 1 and 2
| Group number | Acid scavenger | Extraction agent | Volume of extract | Extract concentration (mg/L) |
| Example 1 | Hydrochloric acid | Ethyl acetate | 3 | 351 |
| Example 2 | Is free of | Ethyl acetate | 1 | 125 |
As can be seen from Table 1, the bacteriostatic substances extracted from the pomace after deacidification were 2.8 times as much as the bacteriostatic substances extracted without deacidification. The olive pomace has good extraction effect after deacidification.
Table 2 minimum inhibitory concentration results for the extract of example 3
| Group number | Standard strains | Sample application material | Sample concentration (mg/m L) | d/mm | Sensitivity of the device |
| Example 3 | Salmonella typhimurium | Ethyl acetate extract | 0.00 | 6 | Is not sensitive |
| Example 3 | Salmonella typhimurium | Ethyl acetate extract | 0.00078125 | 6 | Is not sensitive |
| Example 3 | Salmonella typhimurium | Ethyl acetate extract | 0.0015625 | 6 | Is not sensitive |
| Example 3 | Salmonella typhimurium | Ethyl acetate extract | 0.03125 | 7.02 | Low degree |
| Example 3 | Salmonella typhimurium | Ethyl acetate extract | 0.0625 | 8.70 | Low degree |
| Example 3 | Salmonella typhimurium | Ethyl acetate extract | 0.25 | 12.54 | Of moderate degree |
| Example 3 | Salmonella typhimurium | Ethyl acetate extract | 0.5 | 14.2 | Of moderate degree |
Note: according to the execution standard of the antibacterial drug susceptibility test, the diameter d of the inhibition zone is not sensitive to 6mm, d is less than 10mm and is less sensitive to low degree, d is more than or equal to 10mm and is less than 10mm and is less sensitive to medium degree, d is more than or equal to 15mm and is less than 20mm and is highly sensitive, and d is more than or equal to 20mm and is extremely sensitive.
As can be seen from Table 2, the minimum inhibitory concentration of the extract against Salmonella typhimurium was 0.03125mg/m L.
The embodiment can show that the polyphenol substances can be extracted from the olive pomace by adopting the method provided by the invention, the extract can inhibit salmonella, the additional value of the olive is increased, and the environment pollution is avoided.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (9)
1. A method for extracting a salmonella-inhibiting substance from olive pomace is characterized by comprising the following steps:
1) mixing olive pomace with an acid solution, adjusting the pH value to be neutral, standing, centrifuging the obtained standing matter, and removing an acid layer to obtain an acid-removing solution;
2) mixing the deacidification solution obtained in the step 1) with ethyl acetate, whirling, filtering, and removing the ethyl acetate to obtain the antibacterial substance.
2. The method of claim 1, wherein the conditions of the step 1) centrifugation comprise: the rotating speed of the centrifugation is 3000-5000 rpm, and the time of the centrifugation is 10-30 min.
3. The method of claim 1, wherein the step 1) acid solution comprises a hydrochloric acid solution.
4. The method according to claim 1 or 3, wherein the volume ratio of the mass of the olive pomace to the acid solution in the step 1) is 200-300 g: 200-300 ml.
5. The method according to claim 1, wherein the time of the step 2) vortex is 30-600 s.
6. The method of claim 1, wherein the standing time in the step 2) is 1.5-2.5 h.
7. The method according to claim 1, wherein the mass ratio of the volume of the ethyl acetate in the step 2) to the olive pomace is 450-550 ml: 200-300 g.
8. The method according to claim 1, wherein the method for removing the ethyl acetate in the step 2) comprises distillation, and the temperature of the distillation is 50-60 ℃.
9. Use of the Salmonella inhibitory substance obtained by the method according to any one of claims 1 to 8 for inhibiting Salmonella.
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Effective date of registration: 20220804 Address after: 746000 Xiangyu eco industrial park, Wudu District, Longnan City, Gansu Province Patentee after: LONGNAN XIANGYU OLIVE DEVELOPMENT CO.,LTD. Patentee after: Lanzhou Customs Technical Center Address before: 730010 no.2168, Nanhe Road, Chengguan District, Lanzhou City, Gansu Province Patentee before: Lanzhou Customs Technical Center |
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