CN116870043A - Method for extracting antioxidant from pomegranate seeds - Google Patents
Method for extracting antioxidant from pomegranate seeds Download PDFInfo
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- CN116870043A CN116870043A CN202310988051.9A CN202310988051A CN116870043A CN 116870043 A CN116870043 A CN 116870043A CN 202310988051 A CN202310988051 A CN 202310988051A CN 116870043 A CN116870043 A CN 116870043A
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- 241000219991 Lythraceae Species 0.000 title claims abstract description 152
- 235000014360 Punica granatum Nutrition 0.000 title claims abstract description 152
- 239000003963 antioxidant agent Substances 0.000 title claims abstract description 97
- 230000003078 antioxidant effect Effects 0.000 title claims abstract description 62
- 238000000034 method Methods 0.000 title claims abstract description 29
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 54
- 238000010438 heat treatment Methods 0.000 claims abstract description 44
- 239000002904 solvent Substances 0.000 claims abstract description 43
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 42
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 40
- 238000001035 drying Methods 0.000 claims abstract description 37
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 36
- 238000000605 extraction Methods 0.000 claims abstract description 29
- 239000002131 composite material Substances 0.000 claims abstract description 23
- 238000000227 grinding Methods 0.000 claims abstract description 22
- 210000000582 semen Anatomy 0.000 claims abstract description 17
- 238000001704 evaporation Methods 0.000 claims abstract description 15
- 150000001875 compounds Chemical class 0.000 claims abstract description 10
- 230000008020 evaporation Effects 0.000 claims abstract description 7
- 238000001914 filtration Methods 0.000 claims abstract description 4
- 230000005484 gravity Effects 0.000 claims description 100
- 239000007788 liquid Substances 0.000 claims description 39
- 239000000243 solution Substances 0.000 claims description 33
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 28
- 239000000843 powder Substances 0.000 claims description 27
- 230000001276 controlling effect Effects 0.000 claims description 22
- 238000002156 mixing Methods 0.000 claims description 22
- 238000003756 stirring Methods 0.000 claims description 17
- 239000006228 supernatant Substances 0.000 claims description 15
- 238000010992 reflux Methods 0.000 claims description 14
- 108010059892 Cellulase Proteins 0.000 claims description 12
- 229940106157 cellulase Drugs 0.000 claims description 12
- 238000000967 suction filtration Methods 0.000 claims description 9
- 239000000523 sample Substances 0.000 claims description 8
- 238000004140 cleaning Methods 0.000 claims description 7
- 239000012153 distilled water Substances 0.000 claims description 7
- 239000011259 mixed solution Substances 0.000 claims description 7
- 239000012528 membrane Substances 0.000 claims description 5
- 238000000643 oven drying Methods 0.000 claims description 5
- 238000004321 preservation Methods 0.000 claims description 5
- 230000000630 rising effect Effects 0.000 claims description 5
- 239000013049 sediment Substances 0.000 claims description 5
- 238000000926 separation method Methods 0.000 claims description 5
- 238000005086 pumping Methods 0.000 claims description 3
- 230000001105 regulatory effect Effects 0.000 claims description 2
- 238000005119 centrifugation Methods 0.000 claims 3
- 230000000694 effects Effects 0.000 abstract description 12
- 229930003935 flavonoid Natural products 0.000 abstract description 10
- 235000017173 flavonoids Nutrition 0.000 abstract description 10
- 150000002215 flavonoids Chemical class 0.000 abstract description 10
- 150000008442 polyphenolic compounds Chemical class 0.000 abstract description 10
- 235000013824 polyphenols Nutrition 0.000 abstract description 10
- 239000002699 waste material Substances 0.000 abstract description 7
- 238000013329 compounding Methods 0.000 abstract description 6
- 239000002994 raw material Substances 0.000 abstract description 2
- 238000002835 absorbance Methods 0.000 description 21
- HEILIGJNYTWOHU-UHFFFAOYSA-N ethanol 2-hydroxybenzoic acid Chemical compound CCO.OC(=O)C1=CC=CC=C1O HEILIGJNYTWOHU-UHFFFAOYSA-N 0.000 description 14
- 230000000052 comparative effect Effects 0.000 description 12
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 7
- 235000003891 ferrous sulphate Nutrition 0.000 description 7
- 239000011790 ferrous sulphate Substances 0.000 description 7
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 7
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 7
- 239000000203 mixture Substances 0.000 description 7
- 238000002360 preparation method Methods 0.000 description 6
- 150000003254 radicals Chemical class 0.000 description 5
- 239000012141 concentrate Substances 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 206010046996 Varicose vein Diseases 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 208000027185 varicose disease Diseases 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/13—Preparation or pretreatment of starting material involving cleaning, e.g. washing or peeling
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/17—Preparation or pretreatment of starting material involving drying, e.g. sun-drying or wilting
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
Abstract
The invention provides a method for extracting antioxidants from pomegranate seeds. A method for extracting antioxidant from semen Granati comprises the following steps: drying and grinding the pomegranate seeds; ultrasonic-assisted enzymolysis; adding a compound solvent and reflux-extracting; centrifugally separating and suction-filtering; and heating for evaporation concentration. According to the invention, by taking the pomegranate seeds as raw materials and taking the composite solvent prepared by compounding methanol, ethanol, acetone and hydrochloric acid according to a proportion as the extraction solvent, the antioxidants such as polyphenols, flavonoids and the like in the pomegranate seeds can be efficiently extracted, and the activities of the antioxidants such as polyphenols, flavonoids and the like are higher, so that the effect of fully and effectively utilizing the pomegranate seeds is achieved, and the waste is reduced.
Description
Technical Field
The invention relates to the technical field of extraction of plant active ingredients, in particular to a method for extracting antioxidants from pomegranate seeds.
Background
The pomegranate seeds contain rich polyphenols and flavonoid substances, so that the pomegranate seeds have an antioxidant effect, can remove free radicals in vivo, delay aging, prevent cardiovascular diseases, improve skin and varicose veins, and reduce blood pressure, blood fat and cholesterol.
At present, there are few methods for extracting antioxidants from pomegranate seeds, and people usually do not eat the pomegranate seeds when eating the pomegranate seeds, so that substances such as antioxidants in the pomegranate seeds are wasted greatly.
Therefore, we propose a method for extracting antioxidants from pomegranate seeds with high efficiency to reduce waste of the pomegranate seeds.
Disclosure of Invention
Aiming at the defects existing in the prior art, the invention aims to provide a method for extracting antioxidants from pomegranate seeds.
A method for extracting antioxidant from semen Granati comprises the following steps:
s1: oven drying and grinding semen Granati
Cleaning the pomegranate seeds, drying, adding into a grinder, and grinding to obtain pomegranate seed powder;
s2: ultrasonic-assisted enzymolysis
Stirring and mixing the pomegranate seed powder and distilled water, adding cellulase for enzymolysis, and simultaneously carrying out ultrasonic assistance to obtain an enzymolysis liquid;
s3: adding compound solvent and reflux extracting
Mixing methanol, ethanol, acetone and hydrochloric acid to prepare a composite solvent, adding the composite solvent into the enzymolysis liquid, heating, and carrying out heat preservation reflux extraction to obtain a mixed liquid;
s4: centrifugal separation and suction filtration
The controller controls the centrifugal component of the extractor to start, the mixed liquid is centrifugally separated through the centrifugal component to obtain supernatant and lower sediment, the supernatant is pumped and filtered through the pumping and filtering component, and the supernatant is pumped into the concentrator through the filter membrane to obtain extracting solution;
s5: heating for evaporating and concentrating
And (3) performing preliminary evaporation concentration on the extracting solution by heating, then adjusting the heating temperature, and continuing the evaporation concentration to obtain the pomegranate seed antioxidant.
Further, the step S1 of drying and grinding the pomegranate seeds specifically comprises the following steps:
s1.1: cleaning the pomegranate seeds, putting the cleaned pomegranate seeds into a drying box, starting the drying box, heating and drying until a first gravity sensor in the drying box detects that the gravity of the pomegranate seeds is no longer reduced, and sending a signal to a controller by the first gravity sensor;
s1.2: after receiving the signal sent by the gravity sensor, the controller controls the drying box to stop heating so as to obtain dry pomegranate seeds;
s1.3: then, the controller controls the discharge port of the drying box to be opened, and the dried pomegranate seeds enter the grinder through the discharge port of the drying box;
s1.4: until a second gravity sensor in the grinder detects that the gravity in the grinder is no longer increasing, the second gravity sensor sends a signal to the controller;
s1.5: and after the controller receives the signal sent by the second gravity sensor, controlling the grinder to grind for 2-5 hours to obtain the pomegranate seed powder.
Further, the ultrasonic-assisted enzymolysis in the step S2 specifically comprises the following steps:
s2.1: adding the pomegranate seed powder prepared in the step S1.5 and distilled water into an extractor together until a third gravity sensor in the extractor detects that the gravity in the extractor is not increased any more, and sending a signal to a controller by the third gravity sensor;
s2.2: after the controller receives the signal sent by the third gravity sensor, the stirrer in the extractor is controlled to stir and disperse at the speed of 300-500 r/min;
s2.3: after stirring for 30-40min, the controller controls the feed component of the extractor to be opened, cellulase is added into the extractor through the feed component for enzymolysis, and meanwhile, the controller controls the ultrasonic probe to penetrate into the mixed solution for ultrasonic auxiliary enzymolysis, so that enzymolysis solution is obtained.
Further, the step S3 of adding the compound solvent and extracting by reflux specifically comprises the following steps:
s3.1: adding methanol, ethanol, acetone and hydrochloric acid into a mixing tank together according to the volume ratio of 1-3:2-3:3-5:1-2, and stirring and mixing uniformly to obtain a composite solvent;
s3.2: the enzymolysis liquid is prepared by a hydraulic pump according to the volume ratio step S2.3: the compound solvent=1:30-50 is pressed into the extractor until a liquid level sensor in the extractor detects that the liquid level in the extractor is no longer rising, and the liquid level sensor sends a signal to a controller;
s3.3: after the controller receives the signal sent by the liquid level sensor, controlling a heater in the extractor to heat at a speed of 2-5 ℃/min until a temperature sensor in the extractor detects that the temperature in the extractor is increased to 60-70 ℃, and sending a signal to the controller by the temperature sensor;
s3.4: after the controller receives the signal sent by the temperature sensor, the heater is controlled to stop heating and keep the temperature, and the mixed solution is obtained after heat-preserving reflux extraction for 1-2 h.
Further, the evaporating concentration is performed by heating in the step S5, which specifically comprises the following steps:
s5.1: when the fourth gravity sensor in the concentrator detects that the gravity in the concentrator is not increased any more, the fourth gravity sensor sends a signal to the controller;
s5.2: after receiving the signal sent by the fourth gravity sensor, the controller controls the concentrator to heat at 85-95 ℃, and the extracting solution prepared in the step S4 is evaporated and concentrated, and the composite solvent is recovered through the condenser to obtain concentrated solution;
s5.3: heating for 2-3 hr, regulating the heating temperature of the concentrator to 100-120deg.C, and evaporating and concentrating the concentrated solution to obtain semen Granati antioxidant;
s5.4: and the fourth gravity sensor sends a signal to the controller again until the fourth gravity sensor detects that the gravity in the concentrator is not reduced, and the controller stops heating after receiving the signal sent by the fourth gravity sensor again.
Further, the ultrasonic power is 80-100W.
Further, the centrifugal speed of the centrifugal component is 4000-5000r/min, and the centrifugal duration is 40-50min.
Compared with the prior art, the invention has the advantages that:
1. according to the invention, by taking the pomegranate seeds as raw materials and taking the composite solvent prepared by compounding methanol, ethanol, acetone and hydrochloric acid according to a proportion as the extraction solvent, the antioxidants such as polyphenols, flavonoids and the like in the pomegranate seeds can be efficiently extracted, and the activities of the antioxidants such as polyphenols, flavonoids and the like are higher, so that the effect of fully and effectively utilizing the pomegranate seeds is achieved, and the waste is reduced.
2. According to the invention, by using ultrasonic wave to assist cellulase enzymolysis and then carrying out reflux extraction on the antioxidants in the pomegranate seeds, not only can the extraction rate and the extraction efficiency of the antioxidants be improved, but also the utilization rate of solvent resources can be improved.
3. According to the invention, the extracting solution is heated to 85-90 ℃, so that not only can the composite solvent be removed by evaporation, but also the cellulase can be inactivated, and the quality and purity of the prepared antioxidant are improved.
Drawings
Fig. 1 is a flowchart of a method for extracting antioxidants from pomegranate seeds according to an embodiment of the present invention.
FIG. 2 is a summary of the test results of example 1 and comparative example 1.
FIG. 3 is a summary of the test results of example 1 and comparative example 2.
FIG. 4 is a summary of the test results of example 1 and comparative example 3.
FIG. 5 is a summary of the test results of example 1 and comparative example 4.
FIG. 6 is a summary of the test results of example 1 and comparative example 5.
FIG. 7 is a summary of the test results of example 1 and comparative example 6.
Detailed Description
The present invention will be described in further detail with reference to specific examples.
Example 1
A method for extracting antioxidant from semen Granati, as shown in figures 1-7, comprises the following steps:
s1: oven drying and grinding semen Granati
Cleaning the pomegranate seeds, putting the pomegranate seeds into a drying box, starting the drying box, heating the pomegranate seeds to dry the pomegranate seeds until a first gravity sensor in the drying box detects that the gravity of the pomegranate seeds is not reduced any more, sending a signal to a controller by the first gravity sensor, controlling the drying box to stop heating after receiving the signal sent by the gravity sensor by the controller, obtaining dry pomegranate seeds, then controlling a discharge hole of the drying box to be opened by the controller, enabling the dry pomegranate seeds to enter a grinding machine through the discharge hole of the drying box until a second gravity sensor in the grinding machine detects that the gravity in the grinding machine is not increased any more, sending a signal to the controller by the second gravity sensor, and controlling the grinding machine to grind the pomegranate seeds for 2 hours after receiving the signal sent by the second gravity sensor by the controller, thus obtaining the pomegranate seed powder;
s2: ultrasonic-assisted enzymolysis
Weighing the pomegranate seed powder, marking the weight of the pomegranate seed powder as M1, adding the pomegranate seed powder and distilled water into an extractor, sending a signal to a controller by a third gravity sensor in the extractor until the third gravity sensor in the extractor detects that the gravity in the extractor is not increased any more, after receiving the signal sent by the third gravity sensor, controlling a stirrer in the extractor to stir and disperse at the speed of 300r/min, after stirring for 30min, controlling a feeding component of the extractor to open by the controller, adding cellulase into the extractor through the feeding component for enzymolysis, and simultaneously controlling an ultrasonic probe to probe into the mixed liquid for ultrasonic auxiliary enzymolysis to obtain enzymolysis liquid;
s3: adding compound solvent and reflux extracting
Adding methanol, ethanol, acetone and hydrochloric acid into a mixing tank together according to the volume ratio of 1:2:3:1, stirring and mixing uniformly to obtain a composite solvent, and carrying out enzymolysis on the solution according to the volume ratio by a hydraulic pump: the method comprises the steps that a composite solvent=1:30 is pressed into an extractor until a liquid level sensor in the extractor detects that the liquid level in the extractor is not rising any more, the liquid level sensor sends a signal to a controller, after receiving the signal sent by the liquid level sensor, the controller controls a heater in the extractor to heat at a speed of 2 ℃/min until a temperature sensor in the extractor detects that the temperature in the extractor rises to 60 ℃, the temperature sensor sends a signal to the controller, and after receiving the signal sent by the temperature sensor, the controller controls the heater to stop heating and preserving heat, and the heat preservation and reflux extraction are carried out for 1h, so that mixed liquid is obtained;
s4: centrifugal separation and suction filtration
The controller controls the centrifugal component of the extractor to start, the mixed solution is centrifuged for 40min at the centrifugal rate of 4000r/min through the centrifugal component to obtain supernatant and lower sediment, the supernatant is subjected to suction filtration through the suction filtration component, and the supernatant is pumped into the concentrator through the filter membrane to obtain extract;
s5: heating for evaporating and concentrating
When the fourth gravity sensor in the concentrator detects that the gravity in the concentrator is not increased any more, the fourth gravity sensor sends a signal to the controller, the controller receives the signal sent by the fourth gravity sensor, then controls the concentrator to heat at 85 ℃, evaporates and concentrates the extracting solution, and recovers the compound solvent through the condenser to obtain concentrated solution, after heating for 2 hours, the heating temperature of the concentrator is adjusted to 100 ℃, the concentrated solution is continuously evaporated and concentrated to obtain the pomegranate seed antioxidant, until the fourth gravity sensor detects that the gravity in the concentrator is not reduced any more, the fourth gravity sensor sends a signal to the controller again, and after receiving the signal sent by the fourth gravity sensor again, the controller is controlled to stop heating, and the weight M2 of the pomegranate seed antioxidant is recorded.
Then, each test was performed on the above-mentioned pomegranate seed antioxidants:
firstly, calculating the extraction rate of the pomegranate seed antioxidant to be 9.36% according to a formula by using the weight M1 of the weighed pomegranate seed powder and the weight M2 of the pomegranate seed antioxidant;
then, 2g of pomegranate seed antioxidant is weighed and added into 2mL of 6mmol/L ferrous sulfate, then 2mL of 6mmol/L salicylic acid-ethanol is added, finally 2mL of 6mmol/L hydrogen peroxide solution is added, the mixture is shaken uniformly and then water bath is carried out at 37 ℃ for 0.5h, the absorbance at 510nm is measured and is marked as A1, then equal volume water is used for replacing salicylic acid-ethanol solution, the absorbance at 510nm is measured and is marked as A2, equal volume water is used for replacing the pomegranate seed antioxidant as a blank control, the absorbance at 510nm is marked as A0, then the clearance rate of the pomegranate seed antioxidant to OH free radical is calculated to be 79.8 percent according to the formula of which the clearance rate is = [1- (A1-A2)/A0 ]. Times.100%.
Example 2
A method for extracting antioxidant from semen Granati, as shown in figure 1, comprises the following steps:
s1: oven drying and grinding semen Granati
Cleaning the pomegranate seeds, putting the pomegranate seeds into a drying box, starting the drying box, heating the pomegranate seeds to dry until a first gravity sensor in the drying box detects that the gravity of the pomegranate seeds is not reduced any more, sending a signal to a controller by the first gravity sensor, controlling the drying box to stop heating after receiving the signal sent by the gravity sensor by the controller to obtain the dried pomegranate seeds, then controlling a discharge hole of the drying box to be opened by the controller, enabling the dried pomegranate seeds to enter a grinding machine through the discharge hole of the drying box until a second gravity sensor in the grinding machine detects that the gravity in the grinding machine is not increased any more, sending a signal to the controller by the second gravity sensor, and controlling the grinding machine to grind for 3.5 hours after receiving the signal sent by the second gravity sensor by the controller to obtain the pomegranate seed powder;
s2: ultrasonic-assisted enzymolysis
Weighing the pomegranate seed powder, marking the weight of the pomegranate seed powder as M1, adding the pomegranate seed powder and distilled water into an extractor, sending a signal to a controller by a third gravity sensor in the extractor until the third gravity sensor in the extractor detects that the gravity in the extractor is not increased any more, after receiving the signal sent by the third gravity sensor, controlling a stirrer in the extractor to stir and disperse at the speed of 400r/min, after stirring for 35min, controlling a feeding component of the extractor to open by the controller, adding cellulase into the extractor through the feeding component for enzymolysis, and simultaneously controlling an ultrasonic probe to probe into the mixed liquid for ultrasonic auxiliary enzymolysis to obtain enzymolysis liquid;
s3: adding compound solvent and reflux extracting
Adding methanol, ethanol, acetone and hydrochloric acid into a mixing tank together according to the volume ratio of 2:2.5:4:1.5, stirring and mixing uniformly to obtain a composite solvent, and carrying out enzymolysis on the solution according to the volume ratio by a hydraulic pump: the method comprises the steps that a composite solvent=1:40 is pressed into an extractor until a liquid level sensor in the extractor detects that the liquid level in the extractor is not rising any more, the liquid level sensor sends a signal to a controller, after receiving the signal sent by the liquid level sensor, the controller controls a heater in the extractor to heat at a speed of 3.5 ℃/min until a temperature sensor in the extractor detects that the temperature in the extractor rises to 65 ℃, the temperature sensor sends a signal to the controller, and after receiving the signal sent by the temperature sensor, the controller controls the heater to stop heating and preserving heat, and the heat preservation and reflux extraction are performed for 1.5h, so that mixed liquid is obtained;
s4: centrifugal separation and suction filtration
The controller controls the centrifugal component of the extractor to start, the mixed solution is centrifuged for 45min at a centrifugal speed of 4500r/min through the centrifugal component to obtain supernatant and lower sediment, the supernatant is pumped and filtered through the pumping and filtering component, and the supernatant is pumped into the concentrator through the filter membrane to obtain extracting solution;
s5: heating for evaporating and concentrating
When the fourth gravity sensor in the concentrator detects that the gravity in the concentrator is not increased any more, the fourth gravity sensor sends a signal to the controller, the controller receives the signal sent by the fourth gravity sensor, then controls the concentrator to heat at 90 ℃, evaporates and concentrates the extracting solution, and recovers the composite solvent through the condenser to obtain concentrated solution, after heating for 2.5h, the heating temperature of the concentrator is adjusted to 110 ℃, the concentrated solution is continuously evaporated and concentrated to obtain the pomegranate seed antioxidant, until the fourth gravity sensor detects that the gravity in the concentrator is not reduced any more, the fourth gravity sensor sends a signal to the controller again, and after receiving the signal sent by the fourth gravity sensor again, the controller is controlled to stop heating, and the weight of the pomegranate seed antioxidant is recorded.
Example 3
A method for extracting antioxidant from semen Granati, as shown in figure 1, comprises the following steps:
s1: oven drying and grinding semen Granati
Cleaning the pomegranate seeds, putting the pomegranate seeds into a drying box, starting the drying box, heating the pomegranate seeds to dry the pomegranate seeds until a first gravity sensor in the drying box detects that the gravity of the pomegranate seeds is not reduced any more, sending a signal to a controller by the first gravity sensor, controlling the drying box to stop heating after receiving the signal sent by the gravity sensor by the controller, obtaining dry pomegranate seeds, then controlling a discharge hole of the drying box to be opened by the controller, enabling the dry pomegranate seeds to enter a grinding machine through the discharge hole of the drying box until a second gravity sensor in the grinding machine detects that the gravity in the grinding machine is not increased any more, sending a signal to the controller by the second gravity sensor, and controlling the grinding machine to grind the pomegranate seeds for 5 hours after receiving the signal sent by the second gravity sensor by the controller, thus obtaining the pomegranate seed powder;
s2: ultrasonic-assisted enzymolysis
Weighing the pomegranate seed powder, marking the weight of the pomegranate seed powder as M1, adding the pomegranate seed powder and distilled water into an extractor, sending a signal to a controller by a third gravity sensor in the extractor until the third gravity sensor in the extractor detects that the gravity in the extractor is not increased any more, after receiving the signal sent by the third gravity sensor, controlling a stirrer in the extractor to stir and disperse at the speed of 500r/min, after stirring for 40min, controlling a feeding component of the extractor to open by the controller, adding cellulase into the extractor through the feeding component for enzymolysis, and simultaneously controlling an ultrasonic probe to probe into the mixed liquid for ultrasonic auxiliary enzymolysis to obtain enzymolysis liquid;
s3: adding compound solvent and reflux extracting
Adding methanol, ethanol, acetone and hydrochloric acid into a mixing tank together according to the volume ratio of 3:3:5:2, stirring and mixing uniformly to obtain a composite solvent, and carrying out enzymolysis on the solution according to the volume ratio by a hydraulic pump: the method comprises the steps that a composite solvent=1:50 is pressed into an extractor until a liquid level sensor in the extractor detects that the liquid level in the extractor is not rising any more, the liquid level sensor sends a signal to a controller, after receiving the signal sent by the liquid level sensor, the controller controls a heater in the extractor to heat at a speed of 5 ℃/min until a temperature sensor in the extractor detects that the temperature in the extractor rises to 70 ℃, the temperature sensor sends a signal to the controller, and after receiving the signal sent by the temperature sensor, the controller controls the heater to stop heating and preserving heat, and the heat preservation and reflux extraction are carried out for 2 hours, so that mixed liquid is obtained;
s4: centrifugal separation and suction filtration
The controller controls the centrifugal component of the extractor to start, the mixed solution is centrifuged for 50min at a centrifugal speed of 5000r/min through the centrifugal component to obtain supernatant and lower sediment, the supernatant is subjected to suction filtration through the suction filtration component, and the supernatant is pumped into the concentrator through the filter membrane to obtain extract;
s5: heating for evaporating and concentrating
When the fourth gravity sensor in the concentrator detects that the gravity in the concentrator is not increased any more, the fourth gravity sensor sends a signal to the controller, the controller receives the signal sent by the fourth gravity sensor, then controls the concentrator to heat at 95 ℃, evaporates and concentrates the extracting solution, and recovers the composite solvent through the condenser to obtain concentrated solution, after heating for 3 hours, the heating temperature of the concentrator is adjusted to 120 ℃, the concentrated solution is continuously evaporated and concentrated to obtain the pomegranate seed antioxidant, until the fourth gravity sensor detects that the gravity in the concentrator is not reduced any more, the fourth gravity sensor sends a signal to the controller again, and after receiving the signal sent by the fourth gravity sensor again, the controller is controlled to stop heating, and the weight of the pomegranate seed antioxidant is recorded.
Comparative example 1
A method for extracting antioxidant from semen Granati comprises, referring to the preparation steps of example 1 as shown in fig. 1 and 2, adding methanol, ethanol, acetone and hydrochloric acid into a mixing tank together at a volume ratio of 1:2:3:1 instead of adding methanol and acetone into the mixing tank together at a volume ratio of 1:3 under the same conditions.
Then, each test was performed on the above-mentioned pomegranate seed antioxidants:
firstly, calculating the extraction rate of the pomegranate seed antioxidant to be 7.03% according to a formula by using the weight M1 of the weighed pomegranate seed powder and the weight M2 of the pomegranate seed antioxidant;
then, 2g of pomegranate seed antioxidant is weighed and added into 2mL of 6mmol/L ferrous sulfate, then 2mL of 6mmol/L salicylic acid-ethanol is added, finally 2mL of 6mmol/L hydrogen peroxide solution is added, the mixture is shaken uniformly and then water bath is carried out at 37 ℃ for 0.5h, the absorbance at 510nm is measured and is marked as A1, then equal volume water is used for replacing salicylic acid-ethanol solution, the absorbance at 510nm is measured and is marked as A2, equal volume water is used for replacing the pomegranate seed antioxidant as a blank control, the absorbance at 510nm is marked as A0, then the clearance rate of the pomegranate seed antioxidant is calculated to be 65.2 percent according to the formula of = [1- (A1-A2)/A0 ]. Times.100 percent.
As can be seen from comparing the test results of the above example 1, the composite solvent prepared by compounding methanol, ethanol, acetone and hydrochloric acid in proportion is used as the extraction solvent, which can efficiently extract the antioxidants such as polyphenols, flavonoids and the like in the pomegranate seeds, and the antioxidant has higher activity, thereby achieving the effect of fully and effectively utilizing the pomegranate seeds and reducing waste.
Comparative example 2
A method for extracting antioxidant from semen Granati comprises, referring to the preparation steps of example 1 as shown in fig. 1 and 3, adding methanol, ethanol, acetone and hydrochloric acid into a mixing tank together at a volume ratio of 1:2:3:1 instead of adding ethanol and acetone into the mixing tank together at a volume ratio of 2:3, with the other conditions unchanged.
Then, each test was performed on the above-mentioned pomegranate seed antioxidants:
firstly, calculating the extraction rate of the pomegranate seed antioxidant to be 7.32% according to a formula by using the weight M1 of the weighed pomegranate seed powder and the weight M2 of the pomegranate seed antioxidant;
then, 2g of pomegranate seed antioxidant is weighed and added into 2mL of 6mmol/L ferrous sulfate, then 2mL of 6mmol/L salicylic acid-ethanol is added, finally 2mL of 6mmol/L hydrogen peroxide solution is added, the mixture is shaken uniformly and then water bath is carried out at 37 ℃ for 0.5h, the absorbance at 510nm is measured and is marked as A1, then equal volume water is used for replacing salicylic acid-ethanol solution, the absorbance at 510nm is measured and is marked as A2, equal volume water is used for replacing the pomegranate seed antioxidant as a blank control, the absorbance at 510nm is marked as A0, then the clearance rate of the pomegranate seed antioxidant to OH free radical is calculated to be 63.8 percent according to the formula of which the clearance rate is = [1- (A1-A2)/A0 ]. Times.100%.
As can be seen from comparing the test results of the above example 1, the composite solvent prepared by compounding methanol, ethanol, acetone and hydrochloric acid in proportion is used as the extraction solvent, which can efficiently extract the antioxidants such as polyphenols, flavonoids and the like in the pomegranate seeds, and the antioxidant has higher activity, thereby achieving the effect of fully and effectively utilizing the pomegranate seeds and reducing waste.
Comparative example 3
Referring to the preparation procedure of example 1, as shown in fig. 1 and 4, other conditions are unchanged, and only methanol, ethanol, acetone and hydrochloric acid are added into a mixing tank together according to a volume ratio of 1:2:3:1 instead of ethanol, acetone and hydrochloric acid being added into the mixing tank together according to a volume ratio of 2:3:1.
Then, each test was performed on the above-mentioned pomegranate seed antioxidants:
firstly, calculating the extraction rate of the pomegranate seed antioxidant to be 8.87% according to a formula by using the weight M1 of the weighed pomegranate seed powder and the weight M2 of the pomegranate seed antioxidant;
then, 2g of pomegranate seed antioxidant is weighed and added into 2mL of 6mmol/L ferrous sulfate, then 2mL of 6mmol/L salicylic acid-ethanol is added, finally 2mL of 6mmol/L hydrogen peroxide solution is added, the mixture is shaken uniformly and then water bath is carried out at 37 ℃ for 0.5h, the absorbance at 510nm is measured and is marked as A1, then equal volume water is used for replacing salicylic acid-ethanol solution, the absorbance at 510nm is measured and is marked as A2, equal volume water is used for replacing the pomegranate seed antioxidant as a blank control, the absorbance at 510nm is marked as A0, then the clearance rate of the pomegranate seed antioxidant is calculated to be 74.6 percent according to the formula of = [1- (A1-A2)/A0 ]. Times.100 percent.
As can be seen from comparing the test results of the above example 1, the composite solvent prepared by compounding methanol, ethanol, acetone and hydrochloric acid in proportion is used as the extraction solvent, which can efficiently extract polyphenols and flavonoids antioxidants from the pomegranate seeds, and the polyphenols and flavonoids antioxidants have higher activity, so that the effect of fully and effectively utilizing the pomegranate seeds is achieved, and the waste is reduced.
Comparative example 4
A method for extracting antioxidant from semen Granati comprises, referring to the preparation steps of example 1 as shown in fig. 1 and 4, adding methanol, ethanol, acetone and hydrochloric acid into a mixing tank together at a volume ratio of 1:2:3:1 instead of adding ethanol and ethanol into the mixing tank together at a volume ratio of 1:2 under the same conditions.
Then, each test was performed on the above-mentioned pomegranate seed antioxidants:
firstly, calculating the extraction rate of the pomegranate seed antioxidant to be 6.86% according to a formula by using the weight M1 of the weighed pomegranate seed powder and the weight M2 of the pomegranate seed antioxidant;
then, 2g of pomegranate seed antioxidant is weighed and added into 2mL of 6mmol/L ferrous sulfate, then 2mL of 6mmol/L salicylic acid-ethanol is added, finally 2mL of 6mmol/L hydrogen peroxide solution is added, the mixture is shaken uniformly and then water bath is carried out at 37 ℃ for 0.5h, the absorbance at 510nm is measured and is marked as A1, then equal volume water is used for replacing salicylic acid-ethanol solution, the absorbance at 510nm is measured and is marked as A2, equal volume water is used for replacing the pomegranate seed antioxidant as a blank control, the absorbance at 510nm is marked as A0, then the clearance rate of the pomegranate seed antioxidant is calculated to be 74.3 percent of OH free radical according to the formula of = [1- (A1-A2)/A0 ]. Times.100%.
As can be seen from comparing the test results of the above example 1, the composite solvent prepared by compounding methanol, ethanol, acetone and hydrochloric acid in proportion is used as the extraction solvent, which can efficiently extract the antioxidants such as polyphenols, flavonoids and the like in the pomegranate seeds, and the antioxidant has higher activity, thereby achieving the effect of fully and effectively utilizing the pomegranate seeds and reducing waste.
Comparative example 5
A method for extracting antioxidant from semen Granati comprises, as shown in FIGS. 1 and 5, referring to the preparation step of example 1, other conditions are unchanged, and only the ultrasonic auxiliary extraction step is removed.
Then, each test was performed on the above-mentioned pomegranate seed antioxidants:
firstly, calculating the extraction rate of the pomegranate seed antioxidant to be 7.54% according to a formula by using the weight M1 of the weighed pomegranate seed powder and the weight M2 of the pomegranate seed antioxidant;
then, 2g of pomegranate seed antioxidant is weighed and added into 2mL of 6mmol/L ferrous sulfate, then 2mL of 6mmol/L salicylic acid-ethanol is added, finally 2mL of 6mmol/L hydrogen peroxide solution is added, the mixture is shaken uniformly and then water bath is carried out at 37 ℃ for 0.5h, the absorbance at 510nm is measured and is marked as A1, then equal volume water is used for replacing salicylic acid-ethanol solution, the absorbance at 510nm is measured and is marked as A2, equal volume water is used for replacing the pomegranate seed antioxidant as a blank control, the absorbance at 510nm is marked as A0, then the clearance rate of the pomegranate seed antioxidant to OH free radical is calculated to be 76.8 percent according to the formula of which the clearance rate is = [1- (A1-A2)/A0 ]. Times.100 percent.
As is clear from comparison of the test results of example 1, the extraction rate and extraction efficiency of the antioxidants in the pomegranate seeds can be improved and the utilization rate of the solvent resources can be improved by using ultrasonic-assisted cellulase enzymolysis and then reflux-extracting the antioxidants.
Comparative example 6
A method for extracting antioxidant from semen Granati comprises, as shown in FIGS. 1 and 6, referring to the preparation step of example 1, removing only the step of adding cellulase under the same conditions.
Then, each test was performed on the above-mentioned pomegranate seed antioxidants:
firstly, calculating the extraction rate of the pomegranate seed antioxidant to be 8.03% according to a formula by using the weight M1 of the weighed pomegranate seed powder and the weight M2 of the pomegranate seed antioxidant;
then, 2g of pomegranate seed antioxidant is weighed and added into 2mL of 6mmol/L ferrous sulfate, then 2mL of 6mmol/L salicylic acid-ethanol is added, finally 2mL of 6mmol/L hydrogen peroxide solution is added, the mixture is shaken uniformly and then water bath is carried out at 37 ℃ for 0.5h, the absorbance at 510nm is measured and is marked as A1, then equal volume water is used for replacing salicylic acid-ethanol solution, the absorbance at 510nm is measured and is marked as A2, equal volume water is used for replacing the pomegranate seed antioxidant as a blank control, the absorbance at 510nm is marked as A0, then the clearance rate of the pomegranate seed antioxidant is calculated to be 78.6 percent according to the formula of = [1- (A1-A2)/A0 ]. Times.100 percent.
As is clear from comparison of the test results of example 1, the extraction rate and extraction efficiency of the antioxidants in the pomegranate seeds can be improved and the utilization rate of the solvent resources can be improved by using ultrasonic-assisted cellulase enzymolysis and then reflux-extracting the antioxidants.
The above embodiments are merely illustrative of the principles of the present invention and its effectiveness, and are not intended to limit the invention. Modifications and variations may be made to the above-described embodiments by those skilled in the art without departing from the spirit and scope of the invention. Accordingly, it is intended that all equivalent modifications and variations of the invention be covered by the claims, which are within the ordinary skill of the art, be within the spirit and scope of the present disclosure.
Claims (7)
1. A method for extracting antioxidants from pomegranate seeds is characterized by comprising the following steps:
s1: oven drying and grinding semen Granati
Cleaning the pomegranate seeds, drying, adding into a grinder, and grinding to obtain pomegranate seed powder;
s2: ultrasonic-assisted enzymolysis
Stirring and mixing the pomegranate seed powder and distilled water, adding cellulase for enzymolysis, and simultaneously carrying out ultrasonic assistance to obtain an enzymolysis liquid;
s3: adding compound solvent and reflux extracting
Mixing methanol, ethanol, acetone and hydrochloric acid to prepare a composite solvent, adding the composite solvent into the enzymolysis liquid, heating, and carrying out heat preservation reflux extraction to obtain a mixed liquid;
s4: centrifugal separation and suction filtration
The controller controls the centrifugal component of the extractor to start, the mixed liquid is centrifugally separated through the centrifugal component to obtain supernatant and lower sediment, the supernatant is pumped and filtered through the pumping and filtering component, and the supernatant is pumped into the concentrator through the filter membrane to obtain extracting solution;
s5: heating for evaporating and concentrating
And (3) performing preliminary evaporation concentration on the extracting solution by heating, then adjusting the heating temperature, and continuing the evaporation concentration to obtain the pomegranate seed antioxidant.
2. The method for extracting antioxidants from pomegranate seeds according to claim 1, wherein the step S1 of drying and grinding the pomegranate seeds comprises the steps of:
s1.1: cleaning the pomegranate seeds, putting the cleaned pomegranate seeds into a drying box, starting the drying box, heating and drying until a first gravity sensor in the drying box detects that the gravity of the pomegranate seeds is no longer reduced, and sending a signal to a controller by the first gravity sensor;
s1.2: after receiving the signal sent by the gravity sensor, the controller controls the drying box to stop heating so as to obtain dry pomegranate seeds;
s1.3: then, the controller controls the discharge port of the drying box to be opened, and the dried pomegranate seeds enter the grinder through the discharge port of the drying box;
s1.4: until a second gravity sensor in the grinder detects that the gravity in the grinder is no longer increasing, the second gravity sensor sends a signal to the controller;
s1.5: and after the controller receives the signal sent by the second gravity sensor, controlling the grinder to grind for 2-5 hours to obtain the pomegranate seed powder.
3. The method for extracting antioxidants from pomegranate seeds according to claim 2, wherein the ultrasonic-assisted enzymolysis of step S2 specifically comprises the following steps:
s2.1: adding the pomegranate seed powder prepared in the step S1.5 and distilled water into an extractor together until a third gravity sensor in the extractor detects that the gravity in the extractor is not increased any more, and sending a signal to a controller by the third gravity sensor;
s2.2: after the controller receives the signal sent by the third gravity sensor, the stirrer in the extractor is controlled to stir and disperse at the speed of 300-500 r/min;
s2.3: after stirring for 30-40min, the controller controls the feed component of the extractor to be opened, cellulase is added into the extractor through the feed component for enzymolysis, and meanwhile, the controller controls the ultrasonic probe to penetrate into the mixed solution for ultrasonic auxiliary enzymolysis, so that enzymolysis solution is obtained.
4. The method for extracting antioxidants from pomegranate seeds according to claim 3, wherein the step S3 of adding a complex solvent and reflux-extracting comprises the steps of:
s3.1: adding methanol, ethanol, acetone and hydrochloric acid into a mixing tank together according to the volume ratio of 1-3:2-3:3-5:1-2, and stirring and mixing uniformly to obtain a composite solvent;
s3.2: the enzymolysis liquid is prepared by a hydraulic pump according to the volume ratio step S2.3: the compound solvent=1:30-50 is pressed into the extractor until a liquid level sensor in the extractor detects that the liquid level in the extractor is no longer rising, and the liquid level sensor sends a signal to a controller;
s3.3: after the controller receives the signal sent by the liquid level sensor, controlling a heater in the extractor to heat at a speed of 2-5 ℃/min until a temperature sensor in the extractor detects that the temperature in the extractor is increased to 60-70 ℃, and sending a signal to the controller by the temperature sensor;
s3.4: after the controller receives the signal sent by the temperature sensor, the heater is controlled to stop heating and keep the temperature, and the mixed solution is obtained after heat-preserving reflux extraction for 1-2 h.
5. The method for extracting antioxidants from pomegranate seeds according to claim 1, wherein the heating in step S5 is performed for evaporation concentration, specifically comprising the steps of:
s5.1: when the fourth gravity sensor in the concentrator detects that the gravity in the concentrator is not increased any more, the fourth gravity sensor sends a signal to the controller;
s5.2: after receiving the signal sent by the fourth gravity sensor, the controller controls the concentrator to heat at 85-95 ℃, and the extracting solution prepared in the step S4 is evaporated and concentrated, and the composite solvent is recovered through the condenser to obtain concentrated solution;
s5.3: heating for 2-3 hr, regulating the heating temperature of the concentrator to 100-120deg.C, and evaporating and concentrating the concentrated solution to obtain semen Granati antioxidant;
s5.4: and the fourth gravity sensor sends a signal to the controller again until the fourth gravity sensor detects that the gravity in the concentrator is not reduced, and the controller stops heating after receiving the signal sent by the fourth gravity sensor again.
6. The method for extracting antioxidants from pomegranate seeds of claim 3, wherein the ultrasonic power is 80-100W.
7. The method for extracting antioxidants from pomegranate seeds according to claim 1, wherein the centrifugation rate of the centrifugation assembly is 4000-5000r/min and the centrifugation time is 40-50min.
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