CN108865415A - A kind of extracting method of peony seed oil - Google Patents
A kind of extracting method of peony seed oil Download PDFInfo
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- CN108865415A CN108865415A CN201810957353.9A CN201810957353A CN108865415A CN 108865415 A CN108865415 A CN 108865415A CN 201810957353 A CN201810957353 A CN 201810957353A CN 108865415 A CN108865415 A CN 108865415A
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/02—Pretreatment
- C11B1/04—Pretreatment of vegetable raw material
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/02—Pretreatment
- C11B1/025—Pretreatment by enzymes or microorganisms, living or dead
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B3/00—Refining fats or fatty oils
- C11B3/001—Refining fats or fatty oils by a combination of two or more of the means hereafter
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B3/00—Refining fats or fatty oils
- C11B3/006—Refining fats or fatty oils by extraction
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B3/00—Refining fats or fatty oils
- C11B3/12—Refining fats or fatty oils by distillation
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- Life Sciences & Earth Sciences (AREA)
- Microbiology (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a kind of extracting methods of peony seed oil, and specific step is as follows:1)Peony seeds are immersed in dense common salt aqueous solution;2)Peony seeds are moved into progress explosion puffing drying processing in vacuum tank;3)It is digested after peony seeds after drying are crushed;4)It is cooled to room temperature after enzymatic hydrolysis is expected heating and thermal insulation, supernatant is collected after centrifuge separation, then carried out ultrafiltration membrane extraction, obtain peony seeds crude oil;5)After peony seeds crude oil is mixed with monoethanolamine salt, lipase solution, the peony seeds crude oil after obtaining depickling is added;6)Peony seed oil will can be obtained after the heated refluxing extraction of peony seeds crude oil.The extracting method of peony seed oil provided by the invention, the loss of the nutritional ingredient after mentioning oil can be reduced, improve the nutritive value of peony seed oil, this method can be by the extraction of peony seed oil progress synchronous with degreasing depickling simultaneously, shorten production process, cost is reduced, and recovery rate is high, there is good popularization and application foreground.
Description
Technical field
The invention belongs to tree peony deep process technology fields, and in particular to a kind of extracting method of peony seed oil.
Background technique
Peony seed oil, also known as peony oil are the woody nut plant oils extracted by peony seeds, are that China is distinctive, because of it
It is full of nutrition and unique, and have medical care effect, it is the treasure in vegetable oil that " best oil in the world, " is known as by relevant expert
Product, and the health care edible oil and fat that China is exclusive.Peony seed oil is that human body is most suitable in the grease found so far
The grease of nutrition, is the total nutritive value highest of all edible oil, and constituent structure is most reasonable.
Squeezing method, organic solvent extractionprocess is mostly used to prepare peony seed oil at present, these methods are not only at high cost, but also
The denaturation of protein is easily caused, nutritive value is reduced.Therefore it provides a kind of extracting method being simple and efficient, reduces to nutrition
The damage of ingredient is a urgent problem to be solved.
Summary of the invention
The purpose of the present invention is being directed to existing problem, a kind of extracting method of peony seed oil is provided.
The present invention is achieved by the following technical solutions:
A kind of extracting method of peony seed oil, specific step is as follows:
1)The peony seeds selected are immersed in 3-4h in the common salt aqueous solution that concentration is 10-15%;
2)Peony seeds after immersion are drained away the water and are moved back into vacuum tank, are pressurizeed with the rate of rise of 0.5-1.0MPa/min
To 4-7MPa, HIGH PRESSURE TREATMENT 10-15min, then rapid pressure release to normal pressure, is again heated to 50-70 DEG C, is evacuated to 0.03-
0.05MPa is vacuum-treated 20-25min, the drying peony seeds after obtaining extruding;
3)Peony seeds after drying are smashed it through into 50 meshes, are then immersed in the composite enzyme solution that concentration is 0.2-0.5mg/ml
In, a small amount of citric acid is added, adjusting pH is 4.5-6.5, is heated to 50-60 DEG C, is vibrated under 200-300W ultrasonic wave
Digest 40-50min;
4)Will enzymatic hydrolysis material be heated to 120-150 DEG C, isothermal holding 15-20min, after being cooled to room temperature revolving speed be 5000-
It is centrifugated 15-20min under 6000r/min, collects supernatant, is then heated to 30-35 DEG C, in the case where pressure is 0.2-0.5MPa
Ultrafiltration membrane extraction is carried out, peony seeds crude oil is obtained;
5)After peony seeds crude oil is mixed with monoethanolamine salt, the lipase solution that concentration is 0.1-0.2mg/ml is added, moves to
Vacuum degree is to be heated to 45-50 DEG C, heating reaction 40-60min, after obtaining depickling in the vacuum tank of 0.05-0.07MPa
Peony seeds crude oil;
6)The ethanol solution that concentration is 60-70% is added in peony seeds crude oil, 50-60 DEG C of progress refluxing extraction 50- is heated to
Then 70min recycles ethyl alcohol after being evaporated under reduced pressure, peony seed oil can be obtained.
Preferably, a kind of extracting method of peony seed oil, wherein step 1)In, solid-liquid ratio in peony seeds and common salt aqueous solution
For 1g/10-15ml.
Preferably, a kind of extracting method of peony seed oil, wherein step 2)In, the peony seeds drain away the water as peony seeds
Dry tack free.
Preferably, a kind of extracting method of peony seed oil, wherein step 3)In, the solid-liquid ratio of peony seeds and composite enzyme solution
For 1g/5-10ml, wherein complex enzyme is by cellulase, hemicellulase and pectase according to 1:1:1.5 weight ratio composition
's.
Preferably, a kind of extracting method of peony seed oil, wherein step 4)In, the ultrafiltration membrane is that molecular weight cutoff is 10
Ten thousand hollow fiber ultrafiltration membrane.
Preferably, a kind of extracting method of peony seed oil, wherein step 5)In, the additive amount of single ethanol ammonium salt is peony seeds
The 2-3% of crude oil weight, the additive amount of lipase solution are the 4-7% of peony seeds crude oil weight.
Preferably, a kind of extracting method of peony seed oil, wherein step 6)In, the weight of peony seeds crude oil and ethanol solution
Than being 1:1.5.
The present invention has the following advantages that compared with prior art:
1. the extracting method of peony seed oil provided by the invention can be improved firstly, peony seeds are immersed in common salt aqueous solution
The osmotic pressure of peony seeds promotes the moisture penetration in saline solution into peony seeds, improves the water content of peony seeds, be conducive to improve
Subsequent peony seeds puffed degree can accelerate hydrogen between the cellulosic molecule in peony seeds for peony seeds are pressurized and pressure release is handled
Key fracture, promotes cyto-architectural rupture, is conducive to the progress of subsequent enzymatic hydrolysis, carry out vacuumizing place after then heating peony seeds
Reason, can accelerate the evaporation of moisture in peony seeds, form dry extruding peony seeds;Secondly, carrying out ultrasound under acidic environment
Wave enzymolysis processing can accelerate the removal of the cellulose and pectin in peony seeds, improve the eduction rate of peony seed oil, through centrifugation point
From the impurity that can be removed in enzymatic hydrolysis material, pressurization carries out ultrafiltration membrane extraction, can increase the motive force that solute penetrates ultrafiltration membrane, make
It obtains more impurity molecules denaturation and squeezes through filter membrane, to improve the purity of peony seeds crude oil;Again, the single ethanol ammonium salt of addition can
Amidation process occurs with peony seed oil, the alkanolamide that there is amide groups and hydroxyl etc. to enliven group is obtained, and alkanol acyl
Amine is a kind of special surfactant, can make solution retrogradation, improve the degreasing of lipase, to improve free-fat
The removal efficiency of acid;Finally, ethyl alcohol, which is added, carries out hot reflux, the complex enzyme and lipase in peony seeds crude oil can be removed, is obtained
Peony seed oil after degreasing depickling.
2. the extracting method of peony seed oil provided by the invention can reduce the loss of the nutritional ingredient after mentioning oil, improve
The nutritive value of peony seed oil, while the extraction of peony seed oil progress synchronous with degreasing depickling can be shortened production by this method
Process reduces cost, and recovery rate is high.
Specific embodiment
Below with reference to specific implementation method, the present invention is described further.
Embodiment 1
A kind of extracting method of peony seed oil, specific step is as follows:
1)The peony seeds selected are immersed in 3h in the common salt aqueous solution that concentration is 10%;
2)Peony seeds after immersion are drained away the water and are moved back into vacuum tank, are forced into the rate of rise of 0.5MPa/min
4MPa, HIGH PRESSURE TREATMENT 10min, then rapid pressure release to normal pressure, are again heated to 50 DEG C, are evacuated to 0.03MPa, are vacuum-treated
20min, the drying peony seeds after obtaining extruding;
3)Peony seeds after drying are smashed it through into 50 meshes, are then immersed in the composite enzyme solution that concentration is 0.2mg/ml,
A small amount of citric acid is added, adjusting pH is 4.5, is heated to 50 DEG C, and oscillation enzymatic hydrolysis 40min is carried out under 200W ultrasonic wave;
4)Enzymatic hydrolysis material is heated to 120 DEG C, isothermal holding 15min, the centrifugation point in the case where revolving speed is 5000r/min after being cooled to room temperature
From 15min, supernatant is collected, is then heated to 30 DEG C, carried out ultrafiltration membrane extraction in the case where pressure is 0.2MPa, obtain peony seeds hair
Oil;
5)After peony seeds crude oil is mixed with monoethanolamine salt, the lipase solution that concentration is 0.1mg/ml is added, moves to vacuum
Degree is to be heated to 45 DEG C, heating reaction 40min, the peony seeds crude oil after obtaining depickling in the vacuum tank of 0.05MPa;
6)The ethanol solution that concentration is 60% is added in peony seeds crude oil, is heated to 50 DEG C of progress refluxing extraction 50min, so
By ethyl alcohol is recycled after vacuum distillation, peony seed oil can be obtained.
Preferably, wherein step 1)In, solid-liquid ratio is 1g/10ml in peony seeds and common salt aqueous solution.
Preferably, wherein step 2)In, the peony seeds drain away the water as peony seeds dry tack free.
Preferably, wherein step 3)In, the solid-liquid ratio of peony seeds and composite enzyme solution is 1g/5ml, and wherein complex enzyme is
By cellulase, hemicellulase and pectase according to 1:1:1.5 weight ratio composition.
Preferably, wherein step 4)In, the ultrafiltration membrane is the hollow fiber ultrafiltration membrane that molecular weight cutoff is 100,000.
Preferably, wherein step 5)In, the additive amount of single ethanol ammonium salt is the 2% of peony seeds crude oil weight, and lipase is molten
The additive amount of liquid is the 4% of peony seeds crude oil weight.
Preferably, wherein step 6)In, the weight ratio of peony seeds crude oil and ethanol solution is 1:1.5.
Embodiment 2
A kind of extracting method of peony seed oil, specific step is as follows:
1)The peony seeds selected are immersed in 3.5h in the common salt aqueous solution that concentration is 12%;
2)Peony seeds after immersion are drained away the water and are moved back into vacuum tank, are forced into the rate of rise of 0.7MPa/min
5MPa, HIGH PRESSURE TREATMENT 12min, then rapid pressure release to normal pressure, are again heated to 60 DEG C, are evacuated to 0.04MPa, are vacuum-treated
23min, the drying peony seeds after obtaining extruding;
3)Peony seeds after drying are smashed it through into 50 meshes, are then immersed in the composite enzyme solution that concentration is 0.3mg/ml,
A small amount of citric acid is added, adjusting pH is 5.5, is heated to 55 DEG C, and oscillation enzymatic hydrolysis 45min is carried out under 200W ultrasonic wave;
4)Enzymatic hydrolysis material is heated to 130 DEG C, isothermal holding 17min, the centrifugation point in the case where revolving speed is 5500r/min after being cooled to room temperature
From 17min, supernatant is collected, is then heated to 32 DEG C, carried out ultrafiltration membrane extraction in the case where pressure is 0.3MPa, obtain peony seeds hair
Oil;
5)After peony seeds crude oil is mixed with monoethanolamine salt, the lipase solution that concentration is 0.1mg/ml is added, moves to vacuum
Degree is to be heated to 47 DEG C, heating reaction 50min, the peony seeds crude oil after obtaining depickling in the vacuum tank of 0.06MPa;
6)The ethanol solution that concentration is 65% is added in peony seeds crude oil, is heated to 55 DEG C of progress refluxing extraction 60min, so
By ethyl alcohol is recycled after vacuum distillation, peony seed oil can be obtained.
Preferably, wherein step 1)In, solid-liquid ratio is 1g/13ml in peony seeds and common salt aqueous solution.
Preferably, wherein step 2)In, the peony seeds drain away the water as peony seeds dry tack free.
Preferably, wherein step 3)In, the solid-liquid ratio of peony seeds and composite enzyme solution is 1g/7ml, and wherein complex enzyme is
By cellulase, hemicellulase and pectase according to 1:1:1.5 weight ratio composition.
Preferably, wherein step 4)In, the ultrafiltration membrane is the hollow fiber ultrafiltration membrane that molecular weight cutoff is 100,000.
Preferably, wherein step 5)In, the additive amount of single ethanol ammonium salt is the 2.5% of peony seeds crude oil weight, lipase
The additive amount of solution is the 5% of peony seeds crude oil weight.
Preferably, wherein step 6)In, the weight ratio of peony seeds crude oil and ethanol solution is 1:1.5.
Embodiment 3
A kind of extracting method of peony seed oil, specific step is as follows:
1)The peony seeds selected are immersed in 4h in the common salt aqueous solution that concentration is 15%;
2)Peony seeds after immersion are drained away the water and are moved back into vacuum tank, are forced into the rate of rise of 1.0MPa/min
7MPa, HIGH PRESSURE TREATMENT 15min, then rapid pressure release to normal pressure, are again heated to 70 DEG C, are evacuated to 0.05MPa, are vacuum-treated
25min, the drying peony seeds after obtaining extruding;
3)Peony seeds after drying are smashed it through into 50 meshes, are then immersed in the composite enzyme solution that concentration is 0.5mg/ml,
A small amount of citric acid is added, adjusting pH is 6.5, is heated to 60 DEG C, and oscillation enzymatic hydrolysis 50min is carried out under 300W ultrasonic wave;
4)Enzymatic hydrolysis material is heated to 150 DEG C, isothermal holding 20min, the centrifugation point in the case where revolving speed is 6000r/min after being cooled to room temperature
From 20min, supernatant is collected, is then heated to 35 DEG C, carried out ultrafiltration membrane extraction in the case where pressure is 0.5MPa, obtain peony seeds hair
Oil;
5)After peony seeds crude oil is mixed with monoethanolamine salt, the lipase solution that concentration is 0.2mg/ml is added, moves to vacuum
Degree is to be heated to 50 DEG C, heating reaction 60min, the peony seeds crude oil after obtaining depickling in the vacuum tank of 0.07MPa;
6)The ethanol solution that concentration is 70% is added in peony seeds crude oil, is heated to 60 DEG C of progress refluxing extraction 70min, so
By ethyl alcohol is recycled after vacuum distillation, peony seed oil can be obtained.
Preferably, wherein step 1)In, solid-liquid ratio is 1g/15ml in peony seeds and common salt aqueous solution.
Preferably, wherein step 2)In, the peony seeds drain away the water as peony seeds dry tack free.
Preferably, wherein step 3)In, the solid-liquid ratio of peony seeds and composite enzyme solution is 1g/10ml, wherein complex enzyme
It is by cellulase, hemicellulase and pectase according to 1:1:1.5 weight ratio composition.
Preferably, wherein step 4)In, the ultrafiltration membrane is the hollow fiber ultrafiltration membrane that molecular weight cutoff is 100,000.
Preferably, wherein step 5)In, the additive amount of single ethanol ammonium salt is the 3% of peony seeds crude oil weight, and lipase is molten
The additive amount of liquid is the 7% of peony seeds crude oil weight.
Preferably, wherein step 6)In, the weight ratio of peony seeds crude oil and ethanol solution is 1:1.5.
Comparative example 1:Remove step 1)In saline solution impregnate, remaining is same as Example 1.
Comparative example 2:Remove step 2)In peony seeds expanding treatment, remaining is same as Example 1.
Comparative example 3:Remove step 3)In supersonic oscillations, remaining is same as Example 1.
Comparative example 4:Remove step 5)In single ethanol ammonium salt, remaining is same as Example 1.
Experimental example:The extraction of peony seed oil, control group are carried out using the method that embodiment 1 and comparative example 1-4 are provided respectively
1-2 selects squeezing method and ultrasonic extraction respectively, and experimental result is as shown in Table 1:
Table one
Peony seeds weight(g) | Discharging weight(g) | Recovery rate(%) | Total unsaturated fatty acid(%) | |
Embodiment 1 | 1000 | 237 | 23.7 | 91.8 |
Comparative example 1 | 1000 | 201 | 20.1 | 89.2 |
Comparative example 2 | 1000 | 205 | 20.5 | 88.7 |
Comparative example 3 | 1000 | 200 | 20.0 | 88.9 |
Comparative example 4 | 1000 | 203 | 20.3 | 89.1 |
Control group 1 | 1200 | 224.4 | 18.7 | 83.5 |
Control group 2 | 1200 | 230.4 | 19.2 | 84.7 |
As can be seen from Table I, in embodiment 1, the recovery rate highest of peony seed oil, and unsaturated fatty acid content highest, by
This as it can be seen that the peony seed oil that the present invention improves extracting method, not only recovery rate is high, and can reduce the nutrition after mentioning oil at
The loss divided, improves the nutritive value of peony seed oil.
The above description is merely a specific embodiment, but scope of protection of the present invention is not limited thereto, any
It transforms or replaces, should be covered by the scope of protection of the present invention without what creative work was expected.
Claims (7)
1. a kind of extracting method of peony seed oil, which is characterized in that specific step is as follows:
1)The peony seeds selected are immersed in 3-4h in the common salt aqueous solution that concentration is 10-15%;
2)Peony seeds after immersion are drained away the water and are moved back into vacuum tank, are pressurizeed with the rate of rise of 0.5-1.0MPa/min
To 4-7MPa, HIGH PRESSURE TREATMENT 10-15min, then rapid pressure release to normal pressure, is again heated to 50-70 DEG C, is evacuated to 0.03-
0.05MPa is vacuum-treated 20-25min, the drying peony seeds after obtaining extruding;
3)Peony seeds after drying are smashed it through into 50 meshes, are then immersed in the composite enzyme solution that concentration is 0.2-0.5mg/ml
In, a small amount of citric acid is added, adjusting pH is 4.5-6.5, is heated to 50-60 DEG C, is vibrated under 200-300W ultrasonic wave
Digest 40-50min;
4)Will enzymatic hydrolysis material be heated to 120-150 DEG C, isothermal holding 15-20min, after being cooled to room temperature revolving speed be 5000-
It is centrifugated 15-20min under 6000r/min, collects supernatant, is then heated to 30-35 DEG C, in the case where pressure is 0.2-0.5MPa
Ultrafiltration membrane extraction is carried out, peony seeds crude oil is obtained;
5)After peony seeds crude oil is mixed with monoethanolamine salt, the lipase solution that concentration is 0.1-0.2mg/ml is added, moves to
Vacuum degree is to be heated to 45-50 DEG C, heating reaction 40-60min, after obtaining depickling in the vacuum tank of 0.05-0.07MPa
Peony seeds crude oil;
6)The ethanol solution that concentration is 60-70% is added in peony seeds crude oil, 50-60 DEG C of progress refluxing extraction 50- is heated to
Then 70min recycles ethyl alcohol after being evaporated under reduced pressure, peony seed oil can be obtained.
2. a kind of extracting method of peony seed oil as described in claim 1, which is characterized in that step 1)In, peony seeds and food
Solid-liquid ratio is 1g/10-15ml in saline solution.
3. a kind of extracting method of peony seed oil as described in claim 1, which is characterized in that step 2)In, the peony seeds
It drains away the water as peony seeds dry tack free.
4. a kind of extracting method of peony seed oil as described in claim 1, which is characterized in that step 3)In, peony seeds and multiple
The solid-liquid ratio of synthase solution is 1g/5-10ml, and wherein complex enzyme is by cellulase, hemicellulase and pectase according to 1:1:
1.5 weight ratio composition.
5. a kind of extracting method of peony seed oil as described in claim 1, which is characterized in that step 4)In, the ultrafiltration membrane
The hollow fiber ultrafiltration membrane for being 100,000 for molecular weight cutoff.
6. a kind of extracting method of peony seed oil as described in claim 1, which is characterized in that step 5)In, single ethanol ammonium salt
Additive amount be peony seeds crude oil weight 2-3%, the additive amount of lipase solution is the 4-7% of peony seeds crude oil weight.
7. a kind of extracting method of peony seed oil as described in claim 1, which is characterized in that step 6)In, peony seeds crude oil
Weight ratio with ethanol solution is 1:1.5.
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Cited By (11)
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CN112920886A (en) * | 2021-01-07 | 2021-06-08 | 龙池牡丹实业有限公司 | Enzymolysis preparation method of peony seed micromolecule oil |
CN112980560A (en) * | 2021-03-09 | 2021-06-18 | 东北农业大学 | Method for extracting peony seed oil through ultrasonic-assisted biological enzymolysis |
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CN116240067A (en) * | 2023-03-13 | 2023-06-09 | 菏泽牡丹产业技术研究院 | Peony seed crude oil, refined peony seed oil, and preparation methods and applications thereof |
CN118576680A (en) * | 2024-08-06 | 2024-09-03 | 江西省药品检验检测研究院 | Jindan accessory delay suppository and preparation method thereof |
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