阅读说明：本技术 一种用于山茶籽的提取方法 (Extraction method for camellia seeds ) 是由 李艳 李梓源 章文浩 孙学华 范冠宝 于 2021-08-31 设计创作，主要内容包括：本发明提供一种用于山茶籽的提取方法,包括如下步骤：(1)在山茶籽中加入水浸泡,滤去水,加入乙醇溶液,冷藏浸泡,取山茶籽,烘干后破碎,得破碎物；(2)在破碎物中加入乙醇溶液,搅拌,微波处理,得微波处理物；(3)在微波处理物中加入碱性蛋白酶改性颗粒,搅拌,保温,均质后,调节pH值,加入糖酶溶液,酶解,过滤,得粗油；(4)将粗油进行压榨,离心,取上清液,得山茶籽油。本发明通过特定的制备工艺,可提高破乳率和山茶籽油的出油率,并可延缓脂肪生成脂肪酸的速率,使山茶籽油具有好的氧化稳定性,本发明的山茶籽提取方法简单、易于操控,适合工业化大生产。(The invention provides an extraction method for camellia seeds, which comprises the following steps: (1) soaking semen Camelliae Japonicae in water, filtering off water, adding ethanol solution, cold preserving, soaking, oven drying semen Camelliae Japonicae, and crushing to obtain crushed product; (2) adding ethanol solution into the crushed material, stirring, and performing microwave treatment to obtain a microwave treated material; (3) adding alkaline protease modified particles into the microwave treated product, stirring, keeping the temperature, homogenizing, adjusting the pH value, adding carbohydrase solution, performing enzymolysis, and filtering to obtain crude oil; (4) squeezing the crude oil, centrifuging, and collecting supernatant to obtain oleum Camelliae Japonicae. The camellia seed extraction method disclosed by the invention has the advantages that the demulsification rate and the oil yield of the camellia seed oil can be improved through a specific preparation process, the rate of fatty acid generation by fat can be delayed, and the camellia seed oil has good oxidation stability.)

1. A method for extracting camellia seeds is characterized by comprising the following steps: the method comprises the following steps: taking camellia seeds, soaking and refrigerating the camellia seeds for pretreatment, performing particle modification treatment by using ethanol microwave and alkaline protease to obtain crude oil, and finally squeezing and centrifuging to obtain camellia seed oil;

the alkaline protease modified particles are prepared by adding alkaline protease into nano SiO₂Suspension, feed liquid mass ratio 1: stirring at a speed of 80-90r/min for 25-35min at a speed of 6-8, and spray drying to obtain the product;

the alkaline protease modified particle treatment comprises the following steps: adding alkaline protease modified particles into the camellia seeds subjected to soaking and cold storage pretreatment and ethanol microwave treatment, stirring, heating to 55-65 deg.C, keeping the temperature for 40-60min, homogenizing for 10-15min, adjusting pH to 8-9, and adding carbohydrase solution for enzymolysis.

2. An extraction method for camellia seeds according to claim 1, wherein: the nano SiO₂The suspension is prepared from nano SiO₂The powder comprises the following components in percentage by mass 1: 6-7: 3-4, adding water and sucrose ester, stirring at the speed of 600-50 r/min under ultrasonic treatment for 35-45min, and standing for 40-50 min.

3. An extraction method for camellia seeds according to any one of claims 1 or 2, characterized in that: the method comprises the following steps:

(1) soaking and refrigerating pretreatment: adding water into camellia seeds, soaking for 2-3h, filtering to remove water, adding an ethanol solution with the mass concentration of 35-45%, cold-preserving and soaking for 4-6h at 8-12 ℃, taking the camellia seeds, drying and crushing to obtain crushed substances;

(2) ethanol microwave: adding an ethanol solution with the mass concentration of 60-70% into the crushed object, stirring, treating for 45-55s with the microwave power of 500-;

(3) and (3) treating alkaline protease modified particles: according to the feed liquid mass ratio of 1: 0.5-0.7, adding alkaline protease modified particles into the microwave treated substance, stirring, heating to 55-65 ℃, keeping the temperature for 40-60min, homogenizing at 5000-6000r/min and 60-80MPa for 10-15min, adjusting the pH value to 8-9, adding 0.5-0.7% w/w carbohydrase solution based on the microwave treated substance, carrying out enzymolysis at 35-45 ℃ for 30-40min, and filtering to obtain crude oil;

(4) squeezing and centrifuging: squeezing the crude oil, centrifuging, and collecting supernatant to obtain oleum Camelliae Japonicae.

4. An extraction method for camellia seeds according to claim 3, wherein: in the step (1), the mass ratio of the camellia seeds, water and 35-45% ethanol solution is 1: 3-5: 3 to 5.

5. An extraction method for camellia seeds according to claim 3, wherein: in the step (1), the mixture is refrigerated and soaked for 5 hours at the temperature of 10 ℃.

6. An extraction method for camellia seeds according to claim 3, wherein: in the step (2), the microwave power is 600W for 50s, the microwave is stopped for 2min, and the microwave power is increased to 800W for 3 min.

7. An extraction method for camellia seeds according to claim 3, wherein: in the step (3), the heating rate is 5 ℃/min.

8. An extraction method for camellia seeds according to claim 3, wherein: in the step (3), the homogenizing condition is that the rotating speed is 5500r/min and the pressure is 70 MPa.

9. An extraction method for camellia seeds according to claim 3, wherein: in the step (4), the squeezing conditions are that the temperature is 45-55 ℃, the pressure is 40-50MPa, the pressure is 40-50min, and the pressure is maintained for 40-60 min; centrifuging at 3000-5000r/min for 30-40min after squeezing.

10. An extraction method for camellia seeds according to claim 3, wherein: in the step (4), the squeezing conditions are that the temperature is 50 ℃, the pressure is 45MPa, the pressure is 45min, and the pressure is maintained for 50 min.

Technical Field

The invention relates to the technical field of plant extraction, in particular to an extraction method for camellia seeds.

Background

Camellia seeds are the alias name of Camellia seeds (the academic name Camellia seed) and are the fruits of Camellia trees, and the Camellia trees belong to woody oil trees and are unique woody plant oil resources in China. The camellia seeds are rich in various active substances, more active ingredients in the camellia seeds are extracted, and the utilization value of the camellia seed oil can be fully improved. Because the camellia seeds contain more grease components, emulsion is easily formed in the extraction process, so that the effective components of the camellia seeds are reduced, the difficulty of the subsequent extraction step is increased, and the stability and the quality of the seed oil are also influenced by the existence of emulsified particles. The camellia oil is easy to lose internal effective components through processes such as multiple treatment and the like, so that the nutritive value of a finished product is reduced.

Disclosure of Invention

Accordingly, the present invention is directed to a method for extracting camellia seeds, which solves the above problems.

The technical scheme of the invention is realized as follows:

a method for extracting camellia seeds comprises the following steps: taking camellia seeds, soaking and refrigerating the camellia seeds for pretreatment, performing particle modification treatment by using ethanol microwave and alkaline protease to obtain crude oil, and finally squeezing and centrifuging to obtain camellia seed oil;

wherein the alkaline protease modified particles are prepared by adding alkaline protease into nano SiO₂Suspension, feed liquid mass ratio 1: stirring at a speed of 80-90r/min for 25-35min at a speed of 6-8, and spray drying to obtain the product;

wherein, the alkaline protease modified particles are treated by the following steps: adding alkaline protease modified particles into the camellia seeds subjected to soaking and cold storage pretreatment and ethanol microwave treatment, stirring, heating to 55-65 deg.C, keeping the temperature for 40-60min, homogenizing for 10-15min, adjusting pH to 8-9, and adding carbohydrase solution for enzymolysis;

further, the nano SiO₂The suspension is prepared from nano SiO₂The powder comprises the following components in percentage by mass 1: 6-7: 3-4, adding water and sucrose ester, performing ultrasonic stirring treatment at the temperature of 30-50 ℃ for 35-45min at the stirring speed of 600-;

further, the method for extracting the camellia seeds comprises the following steps:

(1) soaking and refrigerating pretreatment: adding water into camellia seeds, soaking for 2-3h, filtering to remove water, adding an ethanol solution with the mass concentration of 35-45%, cold-preserving and soaking for 4-6h at 8-12 ℃, taking the camellia seeds, drying and crushing to obtain crushed substances;

(2) ethanol microwave: adding an ethanol solution with the mass concentration of 60-70% into the crushed object, stirring, treating for 45-55s with the microwave power of 500-;

(3) and (3) treating alkaline protease modified particles: according to the feed liquid mass ratio of 1: 0.5-0.7, adding alkaline protease modified particles into the microwave treated substance, stirring, heating to 55-65 ℃, keeping the temperature for 40-60min, homogenizing at 5000-6000r/min and 60-80MPa for 10-15min, adjusting the pH value to 8-9, adding 0.5-0.7% w/w carbohydrase solution based on the microwave treated substance, carrying out enzymolysis at 35-45 ℃ for 30-40min, and filtering to obtain crude oil;

(4) squeezing and centrifuging: squeezing the crude oil, centrifuging, and collecting supernatant to obtain oleum Camelliae Japonicae.

Further, in the step (1), the mass ratio of the camellia seeds, water and 35-45% ethanol solution is 1: 3-5: 3-5;

further explaining, in the step (1), the mixture is refrigerated and soaked for 5 hours at the temperature of 10 ℃;

further explaining, in the step (2), the microwave power is 600W for 50s, the microwave is stopped for 2min, and the microwave power is increased to 800W for 3 min;

further explaining, in the step (3), the heating rate is 5 ℃/min;

further explaining, in the step (3), the homogenizing condition is that the rotating speed is 5500r/min and the pressure is 70 MPa;

further explaining, in the step (4), the squeezing conditions are that the temperature is 45-55 ℃, the pressure is 40-50MPa, the pressure is 40-50min, and the pressure is maintained for 40-60 min; centrifuging at 3000-;

further, in the step (4), the squeezing conditions are 50 ℃ of temperature, 45MPa of pressure, 45min of pressurization and 50min of pressure holding.

Compared with the prior art, the invention has the beneficial effects that: according to the invention, through camellia seed pretreatment, microwave treatment, alkaline protease modified particles combined with homogenization and carbohydrase enzymolysis are adopted, so that emulsion formed in the camellia seed extraction process is broken, the content of active ingredients of camellia seed oil is increased, the prepared camellia seed oil has high quality, meets the product quality standard, and has good oxidation stability and oxidation resistance.

The solvent is soaked to enter intracellular components of a soluble part in cells, water in the cells is vaporized by the heat effect of microwaves, micro cracks can be generated on cell walls, small molecules in the partially dissolved cells are dissolved out, the extraction rate is improved, proteins can be prevented from forming a colloidal solution by the heat effect, and the formation of an emulsion is reduced; in the process of rapid temperature rise, the alkaline protease modified particles can disperse sugar molecules and protein molecules by utilizing the heat resistance difference of sugar and protein, so that a suspension system is stable, strong affinity is formed with the protein, the stability of emulsion is damaged, and the nano SiO₂The uniform dispersion degree of components in the system can be improved, the activity of the alkaline protease is kept, and the multi-mesoporous structure on the surface of the alkaline protease has strong adsorption capacity; the homogenization can further atomize the dispersion in the suspension system, so that the system is more stable, the viscosity of the system is reduced, the dissolution of an emulsion interfacial film is facilitated, the strength of the interfacial film is reduced, and the emulsion breaking efficiency is improved.

Detailed Description

In order to better understand the technical content of the invention, specific examples are provided below to further illustrate the invention.

The experimental methods used in the examples of the present invention are all conventional methods unless otherwise specified.

The materials, reagents and the like used in the examples of the present invention can be obtained commercially without specific description.

Example 1

A method for extracting camellia seeds comprises the following steps:

(1) soaking and refrigerating pretreatment: according to the mass ratio of 1: 5: 3, adding water into camellia seeds, soaking for 2 hours, filtering out the water, adding an ethanol solution with the mass concentration of 35%, refrigerating and soaking for 4 hours at 8 ℃, taking the camellia seeds, drying and crushing to obtain crushed substances;

(2) ethanol microwave: adding 60% ethanol solution into the crushed material, stirring, treating with 500W microwave power for 55s, stopping microwave for 2min, and increasing microwave power to 700W for 3min to obtain microwave treated material;

(3) and (3) treating alkaline protease modified particles: according to the feed liquid mass ratio of 1: 0.5, adding alkaline protease modified particles into the microwave treated substance, stirring, heating to 55 ℃ at the speed of 5 ℃/min, keeping the temperature for 60min, homogenizing at the speed of 5000r/min and the pressure of 80MPa for 10min, adjusting the pH value to 8, adding 0.5% w/w carbohydrase solution based on the microwave treated substance, carrying out enzymolysis at the temperature of 35 ℃ for 40min, and filtering to obtain crude oil;

the preparation method of the alkaline protease modified particles comprises the following steps:

(a) according to the mass ratio of 1: 6: 3, in nano SiO₂Adding water and sucrose ester into the powder, stirring at 600r/min at 30 deg.C under ultrasonic condition for 45min, standing for 40min to obtain nanometer SiO₂A suspension;

(b) according to the feed liquid mass ratio of 1: 6, adding alkaline protease into the nano SiO₂Stirring the suspension for 25min at a speed of 80r/min, and spray drying to obtain alkaline protease modified particles;

(4) squeezing and centrifuging: squeezing the crude oil at 45 deg.C under 40MPa for 40min, maintaining the pressure for 40min, centrifuging at 3000r/min for 40min, and collecting supernatant to obtain oleum Camelliae Japonicae.

Example 2

A method for extracting camellia seeds comprises the following steps:

(1) soaking and refrigerating pretreatment: according to the mass ratio of 1: 3: 5, adding water into camellia seeds, soaking for 3 hours, filtering off the water, adding an ethanol solution with the mass concentration of 45%, refrigerating and soaking for 6 hours at 12 ℃, taking the camellia seeds, drying and crushing to obtain crushed substances;

(2) ethanol microwave: adding 70% ethanol solution into the crushed material, stirring, treating with microwave power of 700W for 45s, stopping microwave for 3min, and increasing microwave power to 900W for 2min to obtain microwave treated material;

(3) and (3) treating alkaline protease modified particles: according to the feed liquid mass ratio of 1: 0.7, adding alkaline protease modified particles into the microwave treated substance, stirring, heating to 65 ℃ at the speed of 5 ℃/min, keeping the temperature for 40min, homogenizing at 6000r/min under the pressure of 60MPa for 15min, adjusting the pH value to 9, adding 0.7% w/w carbohydrase solution based on the microwave treated substance, carrying out enzymolysis at the temperature of 45 ℃ for 30min, and filtering to obtain crude oil;

the preparation method of the alkaline protease modified particles comprises the following steps:

(a) according to the mass ratio of 1: 7: 4, in nano SiO₂Adding water and sucrose ester into the powder, stirring at 700r/min at 50 deg.C for 35min while ultrasonic processing, standing for 50min to obtain nanometer SiO₂A suspension;

(b) according to the feed liquid mass ratio of 1: 8, adding alkaline protease into the nano SiO₂Stirring the suspension for 35min at the speed of 90r/min, and spray drying to obtain alkaline protease modified particles;

(4) squeezing and centrifuging: squeezing the crude oil at 55 deg.C under 50MPa for 50min, maintaining the pressure for 60min, centrifuging at 5000r/min for 30min, and collecting supernatant to obtain camellia seed oil.

Example 3

A method for extracting camellia seeds comprises the following steps:

(1) soaking and refrigerating pretreatment: according to the mass ratio of 1: 3: 3, adding water into camellia seeds, soaking for 3 hours, filtering off the water, adding an ethanol solution with the mass concentration of 40%, refrigerating and soaking for 5 hours at 10 ℃, taking the camellia seeds, drying and crushing to obtain crushed substances;

(2) ethanol microwave: adding 65% ethanol solution into the crushed material, stirring, treating with microwave power of 600W for 50s, stopping microwave for 2min, and increasing microwave power to 800W for 3min to obtain microwave treated material;

(3) and (3) treating alkaline protease modified particles: according to the feed liquid mass ratio of 1: 0.6, adding alkaline protease modified particles into the microwave treated substance, stirring, heating to 60 ℃ at the speed of 5 ℃/min, keeping the temperature for 50min, homogenizing for 12min at the pressure of 5500r/min and 70MPa, adjusting the pH value to 8.5, adding 0.6% w/w carbohydrase solution based on the microwave treated substance, carrying out enzymolysis for 35min at the temperature of 40 ℃, and filtering to obtain crude oil;

the preparation method of the alkaline protease modified particles comprises the following steps:

(a) according to the mass ratio of 1: 6: 4, in nano SiO₂Adding water and sucrose ester into the powder, stirring at 650r/min under ultrasonic condition at 40 deg.C for 40min, standing for 45min to obtain nanometer SiO₂A suspension;

(b) according to the feed liquid mass ratio of 1: 7, adding alkaline protease into the nano SiO₂Stirring the suspension for 30min at the speed of 85r/min, and spray drying to obtain alkaline protease modified particles;

(4) squeezing and centrifuging: squeezing the crude oil at 50 deg.C under 45MPa for 45min, maintaining the pressure for 50min, centrifuging at 4000r/min for 35min, and collecting supernatant to obtain oleum Camelliae Japonicae.

Example 4

This example differs from example 3 in that: nano SiO₂The suspensions are prepared differently, namely: according to the mass ratio of 1: 6: 4, in nano SiO₂Adding water and Arabic gum into the powder, stirring at 650r/min under ultrasonic at 40 deg.C for 40min, standing for 45min to obtain nanometer SiO₂And (3) suspension.

Example 5

This example differs from example 3 in that: in the step (3), homogenization is carried out at 3000r/min and 30 MPa.

Example 6

This example differs from example 3 in that: the microwave treatment method is different, namely: adding 65% ethanol solution into the crushed material, stirring, treating with microwave power of 900W for 80s, stopping microwave for 2min, and reducing microwave power to 500W for 5min to obtain microwave treated material.

Comparative example 1

This comparative example differs from example 3 in that: the preparation method of the alkaline protease modified particles is different, namely: according to the feed liquid mass ratio of 1: 10, adding alkaline protease into the nano SiO₂Stirring the suspension for 30min at the speed of 85r/min, and carrying out vacuum freeze drying to obtain alkaline protease modified particles; the rest of the procedure was the same as in example 3.

Comparative example 2

This comparative example differs from example 3 in that: enzyme-modified particles were prepared using carbohydrase instead of alkaline protein, i.e.:

the preparation method of the carbohydrase modified granule comprises the following steps:

(a) according to the mass ratio of 1: 6: 4, in nano SiO₂Adding water and sucrose ester into the powder, stirring at 650r/min under ultrasonic condition at 40 deg.C for 40min, standing for 45min to obtain nanometer SiO₂A suspension;

(b) according to the feed liquid mass ratio of 1: 7, adding carbohydrase into the nano SiO₂Stirring the suspension for 30min at the speed of 85r/min, and spray drying to obtain carbohydrase modified particles;

the processing method of the carbohydrase modified particles comprises the following steps: according to the feed liquid mass ratio of 1: 0.6, adding carbohydrase modified particles into the microwave treated substance, stirring, heating to 75 ℃ at the speed of 5 ℃/min, keeping the temperature for 50min, homogenizing at the speed of 5500r/min and the pressure of 70MPa for 12min, adjusting the pH value to 8.5, adding 0.6% w/w protease solution based on the microwave treated substance, carrying out enzymolysis at the temperature of 40 ℃ for 35min, and filtering to obtain crude oil; the rest of the procedure was the same as in example 3.

Firstly, experimental detection

Centrifuging the product after the enzymolysis of the carbohydrase, respectively collecting a free oil layer and an emulsion layer, centrifuging the camellia seed oil prepared in the embodiments 1-6 and the comparative examples 1-2, collecting the free oil, respectively calculating the fat content of the emulsion and the fat content of the camellia seed raw material according to the determination of fat in national food safety standard (GB 5009.6-2016), and respectively calculating the demulsification rate and the oil yield:

breaking ratio (%) < free oil mass/(emulsion mass x oil content of emulsion) × 100%

The oil yield (%) is free oil quality/oil content of camellia seed material × 100%

The results are shown in Table 1:

item	Percentage of emulsion breaking (%)	Oil yield (%)
			Example 1	94.9	72.5
Example 2	95.6	76.7
			Example 3	96.2	79.3
Example 4	86.2	67.9
			Example 5	90.0	70.2
Example 6	83.7	62.5
			Comparative example 1	85.6	63.6
Comparative example 2	80.4	61.3

From the above table, the demulsification rate of the camellia seed oil is 94.9-96.2%, and the oil yield of the camellia seed oil can reach 79.3%, which shows that the emulsion formed in the camellia seed extraction process can be broken through the microwave treatment and the alkaline protease modified particles combined with the homogenization and the carbohydrase enzymolysis, so that the extraction efficiency and the oil yield are improved. Example 4 reduction of emulsion breaking Rate, indicating that Nano SiO prepared with sucrose ester₂The suspension can make the alkaline protease fully play a role, and is beneficial to improving the demulsification efficiency; example 5 the emulsion breaking rate is reduced, which indicates that homogenization under certain conditions can further homogenize the dispersion, and can remove the existence of emulsified particles in the suspension system, thereby reducing the stability of the emulsion and improving the emulsion breaking rate; comparative example 6 changes the microwave conditions, emulsion is easy to appear too early, the difficulty of subsequent extraction is increased, and the extraction rate is reduced.

Comparative example 1 the alkaline protease-modified particles failed to sufficiently solidify the protein; comparative example 2 the use of carbohydrase modified particles did not reduce the stability of the emulsion in the early stage of extraction, affecting the subsequent extraction efficiency.

Second, Oxidation stability test

The camellia oil prepared in the embodiments 1 to 6 and the comparative examples 1 to 2 of the present invention was sealed and packaged, and then placed in a constant temperature incubator at 65 ℃, protected from light, and subjected to an accelerated oxidation experiment, and observed for one month, and the acid value and the peroxide value were measured according to "measurement of acid value in food safety national standard food" (GB 5009.229-2016) and "measurement of peroxide value in food safety national standard food" (GB 5009.227-2016), respectively, and the results are shown in table 2:

as can be seen from the above table, the camellia oil obtained in the embodiments 1-3 of the present invention has less increase of acid value and peroxide value and less deterioration of oil during the high temperature acceleration experiment, which indicates that the present invention adopts microwave treatment and alkali treatmentThe modified particles of the sexual protease are combined with homogenization and carbohydrase enzymolysis, so that the oxidation resistance of the camellia seed oil can be improved, the hydrolysis process of fat is slowed down, the generation of free fatty acid is reduced during the storage period of the camellia seed oil, and the camellia seed oil has good oxidation stability. Example 4 shows the preparation of sucrose esters to produce nano-SiO₂The suspension has good dispersing power and adsorption capacity, and can enhance the stability of a system after the alkaline protease modified particles are dispersed; example 5 shows that homogenization can further enhance the stability of the suspension system and improve the particle size of the emulsified particles; example 6 could not sufficiently dissolve the intracellular material, emulsion was easily formed during the camellia seed extraction process, and the extraction difficulty was increased.

Comparative examples 2 and 3 did not stabilize the suspension well and did not slow down the formation of oxidation products, thereby reducing the oxidative stability of the camellia oil.

In conclusion, the camellia seed is pretreated, and the microwave treatment, the alkaline protease modified particles, the homogenization and the carbohydrase enzymolysis are combined, so that emulsion formed in the camellia seed extraction process is broken, the emulsion breaking rate and the oil yield of the camellia seed oil are improved, the camellia seed oil has good oxidation stability and oxidation resistance, the extraction method is simple, the process operation is controllable, and the camellia seed oil is suitable for industrial production.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.