阅读说明：本技术 一种油菜花粉提取物的制备方法 (Preparation method of rape pollen extract ) 是由 李元 申佳敏 张薇 林欣怡 苗宁宝 王卫东 于 2021-07-30 设计创作，主要内容包括：本发明提供一种油菜花粉提取物的制备方法,包括以下步骤：S1、制备混合油脂的油菜花粉肽溶液；S2、向S1的混合油脂的油菜花粉肽溶液中加入植物油,振荡,离心,分离出上层富含神经酸和花粉黄酮的油脂,下层溶液经过干燥处理得到油菜花粉肽。该方法通过加入少量的植物油打破乳化状态的临界点,实现一步同时制备出油菜花粉肽、富含神经酸和黄酮的油菜花粉油脂。此法提取神经酸工艺流程简单、省时省力、节约物资,同步提取神经酸和花粉黄酮无任何工业药物的残留,健康无污染,经简单处理后便可投入使用,制备油菜花粉肽粉同时提取神经酸,同步还可以将油菜花中的黄酮类物质提取出来,较大程度上对油菜花进行了充分利用。(The invention provides a preparation method of a rape pollen extract, which comprises the following steps: s1, preparing a rape pollen peptide solution of mixed oil; s2, adding vegetable oil into the rape pollen peptide solution of the mixed oil S1, oscillating, centrifuging, separating oil rich in nervonic acid and pollen flavone at the upper layer, and drying the solution at the lower layer to obtain the rape pollen peptide. The method can break the critical point of emulsified state by adding a small amount of vegetable oil, and can simultaneously prepare rape pollen peptide and rape pollen oil rich in nervonic acid and flavone in one step. The method has the advantages of simple process flow, time and labor saving, material saving, no industrial drug residue in the process of synchronously extracting the nervonic acid and the pollen flavone, health and no pollution, can be put into use after simple treatment, can be used for preparing the rape pollen peptide powder and simultaneously extracting the nervonic acid, can synchronously extract the flavonoid substances in the rape flowers, and fully utilizes the rape flowers to a greater extent.)

1. A preparation method of a rape pollen extract is characterized by comprising the following steps:

s1, preparing a rape pollen peptide solution of mixed oil;

s2, adding vegetable oil into the rape pollen peptide solution of the mixed oil S1, oscillating, centrifuging, separating oil rich in nervonic acid and pollen flavone at the upper layer, and drying the solution at the lower layer to obtain the rape pollen peptide.

2. The method for preparing the canola pollen extract as claimed in claim 1, wherein in S2, the volume ratio of the canola pollen peptide solution of the mixed oil to the vegetable oil is 10-20: 1.

3. the method of claim 1, wherein the vegetable oil is selected from the group consisting of rapeseed oil, sunflower seed oil, corn oil, and soybean oil at S2.

4. The method of claim 1, wherein the rotation speed of the oscillation of S2 is 1000 rpm for 1 hour.

5. The method for preparing rape pollen extract as claimed in claim 1, wherein the rotation speed of S2 is 1000-2000 rpm for 20 min.

6. The method of preparing canola pollen extract of claim 1, wherein the process of preparing the canola pollen peptide solution of mixed oils and fats in S1 is as follows:

breaking cell wall of rape pollen granule for 10min, adding water, mixing, adjusting pH to 5.5 at constant temperature of 55 deg.C, adding cellulase and pectinase, and stirring for 2 hr; then adding papain at the constant temperature of 60 ℃, and stirring for 1 h; and then three kinds of hydrolytic enzymes are inactivated at 100 ℃, supernatant is separated out by a tubular centrifuge at 8000 rpm, and the supernatant is taken and concentrated under reduced pressure to obtain the rape pollen peptide solution of the mixed oil.

7. The method for preparing rape pollen extract as claimed in claim 6, wherein the mass ratio of rape pollen particles to water is 1: 5.

8. the method for preparing rape pollen extract as claimed in claim 6, wherein the mass ratio of rape pollen particles to cellulase, pectinase and papain is 100: 0.5-1: 0.5-1: 0.5 to 1.

Technical Field

The invention relates to the technical field of health-care food, in particular to a preparation method of a rape pollen extract.

Background

A reasonable extraction and separation process can be designed according to the difference of the existing state, polarity, solubility and the like of the effective components in the plants. For example, canola pollen contains canola pollen flavonoids effective in treating prostate enlargement, and its lipids also contain nervonic acid to restore nerve damage. The pollen flavone and the nervonic acid both belong to fat-soluble substances, so the nervonic acid and the flavonoid substances can be extracted by adopting a fat-soluble solvent, such as a method for extracting organic matters such as petroleum ether, ethanol and the like to extract the nervonic acid and the flavonoid substances in the rape flower, but the extraction method has the problems of solvent residue, environmental pollution, large cost, high explosion-proof requirement of a factory building and the like.

On the basis of preparing the pollen peptide, after the pollen protein is degraded by protease, the pollen oil loses the constraint and is dissociated in the pollen peptide solution, so that the key point of the invention is how to realize oil-water separation. However, the oil content of rape pollen is low, so that the oil and peptide powder are in an emulsified state, and in the test process, the method for extracting by using a fat-soluble solvent is found to be difficult to separate the pollen oil from the pollen peptide solution, and the separation effect is poor.

Disclosure of Invention

In order to solve the problems, the invention aims to provide a preparation method of a rape pollen extract, which can simultaneously prepare rape pollen peptide and rape pollen oil rich in nervonic acid and flavone in one step by adding a small amount of vegetable oil to break the critical point of an emulsified state. The method has the advantages of simple process flow, time and labor saving, material saving, no industrial drug residue in the process of synchronously extracting the nervonic acid and the pollen flavone, health and no pollution, can be put into use after simple treatment, can be used for preparing the rape pollen peptide powder and simultaneously extracting the nervonic acid, can synchronously extract the flavonoid substances in the rape flowers, and fully utilizes the rape flowers to a greater extent.

In order to achieve the above object, the technical solution of the present invention is as follows.

A preparation method of rape pollen extract comprises the following steps:

s1, preparing a rape pollen peptide solution of mixed oil;

s2, adding vegetable oil into the rape pollen peptide solution of the mixed oil S1, oscillating, centrifuging, separating oil rich in nervonic acid and pollen flavone at the upper layer, and drying the solution at the lower layer to obtain the rape pollen peptide.

Further, in S2, the volume ratio of the rape pollen peptide solution of the mixed oil to the vegetable oil is 10-20: 1.

further, in S2, the vegetable oil is any one or more of rapeseed oil, sunflower seed oil, corn oil, and soybean oil.

Further, in S2, the rotation speed at the time of oscillation was 1000 revolutions/min, and the time was 1 hour.

Further, in S2, the rotation speed during centrifugation is 1000-2000 rpm, and the time is 20 min.

Further, in S1, the procedure for preparing the oil-and-fat mixed rape pollen peptide solution is as follows:

breaking cell wall of rape pollen granule for 10min, adding water, mixing, adjusting pH to 5.5 at constant temperature of 55 deg.C, adding cellulase and pectinase, and stirring for 2 hr; then adding papain at the constant temperature of 60 ℃, and stirring for 1 h; and then three kinds of hydrolytic enzymes are inactivated at 100 ℃, supernatant is separated out by a tubular centrifuge at 8000 rpm, and the supernatant is taken and concentrated under reduced pressure to obtain the rape pollen peptide solution of the mixed oil.

Furthermore, the mass ratio of the rape pollen particles to the water is 1: 5.

furthermore, the mass ratio of the rape pollen particles to the cellulase, the pectinase and the papain is 100: 0.5-1: 0.5-1: 0.5 to 1.

The invention has the beneficial effects that:

1. the method of the invention realizes the simultaneous preparation of rape pollen peptide and rape pollen grease rich in nervonic acid and flavone in one step by adding a small amount of vegetable oil to break the critical point of the emulsified state. The method has the advantages of simple process flow, time and labor saving, material saving, no industrial drug residue in the process of synchronously extracting the nervonic acid and the pollen flavone, health and no pollution, can be put into use after simple treatment, can be used for preparing the rape pollen peptide powder and simultaneously extracting the nervonic acid, can synchronously extract the flavonoid substances in the rape flowers, and fully utilizes the rape flowers to a greater extent.

2. The oil and the pollen peptide in the rape pollen peptide solution exist in an emulsified state, once the oil and fat emulsifying capacity of the pollen peptide breaks through a critical value, the pollen peptide loses the emulsifying capacity and releases the original bound oil and fat. Nervonic acid is one of the components in rape pollen oil, is synchronously extracted along with the oil, and simultaneously carries out oil-soluble rape pollen flavone. Therefore, the method can also make rape pollen as one of plant resources with the potential of using the extracted nervonic acid.

3. The invention adopts a mode of adding three enzymes step by step for hydrolysis, because the papain is non-specific for degrading protein, the activity of other enzymes can be degraded by one-time addition, the first two enzymes can be prevented from being influenced by the papain by step addition, and the degradation of cellulose and pectin is completed; then adding papain to degrade protein to prepare peptide powder, thereby improving enzymolysis efficiency.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The experimental methods described in the following examples are all conventional methods unless otherwise specified; the reagents and materials are commercially available, unless otherwise specified.

Example 1

A preparation method of rape pollen extract comprises the following steps:

s1, preparing rape pollen peptide solution of mixed oil

S1.1, breaking the wall of the dried pollen particles for 10 minutes by a wall breaking machine.

S1.2, taking a proper amount of the pollen with the wall broken, putting the pollen into an extraction kettle, adding water according to a ratio of 1:5, heating to 55 ℃, adding a small amount of HCl to adjust the pH value to 5.5, and mixing the pollen: cellulase: adding cellulase and pectinase at a ratio of 100:0.5:0.5, and allowing to act for 2 hr.

S1.3, according to the ratio of pollen: adding papain in a ratio of 100:1, and acting at 60 deg.C for 1 hr to degrade into pollen polypeptide.

S1.4, inactivating three hydrolases in the solution of S1.3 at 100 ℃, using a tubular centrifuge at 8000 rpm, and centrifuging to separate out supernatant. And pouring the supernatant into a rotary evaporator, and concentrating by rotary evaporation at 80 ℃ and the rotating speed of 30-40 r/min by 10 times to obtain the concentrated rape pollen peptide solution.

S2, following the concentrated canola pollen peptide solution: adding 100mL of rape pollen peptide concentrated solution and 5mL of vegetable oil in sequence in a ratio of 100:5, stirring for 1 hour at 1000 revolutions per minute, centrifuging for 20 minutes at 1000 revolutions per minute and 2000 revolutions per minute, and collecting upper-layer oil, namely the oil rich in nervonic acid and flavone; and drying the lower layer rape pollen peptide solution obtained by centrifugation by using a spray drying method, wherein the temperature of a spray outlet is 120-160 ℃, and obtaining the rape pollen peptide.

Example 2

A preparation method of rape pollen extract comprises the following steps:

s1, preparing rape pollen peptide solution of mixed oil

S1.1, breaking the wall of the dried pollen particles for 10 minutes by a wall breaking machine.

S1.2, taking a proper amount of the pollen with the wall broken, putting the pollen into an extraction kettle, adding water according to a ratio of 1:5, heating to 55 ℃, adding a small amount of HCl to adjust the pH value to 5.5, and mixing the pollen: cellulase: adding cellulase and pectinase at a ratio of 100:1:1, and acting for 2 hours.

S1.3, according to the ratio of pollen: adding papain in a ratio of 100:1, and acting at 60 deg.C for 1 hr to degrade into pollen polypeptide.

S1.4, inactivating three hydrolases in the solution of S1.3 at 100 ℃, using a tubular centrifuge at 8000 rpm, and centrifuging to separate out supernatant. And pouring the supernatant into a rotary evaporator, and concentrating by rotary evaporation at 80 ℃ and the rotating speed of 30-40 r/min by 10 times to obtain the concentrated rape pollen peptide solution.

S2, following the concentrated canola pollen peptide solution: adding 50mL of rape pollen peptide concentrated solution and 5mL of vegetable oil in sequence in a ratio of 50:5, stirring for 1 hour at 1000 revolutions per minute, centrifuging for 20 minutes at 1000 revolutions per minute and 2000 revolutions per minute, and collecting upper-layer oil, namely the oil rich in nervonic acid and flavone; and drying the lower layer rape pollen peptide solution obtained by centrifugation by using a spray drying method, wherein the temperature of a spray outlet is 120-160 ℃, and obtaining the rape pollen peptide.

Example 3

A preparation method of rape pollen extract comprises the following steps:

s1, preparing rape pollen peptide solution of mixed oil

S1.1, breaking the wall of the dried pollen particles for 10 minutes by a wall breaking machine.

S1.2, taking a proper amount of the pollen with the wall broken, putting the pollen into an extraction kettle, adding water according to a ratio of 1:5, heating to 55 ℃, adding a small amount of HCl to adjust the pH value to 5.5, and mixing the pollen: cellulase: adding cellulase and pectinase at a ratio of 100:0.75:0.75, and allowing to act for 2 hours.

S1.3, according to the ratio of pollen: adding papain at a ratio of 100:0.5, and degrading into pollen polypeptide at 60 deg.C for 1 hr.

S1.4, inactivating three hydrolases in the solution of S1.3 at 100 ℃, using a tubular centrifuge at 8000 rpm, and centrifuging to separate out supernatant. And pouring the supernatant into a rotary evaporator, and concentrating by rotary evaporation at 80 ℃ and the rotating speed of 30-40 r/min by 10 times to obtain the concentrated rape pollen peptide solution.

S2, following the concentrated canola pollen peptide solution: adding 75mL of rape pollen peptide concentrated solution and 5mL of vegetable oil in turn into 75:5 vegetable oil, stirring for 1 hour at 1000 revolutions per minute, centrifuging for 20 minutes at 1000 revolutions per minute and 2000 revolutions per minute, and collecting upper-layer oil, namely the oil rich in nervonic acid and flavone; and drying the lower layer rape pollen peptide solution obtained by centrifugation by using a spray drying method, wherein the temperature of a spray outlet is 120-160 ℃, and obtaining the rape pollen peptide.

Comparative example 1

A method for preparing rape pollen extract, which is substantially the same as that of example 1, except that,

s2, following the concentrated canola pollen peptide solution: vegetable oil 100: 3, sequentially adding 100mL of rape pollen peptide concentrated solution and 3mL of vegetable oil, stirring for 2 hours at 1000 revolutions per minute, centrifuging for 20 minutes at 1000 revolutions per minute and 2000 revolutions per minute, and collecting upper-layer oil, namely the oil rich in nervonic acid and flavone; and drying the lower layer rape pollen peptide solution obtained by centrifugation by using a spray drying method, wherein the temperature of a spray outlet is 120-160 ℃, and obtaining the rape pollen peptide.

Comparative example 2

A method for preparing rape pollen extract, which is substantially the same as that of example 1, except that,

s2, following the concentrated canola pollen peptide solution: vegetable oil 100:1, sequentially adding 100mL of rape pollen peptide concentrated solution and 1mL of vegetable oil, stirring for 2 hours at 1000 revolutions per minute, centrifuging for 20 minutes at 1000 revolutions per minute and 2000 revolutions per minute, and collecting upper-layer oil, namely the oil rich in nervonic acid and flavone; and drying the lower layer rape pollen peptide solution obtained by centrifugation by using a spray drying method, wherein the temperature of a spray outlet is 120-160 ℃, and obtaining the rape pollen peptide.

Comparative example 3

A method for preparing rape pollen extract, which is substantially the same as that of example 1, except that,

s2, following the concentrated canola pollen peptide solution: vegetable oil 100: 10, sequentially adding 100mL of rape pollen peptide concentrated solution and 10mL of vegetable oil, stirring for 1 hour at 1000 revolutions per minute, centrifuging for 20 minutes at 1000 revolutions per minute and 2000 revolutions per minute, and collecting upper-layer oil, namely the oil rich in nervonic acid and flavone; and drying the lower layer rape pollen peptide solution obtained by centrifugation by using a spray drying method, wherein the temperature of a spray outlet is 120-160 ℃, and obtaining the rape pollen peptide.

The results obtained by the methods of examples 1 to 3 are substantially the same, and therefore the results of example 1 are compared with those of comparative examples 1 to 3 to illustrate the effects of the method of the present invention. The measurement results are shown in tables 1 and 2, wherein table 1 shows the extraction conditions and results of example 1 and comparative examples 1 to 3; table 2 shows the results of measuring the contents of the added vegetable oil and the extracted rape pollen oil in example 1.

TABLE 1 extraction conditions and results for example 1 and comparative examples 1-3

As can be seen from Table 1, when the amount of oil used for extracting the rape pollen peptide solution by the vegetable oil extraction method is small, the critical point of emulsification of the oil and the peptide powder cannot be broken, the added oil is emulsified, when the adding ratio is 100:5, the oil extraction amount and the pollen flavone amount are stable, and the time is most suitable for 1 h.

TABLE 2 measurement results of contents of vegetable oil and rape pollen oil in example 1

Wherein, No. 1 is added rape oil. And 2# is extracted rape pollen oil.

As can be seen from the results in Table 2, the pollen oil contains nervonic acid, and the nervonic acid content extracted in the example 1 of the present invention is 1.19%. The combination of high yield of rape flowers can show that the rape pollen oil has excellent development prospect.

In summary, the invention takes rape pollen as raw material, rape pollen peptide is prepared by protease hydrolysis, in the process, the grease in the rape pollen is released, the grease and the peptide are emulsified into a whole, after oil is additionally added, the emulsified state is destroyed, the grease and oil-soluble flavonoid substances are blended into the added oil, and nervonic acid and oil-soluble flavonoid substances in the rape pollen grease are extracted. And the rape flowers are widely planted, the production cost is low, the large-scale production is easy, and the method is more economical and applicable. The method simultaneously synchronously extracts the nervonic acid and the flavone in the rape pollen while preparing the rape pollen peptide, develops high-quality raw materials for preparing functional health-care food, and more effectively utilizes the pollen raw materials.

The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention.