CN112794929A - Highland barley beta-glucan extraction process based on ultrasonic assistance - Google Patents
Highland barley beta-glucan extraction process based on ultrasonic assistance Download PDFInfo
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Abstract
The invention discloses an extraction process of highland barley beta-glucan based on ultrasonic assistance, which comprises the following steps: crushing cleaned highland barley raw materials to 40-60 meshes, degreasing to obtain a sample to be extracted, adding an extracting agent with the pH of 8-10, and stirring, wherein the weight ratio of the sample to be extracted to the extracting agent is 1: 18-23; introducing nitrogen, heating to 55-70 ℃, and carrying out ultrasonic treatment for 18-26 min; carrying out microwave treatment for 15-20 s, carrying out dispersion oscillation for 3-4 h, carrying out ultrasonic treatment for 20-40 min, and carrying out post-treatment; the method has the advantages of simple steps and convenient operation, does not need to add chemical treatment agents of other steps, realizes higher extraction efficiency of the highland barley beta-glucan by physical means, effectively inhibits Maillard reaction which is easy to occur in the extraction process, reduces the browning degree of products, improves the yield of the products, and has the highest purity of 84.68%.
Description
Technical Field
The invention relates to the technical field of beta-glucan extraction. More specifically, the invention relates to an extraction process of highland barley beta-glucan based on ultrasonic assistance.
Background
The highland barley is the crop with the highest beta-glucan content in wheat crops in the world, and the average content of the beta-glucan in the highland barley is detected to be 6.57 percent, the 25 percent of the excellent variety of highland barley can reach 8.62 percent and is 50 times of the average content of wheat.
Beta-glucan prevents colon cancer by reducing the exposure of the intestinal mucosa to carcinogens and indirectly inhibiting carcinogenic microorganisms; prevention of cardiovascular diseases by lowering blood lipids and cholesterol synthesis: can be used for preventing and treating diabetes by controlling blood sugar. Has effects in improving body defense ability and regulating physiological rhythm.
The existing extraction methods of highland barley beta-glucan comprise stir-frying, ultrasonic treatment, microwave treatment, cooking, extrusion puffing, high-pressure treatment and the like. The highest extraction rate of the finally obtained beta-glucan is only 3-4%, the yield is very limited, and a highland barley beta-glucan extraction process which is simple to operate and high in extraction efficiency is urgently needed.
Disclosure of Invention
An object of the present invention is to solve at least the above problems and to provide at least the advantages described later.
The invention also aims to provide an extraction process of highland barley beta-glucan based on ultrasonic assistance, which has simple steps and convenient operation, does not need to add chemical treatment agents of other steps, realizes higher extraction efficiency of highland barley beta-glucan by physical means, effectively inhibits Maillard reaction which is easy to occur in the extraction process, reduces the browning degree of products and improves the yield of products.
To achieve these objects and other advantages in accordance with the present invention, there is provided an ultrasound-assisted highland barley beta-glucan extraction process, comprising the steps of:
step one, crushing cleaned highland barley raw materials to 40-60 meshes, degreasing to obtain a sample to be extracted, adding an extracting agent with the pH of 8-10, and stirring, wherein the weight ratio of the sample to be extracted to the extracting agent is 1: 18-23;
introducing nitrogen, heating to 55-70 ℃, and carrying out ultrasonic treatment for 18-26 min, wherein the ultrasonic power is 600-800W;
step three, performing microwave treatment for 15-20 s, performing dispersion oscillation for 3-4 h, performing ultrasonic treatment for 20-40 min, and performing post-treatment; wherein the microwave frequency is 850-900W, and the ultrasonic power is 600-800W.
Preferably, in the step one, the weight ratio of the sample to be extracted to the extractant is 1: 20.
preferably, in the second step, the heating temperature is 60 ℃.
Preferably, the microwave frequency in step three is 900W, and the microwave treatment time is 18 s.
Preferably, in the second step, the ultrasonic power is 650W, the frequency is 20KHz, and the time is 20 min; in the third step, the ultrasonic power is 800W, the frequency is 20KHz, and the time is 20 min.
Preferably, nitrogen is introduced for 5min before the extractant is added in the step one.
Preferably, the porous carbon spheres with the diameter of 200-300 nm are added in the ultrasonic treatment in the second step, after the reaction is finished, the porous carbon spheres are taken out, the microwave treatment in the third step is carried out, and after the microwave treatment is finished, the taken-out porous carbon spheres are put in again for the rest steps; the weight ratio of the porous carbon balls to the highland barley raw materials is 1: 20.
Preferably, the degreasing treatment is: taking the crushed highland barley powder, adding absolute ethyl alcohol, uniformly stirring, refluxing the highland barley powder and the absolute ethyl alcohol at 76 ℃ for 2h at a mass ratio of 1:4, concentrating and drying.
Preferably, the post-treatment is: taking a mixture obtained after ultrasonic treatment, filtering to obtain a filtrate, adjusting the pH value of the filtrate to 4.5 by using 20% hydrochloric acid, standing for 10 hours at the temperature of 3 ℃, centrifuging to obtain the filtrate, adjusting the pH value of the filtrate to 7, adding medium-temperature alpha-amylase, uniformly stirring the highland barley raw material with the addition of 4u per gram, heating to 85-90 ℃, liquefying for 30 minutes, concentrating to obtain a concentrated solution, adding 95% ethanol into the concentrated solution for settling, centrifuging to obtain a sediment, washing the sediment by using 95% ethanol, and drying.
Preferably, step two and step three are carried out in a double-guide reaction apparatus comprising:
the outer wall of the spherical reaction cavity is provided with a pair of shaft holes which are symmetrical relative to the spherical center of the spherical reaction cavity; the two shaft holes are respectively connected with a left shaft and a right shaft which are coaxial; a coaxial transmission assembly is arranged on the right shaft to drive the spherical reaction cavity to rotate; the spherical reaction cavity is rotatably erected between a pair of vertical bases through the left shaft and the right shaft;
a feed inlet and a material placing port are arranged on the circumference of the highest point of the spherical reaction cavity, and the feed inlet and the material placing port are symmetrical about the spherical center of the spherical reaction cavity;
a plate-shaped partition is fixedly arranged in the spherical reaction cavity in an inclined manner, the edge of the partition is fixedly connected with the inner wall of the spherical reaction cavity, the center of the partition is opposite to the feed inlet, the partition divides the space in the spherical reaction cavity into a microwave treatment chamber and an ultrasonic treatment chamber, the feed inlet is communicated with the microwave treatment chamber, and the material placing port is communicated with the ultrasonic treatment chamber; a first material guide valve is arranged on the separator at the position facing the ultrasonic treatment chamber and close to the lower part of the edge of the separator, and a second material guide valve is arranged on the separator at the position facing the microwave treatment chamber and close to the lower part of the edge of the separator; the first guide valve and the second guide valve penetrate through the partition and are communicated with the microwave treatment chamber and the ultrasonic treatment chamber, and the first guide valve and the second guide valve are electrically controlled valves;
the microwave treatment device comprises a microwave treatment chamber, a microwave generator, an anti-overflow layer and a water tank, wherein the arc-shaped side wall in the microwave treatment chamber is provided with the microwave generator, and the inner side wall of the microwave treatment chamber is also adhered with the anti-overflow layer; an ultrasonic vibration sheet is arranged on the arc-shaped side wall in the ultrasonic treatment chamber, a filter sieve plate is arranged at the part, close to the material placing port, in the ultrasonic treatment chamber, the edge of the filter sieve plate is fixedly connected with the inner wall of the ultrasonic treatment chamber, and the filter sieve plate is positioned below the ultrasonic vibration sheet; the left shaft is of a hollow shaft structure and is provided with a rotary joint, one end of an air duct is connected with a nitrogen bottle outside the spherical reaction cavity, and the other end of the air duct penetrates through the rotary joint of the left shaft and extends into the ultrasonic treatment chamber and below the filter sieve plate; an exhaust port is arranged at the edge of the material placing port and is communicated with an exhaust pipeline, and the exhaust pipeline vertically extends to the upper part of the filter sieve plate and is close to the separator;
the spherical reaction chamber is externally wrapped with an electric heating layer, the electric heating layer is externally wrapped with a heat preservation and insulation layer, and the electric heating layer is electrically connected with an external power supply.
The invention at least comprises the following beneficial effects:
firstly, compounding physical operations according to a certain sequence to increase the dissolution rate and the extraction rate of the beta-glucan, further improving the extraction rate of the beta-glucan by performing the operations according to the sequence, namely crushing, adding an extracting agent, introducing nitrogen, ultrasound, microwave, dispersing and ultrasound, and converting insoluble beta-glucan into water-soluble beta-glucan by a microwave means in the extraction process, wherein the extraction rate is improved by 1.2-1.5 times compared with the existing single extraction operation; meanwhile, the method does not adopt means such as chemical additives and biological enzymes, but adopts a physical method, so that the extraction of the product is more environment-friendly, the production cost is lower, and the method has wide market prospect;
secondly, the porous carbon spheres are added during ultrasonic treatment, so that a multi-channel structure of the porous carbon spheres can be effectively utilized to increase air nuclei and strengthen a cavitation effect during ultrasonic treatment, the plant cell wall breaking amount is increased, and the beta-glucan is dissolved out more completely;
and thirdly, a double-guide reaction device is adopted for reducing and inhibiting Maillard reaction generated in the product extraction process, reducing the browning probability of the product, and the obtained product has good appearance quality and high purity and can be used as a high-quality product for production.
Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention.
Drawings
FIG. 1 is a schematic structural diagram of a double-guide reaction apparatus according to one embodiment of the present invention.
Detailed Description
The present invention is further described in detail below with reference to examples so that those skilled in the art can practice the invention with reference to the description.
< example 1>
Highland barley beta-glucan extraction process based on ultrasonic assistance
Step one, crushing cleaned highland barley raw materials to 40 meshes, degreasing to obtain a sample to be extracted, adding an extracting agent with the pH of 10, and stirring, wherein the weight ratio of the sample to be extracted to the extracting agent is 1: 18;
introducing nitrogen for 5min, heating to 55 ℃, and carrying out ultrasonic treatment for 26min at the power of 600W;
thirdly, processing for 15s by using microwave with the frequency of 900W, performing dispersion oscillation for 3.5h, performing ultrasonic processing with the power of 600W for 22min, and performing post-processing;
the degreasing treatment specifically comprises the following steps: adding absolute ethyl alcohol into the crushed highland barley powder, and uniformly stirring, wherein the mass ratio of the highland barley powder to the absolute ethyl alcohol is 1:4, refluxing for 2 hours at 76 ℃, concentrating and drying;
the post-treatment comprises the following steps: taking a mixture obtained after ultrasonic treatment, filtering to obtain a filtrate, adjusting the pH value of the filtrate to 4.5 by using 20% hydrochloric acid, standing for 10 hours at the temperature of 3 ℃, centrifuging to obtain the filtrate, adjusting the pH value of the filtrate to 7, adding medium-temperature alpha-amylase, heating to 85-90 ℃ for liquefying for 30min after stirring uniformly, concentrating to obtain a concentrated solution, adding 95% ethanol into the concentrated solution for settling, centrifuging to obtain a sediment, washing the sediment by using 95% ethanol, and drying.
The extraction rate of the highland barley beta-glucan obtained by calculation is 4.56 percent, the highland barley beta-glucan is light yellow solid, and the purity is 79.15 percent.
< example 2>
Highland barley beta-glucan extraction process based on ultrasonic assistance
Step one, crushing cleaned highland barley raw materials to 50 meshes, degreasing to obtain a sample to be extracted, adding an extracting agent with the pH of 8, and stirring, wherein the weight ratio of the sample to be extracted to the extracting agent is 1: 23;
introducing nitrogen for 5min, heating to 70 ℃, and carrying out ultrasonic treatment for 18min at the power of 800W;
step three, performing microwave treatment with the frequency of 850W for 20s, performing dispersion oscillation for 4h, performing ultrasonic treatment with the power of 800W for 40min, and performing post-treatment;
the degreasing treatment specifically comprises the following steps: adding absolute ethyl alcohol into the crushed highland barley powder, and uniformly stirring, wherein the mass ratio of the highland barley powder to the absolute ethyl alcohol is 1:4, refluxing for 2 hours at 76 ℃, concentrating and drying;
the post-treatment comprises the following steps: taking a mixture obtained after ultrasonic treatment, filtering to obtain a filtrate, adjusting the pH value of the filtrate to 4.5 by using 20% hydrochloric acid, standing for 10 hours at the temperature of 3 ℃, centrifuging to obtain the filtrate, adjusting the pH value of the filtrate to 7, adding medium-temperature alpha-amylase, heating to 85-90 ℃ for liquefying for 30min after stirring uniformly, concentrating to obtain a concentrated solution, adding 95% ethanol into the concentrated solution for settling, centrifuging to obtain a sediment, washing the sediment by using 95% ethanol, and drying.
The extraction rate of the highland barley beta-glucan obtained by calculation is 4.72 percent, the highland barley beta-glucan is light yellow solid, and the purity is 78.26 percent.
< example 3>
Highland barley beta-glucan extraction process based on ultrasonic assistance
Step one, crushing cleaned highland barley raw materials to 60 meshes, degreasing to obtain a sample to be extracted, adding an extracting agent with the pH of 10, and stirring, wherein the weight ratio of the sample to be extracted to the extracting agent is 1: 20; introducing nitrogen for 5min before adding the extractant;
the degreasing treatment specifically comprises the following steps: adding absolute ethyl alcohol into the crushed highland barley powder, and uniformly stirring, wherein the mass ratio of the highland barley powder to the absolute ethyl alcohol is 1:4, refluxing for 2 hours at 76 ℃, concentrating and drying;
introducing nitrogen for 5min, heating to 60 ℃, and carrying out ultrasonic treatment for 20min at the power of 650W;
step three, microwave treatment with the frequency of 900W is carried out for 18s, dispersion oscillation is carried out for 3h, ultrasonic treatment with the power of 800W is carried out for 20min, and post treatment is carried out;
the post-treatment comprises the following steps: taking a mixture obtained after ultrasonic treatment, filtering to obtain a filtrate, adjusting the pH value of the filtrate to 4.5 by using 20% hydrochloric acid, standing for 10 hours at the temperature of 3 ℃, centrifuging to obtain the filtrate, adjusting the pH value of the filtrate to 7, adding medium-temperature alpha-amylase, heating to 85-90 ℃ for liquefying for 30min after stirring uniformly, concentrating to obtain a concentrated solution, adding 95% ethanol into the concentrated solution for settling, centrifuging to obtain a sediment, washing the sediment by using 95% ethanol, and drying.
The extraction rate of the highland barley beta-glucan obtained by calculation is 5.14%, and the obtained product is light yellow in color and 79.52% in purity.
< example 4>
Highland barley beta-glucan extraction process based on ultrasonic assistance
Step one, crushing cleaned highland barley raw materials to 60 meshes, degreasing to obtain a sample to be extracted, adding an extracting agent with the pH of 10, and stirring, wherein the weight ratio of the sample to be extracted to the extracting agent is 1: 20; introducing nitrogen for 5min before adding the extractant;
the degreasing treatment specifically comprises the following steps: adding absolute ethyl alcohol into the crushed highland barley powder, and uniformly stirring, wherein the mass ratio of the highland barley powder to the absolute ethyl alcohol is 1:4, refluxing for 2 hours at 76 ℃, concentrating and drying;
introducing nitrogen for 5min, heating to 60 ℃, adding porous carbon balls with the diameter of 200-300 nm, wherein the weight ratio of the porous carbon balls to the highland barley raw materials is 1:20, carrying out ultrasonic treatment for 20min at the power of 800W, and taking out the porous carbon spheres after the reaction is finished;
step three, treating the mixture obtained in the step two for 18s under the microwave with the frequency of 900W, putting the porous carbon spheres taken out in the step two after the microwave treatment is finished, performing dispersion oscillation for 3h, performing ultrasonic treatment with the power of 800W for 26min, and performing post-treatment;
the post-treatment comprises the following steps: taking a mixture obtained after ultrasonic treatment, filtering to obtain a filtrate, adjusting the pH value of the filtrate to 4.5 by using 20% hydrochloric acid, standing for 10 hours at the temperature of 3 ℃, centrifuging to obtain the filtrate, adjusting the pH value of the filtrate to 7, adding medium-temperature alpha-amylase, heating to 85-90 ℃ for liquefying for 30min after stirring uniformly, concentrating to obtain a concentrated solution, adding 95% ethanol into the concentrated solution for settling, centrifuging to obtain a sediment, washing the sediment by using 95% ethanol, and drying.
The extraction rate of the highland barley beta-glucan obtained by calculation is 4.87%, the obtained product is white, and the purity is 81.26%.
< example 5>
Highland barley beta-glucan extraction process based on ultrasonic assistance
Step one, crushing cleaned highland barley raw materials to 40 meshes, degreasing to obtain a sample to be extracted, adding an extracting agent with the pH of 10, and stirring, wherein the weight ratio of the sample to be extracted to the extracting agent is 1: 18; introducing nitrogen gas for 5min before adding the extractant
The degreasing treatment specifically comprises the following steps: adding absolute ethyl alcohol into the crushed highland barley powder, and uniformly stirring, wherein the mass ratio of the highland barley powder to the absolute ethyl alcohol is 1:4, refluxing for 2 hours at 76 ℃, concentrating and drying;
introducing nitrogen for 5min, heating to 55 ℃, adding porous carbon balls with the diameter of 200-300 nm, wherein the weight ratio of the porous carbon balls to the highland barley raw materials is 1:20, carrying out ultrasonic treatment for 26min at the power of 600W;
step three, treating the mixture obtained in the step two for 15s under the microwave with the frequency of 900W, putting the porous carbon spheres taken out in the step two after the microwave treatment is finished, performing dispersion oscillation for 3h, performing ultrasonic treatment for 40min with the power of 800W, and performing post-treatment;
wherein, step two and step three adopt the reaction of two conduction formula reaction unit, two conduction formula reaction unit includes:
the spherical reaction cavity 2 is provided with a pair of shaft holes which are symmetrical relative to the sphere center on the outer wall of the spherical reaction cavity 2; the two shaft holes are respectively connected with a left shaft and a right shaft which are coaxial; a coaxial transmission assembly 3 is arranged on the right shaft to drive the spherical reaction cavity 2 to rotate, and meanwhile, the transmission assembly 3 also performs vibration dispersion operation; the spherical reaction cavity 2 is rotatably erected between the pair of vertical bases 1 through a left shaft and a right shaft;
a feeding hole and a material placing hole 505 are arranged on the circumference of the highest point of the spherical reaction cavity 2, the diameter of the circumference is the diameter of the spherical reaction cavity 2, and the feeding hole and the material placing hole 505 are symmetrical about the spherical center of the spherical reaction cavity 2; the feeding hole is used for feeding and discharging; the material placing port 505 is used for placing porous carbon balls;
a plate-shaped partition 6 is obliquely and fixedly arranged in the spherical reaction cavity 2, the edge of the partition 6 is fixedly connected with the inner wall of the spherical reaction cavity 2, the center of the partition 6 is over against the feed inlet, the partition 6 divides the space in the spherical reaction cavity 2 into a microwave treatment chamber 4 and an ultrasonic treatment chamber 5, the feed inlet is communicated with the microwave treatment chamber 4, and the material placing port 505 is communicated with the ultrasonic treatment chamber 5; a first material guide valve 403 is arranged on the separator 6 at the position facing the ultrasonic treatment chamber 5 and close to the lower part of the edge of the separator 6, and a second material guide valve 502 is arranged on the separator 6 at the position facing the microwave treatment chamber 4 and close to the lower part of the edge of the separator 6; the first material guide valve 403 and the second material guide valve 502 both penetrate through the partition 6 and communicate the microwave treatment chamber 4 and the ultrasonic treatment chamber 5, and the first material guide valve 403 and the second material guide valve 502 both are electrically controlled valves;
a microwave generator 401 is arranged on an arc-shaped side wall in the microwave processing chamber 4, and an anti-overflow layer 402 is attached to the inner side wall of the microwave processing chamber 4 and used for isolating microwaves; an ultrasonic vibration sheet 501 is arranged on an arc-shaped side wall in the ultrasonic treatment chamber 5, the ultrasonic vibration sheet is arranged along the circumferential direction of the inner wall of the spherical reaction chamber, a filter sieve plate 503 is arranged at a part, close to the material loading port 505, in the ultrasonic treatment chamber 5, the edge of the filter sieve plate 503 is fixedly connected with the inner wall of the ultrasonic treatment chamber 5, the filter sieve plate 503 is positioned below the ultrasonic vibration sheet 501, and the aperture of the filter sieve plate is smaller than the diameter of the porous carbon ball; the left shaft is of a hollow shaft structure and is provided with a rotary joint 203, one end of an air duct 504 is connected with a nitrogen bottle outside the spherical reaction cavity 2, and the other end of the air duct penetrates through the rotary joint 203 of the left shaft and extends into the ultrasonic treatment chamber 5 and below the filter screen plate 503; an exhaust port is arranged at the edge of the material placing port 505, the exhaust port is communicated with an exhaust pipe 506 channel and used for balancing the air pressure in the ultrasonic processing chamber, and the exhaust pipe 506 channel vertically extends to the upper part of the filter sieve plate 503 and is close to the separator 6;
the spherical reaction chamber 2 is externally wrapped with an electric heating layer 201, the electric heating layer 201 is externally wrapped with a heat insulation layer 202, and the electric heating layer 201 is electrically connected with an external power supply.
The specific using method comprises the following steps: filling the mixture of the sample to be extracted and the extracting solution obtained in the step one from a feeding hole, putting porous carbon balls into the mixture from a material placing hole 505 according to the proportion, and closing the material placing hole 505; opening the first material guide valve 403 to allow the mixture to enter the ultrasonic treatment chamber 5, closing the valve, heating the spherical reaction chamber 2 to an internal temperature of 60 ℃, keeping the temperature, opening the external power supply of the ultrasonic vibration sheet 501, opening the exhaust port, introducing nitrogen for 5min, adjusting the power of the ultrasonic vibration piece 501 to 400W, after ultrasonic treatment for 26min, starting the transmission assembly 3, turning over the spherical reaction cavity 2 until the microwave treatment chamber 4 is positioned below the ultrasonic treatment chamber 5, opening the second material guide valve 502 to enable the mixed liquid after ultrasonic treatment to enter the microwave treatment chamber 4, setting the frequency of the microwave generator 401 to be 900W, performing microwave treatment for 15s, then reversing the spherical reaction cavity 2 to enable the ultrasonic treatment chamber 5 to be positioned at the lower part of the microwave treatment chamber 4, starting the transmission assembly to enable the spherical reaction cavity 2 to disperse and vibrate for 3h, then adjusting the power of the ultrasonic vibration piece 501 to be 600W, and performing ultrasonic treatment for 40 min; after the reaction is finished, the mixture is led out for post-treatment;
the post-treatment comprises the following steps: taking a mixture obtained after ultrasonic treatment, filtering to obtain a filtrate, adjusting the pH value of the filtrate to 4.5 by using 20% hydrochloric acid, standing for 10 hours at the temperature of 3 ℃, centrifuging to obtain the filtrate, adjusting the pH value of the filtrate to 7, adding medium-temperature alpha-amylase, heating to 85-90 ℃ for liquefying for 30min after stirring uniformly, concentrating to obtain a concentrated solution, adding 95% ethanol into the concentrated solution for settling, centrifuging to obtain a sediment, washing the sediment by using 95% ethanol, and drying.
The extraction rate of the highland barley beta-glucan obtained by calculation is 4.92 percent, and a white solid with the purity of 84.68 percent is obtained.
By adopting the technical scheme, the Maillard reaction generated in the product extraction process can be effectively reduced and inhibited, the browning probability of the product is reduced, and the obtained product has good appearance quality and high purity and can be used as a high-quality product for production.
< comparative example 1>
The other steps were the same as in example 3 except that nitrogen gas was not introduced.
< comparative example 2>
The other steps were the same as in example 3, except that the ultrasonic treatment of the second step was not performed.
< comparative example 3>
The other steps were the same as in example 3, except that the microwave treatment of the third step was not performed.
< comparative example 4>
The other steps are the same as in example 3, except that the dispersion oscillation treatment of the third step is not performed.
< comparative example 5>
The other steps were the same as in example 3 except that the ultrasonic treatment of step three was not performed.
< comparative example 6>
The other steps are the same as in example 3 except that the dispersed oscillation operation of step three is located before the microwave treatment.
< comparative example 7>
The other steps are the same as in example 3, except that the microwave treatment in step three is prior to the ultrasonic treatment in step two.
< comparative example 7>
The other steps are the same as in example 3, except that the microwave treatment in step three is prior to the ultrasonic treatment in step two.
< comparative example 8>
The other steps are the same as example 3, except that the nitrogen aeration operation in step two is performed after the sonication in step two.
Examples of the experiments
Respectively calculating the extraction rate of the beta-glucan extracted in the example 3 and the comparative examples 1-8;
the formula for calculating the extraction rate is as follows:
wherein, Y is the yield (%) of the beta-glucan; mtThe mass of the extracted beta-glucan is obtained; m0Is the quality of highland barley raw material; the calculation results are shown in Table 1;
TABLE 1
As can be seen from Table 1, the absence of any step (example 3 and comparative examples 1-5) and the change of the sequence (example 3 and comparative examples 6-8) all had an effect on the extraction yield of the final beta-glucan, with a deviation value ranging from 0.41% to 1.3%;
from the comparison of the extraction rates of example 3 and comparative examples 1 to 5, it can be seen that the microwave treatment can greatly increase the extraction rate of β -glucan, and possible reasons include: after the first ultrasonic action, most of the soluble beta-glucan is dissolved into the extraction solution, and through the microwave thermal effect of 800W, the intermolecular energy of the beta-glucan is changed, so that the partially insoluble beta-glucan is converted into the soluble beta-glucan, the dissolution amount of the soluble beta-glucan is increased, and the extraction rate is improved; meanwhile, the original tissue structure of the plant cell wall is loosened due to the heat effect of the microwave, and the cell wall of the highland barley can be broken through more completely due to the cavitation effect generated during the ultrasonic treatment in the third step, so that the soluble beta-glucan can be dissolved out;
comparative example 4 shows that the mixture is treated by dispersion oscillation between microwave operation and ultrasonic operation, so that the soluble glucose is dispersed more uniformly in the extractant molecules, the dissolution amount of the soluble beta-glucan is effectively increased, and the extraction efficiency of the whole reaction is improved;
comparative example 5 shows that the first ultrasound was not shattered without the ultrasound treatment, and further that the microwave-activated, shatterable soluble β -glucan molecules in the cell wall were dissolved out, and no better effect was achieved on the increase of extraction yield;
comparative example 6 shows that after the microwave reaction, further dispersion oscillation treatment is needed to uniformly disperse the soluble beta-glucan in the converted mixture, so that the soluble beta-glucan is conveniently contacted with solvent molecules and plays a role in assisting dissolution so as to improve the extraction rate;
comparative example 7 it can be seen that the reduction of the extraction rate obtained by setting the microwave operation before the sonication in the second step is also likely to occur because, when the microwave operation is performed first, the probability of converting partially insoluble β -glucan into soluble β -glucan is reduced due to the uneven dispersion of β -glucan, and therefore the extraction rate of β -glucan cannot be improved even if the sonication operation is performed twice subsequently;
comparative example 8 it can be seen that reducing the oxygen content in the extract prior to cell wall disruption by ultrasound can inhibit maillard reactions of the dissolved β -glucan during extraction, which has the effect of increasing extraction yield from the aspect of inhibiting re-reactions, while ensuring a better product appearance.
Secondly, respectively detecting the purity of the beta-glucan extracted in the embodiments 1-5;
purity was measured by UV spectrophotometry
β -glucan (standard): aladdin reagents, Inc.;
congo red dye; disodium hydrogen phosphate; sodium dihydrogen phosphate: analytical grade
Model L5 uv-vis spectrophotometer: shanghai sperm family;
1. preparing 0.1mol/L phosphate buffer solution with pH of 8 from disodium hydrogen phosphate and sodium dihydrogen phosphate, and dissolving 0.0100g Congo red in 100mL0.1mol/L phosphate buffer solution with pH of 8.0 to obtain Congo red solution;
2. preparing 0.2mg/ml beta-glucan standard solution;
determining the maximum absorption wavelength to be 545 nm;
determining the pH value of a proper buffer solution to be 8;
measuring a blank sample;
measuring and drawing a standard curve; taking 0.2mg/ml beta-glucan standard solutions with the numbers of 0-9 and 0-0 blank, and the others being 0.1ml, 0.2ml and 0.3ml … 0.9.9 ml in sequence according to the numbers; supplementing pure water to 2.0ml, adding 4.0ml Congo red solution, shaking, measuring absorbance at 545nm to obtain standard curve, and calculating logarithm to obtain standard curve equation: y is 0.0047x + 0.0711; r2＝0.9995
3. Taking 0.0100g +/-0.0020 g of each of the crude samples obtained in the examples 1-5, using volumetric flasks to fix the volume, preparing 100ml of solution, taking 2.0ml, and adding 4.0ml of Congo red solution respectively; the absorbance of each group measured 3 times is averaged, and the average value is converted into corresponding content, and the purity is calculated, and the result is shown in table 2;
TABLE 2
| Item | Example 1 | Example 2 | Example 3 | Example 4 | Example 5 |
| Purity of | 79.15％ | 78.26％ | 79.52％ | 81.26％ | 84.68％ |
As can be seen from table 2, when nitrogen gas is introduced into the extracting solution, it is obvious that the purity of the product is increased, see example 3 and example 4, and compare example 1 and example 2, wherein the nitrogen gas is introduced into the extracting solution of example 3 and example 4, but the nitrogen gas is not introduced into the extracting solution of example 1 and example 2, because the content of dissolved oxygen in the extracting agent is lower after the nitrogen gas is introduced, the maillard reaction can be inhibited to a certain extent, the browning of the beta-glucan can be effectively prevented, and the purity of the product is reduced; meanwhile, the embodiment 5 adopts a double-guide type reaction device, adopts the same device to extract main steps, simultaneously reduces the contact between the product and oxygen, can effectively reduce and inhibit Maillard reaction generated in the product extraction process, reduces the browning probability of the product, has good appearance quality and high purity of the obtained product, and can be used as a high-quality product to produce
The double-guide reaction device is used for reducing and inhibiting Maillard reaction in the product extraction process, reducing the browning probability of the product, and the obtained product has good appearance quality and high purity and can be used for producing high-quality products.
While embodiments of the invention have been described above, it is not limited to the applications set forth in the description and the embodiments, which are fully applicable in various fields of endeavor to which the invention pertains, and further modifications may readily be made by those skilled in the art, it being understood that the invention is not limited to the details shown and described herein without departing from the general concept defined by the appended claims and their equivalents.
Claims (10)
1. The highland barley beta-glucan extraction process based on ultrasonic assistance is characterized by comprising the following steps of:
step one, crushing cleaned highland barley raw materials to 40-60 meshes, degreasing to obtain a sample to be extracted, adding an extracting agent with the pH of 8-10, and stirring, wherein the weight ratio of the sample to be extracted to the extracting agent is 1: 18-23;
introducing nitrogen, heating to 55-70 ℃, and carrying out ultrasonic treatment for 18-26 min, wherein the ultrasonic power is 600-800W;
step three, performing microwave treatment for 15-20 s, performing dispersion oscillation for 3-4 h, performing ultrasonic treatment for 20-40 min, and performing post-treatment; wherein the microwave frequency is 850-900W, and the ultrasonic power is 600-800W.
2. The ultrasonic-assisted highland barley beta-glucan extraction process based on claim 1, wherein in the first step, the weight ratio of the sample to be extracted to the extracting agent is 1: 20.
3. The highland barley beta-glucan extraction process based on ultrasonic assistance as claimed in claim 1, wherein in the second step, the heating temperature is 60 ℃.
4. The highland barley beta-glucan extraction process based on ultrasonic assistance as claimed in claim 1, wherein the microwave frequency in the third step is 900W, and the microwave treatment time is 18 s.
5. The highland barley beta-glucan extraction process based on ultrasonic assistance as claimed in claim 1, wherein in the second step, the ultrasonic power is 650W, the frequency is 20KHz, and the time is 20 min; in the third step, the ultrasonic power is 800W, the frequency is 20KHz, and the time is 20 min.
6. The highland barley beta-glucan extraction process based on ultrasonic assistance as claimed in claim 1, wherein in the step one, nitrogen is introduced for 5min before the extractant is added.
7. The highland barley beta-glucan extraction process based on ultrasonic assistance as claimed in claim 1, wherein the second step is to add porous carbon spheres with the diameter of 200-300 nm during ultrasonic treatment, after the reaction is finished, the porous carbon spheres are taken out, the microwave treatment of the third step is carried out, and after the microwave treatment is finished, the taken out porous carbon spheres are put in again, and the rest steps are carried out; the weight ratio of the porous carbon balls to the highland barley raw materials is 1: 20.
8. the highland barley beta-glucan extraction process based on ultrasonic assistance as claimed in claim 1, wherein the degreasing treatment is: adding absolute ethyl alcohol into the crushed highland barley powder, and uniformly stirring, wherein the mass ratio of the highland barley powder to the absolute ethyl alcohol is 1:4, refluxing for 2h at 76 ℃, concentrating and drying.
9. The highland barley beta-glucan extraction process based on ultrasonic assistance as claimed in claim 1, wherein the post-treatment is: taking a mixture obtained after ultrasonic treatment, filtering to obtain a filtrate, adjusting the pH value of the filtrate to 4.5 by using 20% hydrochloric acid, standing for 10 hours at the temperature of 3 ℃, centrifuging to obtain the filtrate, adjusting the pH value of the filtrate to 7, adding medium-temperature alpha-amylase, uniformly stirring the highland barley raw material with the addition of 4u per gram, heating to 85-90 ℃, liquefying for 30 minutes, concentrating to obtain a concentrated solution, adding 95% ethanol into the concentrated solution for settling, centrifuging to obtain a sediment, washing the sediment by using 95% ethanol, and drying.
10. The ultrasound-assisted highland barley beta-glucan extraction process as claimed in claim 7, wherein the second step and the third step are carried out in a double-conduction reaction device, the double-conduction reaction device comprises:
the outer wall of the spherical reaction cavity is provided with a pair of shaft holes which are symmetrical relative to the spherical center of the spherical reaction cavity; the two shaft holes are respectively connected with a left shaft and a right shaft which are coaxial; a coaxial transmission assembly is arranged on the right shaft to drive the spherical reaction cavity to rotate; the spherical reaction cavity is rotatably erected between a pair of vertical bases through the left shaft and the right shaft;
a feed inlet and a material placing port are arranged on the circumference of the highest point of the spherical reaction cavity, and the feed inlet and the material placing port are symmetrical about the spherical center of the spherical reaction cavity;
a plate-shaped partition is fixedly arranged in the spherical reaction cavity in an inclined manner, the edge of the partition is fixedly connected with the inner wall of the spherical reaction cavity, the center of the partition is opposite to the feed inlet, the partition divides the space in the spherical reaction cavity into a microwave treatment chamber and an ultrasonic treatment chamber, the feed inlet is communicated with the microwave treatment chamber, and the material placing port is communicated with the ultrasonic treatment chamber; a first material guide valve is arranged on the separator at the position facing the ultrasonic treatment chamber and close to the lower part of the edge of the separator, and a second material guide valve is arranged on the separator at the position facing the microwave treatment chamber and close to the lower part of the edge of the separator; the first guide valve and the second guide valve penetrate through the partition and are communicated with the microwave treatment chamber and the ultrasonic treatment chamber, and the first guide valve and the second guide valve are electrically controlled valves;
the microwave treatment device comprises a microwave treatment chamber, a microwave generator, an anti-overflow layer and a water tank, wherein the arc-shaped side wall in the microwave treatment chamber is provided with the microwave generator, and the inner side wall of the microwave treatment chamber is also adhered with the anti-overflow layer; an ultrasonic vibration sheet is arranged on the arc-shaped side wall in the ultrasonic treatment chamber, a filter sieve plate is arranged at the part, close to the material placing port, in the ultrasonic treatment chamber, the edge of the filter sieve plate is fixedly connected with the inner wall of the ultrasonic treatment chamber, and the filter sieve plate is positioned below the ultrasonic vibration sheet; the left shaft is of a hollow shaft structure and is provided with a rotary joint, one end of an air duct is connected with a nitrogen bottle outside the spherical reaction cavity, and the other end of the air duct penetrates through the rotary joint of the left shaft and extends into the ultrasonic treatment chamber and below the filter sieve plate; an exhaust port is arranged at the edge of the material placing port and is communicated with an exhaust pipeline, and the exhaust pipeline vertically extends to the upper part of the filter sieve plate and is close to the separator;
the spherical reaction chamber is externally wrapped with an electric heating layer, the electric heating layer is externally wrapped with a heat preservation and insulation layer, and the electric heating layer is electrically connected with an external power supply.
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