Method for improving active substances of germinated coix seeds through ultrasonic and CaCl2 pretreatment

Abstract

The invention provides a method for improving active substances of germinated coix seeds. The method comprises the following steps: by taking coix seeds as a raw material, performing ultrasonic treatment at 500W for 15 minutes, soaking in a 0.5 mmol / L CaCl2 solution for 9 hours, and germinating for 36 hours, thereby obtaining the germinated coix seeds rich in active substances. Compared with control germinated coix seeds, the content of GABA, polyphenol and reducing sugar in the germinated coix seeds treated by ultrasonic and CaCl2 is increased from 76-81 mg / 100 g, 220-237 mg / 100 g, 0.87-0.95 g / 100 g to 159-164 mg / 100 g, 265-278 mg / 100 g, 1.54-1.74 g / 100 g and is increased by about 101.38%, 18.71% and 79.35% respectively, the content of GABA is remarkably increased, it is indicated that the activity of GABA branch metabolism key enzymes in the germination process of the coix seeds can be better stimulated through ultrasonic and CaCl2 treatment, and the content of polyphenol and reducing sugar in the germinated coix seeds treated by the ultrasonic and CaCl2 treatment can be better improved. The metabolism and conversion of nutrient substances are enhanced, so that the content of active substances in the germinated coix seeds is remarkably increased.

Description

Technical Field

[0001] The present application relates to the technical field of food processing, and in particular to providing a method for increasing the active substances in germinated coix seeds. Background Art

[0002] Job's tears (Coix lachrymal-jobi L.), also known as coix rice, is a common crop used for both medicine and food with a long history. Job's tears has high nutritional value and is rich in a variety of nutrients, and is known as the "king of the grass family." Job's tears has significant nutritional value, high protein and fat content, and is rich in fatty acids, esters, polysaccharides and other compounds. In recent years, with the deepening of research, the value of coix seeds in food nutrition and medicine has gradually become clear, and has received close attention. Job's tears has shown great potential in the treatment of cancer, hypertension, hyperlipidemia, etc., and it also occupies an important position in traditional Chinese medicine because of its good effect of removing dampness and promoting diuresis.

[0003] During the germination stage of grains, internal molecular enzymes begin to activate and release, thus undergoing complex biological and chemical changes. As an economical means of grain processing, germination has been proven to improve the nutritional value and biological activity of grains. During the germination process, endogenous amylase in grains is activated, starch is decomposed, and the content of reducing sugars is significantly increased. The energy required for the reaction is generated by fat decomposition, the activity of proteases is increased, and proteins are decomposed into amino acids, which in turn provide raw materials for new protein synthesis. As the germination time increases, the content of secondary metabolites such as GABA and polyphenols in grains also increases significantly. After germination technology treatment, the nutritional content and functional properties of grains can be significantly improved, and the anti-nutritional and indigestible components in grains are also reduced with germination. Therefore, sprouted grains can enhance the taste, flavor and nutritional value of food to meet consumers' demand for healthy and delicious food. A large number of studies have confirmed that germination is an economical and effective grain processing method that can significantly improve the nutritional value and biological activity of grains. The germination processing technology of grains such as brown rice, oats, and buckwheat has received widespread attention in research at home and abroad. However, research on the germination characteristics of coix seed and its processing methods is still in its infancy.

[0004] In recent years, the application of ultrasonic technology in the field of food processing has gradually attracted attention. Ultrasonic waves can destroy cell structures and promote material transfer and reactions through their cavitation effect, thermal effect and mechanical effect. In liquid systems, the effect between ultrasonic waves and the medium is mainly cavitation effect. Ultrasonic waves can have a series of effects on seeds, including promoting seed germination and growth, improving seed vitality and stress resistance, and increasing crop yields. Ultrasonic waves can promote the physiological metabolism of seeds, promote the activity of enzymes and material transformation in seeds, leading to the promotion of seed germination. It enables seeds to adapt to the environment faster and grow into healthy plants. Ultrasonic waves can also improve the vitality and stress resistance of seeds. By treating seeds, ultrasonic waves can activate the physiological mechanisms inside seeds, allowing them to better adapt to changes in the external environment, such as temperature, light, soil, etc. It helps to improve the resilience and ability of seeds to adapt to adversity, thereby increasing the survival rate and yield of plants. Ultrasonic waves have a certain relationship with the accumulation of active substances in coix seeds. Ultrasonic treatment can destroy cell walls and accelerate the release of intracellular substances, thereby increasing the active ingredients in coix seeds.

[0005] Ca 2+ It is one of the essential mineral elements for plant growth. It not only regulates plant growth and development and participates in the formation of plant cell walls, but is also an important signal molecule and osmotic protective substance for plant cells. 2+ Increasing the activity of amylase can regulate plant photosynthesis, control the activity of related enzymes in glycolysis and pentose phosphate pathway metabolism, Ca 2+ By binding to phospholipids to maintain the structure of the lipid bilayer, it provides structural integrity for the plasma membrane, improves the permeability of the cell membrane, and reduces the effect of water loss on grain germination. 2+ It can alleviate the adverse effects of salinity during seed germination. As a common inorganic salt, exogenous addition of CaCl can alleviate the inhibitory effect of adverse stress on plant growth by increasing the concentration of free calcium ions in the cytoplasm. During seed germination, the activities of various enzymes change, and these enzymes are essential for the metabolic activities inside the seeds. CaCl treatment can regulate the activity of these enzymes and thus affect the germination process of seeds. Therefore, it is feasible to use ultrasound in conjunction with CaCl2 pretreatment to increase the active substances in germinated coix seeds. Summary of the invention

[0006] Purpose of the invention: To provide a method for increasing the active substances in germinated coix seeds.

[0007] (1) Screening and cleaning of coix seeds: Select coix seeds that are free of mildew, insect damage, full grains, and no damage to the epidermis, soak them in 3 times the volume of sodium hypochlorite solution for disinfection, and rinse them thoroughly three times with deionized water to remove residual disinfectant;

[0008] (2) Soaking coix seeds to absorb water: Add the sterilized coix seeds to deionized water to ensure that the coix seeds fully absorb water, and then soak them in a water bath at a constant temperature and then take them out;

[0009] (3) Ultrasonic assisted CaCl2 pretreatment: The sample is then placed in an ultrasonic generator for ultrasonic treatment. The coix seeds after ultrasonic treatment are soaked in a CaCl2 solution in a water bath at a constant temperature and then taken out;

[0010] (4) Germination culture of coix seed: Drain the surface water of the treated coix seed, place it in a culture dish lined with filter paper, spread it evenly, and place it in a constant temperature and humidity chamber for germination. During the germination period, use a watering can to water it every 12 hours, and regularly clean and inspect the coix seed to prevent bacterial growth;

[0011] The method for preparing sprouted coix seeds rich in active substances according to claim 1, characterized in that the concentration of sodium hypochlorite in step (1) is 2% and the soaking time is 15 minutes.

[0012] Preferably, in step (2), the coix seeds are soaked to absorb water: deionized water is added in a volume ratio of 1:3, and the seeds are soaked in a water bath at a constant temperature of 28° C. for 3 hours.

[0013] Preferably, in step (3), ultrasound-assisted CaCl2 pretreatment is performed: frequency 40 kHz, ultrasonic power 500 W for 15 min, material-water ratio is 1:3, CaCl2 solution concentration is 0.5 mmol / L, and soaking is performed at 28°C for 9 h.

[0014] Preferably, in step (4), the coix seeds are germinated and cultured in a constant temperature and humidity chamber at 28° C. and 90% humidity for 36 hours.

[0015] Technical effects:

[0016] 1. The raw material of the present invention is coix seed, which is rich in vitamins and polyphenols, and has the health functions of lowering blood sugar, lowering blood lipids, lowering blood pressure, preventing cancer and anti-cancer, and anti-oxidation. Long-term consumption has a certain preventive and therapeutic effect on hyperglycemia and hyperlipidemia.

[0017] 2. The present invention has made an innovative attempt. In order to further improve the nutritional value of germinated coix seeds, ultrasound and CaCl2 pretreatment are used to increase the active substances in germinated coix seeds. During the germination stage of grains, endogenous amylase in the grains is activated during the germination process, and starch is decomposed, resulting in a significant increase in the content of reducing sugars. The energy required for the reaction is generated by fat decomposition, the activity of proteases is increased, and proteins are decomposed into amino acids, which in turn provide raw materials for new protein synthesis. As the germination time increases, the content of secondary metabolites such as GABA and polyphenols in grains also increases significantly. Compared with ordinary coix seeds, germinated coix seeds pretreated with ultrasound and CaCl2 contain richer nutrients and can better meet the needs of modern people for a healthy diet.

[0018] 3. The nutritional components of the germinated coix seeds of the present invention are changed. Compared with the germinated coix seeds without pretreatment, the starch, protein and fat contents in the germinated coix seeds treated with ultrasound and CaCl2 are changed. The starch and fat contents are reduced from 58.88-59.01 g / 100 g and 14.07-14.21 g / 100 g to 52.72-52.88 g / 100 g and 13.01-13.53 g / 100 g, respectively, which are reduced by about 10.40% and 10.10%, respectively. However, the protein content does not change significantly. The data show that the synergistic effect of calcium chloride and ultrasound can better stimulate enzyme activity, promote the decomposition and utilization of starch and fat, and provide the required energy for seed germination.

[0019] 4. The active substances in the germinated coix seeds of the present invention are significantly improved. Compared with the control germinated coix seeds, the contents of GABA, polyphenols and reducing sugars in the germinated coix seeds treated with ultrasound and CaCl2 are all improved, from 76-81 mg / 100g, 220-237 mg / 100g, 0.87-0.95 g / 100g to 159-164 mg / 100g, 265-278 mg / 100g, 1.65-1.74 g / 100g, respectively, which are increased by about 101.38%, 18.71% and 79.35%, respectively, among which the GABA content is significantly improved, indicating that the synergistic effect can better stimulate the activity of key enzymes of GABA branch metabolism during the germination of coix seeds. The results showed that ultrasound synergistic CaCl2 pretreatment played an important role in the growth and development of coix seed. The synergistic treatment provided good conditions for coix seed germination, enhanced the metabolism and transformation of nutrients, and significantly increased the content of active substances.

[0020] Index determination method:

[0021] Starch content determination

[0022] Iodine colorimetry was used. Preparation of mother solution: First, accurately weigh 1.0g starch using an electronic balance, dissolve it in 100mL deionized water, and stir continuously in a 95℃ water bath to obtain a clear and transparent gelatinized starch solution, which was fixed to volume in a 250mL volumetric flask. Take 5mL of mother solution and add it to a 100mL volumetric flask. Take 0, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, and 4.0mL of standard starch solution, add 0.2mL of iodine solution, and make up to 10mL with deionized water. After fully mixing, let it stand for 10 minutes, and measure the absorbance value at a wavelength of 660nm using a spectrophotometer.

[0023] Protein content determination

[0024] Use automatic Kjeldahl nitrogen analyzer for determination. Weigh 2.0g (accurate to 0.0001g) of fully mixed coix seed powder and digest it in a digestion furnace. Set the amount of reagents added in the Kjeldahl nitrogen analyzer: 50ml deionized water, 20mL 2% boric acid, 80mL 32% NaOH solution. Connect the hydrochloric acid titrator, adjust the pH meter, adjust the Method to SET-pH to start the experiment, insert the pH meter and the hydrochloric acid tube into the Kjeldahl nitrogen analyzer, click the pre-preparation program and record the hydrochloric acid volume and pH, measure the empty tube twice, and then start the experiment. Record the protein content on the automatic Kjeldahl nitrogen analyzer.

[0025] Fat content determination

[0026] Use the Soxhlet extraction method, refer to GB 5009.6-2016. Accurately weigh 2g to 5g of coix seed powder (accurate to four decimal places), place it in a folded filter paper bag, seal the filter paper tube to prevent the powder from scattering, put it in the extraction tube, add petroleum ether to the round-bottom flask that has been dried to constant weight until the liquid volume in the bottle reaches two-thirds of the total capacity, install a condenser, and place the entire system in a 55°C water bath for heating. During the heating process, the petroleum ether needs to be continuously refluxed to complete the extraction process, and the duration is set to 12h. After the extraction process is completed, remove the round-bottom flask and recover the petroleum ether.

[0027] Take the filter paper bag out of the extraction cylinder, dry it for a while and weigh it. The fat content in coix seed powder is calculated as shown in formula (1):

[0028]

[0029] Wherein: X represents the fat content in coix seed powder (dry basis) (g / 100g); M1 represents the mass of the filter paper bag plus coix seed powder after constant weight (g); M0 represents the mass of the filter paper bag plus coix seed powder before defatting (g); M2 represents the mass of coix seed powder (g); 100 represents the conversion factor.

[0030] Active ingredient extraction

[0031] Polyphenol extraction method: weigh 1g of freeze-dried coix seed powder, add 35mL of 60% ethanol by volume into a centrifuge tube, with a solid-liquid ratio of 1:35, ultrasonicate for 1h, centrifuge the resulting solution at a centrifugal force of 1445.5×g for 10min, collect the supernatant, and store at 4°C for later use.

[0032] GABA and reducing sugar extraction method: weigh 0.5 g of freeze-dried coix seed powder, add 10 mL of deionized water into a centrifuge tube, with a solid-liquid ratio of 1:20, ultrasonicate for 1 h, centrifuge the resulting solution at a centrifugal force of 2469.7 × g for 15 min, collect the supernatant, and store at 4 °C for later use.

[0033] GABA content determination

[0034] GABA extraction and determination: Dilute the standard solution of GABA step by step to 0.01, 0.02, 0.03, 0.04, 0.05 and 0.06 mg / mL. Take 0.5 mL of the dilution, add 0.2 mL of H3BO3 buffer with a pH of 9.0 and a concentration of 0.2 mol / L, 1 mL of C6H5OH solution with a mass fraction of 6%, and 0.4 mL of NaClO solution with a mass fraction of 7%, shake well, heat in a boiling water bath for 10 minutes, and then transfer to an ice water bath and cool until the solution turns blue-green. At this time, add 2 mL of 60% ethanol solution, measure the absorbance at a wavelength of 630 nm, and draw a standard curve. Take the supernatant after centrifugation and measure the absorbance value according to the above operation, and calculate the GABA content according to the standard curve.

[0035] Determination of polyphenol content:

[0036] The free phenol content was determined by the Folin-phenol method. 0, 0.25, 0.50, 0.75, 1.00, 1.25 and 1.50 mL gallic acid standard solution was accurately pipetted into a 25 mL test tube, and 3.0 mL Folin-phenol reagent was added respectively. After shaking and standing for 30 s, 6.0 mL Na2CO3 solution (12%, m / v) was added respectively. The volume was made up to 25 mL, and the mixture was placed at 25 ° C in the dark for 2 h. Sample 0 was used as a blank and the absorbance was measured at a wavelength of 765 nm. A standard curve Y=0.0864X+0.0849 (R2=0.9979) was drawn with the standard solution concentrations of 0, 1, 2, 3, 4, 5 and 6 (μg / mL) as the abscissa and the absorbance at a wavelength of 765 nm as the ordinate. The supernatant after centrifugation was taken and the absorbance value was measured according to the above operation, and the free polyphenol content was determined according to the standard curve.

[0037] Reducing sugar content determination

[0038] Accurately add 0, 0.2, 0.4, 0.6, 0.8, 1.0, and 1.2 mL of 1 mg / mL glucose standard solution to 25 mL test tubes, fill each tube to 2 mL with distilled water, add 1.5 mL of 3,5-dinitrosalicylic acid reagent to each test tube, heat in a boiling water bath for 5 minutes, and then move to a cold water bath to cool to room temperature. Subsequently, distilled water is used to make the volume to 25 mL and stir evenly. At a wavelength of 540 nm, the absorbance of each tube is measured and a standard curve is drawn. Take 3 25 mL test tubes, add 2 mL of the reducing sugar test solution to each tube, and follow the steps for making a standard curve. Measure the absorbance of each test tube and calculate the total amount of reducing sugar by referring to the standard curve.

[0039] Determination of coix seed powder by infrared spectrum after germination

[0040] The infrared spectrum of coix seed powder was measured. An appropriate amount of freeze-dried coix seed powder was weighed and placed on a Fourier infrared spectrometer, and the scanning analysis was performed at a scanning frequency of 32 times, with air as a reference, and at a scanning wavelength of 4000-400 cm-1.

[0041] Analysis of thermal properties of coix seed powder after germination

[0042] Thermal properties analysis of coix seed powder. 5 mg coix seed powder was weighed in a special aluminum crucible, 10 μL deionized water was added, and the crucible was sealed and hydrated at 4°C for 12 hours. The empty crucible was used as a reference, the temperature range was 20°C to 200°C, the heating rate was 10°C / min, and nitrogen protection was used. The thermodynamic parameters, sample temperature Ts, reference temperature Tr, and gelatinization endothermic enthalpy ΔH were analyzed by the TA Universal Analysis software that comes with the differential scanning calorimeter. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1 Flowchart for germination of coix seeds

[0044] Figure 2 A, B, and C are the starch content, protein content, and fat content of Comparative Example 1, Example 1, Example 2, Example 3, and Example 4, respectively.

[0045] Figure 2As shown in A, the starch content of comparative example 1 is 60.79-61.05 g / 100 g, the starch content of coix seed in example 1 is 58.88-59.01 g / 100 g, the starch content of coix seed in example 2 is 57.37-57.53 g / 100 g, and the starch content of example 3 is 56.20-56.76 g / 100 g. By comparing the data, CaCl2 treatment and ultrasonic treatment can both increase the decomposition rate of starch. The study found that low-intensity ultrasonic cavitation and thixotropic effect can increase the permeability of cell walls and cell membranes. It enhances the hydration and enzyme activity of seeds and accelerates the material exchange rate and metabolism inside and outside cells. CaCl2 is CaCl2. 2+ The endogenous amylase in the seeds was activated, the hydrolysis of starch was enhanced, and the amylase was stimulated to decompose starch into small sugar molecules. The starch content of Example 4 was 52.72-52.88 g / 100 g. Compared with the single treatment, the starch hydrolysis effect of the synergistic treatment was more significant, indicating that CaCl2 and ultrasound acted together to activate amylase and promote starch hydrolysis.

[0046] Figure 2 As shown in Figure B, the protein content of Comparative Example 1 is 15.55-16.93 g / 100 g, and the protein content of Example 1 is 16.48-17.78 g / 100 g. This is because the content of essential amino acids increases during the germination process, resulting in an increase in protein content. The protein content of Example 2 is 15.65-17.09 g / 100 g. Compared with the protein content of Comparative Example 1, CaCl2 treatment increases protein consumption. Studies have found that the exogenous application of CaCl2 significantly increases the total calcium and calmodulin content in grain germination. The protein content of Example 3 is 16.51-18.43 g / 100 g, and ultrasonic treatment increases the protein content. The protein content of coix seed in Example 4 is 16.07-17.97 g / 100 g. Compared with the single treatment, the synergistic treatment improves the increase in protein consumption after CaCl2 treatment, making the germination of coix seed more stable.

[0047] Figure 2 As shown in Figure C, the fat content of Comparative Example 1 is 14.13-15.69 g / 100 g, and the fat content of Example 1 is 14.07-14.21 g / 100 g. The data show that the seed germination consumes energy during the germination of 36 h, and the fat is decomposed to provide the energy required for seed germination. The fat content of Example 2 is 13.91-13.97 g / 100 g, and the fat content of Example 3 is 13.69-13.79 g / 100 g. Compared with Example 1, the fat content is reduced. The fat content of Example 4 is 13.01-13.53 g / 100 g, which is lower than that of the single treatment. This shows that ultrasonic treatment accelerates the consumption of fat inside coix seed, while Ca 2+It stimulates physiological and biochemical reactions in cells to consume energy, resulting in a significant effect in depleting fat content.

[0048] Figure 3 A, B, and C are the GABA content, polyphenol content, and reducing sugar content of Comparative Example 1, Example 1, Example 2, Example 3, and Example 4, respectively.

[0049] Figure 3 As shown in A, the GABA content of Comparative Example 1 is 55-61 mg / 100 g, and the GABA content of Example 1 is 76-81 mg / 100 g. The GABA content of Example 2 is 85-90 mg / 100 g. CaCl2 treatment will increase the GABA content in the coix seed germination process. 2+ Enhance the plasma membrane permeability, strengthen the seed vitality, and play an important role in the seed germination process. The GABA content of Example 3 is 101-105 mg / 100 g, and ultrasonic treatment will also increase the GABA content of coix seed during germination. The GABA content of Example 4 is 159-164 mg / 100 g, and the GABA content of coix seed after synergistic treatment exceeds the content of single treatment, indicating that ultrasound and CaCl2 treatment jointly promote GABA synthesis.

[0050] Figure 3 As shown in Figure 2, the polyphenol content of Example 1 is 170-175 mg / 100 g, and the polyphenol content of Example 1 is 220-237 mg / 100 g, and the polyphenol content is significantly increased. The polyphenol content of Example 2 is 238-245 mg / 100 g, and the polyphenol content of Example 3 is 249-262 mg / 100 g. It can be seen that both ultrasonic treatment and CaCl2 treatment can increase the polyphenol content during germination. The polyphenol content of Example 4 is 265-278 mg / 100 g. Compared with the single treatment, the polyphenol content of the synergistic treatment is significantly increased, indicating that the cavitation process of ultrasound changes the permeability of the cell membrane to CaCl2. 2+ The regulation inside the cells creates a favorable environment, and the overlapping effects of ultrasound and CaCl2 form a superimposed effect, so that the polyphenol content of the synergistic treatment increases significantly.

[0051] Figure 3As shown in Figure C, the reducing sugar content of Comparative Example 1 is 0.65-0.73 g / 100 g, and the reducing sugar content of Example 1 is 0.87-0.95 g / 100 g. The starch is decomposed into reducing sugar during the germination process, resulting in an increase in the reducing sugar content. The reducing sugar content of Example 2 is 0.97-0.99 g / 100 g, and the reducing sugar content of Example 3 is 1.36-1.48 g / 100 g. The reducing sugar content of coix seeds treated with ultrasound and CaCl2 during germination is significantly increased, and the reducing sugar content of Example 4 is 1.54-1.74 g / 100 g / 100 g. The synergistic treatment has more reducing sugar content than the single treatment, indicating that the ultrasonic treatment increases the reducing sugar decomposition of starch by enhancing the hydration of the seeds and improving the activity of amylase, while CaCl2 increases the reducing sugar content of coix seeds. 2+ It also increases the activity of amylase and controls the activity of related enzymes in glycolysis and pentose phosphate pathway metabolism, both of which jointly promote the increase in reducing sugar content.

[0052] Figure 4 It is the infrared spectra of Comparative Example 1, Example 1, Example 2, Example 3 and Example 4.

[0053] like Figure 4 As shown, 3500~3000, 2970~2837, 2385~2295, 1766~1480, 1182~882cm -1 The absorption peaks are obvious, and the infrared spectra of the five samples are similar, with similar peak positions and shapes. Except for the smoother spectrum of Comparative Example 1, there is no significant difference in the main characteristic absorption peaks of Example 1, and no new peaks appear. In addition, compared with Example 1, the peaks of Examples 2, 3, and 4 are all higher, and Example 4 is higher than Examples 2 and 3, indicating that the synergistic treatment promotes the decomposition of polysaccharide molecules such as starch, but has no obvious effect on the functional groups of the main components, and no new groups are generated.

[0054] Figure 5 It is the DSC curve of comparative example 1, example 1, example 2, example 3 and example 4.

[0055] As the temperature rises, starch granules disintegrate, and as the energy changes, an endothermic phenomenon is formed on the DSC spectrum peak. Figure 5 As shown, the peak values ​​of Examples 2 and 3 are higher than that of Example 1, indicating that the single treatment promotes starch decomposition, resulting in an inconspicuous endothermic phenomenon during measurement. The peak value of Example 4 is higher than that of Example 1, indicating that the synergistic treatment accelerates the decomposition of starch compared with the single treatment, resulting in an extremely inconspicuous endothermic phenomenon during measurement, indicating that the synergistic treatment promotes the decomposition of starch and is beneficial to the growth and development of coix seeds. DETAILED DESCRIPTION

[0056] The present invention is further explained below in conjunction with specific examples, but the examples do not limit the present invention in any form. The percentages involved in the following examples are weight percentages.

[0057] Example 1

[0058] A method for preparing germinated coix seeds, characterized in that the method comprises the following steps in sequence:

[0059] (1) Screening and cleaning of coix seeds: Select coix seeds that are free of mold, insect damage, full grains, and no damage to the epidermis, and soak them in 3 times the volume of 2% sodium hypochlorite solution for 15 min for disinfection, and rinse them thoroughly three times with deionized water to remove residual disinfectant;

[0060] (2) Soaking coix seeds to absorb water: Add deionized water to the sterilized coix seeds at a volume ratio of 1:3, and soak them in a water bath at 28°C for 3 h to ensure that the coix seeds fully absorb water. Then, soak them in a water bath at a constant temperature and take them out.

[0061] (4) Germination culture of coix seed: The surface water of coix seed was drained, and the seeds were placed in a culture dish lined with filter paper, spread evenly, and germinated in a constant temperature and humidity chamber at 28°C and 90% humidity for 36 hours. During the germination period, water was used to soak the seeds every 12 hours using a watering can, and the coix seed seeds were cleaned and inspected regularly to prevent bacterial growth.

[0062] Example 2

[0063] A method for preparing germinated coix seeds pretreated with CaCl2, characterized in that the method comprises the following steps in sequence:

[0064] (1) Screening and cleaning of coix seeds: Select coix seeds that are free of mold, insect damage, full grains, and no damage to the epidermis, and soak them in 3 times the volume of 2% sodium hypochlorite solution for 15 min for disinfection, and rinse them thoroughly three times with deionized water to remove residual disinfectant;

[0065] (2) Soaking coix seeds to absorb water: Add deionized water to the sterilized coix seeds at a volume ratio of 1:3, and soak them in a water bath at 28°C for 3 h to ensure that the coix seeds fully absorb water. Then, soak them in a water bath at a constant temperature and take them out.

[0066] (3) CaCl2 pretreatment: Then, the coix seeds were soaked in a CaCl2 solution with a material-water ratio of 1:3 and placed in a water bath for constant temperature soaking before being taken out;

[0067] (4) Germination culture of coix seed: The surface moisture of the treated coix seed was then drained, placed in a culture dish lined with filter paper, spread evenly, and germinated in a constant temperature and humidity chamber at 28°C and 90% humidity for 36 hours. During the germination period, water was used to soak the coix seed every 12 hours using a watering can, and the coix seed was cleaned and inspected regularly to prevent bacterial growth.

[0068] Example 3

[0069] A method for preparing ultrasonically pretreated germinated coix seeds, characterized in that the method comprises the following steps in sequence:

[0070] (1) Screening and cleaning of coix seeds: Select coix seeds that are free of mold, insect damage, full grains, and no damage to the epidermis, and soak them in 3 times the volume of 2% sodium hypochlorite solution for 15 min for disinfection, and rinse them thoroughly three times with deionized water to remove residual disinfectant;

[0071] (2) Soaking coix seeds to absorb water: Add deionized water to the sterilized coix seeds at a volume ratio of 1:3, and soak them in a water bath at 28°C for 3 h to ensure that the coix seeds fully absorb water. Then, soak them in a water bath at a constant temperature and take them out.

[0072] (3) Ultrasonic pretreatment: The sample was then placed in an ultrasonic generator with a frequency of 40 kHz and an ultrasonic power of 500 W for 15 min;

[0073] (4) Germination culture of coix seed: The surface moisture of the treated coix seed was then drained, placed in a culture dish lined with filter paper, spread evenly, and germinated in a constant temperature and humidity chamber at 28°C and 90% humidity for 36 hours. During the germination period, water was used to soak the coix seed every 12 hours using a watering can, and the coix seed was cleaned and inspected regularly to prevent bacterial growth.

[0074] Example 4

[0075] A method for preparing germinated coix seeds rich in active substances, characterized in that the method comprises the following steps in sequence:

[0076] (1) Screening and cleaning of coix seeds: Select coix seeds that are free of mold, insect damage, full grains, and no damage to the epidermis, and soak them in 3 times the volume of 2% sodium hypochlorite solution for 15 min for disinfection, and rinse them thoroughly three times with deionized water to remove residual disinfectant;

[0077] (2) Soaking coix seeds to absorb water: Add deionized water to the sterilized coix seeds at a volume ratio of 1:3, and soak them in a water bath at 28°C for 3 h to ensure that the coix seeds fully absorb water. Then, soak them in a water bath at a constant temperature and take them out.

[0078] (3) Ultrasonic assisted CaCl2 pretreatment: The sample was then placed in an ultrasonic generator with a frequency of 40 kHz and an ultrasonic power of 500 W for 15 min. The coix seeds after ultrasonic treatment were soaked in a CaCl2 solution with a material-water ratio of 1:3 and placed in a water bath for constant temperature soaking before being taken out;

[0079] (4) Germination culture of coix seed: The surface moisture of the treated coix seed was then drained, placed in a culture dish lined with filter paper, spread evenly, and germinated in a constant temperature and humidity chamber at 28°C and 90% humidity for 36 hours. During the germination period, water was used to soak the coix seed every 12 hours using a watering can, and the coix seed was cleaned and inspected regularly to prevent bacterial growth.

[0080] Comparative Example 1

[0081] The preparation of untreated coix seeds is characterized by being prepared by the following steps in sequence:

[0082] (1) Screening of coix seeds: Select coix seeds that are free of mold, insect damage, full grains, and no damage to the surface for processing;

[0083] (2) Cleaning: Soak in 3 volumes of 2% sodium hypochlorite solution for 15 min for disinfection, and rinse thoroughly three times with deionized water to remove residual disinfectant.

Claims

1. A method for preparing active substances in germinated coix seeds, characterized in that: Prepare as follows: (1) Screening and cleaning of coix seeds: Select coix seeds that are free of mildew, insect damage, full grains, and no damage to the epidermis, soak them in 3 times the volume of sodium hypochlorite solution for disinfection, and rinse them thoroughly three times with deionized water to remove residual disinfectant; (2) Soaking coix seeds to absorb water: Add the sterilized coix seeds to deionized water to ensure that the coix seeds fully absorb water, and then soak them in a water bath at a constant temperature and then take them out; (3) Ultrasonic assisted CaCl2 pretreatment: The sample is then placed in an ultrasonic generator for ultrasonic treatment. The coix seeds after ultrasonic treatment are soaked in a CaCl2 solution in a water bath at a constant temperature and then taken out; (4) Germination culture of coix seed: The surface water of the treated coix seed was then drained, placed in a culture dish lined with filter paper, spread evenly, and placed in a constant temperature and humidity chamber for germination. During the germination period, water was used to soak the coix seed every 12 hours using a watering can, and the coix seed was cleaned and inspected regularly to prevent bacterial growth.

2. The method for preparing germinated coix seeds rich in active substances according to claim 1, characterized in that: The sodium hypochlorite concentration in step (1) is 2%, and the soaking time is 15 minutes.

3. The method for preparing germinated coix seeds rich in active substances according to claim 1, characterized in that: Step (2) soaking the coix seeds to absorb water: add deionized water in a volume ratio of 1:3, and soak in a water bath at a constant temperature of 28°C for 3 hours.

4. The method for preparing germinated coix seeds rich in active substances according to claim 1, characterized in that: Step (3) Ultrasonic assisted CaCl2 pretreatment: frequency 40 kHz, ultrasonic power 500 W for 15 min, material-water ratio of 1:3, CaCl2 solution concentration of 0.5 mmol / L, soaking at 28°C for 9 h.

5. The method for preparing germinated coix seeds rich in active substances according to claim 1, characterized in that: Step (4) Germination culture of coix seeds: Germination for 36 hours in a constant temperature and humidity chamber at 28° C. and 90% humidity.

6. A method for preparing sprouted coix seeds with high active substance content according to the preparation method of claims 1 to 5, characterized in that: Ultrasound combined with CaCl2 treatment provided good conditions for coix seed germination, enhanced the metabolism and transformation of nutrients, and significantly increased the content of active substances.

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