CN111265565A - A kind of Scutellaria baicalensis dregs semi-solid fermentation technology method and its application - Google Patents

Abstract

The invention provides a technical method for semi-solid fermentation of Scutellaria baicalensis dregs, belonging to the technical field of semi-solid fermentation. The technical scheme is as follows: taking the scutellaria dregs, pulverizing the scutellaria dregs to less than 20 mm, to obtain the scutellaria dregs powder, adding a variety of compound enzyme mixtures to the scutellaria dregs powder, fully mixing them evenly, and drying the fully mixed mixture , drying the mixture, fermenting the drying mixture under vacuum conditions and keeping the temperature at 37° C., and obtaining Scutellaria baicalensis dregs fermentation product after the fermentation is completed and vacuum-packing. The beneficial effects of the invention are as follows: the semi-solid fermentation method is used to process the scutellaria baicalensis dregs, and the liquid Lactobacillus plantarum and the compound enzyme used can decompose the components such as fiber and pectin in the scutellaria baicalensis dregs, combined with the flavonoids in the scutellaria baicalensis. The interaction of various probiotics such as lactic acid bacteria can not only realize the utilization of medicinal residues under milder conditions than traditional physical and chemical methods, but also moderately adjust their medicinal properties and expand the scope of medication.

Description

A kind of Scutellaria baicalensis dregs semi-solid fermentation technology method and its application

technical field

The invention relates to the technical field of semi-solid fermentation, in particular to a technical method for semi-solid fermentation of Scutellaria baicalensis dregs and its application.

Background technique

The traditional Chinese medicine Scutellaria baicalensis is the dry root of the perennial herb Scutellaria baicalensis, mainly produced in Heilongjiang, Jilin, Liaoning, Hebei, Shandong, Shaanxi, Gansu, Inner Mongolia and other places. Scutellaria baicalensis has the functions of clearing away heat and dampness, purging fire and detoxifying, hemostasis and toxin. , Carbuncle swollen sore, fetal movement restless. Scutellaria baicalensis mainly contains baicalin, baicalein, wogonin, wogonin, in addition to baicalein-7-O-glycoside, 5,7-dihydroxy-6-methoxydihydroflavonoid, chrysanthin and Its glycosides, wood butterfly A and its glucuronide and other components. Scutellaria baicalensis belongs to a large number of commonly used medicinal materials. According to relevant statistics, 70% of Chinese patent medicines on the market currently contain Scutellaria baicalensis, such as commonly used Chinese patent medicines Qingkailing oral liquid, Shuanghuanglian granules/oral liquid/freeze-dried powder injection, Yinhuang tablet/silver Huang Granules/Yinhuang Oral Liquid, Xiao Chai Hu Capsules/ Xiao Chai Hu Tablets/ Xiao Chai Hu Granules, Longdan Xiegan Pills, Gujing Pills, etc. For Scutellaria baicalensis with an annual dosage of 16,000 to 20,000 tons, the output of Scutellaria baicalensis dregs is also very amazing. With the vigorous development of the pharmaceutical industry, the emission of Scutellaria baicalensis dregs is also increasing. Most of the traditional methods adopted, such as incineration, fixed area stacking, landfill, etc., not only cause huge damage to the environment, but also cause waste of resources. According to relevant researches, the scutellaria baicalensis dregs obtained after extraction of baicalin still contain flavonoids, and the dregs still have the functions of antibacterial, antibacterial, antiviral, and antioxidant.

SUMMARY OF THE INVENTION

The object of the present invention is to provide a method of utilizing semi-solid fermentation to process Scutellaria baicalensis dregs, the microorganisms used and composite enzymes can decompose the components such as fiber and pectin in the Scutellaria baicalensis dregs, combined with flavonoids in Scutellaria baicalensis effective ingredients and lactic acid bacteria etc. The interaction of various probiotics can not only realize the utilization of medicinal residues under milder conditions than traditional physical and chemical methods, but also moderately adjust the medicinal properties and expand the scope of medicine. method.

The present invention is achieved by the following measures:

A technical method for semi-solid fermentation of Scutellaria baicalensis dregs, characterized in that the technical method for semi-solid fermentation comprises the following steps:

S1, get Scutellaria baicalensis dregs, and Scutellaria baicalensis medicinal dregs are squeezed to obtain the Scutellaria baicalensis medicinal dregs with a moisture content of 30% to 70%;

S2, pulverizing the Scutellaria baicalensis dregs with a moisture content of 30% to 70% obtained in step S1 to below 20 mm, and press-filtering the pulverized Scutellaria baicalensis dregs to obtain Scutellaria baicalensis dregs powder with a moisture content of 30% to 60%;

S3, adding 0.3% to 0.5% by weight of liquid Lactobacillus plantarum and 0.3% to 0.5% by weight of multiple complex enzyme mixtures to the Scutellaria baicalensis dregs powder with a moisture content of 30% to 60% obtained in step S2, and fully well mixed;

S4. The mixture finally obtained in step S3 is fermented under vacuum conditions and the temperature is kept at 37° C. for 7 days, and after the fermentation is completed, the fermentation product of Scutellaria baicalensis dregs is obtained and vacuum-packed.

The specific features of the present invention also include:

The semi-solid fermentation technology method comprises the following steps:

S1, get Scutellaria baicalensis dregs 1kg, Scutellaria baicalensis medicinal dregs are squeezed, obtain the Scutellaria baicalensis medicinal dregs with a moisture content of 35%;

S2, the Scutellaria baicalensis dregs with a moisture content of 35% obtained in step S1 is pulverized to less than 20 mm, and the pulverized Scutellaria baicalensis dregs are press-filtered to obtain Scutellaria baicalensis dregs powder with a moisture content of 30%;

S3, adding 0.3% by weight of liquid Lactobacillus plantarum and 0.3% by weight of multiple composite enzyme mixtures in the Scutellaria baicalensis dregs powder with a moisture content of 30% obtained in step S2, and fully mixed;

S4. The mixture finally obtained in step S3 is fermented under vacuum conditions and the temperature is kept at 37° C. for 7 days, and after the fermentation is completed, the fermentation product of Scutellaria baicalensis dregs is obtained and vacuum-packed.

The multiple composite enzyme mixture in the step S2 is composed of cellulase, pectinase and xylanase, and the ratio of cellulase, pectinase and xylanase is: 2:1:1.

The Scutellaria baicalensis dregs fermentation product is used for the application of antibacterial and bacteriostasis.

The beneficial effects of the invention are as follows: using the method of semi-solid fermentation to process the Scutellaria baicalensis dregs, the microorganisms and compound enzymes used can decompose the components such as fiber, pectin and the like in the Scutellaria baicalensis dregs, combined with the flavonoids in the Scutellaria baicalensis effective ingredients and lactic acid bacteria and other components. The interaction of various probiotics can not only realize the utilization of medicinal residues under milder conditions than traditional physical and chemical methods, but also moderately adjust their medicinal properties and expand the scope of medication.

Detailed ways

In order to clearly illustrate the technical features of the solution, the solution will be described below through specific implementations.

Example 1:

A technical method for semi-solid fermentation of Scutellaria baicalensis dregs, the technical method for semi-solid fermentation comprises the following steps:

S1, get Scutellaria baicalensis dregs 1kg, Scutellaria baicalensis medicinal dregs are squeezed, obtain the Scutellaria baicalensis medicinal dregs with a moisture content of 35%;

S2, the Scutellaria baicalensis dregs with a moisture content of 35% obtained in step S1 is pulverized to less than 20 mm, and the pulverized Scutellaria baicalensis dregs are press-filtered to obtain Scutellaria baicalensis dregs powder with a moisture content of 30%;

S3, adding 0.3% by weight of liquid Lactobacillus plantarum and 0.3% by weight of multiple compound enzyme mixtures in the Scutellaria baicalensis dregs powder with a moisture content of 30% obtained in step S2, and fully mixed;

S4. The mixture finally obtained in step S3 is fermented under vacuum conditions and the temperature is kept at 37° C. for 7 days, and after the fermentation is completed, the fermentation product of Scutellaria baicalensis dregs is obtained and vacuum-packed.

During the storage period of 12 months, the Scutellaria baicalensis slag fermented product prepared by this semi-solid fermentation technology has no obvious changes in appearance, identification, inspection and content inspection indicators, and meets the requirements of preclinical drug quality standards. Basically stable.

Example 2:

A technical method for semi-solid fermentation of Scutellaria baicalensis dregs, the technical method for semi-solid fermentation comprises the following steps:

S1, get Scutellaria baicalensis dregs 1kg, Scutellaria baicalensis medicinal dregs are extruded, obtain the Scutellaria baicalensis medicinal dregs with a moisture content of 70%;

S2, the Scutellaria baicalensis dregs with a moisture content of 70% obtained in the step S1 is pulverized to less than 20 mm, and the pulverized Scutellaria baicalensis dregs are press-filtered to obtain the Scutellaria baicalensis dregs powder with a moisture content of 60%;

S3, adding the liquid Lactobacillus plantarum of 0.5% by weight and the multiple compound enzyme mixture of 0.5% by weight in the Scutellaria baicalensis dregs powder with a moisture content of 60% obtained in step S2, and fully mixing;

S4. The mixture finally obtained in step S3 is fermented under vacuum conditions and the temperature is kept at 37° C. for 7 days, and after the fermentation is completed, the fermentation product of Scutellaria baicalensis dregs is obtained and vacuum-packed.

Example 3:

A technical method for semi-solid fermentation of Scutellaria baicalensis dregs, the technical method for semi-solid fermentation comprises the following steps:

S1, get Scutellaria baicalensis dregs 1kg, Scutellaria baicalensis medicinal dregs are extruded, obtain the Scutellaria baicalensis medicinal dregs with a moisture content of 50%;

S2, the Scutellaria baicalensis dregs with a moisture content of 50% obtained in step S1 is pulverized to less than 20 mm, and the pulverized Scutellaria baicalensis dregs are pressure-filtered to obtain a Scutellaria baicalensis dregs powder with a moisture content of 45%;

S3, adding the liquid Lactobacillus plantarum of 0.4% by weight and the multiple compound enzyme mixture of 0.4% by weight in the Scutellaria baicalensis dregs powder with a moisture content of 45% obtained in step S2, and fully mixing;

S4. The mixture finally obtained in step S3 is fermented under vacuum conditions and the temperature is kept at 37° C. for 7 days, and after the fermentation is completed, the fermentation product of Scutellaria baicalensis dregs is obtained and vacuum-packed.

The inventor of this Scutellaria baicalensis dregs semi-solid fermentation technology method has carried out a series of experimental studies on the fermentation technology method and its application, and has determined this Scutellaria baicalensis dregs semi-solid fermentation technical method, which has the advantages of higher efficiency than the prior art. Obvious outstanding therapeutic advantage.

Experiment 1. Determination of flavonoids in semi-solid fermentation of Scutellaria baicalensis residues

1. Materials

1.1. Sample: Scutellaria baicalensis dregs fermented product prepared in Example 1, Example 2 and Example 3 above.

1.2. Control:

(1), Scutellaria baicalensis dregs;

(2), fermenting the mixture finally obtained in step S3 in Example 1 under vacuum conditions and maintaining the temperature at 37° C. for 7 days, obtaining Scutellaria baicalensis slag fermented product after the fermentation is completed and vacuum-packing to obtain a control fermented product.

2. Test method

2.1. Determination method: Determination by high performance liquid chromatography (general rule 0512)

2.1.1. Chromatographic conditions

Chromatographic column: use octadecylsilane-bonded silica gel as filler; use methanol-0.1% phosphoric acid as mobile phase, and carry out gradient elution as specified in Table 1; detection wavelength is 275 nm. The number of theoretical plates should not be less than 2500 according to the baicalin peak.

Table 1

2.2. Solution preparation

Preparation of reference solution: take appropriate amount of baicalin, wogonin, baicalein, and wogonin, add methanol to make a solution of 100 μg/ml.

Preparation of the test solution:

Accurately weigh 2 g of Scutellaria baicalensis dregs powder, accurately add 25 ml of 60% methanol, ultrasonically treat for 10 min, and take the filtrate to obtain;

Take 2 g each of the Scutellaria baicalensis slag fermented products prepared in Example 1, Example 2 and Example 3, accurately weigh, accurately weigh, place in a stoppered conical flask, accurately add 25 ml of 60% methanol, close the stopper, and weigh. weight, ultrasonically treated for 10 minutes, allowed to cool, weighed again, used 60% methanol to make up for the lost weight, shaken well, filtered, and the subsequent filtrate was obtained.

2.3. Detection method

Precisely measure the same amount of the above-mentioned reference substance and the test solution solution, inject it into a high performance liquid chromatograph, and detect it. With reference to the reference substance solution, the external standard method is used to obtain the detection results, and the results are shown in Table 2.

Table 2 Contents of flavonoids

3. Conclusion

As can be seen from the above table 2, the effects of Examples 1, 2 and 3 are better, the content of flavonoids is increased by 2 times, especially the content of baicalein has been greatly improved, and baicalein can reduce cerebral vascular resistance, improve brain Blood circulation, increase cerebral blood flow and anti-platelet aggregation. Clinically used for the treatment of paralysis after cerebrovascular disease. After baicalein enters the animal body, it is rapidly converted into baicalin and other metabolites in the blood. Compared with Example 1, the total content of flavonoids in the control fermented sample fermented for 10 days is lower than that of the sample fermented for 7 days in Example 1. Therefore, from the perspective of cost saving, it is considered that fermentation for 7 days is sufficient.

Experiment 2. Antibacterial experiment of Scutellaria baicalensis dregs semi-solid fermentation product in the present invention

1 sample

1.1 Strains

Escherichia coli O78 standard strain (E. coli O78, number CVCC1418); Staphylococcus aureus strain ATCC29213.

1.2 Samples and Reagents

Samples: Scutellaria baicalensis dregs fermentation product and Scutellaria baicalensis dregs samples prepared in Example 1, Example 2 and Example 3.

Reagents: Nutrient broth medium, MacConkey medium, common nutrient agar medium, ethanol and other reagents are of analytical grade.

1.3 Instruments

Circulating vacuum water pump (SHZ-3 type), vacuum drying oven (ZK-82B type), electronic temperature control electric heater, high pressure steam sterilizer, electronic analytical balance (FA1004 type), reflux condensing device, distillation device, etc.

2. Method

2.1 Preparation of bacterial solution

Standard Escherichia coli and Staphylococcus aureus were streaked on the solid medium and cultured at 37°C for 18-24h. A single colony was picked and inoculated into the nutrient broth medium and cultured at 37°C for 18h. Determine the concentration of the bacterial solution by the plate dilution method, and adjust the bacterial concentration to 1.5×10 ⁸ CFU/ml for later use.

2.2 Preparation of test samples

Take the Scutellaria baicalensis dregs fermentation product and Scutellaria baicalensis dregs samples prepared in Examples 1, 2, and 3 for decoction extraction, add 10 times the amount of water for extraction for 1 h, concentrate to a solution containing 1 g/ml of drug, and place in a vial , 120 ℃ high pressure sterilization 20min, set aside 4 ℃ refrigerator.

2.3 Determination of the minimum inhibitory concentration MIC value

Four experimental groups (including Ⅰ, Ⅱ, Ⅲ, Ⅳ groups) were set up, and 12 small test tubes were taken and numbered in each experimental group; 1 ml of ordinary nutrient broth medium was added to each test tube with a sterile pipette; Add 1ml of the test drug to the first tube under aseptic conditions, shake and mix; draw 1ml of the test drug and add it to the second tube, shake and mix, and so on to the 11th tube, draw 1ml and discard (guarantee the volume of the solution in the 12 test tubes). The same), the 12th tube was not added as a positive control. Add 1 ml of the above-prepared bacterial solution to each tube, set it in a 37°C incubator, and observe the test results after culturing for 18 hours. The previous tube of the growth tube is the MIC of the sample. The test was repeated 3 times and the average value was taken as the MIC value of each sample;

Wherein, the test drug prepared from the fermentation product of Scutellaria baicalensis in Example 1 was used in group I, the test drug prepared from the fermentation product of Scutellaria baicalensis in Example 2 was used in group II, and the test drug prepared from the fermentation product of Scutellaria baicalensis in Example 3 was used in group III. The experimental drug prepared from the scutellaria baicalensis drug residue was used in group IV.

3. Results

The in vitro antibacterial effects of experimental groups I, II, III, and IV on Escherichia coli and Staphylococcus aureus are shown in Table 3:

Table 3 In vitro antibacterial effect

It can be seen from Table 3 that groups I, II, III and IV have certain bacteriostatic effects on gram-negative bacteria such as Escherichia coli and gram-positive bacteria such as Staphylococcus aureus. Among them, the antibacterial effect of group I was the best, and the minimum inhibitory concentration could reach 156 mg/ml; the second was group III, which had better antibacterial effect, with an inhibitory concentration of 225 mg/ml; the antibacterial effect of groups II and IV was the worst. The inhibitory concentration was more than 1000mg/ml.

Experiment 3. Acute toxicity evaluation experiment of Scutellaria baicalensis dregs semi-solid fermentation product in the present invention

Take Examples 1, 2, 3 and Scutellaria baicalensis powder, decoct and extract in water, add 10 times the amount of water to extract for 1 hour, and then concentrate to a solution with a drug content of 200 mg/ml. Each mouse was given each sample by gavage at 5g/kg b.wt. once, and the gavage volume was 0.2ml/10g. After administration, the mice were continuously observed for 4 hours and then for 7 days. During the observation period, the mental state, There was no abnormality in the general state of coat color, autonomous activity, breathing, diet, feces, oral and nasal secretions, and no animal died, and the body weight increased naturally. After the experiment, the mice were killed by dislocation. There were no visible lesions in the main organs, and no obvious poisoning or adverse reactions. It shows that the LD ₅₀ of the fermentation product to Kunming mice is greater than 5g/kg. According to the WTO classification standard of acute toxicity of exogenous chemicals, the test results show that the fermentation product is actually non-toxic.

The undescribed technical features of the present invention can be realized by or using the existing technology, and will not be repeated here. Of course, the above description is not a limitation of the present invention, and the present invention is not limited to the above examples. Those skilled in the art Changes, modifications, additions or substitutions made within the essential scope of the present invention shall also belong to the protection scope of the present invention.

Claims (4)

1. a semi-solid fermentation technical method for Scutellaria baicalensis dregs, is characterized in that, described semi-solid fermentation technical method comprises the following steps: S1, get Scutellaria baicalensis dregs, and Scutellaria baicalensis medicinal dregs are squeezed to obtain the Scutellaria baicalensis medicinal dregs with a moisture content of 30% to 70%; S2, pulverizing the Scutellaria baicalensis dregs with a moisture content of 30% to 70% obtained in step S1 to less than 20 mm, and press-filtering the pulverized Scutellaria baicalensis dregs to obtain Scutellaria baicalensis dregs powder with a moisture content of 30% to 60%; S3, adding 0.3%-0.5% by weight of liquid Lactobacillus plantarum and 0.3%-0.5% by weight of the multi-enzyme mixture in the Scutellaria baicalensis dregs powder with a moisture content of 30%-60% obtained in step S2, and fully well mixed; S4, fermenting the mixture finally obtained in the step S3 under vacuum conditions and maintaining the temperature at 37° C. for 7 days, and after the fermentation is completed, the Scutellaria baicalensis dregs fermentation product is obtained and vacuum-packed. 2. according to the described semi-solid fermentation technical method of Scutellaria baicalensis dregs of claim 1, described semi-solid fermentation technical method comprises the following steps: S1, get Scutellaria baicalensis dregs 1kg, Scutellaria baicalensis medicinal dregs are squeezed, obtain the Scutellaria baicalensis medicinal dregs with a moisture content of 35%; S2, the Scutellaria baicalensis dregs with a moisture content of 35% obtained in step S1 is pulverized to less than 20 mm, and the pulverized Scutellaria baicalensis dregs is press-filtered to obtain the Scutellaria baicalensis dregs powder with a moisture content of 30%; S3, adding the liquid Lactobacillus plantarum of 0.3% by weight and the multiple compound enzyme mixture of 0.3% by weight in the Scutellaria baicalensis dregs powder with a moisture content of 30% in step S2, and fully mixing; S4, fermenting the mixture finally obtained in the step S3 under vacuum conditions and maintaining the temperature at 37° C. for 7 days, and obtaining the Scutellaria baicalensis dregs fermentation product after the fermentation is completed, and performing vacuum packaging. 3. Scutellaria baicalensis dregs semi-solid fermentation technology method according to claim 2, is characterized in that, the described multiple compound enzyme mixture in described S2 step is made up of cellulase, pectinase and xylanase mixed, The ratio of the cellulase, the pectinase and the xylanase is 2:1:1. 4. The method for semi-solid fermentation of Scutellaria baicalensis dregs according to claim 1, wherein the fermentation product of Scutellaria baicalensis dregs is used for the application of antibacterial and bacteriostatic.