CN111265565A - Scutellaria baicalensis dreg semisolid fermentation technical method and application thereof - Google Patents
Scutellaria baicalensis dreg semisolid fermentation technical method and application thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
- A61K36/539—Scutellaria (skullcap)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
Abstract
The invention provides a semisolid fermentation technical method of scutellaria baicalensis dreg, belonging to the technical field of semisolid fermentation. The technical scheme is as follows: pulverizing Scutellariae radix residue to below 20mm to obtain Scutellariae radix residue powder, adding multiple complex enzymes into Scutellariae radix residue powder, mixing, oven drying, fermenting at 37 deg.C under vacuum condition, and vacuum packaging. The invention has the beneficial effects that: the scutellaria baicalensis residue is treated by a semisolid fermentation method, the used liquid lactobacillus plantarum and complex enzyme can decompose fiber, pectin and other components in the scutellaria baicalensis residue, and the interaction of flavonoid active ingredients in scutellaria baicalensis and various probiotics such as lactic acid bacteria is combined, so that the residue can be utilized under a milder condition than the traditional physical and chemical method, the drug property of the scutellaria baicalensis residue can be properly adjusted, and the application range is enlarged.
Description
Technical Field
The invention relates to the technical field of semisolid fermentation, in particular to a technical method for semisolid fermentation of scutellaria baicalensis dreg and application thereof.
Background
The traditional Chinese medicine scutellaria is the dry root of a perennial herb scutellaria in the family of labiatae, and is mainly produced in Heilongjiang, Jilin, Liaoning, Hebei, Shandong, Shanxi, Gansu, inner Mongolia and other places. The scutellaria has the efficacies of clearing heat and drying dampness, purging fire and removing toxicity, and stopping bleeding and preventing abortion, and is mainly used for treating damp warmth, summer dampness, chest distress and nausea, damp-heat fullness, dysentery, jaundice, lung heat cough, high fever polydipsia, blood heat hematemesis and epistaxis, carbuncle swelling and sore toxicity, and threatened abortion. The Scutellariae radix mainly contains baicalin, baicalein, wogonin, baicalein-7-O-glycoside, 5, 7-dihydroxy-6-methoxy dihydroflavone, chrysin and its glycoside, oroxylin A and its glucuronide, etc. The scutellaria belongs to a large amount of common medicinal materials, and according to statistics of related data, 70 percent of Chinese patent medicines in the market at present contain the scutellaria, such as common Chinese patent medicines of qingkailing oral liquid, Shuanghuanglian granules/oral liquid/freeze-dried powder injection, Yinhuang tablets/Yinhuang granules/Yinhuang oral liquid, Xiaochaihu capsules/Xiaochaihu tablets/Xiaochaihu granules, gentian liver-fire-purging pills, Gujing pills and the like. For scutellaria baicalensis with annual dosage of 1.6 to 2 ten thousand tons, the yield of scutellaria baicalensis dregs is also extremely remarkable, the emission of the scutellaria baicalensis dregs is increased with the rapid development of the pharmaceutical industry, but the traditional methods for treating the traditional Chinese medicine dregs such as burning, stacking in a fixed area, burying and the like not only greatly destroy the environment, but also cause resource waste. According to related research and determination, the scutellaria residue obtained after extracting the baicalin still contains flavonoid components, and the residue still has the effects of resisting bacteria, resisting viruses, resisting oxidation and the like.
Disclosure of Invention
The invention aims to provide a scutellaria baicalensis dreg semisolid fermentation technical method for treating scutellaria baicalensis dreg by using a semisolid fermentation method, wherein the used microorganisms and complex enzyme can decompose fiber, pectin and other components in the scutellaria baicalensis dreg, and the interaction of flavonoid active ingredients in scutellaria baicalensis and various probiotics such as lactic acid bacteria is combined, so that the scutellaria baicalensis dreg semisolid fermentation technical method can be used for realizing the utilization of the dreg under the milder condition than the traditional physicochemical method, properly adjusting the drug property and expanding the application range.
The invention is realized by the following measures:
a technical method for semisolid fermentation of scutellaria baicalensis dreg is characterized by comprising the following steps:
s1, taking the scutellaria baicalensis dregs, and extruding the scutellaria baicalensis dregs to obtain the scutellaria baicalensis dregs with the water content of 30-70%;
s2, crushing the scutellaria baicalensis dregs with the water content of 30-70% obtained in the step S1 to be less than 20mm, and carrying out filter pressing on the crushed scutellaria baicalensis dregs to obtain scutellaria baicalensis dregs powder with the water content of 30-60%;
s3, adding 0.3-0.5 wt% of liquid lactobacillus plantarum and 0.3-0.5 wt% of a mixture of a plurality of complex enzymes into the scutellaria baicalensis dreg powder with the water content of 30-60% obtained in the step S2, and fully and uniformly mixing;
and S4, fermenting the mixture finally obtained in the step S3 for 7 days under vacuum condition at 37 ℃, obtaining a scutellaria baicalensis dreg fermented product after fermentation is finished, and carrying out vacuum packaging.
The invention has the following specific characteristics:
the semi-solid fermentation technical method comprises the following steps:
s1, taking 1kg of scutellaria baicalensis dregs, and extruding the scutellaria baicalensis dregs to obtain scutellaria baicalensis dregs with the water content of 35%;
s2, crushing the scutellaria baicalensis residue with the water content of 35% obtained in the step S1 to be less than 20mm, and performing pressure filtration on the crushed scutellaria baicalensis residue to obtain scutellaria baicalensis residue powder with the water content of 30%;
s3, adding 0.3 weight percent of liquid lactobacillus plantarum and 0.3 weight percent of a mixture of various complex enzymes into the scutellaria baicalensis residue powder with the water content of 30 percent obtained in the step S2, and fully and uniformly mixing;
and S4, fermenting the mixture finally obtained in the step S3 for 7 days under vacuum condition at 37 ℃, obtaining a scutellaria baicalensis dreg fermented product after fermentation is finished, and carrying out vacuum packaging.
The complex enzyme mixture in the step S2 is formed by mixing cellulase, pectinase and xylanase, wherein the ratio of the cellulase to the pectinase to the xylanase is as follows: 2:1:1.
The scutellaria baicalensis dreg fermented product is applied to antibiosis and bacteriostasis.
The invention has the beneficial effects that: the method for treating the scutellaria baicalensis dregs by using the semisolid fermentation method has the advantages that the used microorganisms and complex enzyme can decompose the fiber, pectin and other components in the scutellaria baicalensis dregs, and the interaction of flavonoid active ingredients in scutellaria baicalensis and various probiotics such as lactic acid bacteria is combined, so that the utilization of the dregs can be realized under the milder condition than the traditional physical and chemical method, the medicine property of the dregs can be properly adjusted, and the application range is enlarged.
Detailed Description
In order to clearly illustrate the technical features of the present solution, the present solution is explained below by way of specific embodiments.
Example 1:
a semisolid fermentation technical method of scutellaria baicalensis dreg comprises the following steps:
s1, taking 1kg of scutellaria baicalensis dregs, and extruding the scutellaria baicalensis dregs to obtain scutellaria baicalensis dregs with the water content of 35%;
s2, crushing the scutellaria baicalensis residue with the water content of 35% obtained in the step S1 to be less than 20mm, and performing pressure filtration on the crushed scutellaria baicalensis residue to obtain scutellaria baicalensis residue powder with the water content of 30%;
s3, adding 0.3 weight percent of liquid lactobacillus plantarum and 0.3 weight percent of a mixture of various complex enzymes into the scutellaria baicalensis residue powder with the water content of 30 percent obtained in the step S2, and fully and uniformly mixing;
and S4, fermenting the mixture finally obtained in the step S3 for 7 days under vacuum condition at 37 ℃, obtaining a scutellaria baicalensis dreg fermented product after fermentation is finished, and carrying out vacuum packaging.
The radix scutellariae dregs fermented product prepared by the semi-solid fermentation technical method has no obvious change in appearance, identification, inspection and content inspection indexes during 12 months of storage, meets all regulations of quality standards of clinical prodrugs, and has basically stable quality.
Example 2:
a semisolid fermentation technical method of scutellaria baicalensis dreg comprises the following steps:
s1, taking 1kg of scutellaria baicalensis dregs, and extruding the scutellaria baicalensis dregs to obtain scutellaria baicalensis dregs with the water content of 70%;
s2, crushing the scutellaria baicalensis residue with the water content of 70% obtained in the step S1 to be less than 20mm, and carrying out pressure filtration on the crushed scutellaria baicalensis residue to obtain scutellaria baicalensis residue powder with the water content of 60%;
s3, adding 0.5 weight percent of liquid lactobacillus plantarum and 0.5 weight percent of a plurality of complex enzymes into the scutellaria baicalensis dreg powder with the water content of 60 percent obtained in the step S2, and fully and uniformly mixing;
and S4, fermenting the mixture finally obtained in the step S3 for 7 days under vacuum condition at 37 ℃, obtaining a scutellaria baicalensis dreg fermented product after fermentation is finished, and carrying out vacuum packaging.
Example 3:
a semisolid fermentation technical method of scutellaria baicalensis dreg comprises the following steps:
s1, taking 1kg of scutellaria baicalensis dregs, and extruding the scutellaria baicalensis dregs to obtain scutellaria baicalensis dregs with the water content of 50%;
s2, crushing the scutellaria baicalensis residue with the water content of 50% obtained in the step S1 to be less than 20mm, and carrying out pressure filtration on the crushed scutellaria baicalensis residue to obtain scutellaria baicalensis residue powder with the water content of 45%;
s3, adding 0.4 wt% of liquid lactobacillus plantarum and 0.4 wt% of a mixture of a plurality of complex enzymes into the scutellaria baicalensis residue powder with the water content of 45% obtained in the step S2, and fully and uniformly mixing;
and S4, fermenting the mixture finally obtained in the step S3 for 7 days under vacuum condition at 37 ℃, obtaining a scutellaria baicalensis dreg fermented product after fermentation is finished, and carrying out vacuum packaging.
The inventor of the scutellaria baicalensis residue semisolid fermentation technical method conducts a series of experimental researches on the fermentation technical method and the application thereof, and determines that the scutellaria baicalensis residue semisolid fermentation technical method has obvious outstanding curative effect advantages compared with the prior art.
Experiment I, measuring content of flavonoid component in semisolid fermentation product of scutellaria baicalensis dreg
1. Material
1.1, sample: the fermented products of scutellaria baicalensis pomace prepared in the above-mentioned example 1, example 2 and example 3.
1.2, control:
(1) and radix Scutellariae residue;
(2) and fermenting the mixture finally obtained in the step of S3 in example 1 at 37 ℃ for 7 days under vacuum condition, obtaining scutellaria baicalensis dreg fermented product after fermentation is finished, and carrying out vacuum packaging to obtain the control fermented product.
2. Test method
2.1, a measuring method: high performance liquid chromatography (general rule 0512) determination
2.1.1 chromatographic conditions
A chromatographic column: octadecylsilane chemically bonded silica is used as a filling agent; gradient elution was carried out using methanol-0.1% phosphoric acid as mobile phase according to the specifications of table 1 below; the detection wavelength was 275 nm. The number of theoretical plates is not less than 2500 calculated by baicalin peak.
TABLE 1
2.2 solution preparation
Preparation of control solutions: taking appropriate amount of baicalin, wogonoside, baicalein and wogonin, adding methanol to obtain 100 μ g/ml solution.
Preparation of a test solution:
precisely weighing 2g of Scutellariae radix residue powder, precisely adding 25ml of 60% methanol, performing ultrasonic treatment for 10min, and collecting filtrate;
2g of each of the scutellaria baicalensis residue fermentations prepared in the embodiment 1, the embodiment 2 and the embodiment 3 are taken, precisely weighed and precisely weighed, the mixture is placed in a conical flask with a plug, 25ml of 60% methanol is precisely added, the plug is sealed, the weight is weighed, ultrasonic treatment is carried out for 10 minutes, the mixture is cooled and then weighed, the weight loss is compensated by 60% methanol, the mixture is shaken up and filtered, and the subsequent filtrate is taken, so that the scutellaria baicalensis residue fermentation liquid is obtained.
2.3 detection method
Precisely measuring the reference substance and the sample solution, respectively, injecting into a high performance liquid chromatograph, detecting, and obtaining detection results by external standard method with reference to the reference substance solution, wherein the results are shown in Table 2.
TABLE 2 flavonoid content
3. Conclusion
As can be seen from table 2 above, examples 1,2 and 3 have better effects, the content of flavonoid components is increased by 2 times, especially the content of baicalein is greatly increased, and baicalein has the effects of reducing cerebrovascular resistance, improving cerebral blood circulation, increasing cerebral blood flow and resisting platelet aggregation. The traditional Chinese medicine composition is clinically used for treating paralysis caused by cerebrovascular diseases. After entering animal body, baicalein is converted into baicalin and other metabolites in blood. The control fermented product sample fermented for 10 days was found to have a lower total flavonoid content than the sample fermented for 7 days in example 1, compared to example 1, and thus it was considered that the fermentation time was 7 days from the viewpoint of cost saving.
Experiment II, bacteriostasis experiment of scutellaria baicalensis dreg semisolid fermentation product in invention
1 sample of
1.1 strains
Escherichia coli O78 standard strain (e.coli O78, accession number CVCC 1418); staphylococcus aureus strain ATCC 29213.
1.2 samples and reagents
Sample preparation: scutellaria residue fermentation products and scutellaria residue samples prepared in example 1, example 2 and example 3.
Reagent: the nutrient broth culture medium, the MacconKa culture medium, the common nutrient agar culture medium, the ethanol and other reagents are analytically pure.
1.3 instruments
A circulating vacuum water pump (SHZ-3 type), a vacuum drying oven (ZK-82B type), an electronic temperature control electric heater, an autoclave, an electronic analytical balance (FA1004 type), a reflux condenser, a distiller, etc.
2. Method of producing a composite material
2.1 preparation of bacterial liquid
Standard colibacillus and staphylococcus aureus are streaked on a solid culture medium and cultured for 18-24 h at 37 ℃. Individual colonies were picked and inoculated into nutrient broth and cultured at 37 ℃ for 18 h. Measuring the concentration of bacterial liquid by plate dilution method, adjusting the bacterial concentration to 1.5 × 108CFU/ml for use.
2.2 preparation of test specimens
Decocting Scutellariae radix residue fermented product and Scutellariae radix residue sample prepared in examples 1,2 and 3 with water, extracting with 10 times of water for 1 hr, concentrating to obtain solution with drug content of 1g/ml, placing into small bottle, autoclaving at 120 deg.C for 20min, and placing in refrigerator at 4 deg.C for use.
2.3 determination of MIC value of minimum inhibitory concentration
Four groups of experimental groups (including I, II, III and IV groups) are arranged, and 12 small test tubes are taken from each experimental group and numbered; adding 1ml of common nutrient broth culture medium into each test tube by using a sterile pipette; adding 1ml of test medicine into the first tube under the aseptic condition, shaking and uniformly mixing; sucking 1ml of test drug, adding the test drug into the second tube, shaking and mixing uniformly, repeating the process until the 11 th tube is reached, sucking 1ml and discarding (ensuring the volume of the solution in 12 test tubes to be the same), and not adding the test drug into the 12 th tube as a positive control. Adding 1ml of the prepared bacterial liquid into each tube, placing the tube in an incubator at 37 ℃, and observing the test result after culturing for 18h, wherein the front tube of the growth tube is the MIC of the sample. The test is repeated for 3 times, and the average value is taken as the MIC value of each sample;
wherein, the test drug prepared from the scutellaria residue fermented product of example 1 was used in group i, the test drug prepared from the scutellaria residue fermented product of example 2 was used in group ii, the test drug prepared from the scutellaria residue fermented product of example 3 was used in group iii, and the test drug prepared from the scutellaria residue sample was used in group iv.
3. Results
The in vitro bacteriostatic effect of experimental groups i, ii, iii, iv on escherichia coli and staphylococcus aureus is shown in table 3:
TABLE 3 in vitro bacteriostatic effects
As can be seen from Table 3, groups I, II, III, and IV all have certain bacteriostatic effects on gram-negative bacteria such as Escherichia coli and gram-positive bacteria such as Staphylococcus aureus. Wherein, the bacteriostatic effect of the I is best, and the lowest bacteriostatic concentration can reach 156 mg/ml; secondly, the group III has better bacteriostatic effect, and the bacteriostatic concentration is 225 mg/ml; the groups II and IV have the worst bacteriostatic effect, and the bacteriostatic concentration of the groups II and IV exceeds 1000 mg/ml.
Experiment III, evaluation experiment of acute toxicity of semisolid fermentation product of scutellaria baicalensis dreg
Decocting the powder of the residue of the scutellaria baicalensis and the powder of the examples 1,2 and 3 with water, adding 10 times of water for 1 hour, and concentrating to obtain a solution with the drug content of 200 mg/ml. Each mouse is subjected to gavage administration for 1 time according to the weight of 5g/kg b.wt. the gavage volume is 0.2ml/10g, the mice are continuously observed for 4 hours after administration, and then are continuously observed for 7 days, and the general states of mental state, hair color, autonomic activity, respiration, diet, defecation, oral and nasal secretion and the like of each animal are not abnormal in the observation period, no animal dies, and the weight of the animals naturally increases. After the experiment is finished, the mice are dislocated and killed, and no pathological changes can be seen by naked eyes in main organs, and no obvious poisoning expression or adverse reaction exists. Indicating that the fermentation product has LD for Kunming mouse50And the content is more than 5g/kg, evaluation is carried out according to the WTO related exogenous chemical acute toxicity grading standard, and test results show that the fermentation product is actually nontoxic.
The technical features of the present invention which are not described in the above embodiments may be implemented by or using the prior art, and are not described herein again, of course, the above description is not intended to limit the present invention, and the present invention is not limited to the above examples, and variations, modifications, additions or substitutions which may be made by those skilled in the art within the spirit and scope of the present invention should also fall within the protection scope of the present invention.
Claims (4)
1. A technical method for semisolid fermentation of scutellaria baicalensis dreg is characterized by comprising the following steps:
s1, taking the scutellaria baicalensis dregs, and extruding the scutellaria baicalensis dregs to obtain the scutellaria baicalensis dregs with the water content of 30-70%;
s2, crushing the scutellaria baicalensis dregs with the water content of 30-70% obtained in the step S1 to be less than 20mm, and carrying out filter pressing on the crushed scutellaria baicalensis dregs to obtain scutellaria baicalensis dregs powder with the water content of 30-60%;
s3, adding 0.3-0.5 wt% of liquid lactobacillus plantarum and 0.3-0.5 wt% of a mixture of a plurality of complex enzymes into the scutellaria baicalensis dreg powder with the water content of 30-60% obtained in the step S2, and fully and uniformly mixing;
and S4, fermenting the mixture finally obtained in the step S3 for 7 days under vacuum condition at 37 ℃, obtaining a scutellaria baicalensis dreg fermented product after fermentation is finished, and carrying out vacuum packaging.
2. The scutellaria baicalensis residue semisolid fermentation technical method according to claim 1, which comprises the following steps:
s1, taking 1kg of scutellaria baicalensis dregs, and extruding the scutellaria baicalensis dregs to obtain scutellaria baicalensis dregs with the water content of 35%;
s2, crushing the scutellaria baicalensis residue with the water content of 35% obtained in the step S1 to be less than 20mm, and performing pressure filtration on the crushed scutellaria baicalensis residue to obtain scutellaria baicalensis residue powder with the water content of 30%;
s3, adding 0.3 weight percent of liquid lactobacillus plantarum and 0.3 weight percent of a mixture of various complex enzymes into the scutellaria baicalensis residue powder with the water content of 30 percent obtained in the step S2, and fully and uniformly mixing;
and S4, fermenting the mixture finally obtained in the step S3 for 7 days under vacuum condition at 37 ℃, obtaining a scutellaria baicalensis dreg fermented product after fermentation is finished, and carrying out vacuum packaging.
3. The scutellaria baicalensis residue semisolid fermentation technical method according to claim 2, wherein the multiple complex enzyme mixture in the step S2 is formed by mixing cellulase, pectinase and xylanase, and the ratio of the cellulase, the pectinase and the xylanase is as follows: 2:1:1.
4. The scutellaria baicalensis residue semisolid fermentation technical method according to claim 1, which is characterized in that the scutellaria baicalensis residue fermentation product is applied to antibiosis and bacteriostasis.
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| CN111024853B (en) * | 2019-12-29 | 2022-07-29 | 山西中医药大学 | Fermentation method for quickly converting baicalin into baicalein |
| CN113215024B (en) * | 2021-03-08 | 2023-07-04 | 黑龙江省科学院微生物研究所 | Lactobacillus plantarum and method for fermenting baical skullcap root dregs |
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| 冷桂华: "黄芩及其提取药渣黄芩苷含量的比较", 《安徽农业科学》 * |
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| CN112931690A (en) * | 2021-03-08 | 2021-06-11 | 黑龙江省科学院微生物研究所 | Piglet feed additive and preparation method thereof |
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