Preparation method and application of low-coumarin cinnamon extract

Abstract

The invention belongs to the technical field of low-coumarin cinnamon extraction, and particularly relates to a preparation method and application of a low-coumarin cinnamon extract. The method comprises the following steps: (1) coarsely crushing cortex cinnamomi, adding the crushed cortex cinnamomi into water at 60-90 DEG C for extraction, and filtering and collecting a first feed liquid; (2) continuing to add water at 60-90 DEG C into the filter residue for extraction, and filtering and collecting a second feed liquid; (3) combining the two filtrates, concentrating, adding ethyl acetate, stirring, standing, collecting an ethyl acetate layer, adding water, and concentrating until no solvent smell exists; (4) cooling the concentrated solution to room temperature, adding activated carbon, stirring, and filtering to remove the activated carbon; and (5) concentrating the filtrate, and performing spray drying to obtain the low-coumarin cinnamon extract. The low-coumarin cinnamon extract can be applied to health care products with an intestinal mucosa protection effect or medicines for treating intestinal mucosa injury, and can meet large-scale industrial production required by the market.

Description

Technical Field

[0001] The invention belongs to the technical field of low-coumarin cinnamon extraction, and particularly relates to a preparation method and application of a low-coumarin cinnamon extract. Background Art

[0002] The intestine is the main organ for absorbing nutrients and water, and its mucosal barrier is also the greatest guarantee to prevent the displacement of bacteria and endotoxins in the intestinal cavity, so that the internal environment of the body remains relatively stable and maintains the normal life activities of the body. As an important filter for regulating the balance of the intestinal cavity and the body, the intestinal mucosa is composed of mechanical barriers such as intestinal epithelial cells and tight junctions between cells, chemical barriers such as the mucus layer covering the surface of epithelial cells, digestive juices and antimicrobial peptides, biological barriers composed of intestinal microorganisms, and immune barriers composed of immune cells and antibodies. Therefore, the integrity of the intestinal mucosal barrier is crucial to maintaining intestinal homeostasis. When the intestinal mucosal barrier is damaged, some inflammatory factors change and can pass through the intestine to affect other organs and develop into chronic diseases.

[0003] The structural basis of the intestinal mucosal barrier is intact intestinal epithelial cells (IEC) and tight junctions (TJ) between adjacent intestinal epithelia. The intestinal mucosal barrier is a barrier system with efficient and selective functions existing in the intestine. Therefore, an intact intestinal barrier is crucial for maintaining health and preventing a variety of diseases such as tissue damage. Clinically, the treatment drugs for alcoholic intestinal mucosal damage include rabeprazole, omeprazole enteric-coated capsules, etc., but the drugs are expensive and have many side effects when treating any disease. Therefore, the potential of plant-derived health products and foods in promoting health and preventing diseases has attracted more and more attention in recent years.

[0004] Coumarin can mainly cause long-term coma or death in animals. Common symptoms include liver degeneration and necrosis, vascular changes and cavernous hemangiomas. Coumarin was listed as a Class III carcinogen by the World Health Organization in 2000. Studies have shown that coumarin-mediated hepatotoxicity in rats is caused by factors such as increased liver mass, lipid metabolism and carbohydrate metabolism disorders, inhibition of monooxygenase system enzyme activity and cytochrome P450 concentration in liver tissue, and enhanced mitochondrial phosphorylation in hepatocytes. Except for inflammation of the gastric and small intestinal mucosa, no significant morphological and functional changes were found in other internal organs. The maximum daily intake of coumarin by humans from the drink is 0.02 mg kg -1 Many countries, regions and relevant agencies have set regulations on the coumarin content in food, see Table 1.

[0005] Table 1 Regulations on coumarin content in food by many countries, regions and relevant agencies

[0006]

[0007] Patent CN201780083157.7 discloses a method for preparing a composition for preventing or treating gastritis or peptic ulcers, which uses cinnamon for pretreatment, then extracts, concentrates and dries, and then adds the composition. Experiments on rats have shown that it has the functions of preventing, improving or treating gastritis or gastric ulcers. Patent CN201710129459.5 discloses a method for preparing a low-coumarin cinnamon extract, which uses cinnamon as a raw material and adopts supercritical CO 2 The extraction technology removes coumarin to obtain a low-coumarin cinnamon raw material, and the polyphenol content of the extract obtained after preparation is 20% to 40%. In the relevant literature and patents, how to use cinnamon to prepare cinnamon extracts with the function of protecting intestinal mucosa, how to remove and remove potentially harmful coumarins, and how to increase the content of effective substances, etc., have not been specifically solved. Therefore, it is necessary to develop a method for preparing a low-coumarin cinnamon extract that can be used as a drug for treating intestinal mucosal damage. Summary of the invention

[0008] The purpose of the present invention is to provide a preparation method of a low-coumarin cinnamon extract and its application. The present invention extracts cinnamon at a suitable temperature, removes coumarin by activated carbon, obtains a low-coumarin cinnamon extract, and discloses its application in a health product with intestinal mucosal protection effect or a drug for treating intestinal mucosal damage.

[0009] The implementation process of the present invention is as follows:

[0010] A method for preparing a low-coumarin cinnamon extract comprises the following steps:

[0011] (1) Crush cinnamon into powder and add it to 60-90° C. water for extraction, and collect the first feed liquid by filtration;

[0012] (2) adding 60-90°C water to the filter residue for extraction, and filtering and collecting the second feed liquid;

[0013] (3) The two filtrates were combined and concentrated, ethyl acetate was added, stirred and allowed to stand, the ethyl acetate layer was collected, and water was added to concentrate until there was no solvent smell;

[0014] (4) The concentrated solution is cooled to room temperature, activated carbon is added, and the activated carbon is removed by filtering after stirring;

[0015] (5) The filtrate is concentrated and spray-dried to obtain a low-coumarin cinnamon extract.

[0016] Furthermore, in step (1), the extraction time is 1 to 3 hours; in step (2), the extraction time is 1 to 2 hours.

[0017] Furthermore, in step (3), the volume of the filtrate is concentrated to 1.8 to 2 times the amount of cinnamon.

[0018] Furthermore, in step (3), the volume of ethyl acetate is equal to the volume of the filtrate after concentration.

[0019] Furthermore, in step (3), the stirring time is 1 to 10 minutes.

[0020] Furthermore, in step (3), the standing time is 10 to 50 minutes.

[0021] Furthermore, in step (4), the mass of the activated carbon is 0.4 to 0.6% of the mass of cinnamon.

[0022] Furthermore, in step (4), the mesh size of the activated carbon is 100 to 140 meshes.

[0023] Furthermore, in step (4), the stirring time is 10 to 40 minutes.

[0024] The low-coumarin cinnamon extract obtained by the method is used in health products with intestinal mucosa protection effect or drugs for treating intestinal mucosa damage.

[0025] The concept and verification results of the present invention:

[0026] The method of the present invention extracts cinnamic acid from cinnamon by adding water at a suitable temperature, reduces the dissolution of other components, and then adds coconut shell activated carbon for treatment to adsorb oil impurities and minimize harmful substances. Among them, the suitable temperature can effectively extract cinnamic acid while extracting coumarin, cinnamaldehyde and 2-methoxycinnamaldehyde to a minimum. Coconut shell activated carbon will adsorb non-polar impurities, and the cinnamic acid we need is a medium-polar substance, and the activated carbon has a weak adsorption capacity for it. At the same time, coumarin is a bicyclic compound, which is adsorbed in the mesopores (2 to 50 nm) of the activated carbon, and generates van der Waals forces on the surface of the activated carbon cavities, thereby forming physical adsorption.

[0027] The present invention provides an ethanol-induced enteritis model to 5dpf wild-type AB strain zebrafish, adds cinnamon extracts of different concentrations for treatment, collects intestinal images under a dissecting microscope, calculates the number of goblet cells in the intestinal cavity by using image processing software, and evaluates the regulating effect of the cinnamon extract on intestinal mucus secretion according to the number of goblet cells to verify the application of the low-coumarin cinnamon extract in a health product with intestinal mucosa protection efficacy or a drug for treating intestinal mucosa injury.

[0028] Positive effects of the present invention:

[0029] (1) The method of the present invention uses low-cost cinnamon as a raw material to develop a low-coumarin cinnamon extract with intestinal mucosal protective function, which can meet the large-scale industrial production required by the market.

[0030] (2) The method of the present invention controls the temperature during the extraction step, which can improve the dissolution of effective substances and reduce the difficulty of purification.

[0031] (3) The present invention obtains cinnamon extract by one-step extraction and filtering with activated carbon, thereby obtaining not only cinnamon extract with low coumarin content, but also has the function of protecting the intestinal mucosa from alcoholic damage as proved by zebrafish experiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 HPLC spectrum of low-coumarin cinnamon extract prepared in Example 1;

[0033] Figure 2 This is a phenotype diagram of the regulatory effect of low-coumarin cinnamon extract on zebrafish intestinal mucus secretion, where the dotted area is the goblet cells in the intestine, a is the normal control group, b is the model control group, c is RU-21 150μg / mL, d is low-coumarin cinnamon extract 125μg / mL, and e is low-coumarin cinnamon extract 250μg / mL;

[0034] Figure 3 The regulatory effect of low-coumarin cinnamon extract on the number of goblet cells; compared with the model control group, *p<0.05, ***p<0.001;

[0035] Figure 4 The results show that low-coumarin cinnamon extract regulates intestinal mucus secretion; compared with the model control group, *p<0.05, ***p<0.001. DETAILED DESCRIPTION

[0036] The present invention will be further described below in conjunction with the embodiments.

[0037] The present invention adopts a suitable temperature in the preparation process to maximize the dissolution of effective substances, and adds activated carbon treatment in the preparation process to adsorb non-polar substances, remove coumarin, and slightly increase the effective active content. The low-coumarin cinnamon extract obtained has the characteristics of being non-toxic and edible for a long time. The present invention uses ethanol to establish a zebrafish intestinal mucosal injury model, which has no direct correlation with the improvement of gastritis or gastric ulcer in the prior art, and is a different disease type from the rat model.

[0038] The method for preparing the low-coumarin cinnamon extract of the present invention comprises the following steps:

[0039] (1) Crush cinnamon into powder and add it to 60-90° C. water for extraction for 1-3 h, and collect the first feed liquid by filtration;

[0040] (2) Add 60-90°C water to the filter residue and extract for 1-2 hours, then filter and collect the second feed liquid;

[0041] (3) combining the two filtrates and concentrating the filtrate to a volume of 1.8 to 2 times the volume of cinnamon, adding ethyl acetate in an amount equal to the volume of the concentrated filtrate, stirring for 1 to 10 minutes and standing for 10 to 50 minutes, collecting the ethyl acetate layer, and adding water to concentrate until there is no solvent smell;

[0042] (4) the concentrated solution is cooled to room temperature, activated carbon of 0.4-0.6% of the mass of cinnamon is added, the mesh size of the activated carbon is 100-140 mesh, and the activated carbon is removed by filtering after stirring for 10-40 minutes;

[0043] (5) The filtrate is concentrated and spray-dried to obtain a low-coumarin cinnamon extract.

[0044] In step (1), step (2) and step (3), the amount of water is not limited, and a person skilled in the art can add water according to specific experimental needs, as long as the amount of water can cover the cinnamon solid.

[0045] In step (3), the stirring time and the standing time are not limited. Those skilled in the art can set the time according to the specific needs of the experiment. The preferred stirring time of the present invention is 1 to 10 minutes; the standing time is 10 to 50 minutes.

[0046] The stirring time in step (4) is not limited, and those skilled in the art can set the time according to the specific needs of the experiment. The preferred stirring time of the present invention is 10 to 40 minutes.

[0047] The parameters of spray drying in step (5) are not limited. Those skilled in the art can set the parameters according to the specific needs of the experiment. In the present invention, the inlet temperature is preferably 200-210° C. and the outlet temperature is preferably 90-95° C.

[0048] Example 1

[0049] The method for preparing the low-coumarin cinnamon extract of the present invention comprises the following steps:

[0050] (1) Cinnamon is cut into 1-2 cm thick pieces, then added into 70° C. water for extraction for 2 h, and the first feed liquid is collected by filtration;

[0051] (2) Add 70°C water to the residue and extract for 1.5 hours, then filter and collect the second feed liquid;

[0052] (3) The two filtrates were combined and concentrated to a volume of 1.8 times the volume of cinnamon, and ethyl acetate was added in an amount equal to the volume of the concentrated filtrate. The mixture was stirred for 5 min and allowed to stand for 30 min, after which the ethyl acetate layer was collected and concentrated with water until there was no solvent odor.

[0053] (4) the concentrated solution was cooled to room temperature, activated carbon (0.4% by weight of cinnamon) was added, the mesh size of the activated carbon was 120 mesh, and the solution was stirred for 20 minutes and then filtered to remove the activated carbon;

[0054] (5) The filtrate is concentrated and spray-dried (the inlet temperature of the spray-dryer is 200° C. and the outlet temperature is 90° C.) to obtain a low-coumarin cinnamon extract in the form of a brown powder.

[0055] Example 2

[0056] The method for preparing the low-coumarin cinnamon extract of the present invention comprises the following steps:

[0057] (1) Cinnamon is cut into 1-2 cm thick pieces, then added into 60° C. water for extraction for 1 h, and the first feed liquid is collected by filtration;

[0058] (2) Add 60°C water to the residue and extract for 2 h, then filter and collect the second feed liquid;

[0059] (3) The two filtrates were combined and concentrated to a volume of twice that of the cinnamon, and ethyl acetate was added in an amount equal to the volume of the concentrated filtrate. The mixture was stirred for 10 min and allowed to stand for 50 min, after which the ethyl acetate layer was collected and concentrated with water until there was no solvent odor.

[0060] (4) the concentrated solution was cooled to room temperature, activated carbon (0.6% of the mass of cinnamon) was added, the mesh size of the activated carbon was 100 mesh, and the activated carbon was removed by filtering after stirring for 10 minutes;

[0061] (5) The filtrate is concentrated and spray-dried (the inlet temperature of the spray-dryer is 210° C. and the outlet temperature is 95° C.) to obtain a low-coumarin cinnamon extract in the form of a brown powder.

[0062] Example 3

[0063] The method for preparing the low-coumarin cinnamon extract of the present invention comprises the following steps:

[0064] (1) Cinnamon is cut into 1-2 cm thick pieces, then added into 90° C. water for extraction for 3 h, and the first feed liquid is collected by filtration;

[0065] (2) Add 90°C water to the residue and extract for 1 hour, then filter and collect the second feed liquid;

[0066] (3) The two filtrates were combined and concentrated to a volume of 1.8 times the amount of cinnamon, and ethyl acetate was added in an amount equal to the volume of the concentrated filtrate. The mixture was stirred for 1 min and allowed to stand for 10 min, after which the ethyl acetate layer was collected and concentrated with water until there was no solvent odor.

[0067] (4) the concentrated solution is cooled to room temperature, activated carbon of 0.5% of the mass of cinnamon is added, the mesh size of the activated carbon is 140 mesh, and the activated carbon is removed by filtering after stirring for 40 minutes;

[0068] (5) The filtrate is concentrated and spray-dried (the inlet temperature of the spray-dryer is 210° C. and the outlet temperature is 95° C.) to obtain a low-coumarin cinnamon extract in the form of a brown powder.

[0069] Example 4

[0070] The method for preparing the low-coumarin cinnamon extract of the present invention comprises the following steps:

[0071] (1) Cinnamon is cut into 1-2 cm thick pieces, then added into 85°C water for extraction for 1 h, and the first feed liquid is collected by filtration;

[0072] (2) Add 85°C water to the residue and extract for 2 h, then filter and collect the second feed liquid;

[0073] (3) The two filtrates were combined and concentrated to a volume of 1.8 times the amount of cinnamon, and ethyl acetate was added in an amount equal to the volume of the concentrated filtrate. The mixture was stirred for 5 min and allowed to stand for 40 min, and the ethyl acetate layer was collected. Water was added and concentrated until there was no solvent smell.

[0074] (4) the concentrated solution was cooled to room temperature, activated carbon (0.5% of the mass of cinnamon) was added, the mesh size of the activated carbon was 130 mesh, and the activated carbon was removed by filtering after stirring for 30 minutes;

[0075] (5) The filtrate is concentrated and spray-dried (the inlet temperature of the spray-dryer is 200° C. and the outlet temperature is 90° C.) to obtain a low-coumarin cinnamon extract in the form of a brown powder.

[0076] Comparative Example 1: Adjust the solvent to 95% ethanol

[0077] (1) Cinnamon is cut into 1-2 cm thick, then added to 70° C. 95% ethanol for extraction for 2 h, and the first feed liquid is collected by filtration;

[0078] (2) Add 70°C 95% ethanol to the residue and extract for 1.5 h, then filter and collect the second feed liquid;

[0079] (3) The two filtrates were combined and concentrated to a volume of 1.8 times the amount of cinnamon, and ethyl acetate was added in an amount equal to the volume of the concentrated filtrate. The mixture was stirred for 5 min and allowed to stand for 40 min, and the ethyl acetate layer was collected. Water was added and concentrated until there was no solvent smell.

[0080] (4) the concentrated solution was cooled to room temperature, activated carbon (0.5% of the mass of cinnamon) was added, the mesh size of the activated carbon was 130 mesh, and the activated carbon was removed by filtering after stirring for 30 minutes;

[0081] (5) The filtrate is concentrated and spray-dried (the inlet temperature of the spray-dryer is 200° C. and the outlet temperature is 90° C.) to obtain a cinnamon extract in the form of a brown powder.

[0082] Comparative Example 2: Adjust the temperature to 100°C + activated carbon

[0083] The preparation method of the cinnamon extract of this comparative example comprises the following steps:

[0084] (1) Cinnamon is cut into pieces with a thickness of 1-2 cm, then added into 100° C. water for extraction for 2 h, and the first feed liquid is collected by filtration;

[0085] (2) Add 100°C water to the filter residue and extract for 1.5 hours, and filter and collect the second feed liquid;

[0086] (3) The two filtrates were combined and concentrated to a volume of 1.8 times the volume of cinnamon, and ethyl acetate was added in an amount equal to the volume of the concentrated filtrate. The mixture was stirred for 5 min and allowed to stand for 30 min, after which the ethyl acetate layer was collected and concentrated with water until there was no solvent odor.

[0087] (4) the concentrated solution was cooled to room temperature, activated carbon (0.4% by weight of cinnamon) was added, the mesh size of the activated carbon was 120 mesh, and the solution was stirred for 20 minutes and then filtered to remove the activated carbon;

[0088] (5) The filtrate is concentrated and spray-dried (the inlet temperature of the spray-dryer is 200° C. and the outlet temperature is 90° C.) to obtain a cinnamon extract in the form of a brown powder.

[0089] Comparative Example 3: Adjust the temperature to 50°C + activated carbon

[0090] The preparation method of the cinnamon extract of this comparative example comprises the following steps:

[0091] (1) Cinnamon is cut into 1-2 cm thick pieces, then added into 50° C. water for extraction for 2 h, and the first feed liquid is collected by filtration;

[0092] (2) Add 50°C water to the residue and extract for 1.5 hours, then filter and collect the second feed liquid;

[0093] (3) The two filtrates were combined and concentrated to a volume of 1.8 times the volume of cinnamon, and ethyl acetate was added in an amount equal to the volume of the concentrated filtrate. The mixture was stirred for 5 min and allowed to stand for 30 min, after which the ethyl acetate layer was collected and concentrated with water until there was no solvent odor.

[0094] (4) the concentrated solution was cooled to room temperature, activated carbon (0.4% by weight of cinnamon) was added, the mesh size of the activated carbon was 120 mesh, and the solution was stirred for 20 minutes and then filtered to remove the activated carbon;

[0095] (5) The filtrate is concentrated and spray-dried (the inlet temperature of the spray-dryer is 200° C. and the outlet temperature is 90° C.) to obtain a cinnamon extract in the form of a brown powder.

[0096] Comparative Example 4: No activated carbon added

[0097] The preparation method of the cinnamon extract of this comparative example comprises the following steps:

[0098] (1) Cinnamon is cut into 1-2 cm thick pieces, then added into 50° C. water for extraction for 2 h, and the first feed liquid is collected by filtration;

[0099] (2) Add 50°C water to the residue and extract for 1.5 hours, then filter and collect the second feed liquid;

[0100] (3) The two filtrates were combined and concentrated to a volume of 1.8 times the volume of cinnamon, and ethyl acetate was added in an amount equal to the volume of the concentrated filtrate. The mixture was stirred for 5 min and allowed to stand for 30 min, after which the ethyl acetate layer was collected and concentrated with water until there was no solvent odor.

[0101] (4) The filtrate is concentrated and spray-dried (the inlet temperature of the spray-dryer is 200° C. and the outlet temperature is 90° C.) to obtain a cinnamon extract in the form of a brown powder.

[0102] Performance Test:

[0103] (I) Using HPLC technology to detect the cinnamic acid and coumarin content of Example 1 and Comparative Examples 1-4

[0104] (I) Detection Method

[0105] Mobile phase: A: acetonitrile (0.2% trifluoroacetic acid), B: purified water, elution program is shown in Table 1 below;

[0106] Chromatographic column: Kromasil 100-5C18 (4.6 mm × 150 mm, 5.0 μm)

[0107] Column temperature: 35 °C;

[0108] Wavelength: 280nm;

[0109] Flow rate: 0.8 mL / min;

[0110] Elution procedure:

[0111] Table 2 Parameter settings for elution program

[0112]

[0113] (II) Test results:

[0114] As shown in Table 3 and Figure 1 As shown, the suitable extraction temperature of the raw material cinnamon in the method of the present invention, the selection of water as a solvent and the treatment with activated carbon can significantly increase the cinnamic acid content and promote the removal of coumarin and cinnamaldehyde.

[0115] Table 3 Analysis of the content of each substance (mass percentage %) in Example 1 and Comparative Examples 1-4

[0116]

[0117]

[0118] (II) Test of Maximum Detectable Concentration (MTC) of Low-Coumarin Cinnamon Extract Prepared in Example 1

[0119] (1) Experimental subjects

[0120] Wild-type AB strain zebrafish were bred by natural pair mating. The age was 5 days after fertilization (5 dpf).

[0121] (2) Experimental groups

[0122] Experimental group 1: Normal control group (zebrafish treated with fish farming water)

[0123] Experimental 2: Model control group (1.2% ethanol)

[0124] Experimental group 3 Cinnamon extract group 1 (1.2% ethanol + low-coumarin cinnamon extract 62.5 μg / mL)

[0125] Experimental 4 groups Cinnamon extract 2 groups (1.2% ethanol + low-coumarin cinnamon extract 125 μg / mL)

[0126] Experimental 5 groups Cinnamon extract 3 groups (1.2% ethanol + low coumarin cinnamon extract 250 μg / mL)

[0127] Experimental 6 groups 4 groups of cinnamon extract (1.2% ethanol + low-coumarin cinnamon extract 500μg / mL)

[0128] Experimental 7 groups Cinnamon extract 5 groups (1.2% ethanol + low coumarin cinnamon extract 1000 μg / mL)

[0129] (3) Experimental methods

[0130] 5dpf wild-type AB strain zebrafish were randomly selected in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). The concentrations of water-soluble low-coumarin cinnamon extract were 62.5μg / mL, 125μg / mL, 250μg / mL, 500μg / mL, and 1000μg / mL, respectively. A normal control group and a model control group were set up at the same time, and the volume of each well was 3mL. Except for the normal control group, the other experimental groups were given ethanol to establish an intestinal injury model. After 20h of treatment, the MTC of cinnamon extract on model zebrafish was determined.

[0131] The process of establishing the intestinal injury model of zebrafish was to fast for 24 hours before the experiment, during which time the fish were allowed to drink water freely. 0.6 ml of 1.2% ethanol was orally infused per fish to establish the alcohol burn intestinal injury model.

[0132] (4) Experimental results

[0133] Under the experimental conditions, the MTC of the protective effect of low-coumarin cinnamon extract on intestinal mucosa was 250 μg / mL (Table 4).

[0134] Table 4 Experimental results (n=30)

[0135]

[0136]

[0137] (III) Evaluation of the intestinal protective efficacy of the low-coumarin cinnamon extract prepared in Example 1

[0138] (1) Experimental subjects

[0139] Wild-type AB strain zebrafish were bred by natural pair mating. The age was 5 days after fertilization (5 dpf).

[0140] (2) Experimental groups

[0141] Experimental group 1: Normal control group (zebrafish treated with fish farming water)

[0142] Experimental 2: Model control group (1.2% ethanol)

[0143] Experimental group 3 RU-21 group (1.2% ethanol + RU-21 150 μg / mL)

[0144] Experimental group 4 Cinnamon extract group 1 (1.2% ethanol + low-coumarin cinnamon extract 125 μg / mL)

[0145] Experimental 5 groups Cinnamon extract 2 groups (1.2% ethanol + low-coumarin cinnamon extract 250 μg / mL)

[0146] (3) Experimental methods

[0147] Wild-type AB zebrafish of 5 days post-fertilization (5dpf) with healthy development and consistent stages were selected under a microscope and placed in a six-well plate with a capacity of 3mL per well. The zebrafish enteritis model was induced with ethanol. Different doses of low-coumarin cinnamon extract were given in water, and the positive control drug RU-21 was 150μg / mL. A normal control group and a model control group were set up at the same time. 30 zebrafish were treated in each experimental group. After the treatment, 10 zebrafish were randomly selected from each group and stained with Alcian blue. Intestinal images were collected under a dissecting microscope, and the number of goblet cells in the intestinal cavity (N) was calculated using image processing software. The regulatory effect of low-coumarin cinnamon extract on intestinal mucus secretion was evaluated based on the number of goblet cells. The calculation formula for the regulatory effect of low-coumarin cinnamon extract on goblet cells is as follows:

[0148]

[0149] Statistical analysis was performed using analysis of variance and Dunnett's T-test, and p < 0.05 indicated significant differences.

[0150] The process of establishing the intestinal injury model of zebrafish was to fast for 24 hours before the experiment, during which time the fish were allowed to drink water freely. 0.6 ml of 1.2% ethanol was orally infused per fish to establish the alcohol burn intestinal injury model.

[0151] (4) Experimental results

[0152] from Figure 2-Figure 4 The results in Table 5 show that the number of goblet cells in the zebrafish intestine in the model control group (57) was compared with the normal control group (91), p<0.001, indicating that the model was successfully established; the number of goblet cells in the positive control drug RU-21 150μg / mL concentration group was 72, p<0.001 compared with the model control group, and the regulatory effect on intestinal secretion was 29%, indicating that RU-21 has a significant regulatory effect on intestinal mucus secretion. The number of goblet cells in the zebrafish intestine in the low-coumarin cinnamon extract 125μg / mL and 250μg / mL concentration groups were 92 and 98, respectively, and each group was compared with the model control group. P<0.001, and the regulatory effect on intestinal secretion was 89% and 116%, respectively, indicating that the low-coumarin cinnamon extract has a significant regulatory effect on intestinal mucus secretion at this experimental concentration.

[0153] Table 5 Regulatory effect of low-coumarin cinnamon extract on intestinal mucus secretion in zebrafish (n=10)

[0154]

[0155] Note: Compared with the model control group, *p<0.05, ***p<0.001.

[0156] The above contents are further detailed descriptions of the present invention in combination with specific preferred embodiments, and the specific implementation of the present invention cannot be limited to these descriptions. For ordinary technicians in the field to which the present invention belongs, several simple deductions or substitutions can be made without departing from the concept of the present invention, which should be regarded as falling within the protection scope of the present invention.

Claims

1. A method for preparing a low-coumarin cinnamon extract, characterized in that: The steps include: (1) Crush cinnamon into small pieces and add it to 60-90°C water for extraction, and collect the first liquid by filtration; (2) Continue to add 60-90°C water to the filter residue for extraction, and filter and collect the second feed liquid; (3) Combine the two filtrates and concentrate, add ethyl acetate, stir and let stand, collect the ethyl acetate layer, add water and concentrate until there is no solvent smell; (4) The concentrated solution is cooled to room temperature, activated carbon is added, stirred, and filtered to remove the activated carbon; (5) The filtrate is concentrated and spray-dried to obtain a low-coumarin cinnamon extract.

2. The method for preparing the low-coumarin cinnamon extract according to claim 1, characterized in that: In step (1), the extraction time is 1 to 3 hours; in step (2), the extraction time is 1 to 2 hours.

3. The method for preparing the low-coumarin cinnamon extract according to claim 1, characterized in that: In step (3), the volume of the filtrate is concentrated to 1.8 to 2 times the amount of cinnamon.

4. The method for preparing the low-coumarin cinnamon extract according to claim 1, characterized in that: In step (3), the volume of ethyl acetate is equal to the volume of the filtrate after concentration.

5. The method for preparing the low-coumarin cinnamon extract according to claim 1, characterized in that: In step (3), the stirring time is 1 to 10 min.

6. The method for preparing the low-coumarin cinnamon extract according to claim 1, characterized in that: In step (3), the standing time is 10 to 50 minutes.

7. The method for preparing the low-coumarin cinnamon extract according to claim 1, characterized in that: In step (4), the mass of the activated carbon is 0.4-0.6% of the mass of cinnamon.

8. The method for preparing the low-coumarin cinnamon extract according to claim 1, characterized in that: In step (4), the mesh size of the activated carbon is 100 to 140 meshes.

9. The method for preparing the low-coumarin cinnamon extract according to claim 1, characterized in that: In step (4), the stirring time is 10 to 40 min.

10. Use of the low-coumarin cinnamon extract obtained by the method of claim 1 in health products with intestinal mucosal protective efficacy or drugs for treating intestinal mucosal damage.

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