CN103926375B - A kind of device and using method simulating human consumption's glycolysis system - Google Patents

Abstract

A kind of simulation human consumption's glycolysis system and device and using method, comprise oral digestion district, peptic digest district, small intestine digestion zone, large intestine leading portion glycolysis district, large intestine back segment glycolysis district.Add determinand to the import of device top, open whole water bath with thermostatic control system and keep 37 DEG C of constant temperature, opening whole constant flow pump and diverting valve.Determinand realizes full dynamic flow and experiences oral digestion, peptic digest, little intestinal digestion, large intestine leading portion and the glycolysis of large intestine back segment according to this in this digestive system.Simulation digestion glycolysis process in triplicate, is collected the digestion glycolysis product at each position, is detected the digestion glycolysis situation of test substance.The present invention is undertaken by human consumption's glycolysis process and physiological environment completely for the digestion glycolysis simulation at each position first, each digests the full dynamic similation of the process implementation at glycolysis position, can observe various activity or the nutriment digestion glycolysis situation after human body that enters, the physiological environment of the real people that fits.

Description

Device and method for simulating human digestive and fermentation system

technical field

The invention relates to a device for simulating a human body's digestion and fermentation system and a method for using it.

Background technique

The world's authoritative journals stated that it is very important to study the effects of food or active substances in the human body and their mechanisms of action, because the elucidation of the mechanism of action in vivo can provide a research basis for maximizing the use of food or active substances in the human body. It can exert its physiological efficacy more effectively, and at the same time, it has guiding significance for avoiding its possible adverse side effects. The actual physiological effects of a food or active substance largely depend on what is the product of its digestion and fermentation by the human body. For the consideration of ethics in human body research, the in vitro simulation of human digestion and fermentation has become the technical key to the research on the mechanism of action of food or active substances in vivo. Dynamic simulation is carried out by controlling the key points of the human body's digestion and fermentation process, such as temperature and digestive juice. At the same time, compared with the human body's digestive and zymolytic system, the simulated digestive glycolysis system has better accuracy, good reproducibility, and is easy to control. It can collect the digestion or glycolysis products of various parts at any time. When the experiment involves In the case of toxic chemical substances, they will not be bound by morality, which will bring convenience to the study of the mechanism of action of active substances. Therefore, it is crucial to simulate digestive glycolysis in vitro.

Different foods or active nutrients have different digestion and fermentation conditions in the human body, which makes it very meaningful to study the process of digestion and fermentation. It provides data for the study of digestion, and is of great significance for understanding and exploring the digestion and fermentation methods of different substances. In recent years, related patents related to digestion and fermentation simulation include: "Intelligent organism gastrointestinal tract digestive system simulation control device" (application number: 201220211927.6); "animal gastrointestinal tract simulation digester" (application number: 200920105937.X) ; "Monogastric Animal Biomimetic Digestion System and Method for Simulating Monogastric Animal Digestion Based on the System" (Application No.: 200910078147.1); "In vitro Digestion Simulator for Broilers (200810100818.5); "A Digestive Tract System Simulator" (Application No. : 200710078118.6). These patent reports have greatly enriched the devices and methods of use suitable for digestive fermentation, but so far, these patents have not detailed the specific digestion of the simulated oral cavity. Secondly, these patents are for the simulation of the stomach There is no detailed simulation and description of the gastric emptying process. At the same time, these patents are not detailed enough to control the parameters in the small intestine simulation system and large intestine glycolysis system. Some of these patents are only based on animal-related parameters and have no relevance to humans. In this patent, for the first time, the digestion and fermentation simulation of each part is completely carried out according to the human body's digestion and fermentation process and physiological environment, and each digestion and fermentation part is described in detail. Dynamic simulation fits the real human physiological environment.

Contents of the invention

The object of the present invention is to provide a device for simulating the digestive and zymolytic system of a human body and a method for using it. The device and the use method can observe the digestion and fermentation conditions of various active or nutrient substances after entering the human body, and help to explore the digestion and fermentation conditions of various active or nutrient substances after entering the human body.

The present invention is achieved through the following technical solutions.

The device for simulating the digestive and glycolysis system of the human body according to the present invention includes a device including oral cavity digestion area (A), stomach digestion area (B), small intestine digestion area (C), front large intestine fermentation area (D), and rear large intestine fermentation area. Solution area (E). Specifically includes sample inlet (1), oral cavity digestion chamber (2), stomach digestion chamber (3), small intestine digestion chamber (4), front large intestine fermentation chamber (5), rear large intestine fermentation chamber (6), saliva dish (7), gastric juice dish (8), small intestine liquid dish (9), fermentation culture dish for the front part of the large intestine (10), fermentation culture dish for the back part of the large intestine (11), the first dialysis room (12), the second Dialysis chamber (13), airtight first filter screen (14), airtight second filter screen (15), mouth digest product dish (16), stomach digest product dish (17), small intestine digest product dish (18), large intestine The anaerobic dish for the fermentation product of the front section (19), the anaerobic dish for the fermentation product of the rear section of the large intestine (20), and the recovery dish for the remaining fermentation product (21).

The connection relationship of each component.

Oral digestion area (A): The sample inlet (1) is connected to the right end of the oral digestion chamber (2) through the connecting valve and the diverter valve, and the right end of the oral digestion chamber (2) leads directly to the interior of the oral digestion chamber (2). Simultaneously, a constant temperature water bath circulation system is installed on the entire outside of the oral cavity digestion chamber (2). The left end of the oral cavity digestion chamber (2) is connected with the saliva dish (7) through a drainage device and a constant flow pump. The outlet pipe at the left end of the oral digestion chamber (2) is connected with the oral digestion product dish (16) through a diverter valve, a connecting valve and a constant flow pump. The middle part of the outlet pipe at the left end of the oral cavity digestion chamber (2) is connected with the sample injection pipe at the left end of the stomach digestion chamber (3) through an emptying delivery pipe and a connecting valve.

Stomach digestion area (B): The injection tube at the left end of the stomach digestion chamber (3) leads directly into the stomach digestion chamber (3). The gastric digestion chamber (3) is divided into two sub-chambers, and the outside of the two sub-chambers is equipped with a constant temperature water bath circulation system, and the two sub-chambers are fixedly connected by two fixed valves. At the junction of the two sub-chambers, it is connected with the gastric juice dish (8) through a drainage device and a constant flow pump. The outlet pipe at the right end of the gastric digestion chamber (3) is connected to a conduit through a diverter valve, and the other end of the conduit is connected to the gastric digestion product dish (17) through a connecting valve and a constant flow pump. The junction of the diverter valve is also connected to the right-side sampling tube of the digestive chamber of the small intestine (4) through an emptying delivery tube.

Small intestine digestion area (C): The injection tube at the right end of the small intestine digestion chamber (4) leads directly into the small intestine digestion chamber (4). The digestive chamber of the small intestine (4) is divided into two sub-chambers, and the outside of the two sub-chambers is equipped with a constant temperature water bath circulation system, and the two sub-chambers are fixedly connected by two fixed valves. At the junction of the two sub-chambers, it is connected with the small intestine liquid dish (9) through a drainage device and a constant flow pump. The outlet pipe at the left end of the small intestine digestion chamber (4) is connected to a conduit through a diverter valve, and the other end of the conduit is connected to the small intestine digestion product dish (18) through a connection valve and a constant flow pump. The junction of the shunt valve is also connected with the sampling tube at the left end of the fermentation chamber (5) in the front section of the large intestine through the emptying delivery tube.

Front large intestine fermentation area (D): The left end of the large intestine fermentation chamber (5) leads directly to the inside of the front large intestine fermentation chamber (5). The fermentation room (5) in the front part of the large intestine is divided into two sub-chambers, and the outside of the two sub-chambers is equipped with a constant temperature water bath circulation system, and the two sub-chambers are fixedly connected by two fixed valves. The junction of the two sub-chambers is connected with the fermentation culture dish (10) of the front section of the large intestine through a drainage device and a constant flow pump. The outlet pipe at the right end of the fermentation chamber (5) in the front part of the large intestine is connected to one end of the first airtight filter screen (14) through a diverter valve and a catheter, and the other end of the airtight first filter screen (14) is connected to the first dialysis chamber (12) through a catheter One end is connected, and the other end of the first dialysis chamber (12) is connected to the anaerobic dish (19) of the glycolysis product in the front section of the large intestine through a constant flow pump. The junction of the shunt valve is also connected to the right-end sampling tube of the fermentation chamber (6) in the back section of the large intestine through the emptying delivery tube.

Rear large intestine fermentation area (E): The left end of the large intestine fermentation chamber (6) leads directly to the interior of the large intestine fermentation chamber (6). The fermentation chamber (6) in the rear part of the large intestine is divided into two sub-chambers, and the outside of the two sub-chambers is equipped with a constant temperature water bath circulation system, and the two sub-chambers are fixedly connected by two fixed valves. The junction of the two sub-chambers is connected with the fermentation culture dish (11) of the rear segment of the large intestine through a drainage device and a constant flow pump. The outlet pipe at the left end of the fermentation chamber (5) in the rear part of the large intestine is connected to one end of the closed second filter screen (15) through a diverter valve and a catheter, and the other end of the closed second filter screen (15) is connected to the second dialysis chamber (13) through a catheter ) is connected at one end, and the other end of the second dialysis chamber (13) is connected to the anaerobic dish (20) of glycolysis products in the rear section of the large intestine through a constant flow pump. The junction of the diverter valve is also connected to the remaining fermentation product recovery dish (21) through an emptying delivery pipe.

How to use: Dissolve (or disperse) the substance to be digested in distilled water at a certain concentration to make a solution or dispersion of the substance to be tested. Add the prepared saliva, gastric juice, small intestinal juice, zymolysis medium for the front part of the large intestine, and zymolysis medium for the back part of the large intestine to the saliva dish (7), gastric juice dish (8), small intestine liquid dish (9), and front part of the large intestine, respectively. In the fermentation culture dish (10) and the fermentation culture dish (11) of the posterior segment of the large intestine. Turn on all constant temperature water bath systems and maintain a constant temperature of 37°C. Turn on all constant flow pumps and diverter valves. The speed of all constant flow pumps was controlled at 4-6ml/min. Add the solution or dispersion of the analyte from the uppermost sample inlet (1) of the digestive fermentation system device, and add saliva from the saliva dish (7) into the oral cavity digestion chamber (2) through a constant flow pump and a drainage device for oral cavity digestion. Digestion simulation, after 10-15 minutes of oral digestion, part of the digested products are collected in the oral digestion product dish (16), and the other part of the digested products enters the gastric digestion chamber (3) in the gastric digestion area (B) along with the emptying delivery tube middle. Gastric juice is added from the gastric juice dish (8) into the gastric digestion chamber (3) through a constant flow pump and drainage device to simulate gastric digestion. After gastric digestion for 5-6 hours, a part of the digested products are collected in the gastric digestion product dish (17) , another part of the digested product enters the small intestine digestion chamber (4) in the small intestine digestion zone (C) along with the emptying duct. The small intestine juice is added to the small intestine digestion chamber (4) through the small intestine liquid dish (9) through a constant flow pump and a drainage device. After the small intestine is digested for 5-6 hours, part of the digested products are collected in the small intestine digestion product dish (18), and the other part is The digested product enters the fermentation chamber (5) of the front section of the large intestine (D) along with the emptying delivery tube. The fermentation medium of the front part of the large intestine is added to the fermentation chamber of the front part of the large intestine (5) from the fermentation culture dish of the front part of the large intestine (10) through a constant flow pump and a drainage device. After 10-12 hours of fermentation of the front part of the large intestine, part of the The product is collected in the anaerobic dish (19) of the fermentation product in the front part of the large intestine through the airtight first filter screen (14) and the first dialysis chamber (12), and the other part of the fermentation product enters the rear part of the large intestine for fermentation along with the emptying delivery tube Area (E) in the fermentation compartment (6) of the posterior large intestine. The fermentation medium of the posterior segment of the large intestine is added from the fermentation culture dish of the posterior segment of the large intestine (11) into the fermentation chamber of the posterior segment of the large intestine (6) through a constant flow pump and a drainer. After 10-12 hours of fermentation of the posterior segment of the large intestine, part The glycolysis product is collected in the anaerobic dish (20) for the glycolysis product in the rear part of the large intestine through the airtight second filter screen (15) and the second dialysis chamber (13), and another part of the glycolysis product enters with the emptying delivery tube Remaining zymolysis product reclaims in the dish (21). The simulated digestion and fermentation process was repeated 3 times.

The oral digestion product dish (16) is placed in a boiling water bath (prepared in advance) for several minutes to remove the activity of salivary amylase, and then the oral digestion of the test substance in the oral digestion product dish (16) is detected by the liquid phase method. The gastric digestion product dish (17) was placed in a boiling water bath (prepared in advance) for several minutes to remove pepsin activity, and then the gastric digestion of the test substance in the gastric digestion product dish (17) was detected by liquid phase method. The small intestine digestion product dish (18) was placed in a boiling water bath (prepared in advance) for several minutes to remove the activity of trypsin (including pancreatic amylase and pancreatic lipase) and trypsin, and then the small intestine digestion product dish (18 ) of the small intestine digestion of the analyte. The anaerobic dish (19) of the fermentation product of the first section of the large intestine was placed in a boiling water bath (prepared in advance) for 30-40 minutes to remove the activity of the remaining flora, and the fermentation product of the first section of the large intestine was detected by the gas phase method in the anaerobic dish (19). solution product. The anaerobic dish (20) of the zymolyzed product of the rear section of the large intestine is placed in a boiling water bath (prepared in advance) for 30-40 minutes to remove the activity of the remaining flora, and the large intestine in the anaerobic dish (20) of the zymolyzed product of the rear section of the large intestine is detected by gas phase method Subsequent fermentation products.

The technical feature of the device and method of simulating the human digestive and glycolysis system is that the device and the method of use are for the first time simulating the digestion and glycolysis of each part completely according to the human body's digestion and zymolysis process and physiological environment, and the various parts of digestion and glycolysis are simulated. In detail, at the same time, the process of each digestion and fermentation part has realized a full dynamic simulation, which fits the real human physiological environment.

The technical effect of the device and method of use for simulating the human digestive and glycolysis system is that the device and the method of use can observe the digestive and zymolytic conditions of various active or nutrient substances after they enter the human body, and help to explore various active or nutrient substances that enter the human body. Digestive glycolysis after human body.

Description of drawings

Fig. 1 is a schematic diagram of a device for simulating a human digestion and fermentation system. Among them, A is the digestive area of the oral cavity; B is the digestive area of the stomach; C is the digestive area of the small intestine; D is the fermentation area of the front part of the large intestine; E is the fermentation area of the rear part of the large intestine. 1 is the sample inlet, 2 is the mouth digestion chamber, 3 is the stomach digestion chamber, 4 is the small intestine digestion chamber, 5 is the fermentation chamber of the front part of the large intestine, 6 is the fermentation chamber of the rear part of the large intestine, 7 is the saliva dish, 8 is the gastric juice dish, 9 is the liquid dish of the small intestine, 10 is the fermentation culture dish of the front part of the large intestine, 11 is the fermentation culture dish of the rear part of the large intestine, 12 is the first dialysis chamber, 13 is the second dialysis chamber, 14 is the airtight first filter screen, 15 is a closed second filter sieve, 16 is a dish for oral digestion products, 17 is a dish for gastric digestion products, 18 is a dish for small intestine digestion products, 19 is an anaerobic dish for fermentation products of the front part of the large intestine, and 20 is an anaerobic dish for fermentation products of the rear part of the large intestine. Oxygen dish, 21 is the recovery dish of remaining glycolysis product.

FIG. 2 is an enlarged view of the upper half of F-F in the schematic diagram of the device for simulating the human digestion and fermentation system in FIG. 1 .

FIG. 3 is an enlarged view of the lower half of F-F in the schematic diagram of the device for simulating the human digestion and fermentation system in FIG. 1 .

detailed description

Example 1.

Human digestive glycolysis simulation of plant polysaccharides.

1. Raw material preparation.

(1) Saliva.

Fresh clarified saliva samples were collected from a healthy volunteer. The volunteer had no chronic diseases and had not used antibiotics for at least 3 months. Before saliva collection, the volunteer was restricted from eating and drinking. Collection is by direct spitting. The collected saliva was centrifuged at 1500×g for 10 minutes to remove cells, and the supernatant was stored at -20°C until use.

(2) Gastric juice.

Gastric juice is obtained by weighing 30mg of gastric lipase and 30mg of pepsin into 150g of gastric electrolyte (preparation of gastric electrolyte solution: weigh 3gNaCl, 0.5gKCl, 0.1gCaCl ₂ , 0.5gNaHCO ₃ , and dilute to 1L with deionized water In the bottle, adjust the pH of the electrolyte solution to 3) with 0.1M HCl, and then add 1mL CH ₃ COONa (1M, pH 5). Stir magnetically at room temperature for 10 min, adjust the pH of the solution to 3 with 0.1 M HCl, and store in the refrigerator for later use.

(3) small intestinal fluid.

The small intestine juice is obtained by weighing 100g intestinal electrolyte solution (preparation of intestinal electrolyte solution: weigh 5gNaCl, 0.5gKCl, _0.3gCaCl2 , dissolve them in a 1L volumetric flask with deionized water, and adjust the pH of the solution to 7 with 0.1M NaOH ), 100g (7%, w/w) trypsin solution, 12mg of trypsin, 180g of water. The pH of the mixed solution was adjusted to 7 with 0.1 M NaOH. At the same time, bile powder (2%, w/v) was added to the solution to make small intestinal juice.

(4) Fermentation medium for the front part of the large intestine.

1L fermentation medium contains: 4gNaCl, 4gKCl, 1gNaHCO ₃ , 0.6gMgSO ₄ ·H ₂ O, 0.5gKH ₂ PO ₄ , 0.5gK ₂ HPO ₄ , 0.05gCaCl ₂ , 0.005gFeSO ₄ ·7H ₂ O, 1ml Tween 80 and 3ml resin azure solution (0.025%, w/v, anaerobic indicator). The medium was sterilized at 121°C for 15 minutes before use. Then 150 g of human feces were added to the fermentation medium (volunteers with feces usually ate a normal diet, had no digestive diseases, and had not taken antibiotics for at least 3 months).

(5) Fermentation medium for the posterior segment of the large intestine.

1L fermentation medium contains: 4gNaCl, 4gKCl, 1gNaHCO ₃ , 0.6gMgSO ₄ ·H ₂ O, 0.7gL-cysteine HCl·H ₂ O, 0.5gKH ₂ PO ₄ , 0.5gK ₂ HPO ₄ , 0.4g Bile salts, 0.05g CaCl ₂ , 0.005g FeSO ₄ ·7H ₂ O, 1ml Tween 80 and 3ml resin azure solution (0.025%, w/v, anaerobic indicator). The medium was sterilized at 121°C for 15 minutes before use. Then 250 g of human feces were added to the fermentation medium (volunteers with feces usually ate a normal diet, had no digestive diseases, and had not taken antibiotics for at least 3 months).

2. Simulate the process of human digestive fermentation system.

The schematic diagram of the device for simulating the human digestion and fermentation system is shown in Figure 1. The device includes mouth digestion area A, stomach digestion area B, small intestine digestion area C, front large intestine fermentation area D, and rear large intestine fermentation area E. Specifically, it includes sample inlet 1, oral cavity digestion chamber 2, stomach digestion chamber 3, small intestine digestion chamber 4, front large intestine fermentation chamber 5, large intestine fermentation chamber 6, saliva dish 7, gastric juice dish 8, small intestine liquid dish 9, The fermentation culture dish 10 of the front section of the large intestine and the fermentation culture dish 11 of the rear section of the large intestine are the first dialysis chamber 12, the second dialysis chamber 13, the first airtight filter screen 14, the airtight second filter screen 15, and oral digestion Product dish 16 , gastric digestion product dish 17 , small intestine digestion product dish 18 , anaerobic dish 19 for glycolysis product in the front part of the large intestine, anaerobic dish 20 for zymolysis product in the rear part of the large intestine, and a dish 21 for recovering the remaining zymolysis product.

The connection relationship of each component.

Oral digestion area A: The sample inlet 1 is connected to the sampling pipe at the right end of the oral digestion chamber 2 through the connection valve and the diverter valve, and the sampling pipe at the right end of the oral digestion chamber 2 is directly connected to the interior of the oral digestion chamber 2. Simultaneously, a constant temperature water bath circulation system is installed on all outsides of the oral cavity digestion chamber 2. The left end of the oral cavity digestion chamber 2 is connected with the saliva dish 7 through a drainage device and a constant flow pump. The outlet pipe at the left end of the mouth digestion chamber 2 is connected with the mouth digestion product dish 16 through a diverter valve, a connecting valve and a constant flow pump. The middle part of the outlet pipe at the left end of the oral cavity digestion chamber 2 is connected with the sampling pipe at the left end of the stomach digestion chamber 3 through an emptying delivery pipe and a connecting valve.

Stomach digestion area B: the sampling tube at the left end of the stomach digestion chamber 3 is directly connected to the inside of the stomach digestion chamber 3 . The gastric digestion chamber 3 is divided into two sub-chambers, and the outside of the two sub-chambers is equipped with a constant temperature water bath circulation system, and the two sub-chambers are fixedly connected by two fixed valves. The junction of the two sub-chambers is connected with the gastric juice dish 8 through a flow guide and a constant flow pump. The outlet pipe at the right end of the gastric digestion chamber 3 is connected to a conduit through a diverter valve, and the other end of the conduit is connected to a gastric digestion product vessel 17 through a connecting valve and a constant flow pump. The junction of the shunt valve is also connected to the sampling tube at the right end of the small intestine digestion chamber 4 through an emptying delivery tube.

Small intestine digestion area C: the sampling tube at the right end of the small intestine digestion chamber 4 leads directly to the interior of the small intestine digestion chamber 4 . The small intestine digestion chamber 4 is divided into two sub-chambers, and the outside of the two sub-chambers is equipped with a constant temperature water bath circulation system, and the two sub-chambers are fixedly connected by two fixed valves. The junction of the two sub-chambers is connected with the small intestine liquid dish 9 through a flow diverter and a constant flow pump. The outlet pipe at the left end of the small intestine digestion chamber 4 is connected to a conduit through a diverter valve, and the other end of the conduit is connected to the small intestine digestion product vessel 18 through a connection valve and a constant flow pump. The junction of the shunt valve is also connected with the sampling tube at the left end of the fermentation chamber 5 in the front section of the large intestine through an emptying delivery tube.

Front large intestine fermentation area D: The left end of the fermentation chamber 5 at the front of the large intestine has a sample tube that leads directly to the interior of the fermentation chamber 5 at the front large intestine. The fermentation chamber 5 in the front part of the large intestine is divided into two sub-chambers, and the outside of the two sub-chambers is equipped with a constant temperature water bath circulation system, and the two sub-chambers are fixedly connected by two fixed valves. The junction of the two sub-chambers is connected with the fermentation culture vessel 10 of the front section of the large intestine through a flow diverter and a constant flow pump. The outlet pipe at the right end of the fermentation chamber 5 in the front part of the large intestine is connected to one end of the airtight first filter screen 14 through a diverter valve and a conduit, and the other end of the airtight first filter screen 14 is connected to one end of the first dialysis chamber 12 through a conduit, and the first dialysis The other end of the chamber 12 passes through a constant flow pump and an anaerobic dish 19 for the fermentation product of the front section of the large intestine. The junction of the shunt valve is also connected to the right end sampling tube of the rear section of the large intestine fermentation chamber 6 through an emptying delivery tube.

Fermentation area E of the rear section of the large intestine: the sampling tube at the left end of the fermentation chamber 6 of the rear section of the large intestine leads directly to the interior of the fermentation chamber 6 of the rear section of the large intestine. The fermentation chamber 6 in the rear part of the large intestine is divided into two sub-chambers, and the outside of the two sub-chambers is equipped with a constant temperature water bath circulation system, and the two sub-chambers are fixedly connected by two fixed valves. The joint of the two sub-chambers is connected with the zymolysis culture vessel 11 of the rear section of the large intestine through a flow diverter and a constant flow pump. The outlet pipe at the left end of the fermentation chamber 5 in the back section of the large intestine is connected to one end of the second airtight filter screen 15 through a diverter valve and a conduit, and the other end of the airtight second filter screen 15 is connected to one end of the second dialysis chamber 13 through a conduit, and the second dialysis chamber 13 The other end passes through a constant flow pump and the anaerobic dish 20 of the fermentation product of the rear section of the large intestine. The junction of the shunt valve is also connected to the remaining fermentation product recovery vessel 21 through an emptying delivery pipe.

How to use: Plant polysaccharide solution made into a certain concentration solution (5mg/ml). Add the prepared saliva, gastric juice, small intestinal juice, zymolysis medium for the first part of the large intestine, and zymolysis medium for the back part of the large intestine to the saliva dish 7, the gastric juice dish 8, the small intestine juice dish 9, and the zymolysis culture dish 10 for the first part of the large intestine. , in the fermentation culture dish 11 of the posterior segment of the large intestine. Turn on all constant temperature water bath systems and maintain a constant temperature of 37°C. Turn on all constant flow pumps and diverter valves. The speed of all constant flow pumps was controlled at 5ml/min. The plant polysaccharide solution is added from the sample inlet 1 at the top of the digestion and fermentation system device, and the saliva is added from the saliva dish 7 to the oral digestion chamber 2 through a constant flow pump and a drainage device to simulate oral digestion. After 10 minutes of oral digestion, part of the digestion The final product is collected in the oral cavity digestion product dish 16 to obtain the plant polysaccharide oral cavity digestion product, and another part of the digestion product enters the gastric digestion chamber 3 in the gastric digestion area B along with the emptying delivery pipe. Gastric juice is added from the gastric juice dish 8 into the gastric digestion chamber 3 through a constant flow pump and a drainage device to perform gastric digestion simulation. After 6 hours of gastric digestion, a part of the digested products are collected in the gastric digest product dish 17 to obtain plant polysaccharide gastric digest products. Another part of the digested product enters the small intestine digestion chamber 4 in the small intestine digestion zone C along with the emptying conveying pipe. The small intestine juice is added to the small intestine digestion chamber 4 through the constant flow pump and drainage device from the small intestine liquid dish 9. After the small intestine is digested for 6 hours, part of the digested products are collected in the small intestine digest product dish 18 to obtain the plant polysaccharide small intestine digest product, and the other part of the digest product Enter the fermentation chamber 5 of the front section of the large intestine in the fermentation zone D of the front section of the large intestine along with the emptying delivery tube. The fermentation medium of the front part of the large intestine is added into the fermentation chamber 5 of the front part of the large intestine from the fermentation culture dish 10 of the front part of the large intestine through a constant flow pump and a diverter. The filter screen 14 and the first dialysis chamber 12 are collected in the anaerobic dish 19 for the glycolysis product of the front section of the large intestine to obtain the fermentation product of the front section of the large intestine of plant polysaccharides, and the other part of the glycolysis product enters the fermentation area E of the back section of the large intestine along with the emptying delivery tube. In the zymolysis chamber 6 of the posterior segment of the large intestine. The zymolysis medium of the rear section of the large intestine is added into the fermentation room 6 of the rear section of the large intestine from the zymolysis culture dish 11 of the rear section of the large intestine through a constant flow pump and a drainage device. The airtight second filter screen 15 and the second dialysis chamber 13 are collected in the anaerobic dish 20 for the fermentation product of the rear section of the large intestine to obtain the fermentation product of the rear section of the large intestine of plant polysaccharides. The solution product is recovered in the dish 21. The simulated digestion and fermentation process was repeated 3 times.

The oral digestion product dish 16 was placed in a boiling water bath (prepared in advance) for 5 minutes to remove the activity of salivary amylase, and then the oral digestion of plant polysaccharides in the oral digestion product dish 16 was detected (molecular weight change) by liquid phase method. Stomach digestion product plate 17 was placed in a boiling water bath (prepared in advance) for 5 minutes to remove the activity of pepsin, and then the gastric digestion of plant polysaccharides (molecular weight change) in gastric digestion product plate 17 was detected by liquid phase method, and dinitro water Salicylic acid method was used to detect the change of reducing sugar. The small intestine digestion product dish 18 was placed in a boiling water bath (prepared in advance) for 5 minutes to remove the activity of trypsin (including pancreatic amylase and pancreatic lipase) and trypsin, and then the plant polysaccharides in the small intestine digestion product dish 18 were detected by liquid phase method The small intestine digestion (molecular weight change), and the change of reducing sugar was detected by dinitrosalicylic acid method. The anaerobic dish 19 of the fermentation product of the first section of the large intestine was placed in a boiling water bath (prepared in advance) for 40 minutes to remove the activity of the remaining flora, and the fermentation product of the first section of the large intestine of the plant polysaccharides in the anaerobic dish 19 of the fermentation product of the front section of the large intestine was detected by the gas phase method. The post-large intestine fermentation product anaerobic dish 20 was placed in a boiling water bath (prepared in advance) for 40 minutes to remove the activity of the remaining flora, and the post-colon fermentation of plant polysaccharides in the rear large intestine fermentation product anaerobic dish 20 was detected by gas phase method solution product.

Test result: The plant polysaccharide will not be decomposed in human oral digestion. After simulated digestion in the stomach, the molecular weight decreased significantly, and the amount of reducing sugar increased significantly, indicating that the plant polysaccharide was decomposed during the digestion of the human stomach, and at the same time, some glycosidic bonds were interrupted. After simulated digestion in the small intestine, the molecular weight decreased slightly, and the amount of reducing sugar increased to a certain extent, indicating that the plant polysaccharide was decomposed in a small amount and a small amount of glycosidic bonds were interrupted during the digestion of human small intestine juice. After zymolysis in the front part of the large intestine and fermentation in the back part of the large intestine, a large amount of short-chain fatty acids can be produced, and the amount of acetic acid, propionic acid and butyric acid can be significantly increased, which will benefit the health of the human large intestine.

Example 2.

Human digestive glycolysis simulation of cornstarch.

1. Raw material preparation.

(1) Saliva.

Fresh clarified saliva samples were collected from a healthy volunteer. The volunteer had no chronic diseases and had not used antibiotics for at least 3 months. Before saliva collection, the volunteer was restricted from eating and drinking. Collection is by direct spitting. The collected saliva was centrifuged at 1500×g for 10 minutes to remove cells, and the supernatant was stored at -20°C until use.

(2) Gastric juice.

Gastric juice is obtained by weighing 30mg of gastric lipase and 30mg of pepsin into 150g of gastric electrolyte (preparation of gastric electrolyte solution: weigh 3gNaCl, 0.5gKCl, 0.1gCaCl ₂ , 0.5gNaHCO ₃ , and dilute to 1L with deionized water In the bottle, adjust the pH of the electrolyte solution to 3) with 0.1M HCl, and then add 1mL CH ₃ COONa (1M, pH 5). Stir magnetically at room temperature for 10 min, adjust the pH of the solution to 3 with 0.1 M HCl, and store in the refrigerator for later use.

(3) small intestinal fluid.

The small intestine juice is obtained by weighing 100g intestinal electrolyte solution (preparation of intestinal electrolyte solution: weigh 5gNaCl, 0.5gKCl, _0.3gCaCl2 , dissolve them in a 1L volumetric flask with deionized water, and adjust the pH of the solution to 7 with 0.1M NaOH ), 100g (7%, w/w) trypsin solution, 12mg of trypsin, 180g of water. The pH of the mixed solution was adjusted to 7 with 0.1 M NaOH. At the same time, bile powder (2%, w/v) was added to the solution to make small intestinal fluid.

(4) Fermentation medium for the front part of the large intestine.

1L fermentation medium contains: 4gNaCl, 4gKCl, 1gNaHCO ₃ , 0.6gMgSO ₄ ·H ₂ O, 0.5gKH ₂ PO ₄ , 0.5gK ₂ HPO ₄ , 0.05gCaCl ₂ , 0.005gFeSO ₄ ·7H ₂ O, 1ml Tween 80 and 3ml resin azure solution (0.025%, w/v, anaerobic indicator). The medium was sterilized at 121°C for 15 minutes before use. Then 150 g of human feces were added to the fermentation medium (volunteers with feces usually ate a normal diet, had no digestive diseases, and had not taken antibiotics for at least 3 months).

(5) Fermentation medium for the posterior segment of the large intestine.

1L fermentation medium contains: 4gNaCl, 4gKCl, 1gNaHCO ₃ , 0.6gMgSO ₄ ·H ₂ O, 0.7gL-cysteine HCl·H ₂ O, 0.5gKH ₂ PO ₄ , 0.5gK ₂ HPO ₄ , 0.4g Bile salts, 0.05g CaCl ₂ , 0.005g FeSO ₄ ·7H ₂ O, 1ml Tween 80 and 3ml resin azure solution (0.025%, w/v, anaerobic indicator). The medium was sterilized at 121°C for 15 minutes before use. Then 250 g of human feces were added to the fermentation medium (volunteers with feces usually ate a normal diet, had no digestive diseases, and had not taken antibiotics for at least 3 months).

2. Simulate the process of human digestive fermentation system.

The schematic diagram of the device for simulating the human digestion and fermentation system is shown in Figure 1. The device includes mouth digestion area A, stomach digestion area B, small intestine digestion area C, front large intestine fermentation area D, and rear large intestine fermentation area E. Specifically, it includes sample inlet 1, oral cavity digestion chamber 2, stomach digestion chamber 3, small intestine digestion chamber 4, front large intestine fermentation chamber 5, large intestine fermentation chamber 6, saliva dish 7, gastric juice dish 8, small intestine liquid dish 9, The fermentation culture dish 10 of the front section of the large intestine and the fermentation culture dish 11 of the rear section of the large intestine are the first dialysis chamber 12, the second dialysis chamber 13, the first airtight filter screen 14, the airtight second filter screen 15, and oral digestion Product dish 16 , gastric digestion product dish 17 , small intestine digestion product dish 18 , anaerobic dish 19 for glycolysis product in the front part of the large intestine, anaerobic dish 20 for zymolysis product in the rear part of the large intestine, and a dish 21 for recovering the remaining zymolysis product.

The connection relationship of each component.

Oral digestion area A: The sample inlet 1 is connected to the sampling pipe at the right end of the oral digestion chamber 2 through the connection valve and the diverter valve, and the sampling pipe at the right end of the oral digestion chamber 2 is directly connected to the interior of the oral digestion chamber 2. Simultaneously, a constant temperature water bath circulation system is installed on all outsides of the oral cavity digestion chamber 2. The left end of the oral cavity digestion chamber 2 is connected with the saliva dish 7 through a drainage device and a constant flow pump. The outlet pipe at the left end of the mouth digestion chamber 2 is connected with the mouth digestion product dish 16 through a diverter valve, a connecting valve and a constant flow pump. The middle part of the outlet pipe at the left end of the oral cavity digestion chamber 2 is connected with the sampling pipe at the left end of the stomach digestion chamber 3 through an emptying delivery pipe and a connecting valve.

Stomach digestion area B: the sampling tube at the left end of the stomach digestion chamber 3 is directly connected to the inside of the stomach digestion chamber 3 . The gastric digestion chamber 3 is divided into two sub-chambers, and the outside of the two sub-chambers is equipped with a constant temperature water bath circulation system, and the two sub-chambers are fixedly connected by two fixed valves. The junction of the two sub-chambers is connected with the gastric juice dish 8 through a flow guide and a constant flow pump. The outlet pipe at the right end of the gastric digestion chamber 3 is connected to a conduit through a diverter valve, and the other end of the conduit is connected to a gastric digestion product vessel 17 through a connecting valve and a constant flow pump. The junction of the shunt valve is also connected to the sampling tube at the right end of the small intestine digestion chamber 4 through an emptying delivery tube.

Small intestine digestion area C: the sampling tube at the right end of the small intestine digestion chamber 4 leads directly to the interior of the small intestine digestion chamber 4 . The small intestine digestion chamber 4 is divided into two sub-chambers, and the outside of the two sub-chambers is equipped with a constant temperature water bath circulation system, and the two sub-chambers are fixedly connected by two fixed valves. The junction of the two sub-chambers is connected with the small intestine liquid dish 9 through a flow diverter and a constant flow pump. The outlet pipe at the left end of the small intestine digestion chamber 4 is connected to a conduit through a diverter valve, and the other end of the conduit is connected to the small intestine digestion product vessel 18 through a connection valve and a constant flow pump. The junction of the shunt valve is also connected with the sampling tube at the left end of the fermentation chamber 5 in the front section of the large intestine through an emptying delivery tube.

Front large intestine fermentation area D: The left end of the fermentation chamber 5 at the front of the large intestine has a sample tube that leads directly to the interior of the fermentation chamber 5 at the front large intestine. The fermentation chamber 5 in the front part of the large intestine is divided into two sub-chambers, and the outside of the two sub-chambers is equipped with a constant temperature water bath circulation system, and the two sub-chambers are fixedly connected by two fixed valves. The junction of the two sub-chambers is connected with the fermentation culture vessel 10 of the front section of the large intestine through a flow diverter and a constant flow pump. The outlet pipe at the right end of the fermentation chamber 5 in the front part of the large intestine is connected to one end of the airtight first filter screen 14 through a diverter valve and a conduit, and the other end of the airtight first filter screen 14 is connected to one end of the first dialysis chamber 12 through a conduit, and the first dialysis The other end of the chamber 12 passes through a constant flow pump and an anaerobic dish 19 for the fermentation product of the front section of the large intestine. The junction of the shunt valve is also connected to the right end sampling tube of the rear section of the large intestine fermentation chamber 6 through an emptying delivery tube.

Fermentation area E of the rear section of the large intestine: the sampling tube at the left end of the fermentation chamber 6 of the rear section of the large intestine leads directly to the interior of the fermentation chamber 6 of the rear section of the large intestine. The fermentation chamber 6 in the rear part of the large intestine is divided into two sub-chambers, and the outside of the two sub-chambers is equipped with a constant temperature water bath circulation system, and the two sub-chambers are fixedly connected by two fixed valves. The joint of the two sub-chambers is connected with the zymolysis culture vessel 11 of the rear section of the large intestine through a flow diverter and a constant flow pump. The outlet pipe at the left end of the fermentation chamber 5 in the back section of the large intestine is connected to one end of the second airtight filter screen 15 through a diverter valve and a conduit, and the other end of the airtight second filter screen 15 is connected to one end of the second dialysis chamber 13 through a conduit, and the second dialysis chamber 13 The other end passes through a constant flow pump and the anaerobic dish 20 of the fermentation product of the rear section of the large intestine. The junction of the shunt valve is also connected to the remaining fermentation product recovery vessel 21 through an emptying delivery pipe.

How to use: Corn starch solution is made into a certain concentration solution (5mg/ml). Add the prepared saliva, gastric juice, small intestinal juice, zymolysis medium for the first part of the large intestine, and zymolysis medium for the back part of the large intestine to the saliva dish 7, the gastric juice dish 8, the small intestine juice dish 9, and the zymolysis culture dish 10 for the first part of the large intestine. , in the fermentation culture dish 11 of the posterior segment of the large intestine. Turn on all constant temperature water bath systems and maintain a constant temperature of 37°C. Turn on all constant flow pumps and diverter valves. The speed of all constant flow pumps was controlled at 6ml/min. Add the cornstarch solution from the uppermost sample inlet 1 of the digestion and fermentation system device, and add the saliva from the saliva dish 7 into the oral digestion chamber 2 through the constant flow pump and drainage device to perform oral digestion simulation. After 15 minutes of oral digestion, part of the digestion The final product is collected in the oral cavity digestion product dish 16 to obtain the cornstarch oral cavity digestion product, and another part of the digestion product enters the stomach digestion chamber 3 in the stomach digestion zone B along with the emptying delivery pipe. Gastric juice is added from the gastric juice dish 8 into the gastric digestion chamber 3 through a constant flow pump and a drainage device to perform gastric digestion simulation. After 6 hours of gastric digestion, a part of the digested products are collected in the gastric digest product dish 17 to obtain cornstarch gastric digest products. Another part of the digested product enters the small intestine digestion chamber 4 in the small intestine digestion zone C along with the emptying conveying pipe. The small intestine juice is added to the small intestine digestion chamber 4 through the constant flow pump and drainage device from the small intestine liquid dish 9. After the small intestine is digested for 6 hours, a part of the digested products are collected in the small intestine digestion product dish 18 to obtain the cornstarch small intestine digestion product, and the other part of the digestion product is Enter the fermentation chamber 5 of the front section of the large intestine in the fermentation zone D of the front section of the large intestine along with the emptying delivery tube. The fermentation medium of the front part of the large intestine is added into the fermentation chamber 5 of the front part of the large intestine from the fermentation culture dish 10 of the front part of the large intestine through a constant flow pump and a diverter. The filter screen 14 and the first dialysis chamber 12 are collected in the anaerobic dish 19 for the fermentation product of the front section of the large intestine to obtain the fermentation product of the front section of the large intestine of cornstarch, and the other part of the fermentation product enters the fermentation zone E of the rear section of the large intestine along with the emptying delivery pipe. In the zymolysis chamber 6 of the posterior segment of the large intestine. The zymolysis medium of the rear section of the large intestine is added into the fermentation room 6 of the rear section of the large intestine from the zymolysis culture dish 11 of the rear section of the large intestine through a constant flow pump and a drainage device. The airtight second filter screen 15 and the second dialysis chamber 13 are collected in the anaerobic dish 20 for the fermentation product of the rear section of the large intestine to obtain the fermentation product of the rear section of the large intestine of corn starch, and the other part of the fermentation product enters the remaining fermentation product along with the emptying delivery pipe 15. The solution product is recovered in the dish 21. The simulated digestion and fermentation process was repeated 3 times.

The oral digestion product dish 16 was placed in a boiling water bath (prepared in advance) for 5 minutes to remove the activity of salivary amylase, and then the oral digestion (molecular weight change) of cornstarch in the oral digestion product dish 16 was detected by liquid phase method. Stomach digestion product plate 17 was placed in a boiling water bath (prepared in advance) for 5 minutes to remove the activity of pepsin, and then the gastric digestion of cornstarch (molecular weight change) in gastric digestion product plate 17 was detected by liquid phase method, and dinitro water Salicylic acid method was used to detect the change of reducing sugar. The small intestine digestion product dish 18 was placed in a boiling water bath (prepared in advance) for 5 minutes to remove the activity of trypsin (including pancreatic amylase and pancreatic lipase) and trypsin, and then the cornstarch in the small intestine digestion product dish 18 was detected by liquid phase method The small intestine digestion (molecular weight change), and the change of reducing sugar was detected by dinitrosalicylic acid method. The anaerobic dish 19 of the fermentation product of the first section of the large intestine was placed in a boiling water bath (prepared in advance) for 30 minutes to remove the activity of the remaining flora, and the fermentation product of the first section of the large intestine of cornstarch in the anaerobic dish 19 of the fermentation product of the front section of the large intestine was detected by gas phase method. The fermentation product of the rear part of the large intestine anaerobic dish 20 was placed in a boiling water bath (prepared in advance) for 30 minutes to remove the activity of the remaining flora, and the rear part of the large intestine fermentation of cornstarch in the anaerobic dish 20 of the fermentation product of the rear part of the large intestine was detected by gas phase method solution product.

Test results: After the corn starch was digested in the oral cavity, the molecular weight decreased significantly, and the amount of reducing sugar increased significantly, indicating that the corn starch was decomposed during oral digestion, and part of the glycosidic bonds were interrupted. After simulated digestion in the stomach, the molecular weight was partially reduced, and the amount of reducing sugar increased to a certain extent, indicating that the corn starch was decomposed in a small amount during the digestion of the human stomach, and a small amount of glycosidic bonds were interrupted at the same time. After simulated digestion in the small intestine, the molecular weight decreased significantly, and the amount of reducing sugar increased significantly, indicating that the corn starch was decomposed in large quantities during digestion in the small intestine of the human body, and a large number of glycosidic bonds were interrupted at the same time. Finally, after the fermentation of the front and back sections of the large intestine, a large amount of short-chain fatty acids can be produced, and the amount of acetic acid, propionic acid and butyric acid can be significantly increased.

Example 3.

Human digestive glycolysis simulation of soybean protein.

1. Raw material preparation.

(1) Saliva.

Fresh clarified saliva samples were collected from a healthy volunteer. The volunteer had no chronic diseases and had not used antibiotics for at least 3 months. Before saliva collection, the volunteer was restricted from eating and drinking. Collection is by direct spitting. The collected saliva was centrifuged at 1500×g for 10 minutes to remove cells, and the supernatant was stored at -20°C until use.

(2) Gastric juice.

Gastric juice is obtained by weighing 30mg of gastric lipase and 30mg of pepsin into 150g of gastric electrolyte (preparation of gastric electrolyte solution: weigh 3gNaCl, 0.5gKCl, 0.1gCaCl ₂ , 0.5gNaHCO ₃ , and dilute to 1L with deionized water In the bottle, adjust the pH of the electrolyte solution to 3) with 0.1M HCl, and then add 1mL CH ₃ COONa (1M, pH 5). Stir magnetically at room temperature for 10 min, adjust the pH of the solution to 3 with 0.1 M HCl, and store in the refrigerator for later use.

(3) small intestinal fluid.

The small intestine juice is obtained by weighing 100g intestinal electrolyte solution (preparation of intestinal electrolyte solution: weigh 5gNaCl, 0.5gKCl, _0.3gCaCl2 , dissolve them in a 1L volumetric flask with deionized water, and adjust the pH of the solution to 7 with 0.1M NaOH ), 100g (7%, w/w) trypsin solution, 12mg of trypsin, 180g of water. The pH of the mixed solution was adjusted to 7 with 0.1 M NaOH. At the same time, bile powder (2%, w/v) was added to the solution to make small intestinal fluid.

(4) Fermentation medium for the front part of the large intestine.

1L fermentation medium contains: 4gNaCl, 4gKCl, 1gNaHCO ₃ , 0.6gMgSO ₄ ·H ₂ O, 0.5gKH ₂ PO ₄ , 0.5gK ₂ HPO ₄ , 0.05gCaCl ₂ , 0.005gFeSO ₄ ·7H ₂ O, 1ml Tween 80 and 3ml resin azure solution (0.025%, w/v, anaerobic indicator). The medium was sterilized at 121°C for 15 minutes before use. Then 150 g of human feces were added to the fermentation medium (volunteers with feces usually ate a normal diet, had no digestive diseases, and had not taken antibiotics for at least 3 months).

(5) Fermentation medium for the posterior segment of the large intestine.

1L fermentation medium contains: 4gNaCl, 4gKCl, 1gNaHCO ₃ , 0.6gMgSO ₄ ·H ₂ O, 0.7gL-cysteine HCl·H ₂ O, 0.5gKH ₂ PO ₄ , 0.5gK ₂ HPO ₄ , 0.4g Bile salts, 0.05g CaCl ₂ , 0.005g FeSO ₄ ·7H ₂ O, 1ml Tween 80 and 3ml resin azure solution (0.025%, w/v, anaerobic indicator). The medium was sterilized at 121°C for 15 minutes before use. Then 250 g of human feces were added to the fermentation medium (volunteers with feces usually ate a normal diet, had no digestive diseases, and had not taken antibiotics for at least 3 months).

2. Simulate the process of human digestive fermentation system.

The schematic diagram of the device for simulating the human digestion and fermentation system is shown in Figure 1. The device includes mouth digestion area A, stomach digestion area B, small intestine digestion area C, front large intestine fermentation area D, and rear large intestine fermentation area E. Specifically, it includes sample inlet 1, oral cavity digestion chamber 2, stomach digestion chamber 3, small intestine digestion chamber 4, front large intestine fermentation chamber 5, large intestine fermentation chamber 6, saliva dish 7, gastric juice dish 8, small intestine liquid dish 9, The fermentation culture dish 10 of the front section of the large intestine and the fermentation culture dish 11 of the rear section of the large intestine are the first dialysis chamber 12, the second dialysis chamber 13, the first airtight filter screen 14, the airtight second filter screen 15, and oral digestion Product dish 16 , gastric digestion product dish 17 , small intestine digestion product dish 18 , anaerobic dish 19 for glycolysis product in the front part of the large intestine, anaerobic dish 20 for zymolysis product in the rear part of the large intestine, and a dish 21 for recovering the remaining zymolysis product.

The connection relationship of each component.

Oral digestion area A: The sample inlet 1 is connected to the sampling pipe at the right end of the oral digestion chamber 2 through the connection valve and the diverter valve, and the sampling pipe at the right end of the oral digestion chamber 2 is directly connected to the interior of the oral digestion chamber 2. Simultaneously, a constant temperature water bath circulation system is installed on all outsides of the oral cavity digestion chamber 2. The left end of the oral cavity digestion chamber 2 is connected with the saliva dish 7 through a drainage device and a constant flow pump. The outlet pipe at the left end of the mouth digestion chamber 2 is connected with the mouth digestion product dish 16 through a diverter valve, a connecting valve and a constant flow pump. The middle part of the outlet pipe at the left end of the oral cavity digestion chamber 2 is connected with the sampling pipe at the left end of the stomach digestion chamber 3 through an emptying delivery pipe and a connecting valve.

Stomach digestion area B: the sampling tube at the left end of the stomach digestion chamber 3 is directly connected to the inside of the stomach digestion chamber 3 . The gastric digestion chamber 3 is divided into two sub-chambers, and the outside of the two sub-chambers is equipped with a constant temperature water bath circulation system, and the two sub-chambers are fixedly connected by two fixed valves. The junction of the two sub-chambers is connected with the gastric juice dish 8 through a flow guide and a constant flow pump. The outlet pipe at the right end of the gastric digestion chamber 3 is connected to a conduit through a diverter valve, and the other end of the conduit is connected to a gastric digestion product vessel 17 through a connecting valve and a constant flow pump. The junction of the shunt valve is also connected to the sampling tube at the right end of the small intestine digestion chamber 4 through an emptying delivery tube.

Small intestine digestion area C: the sampling tube at the right end of the small intestine digestion chamber 4 leads directly to the interior of the small intestine digestion chamber 4 . The small intestine digestion chamber 4 is divided into two sub-chambers, and the outside of the two sub-chambers is equipped with a constant temperature water bath circulation system, and the two sub-chambers are fixedly connected by two fixed valves. The junction of the two sub-chambers is connected with the small intestine liquid dish 9 through a flow diverter and a constant flow pump. The outlet pipe at the left end of the small intestine digestion chamber 4 is connected to a conduit through a diverter valve, and the other end of the conduit is connected to the small intestine digestion product vessel 18 through a connection valve and a constant flow pump. The junction of the shunt valve is also connected with the sampling tube at the left end of the fermentation chamber 5 in the front section of the large intestine through an emptying delivery tube.

Front large intestine fermentation area D: The left end of the fermentation chamber 5 at the front of the large intestine has a sample tube that leads directly to the interior of the fermentation chamber 5 at the front large intestine. The fermentation chamber 5 in the front part of the large intestine is divided into two sub-chambers, and the outside of the two sub-chambers is equipped with a constant temperature water bath circulation system, and the two sub-chambers are fixedly connected by two fixed valves. The junction of the two sub-chambers is connected with the fermentation culture vessel 10 of the front section of the large intestine through a flow diverter and a constant flow pump. The outlet pipe at the right end of the fermentation chamber 5 in the front part of the large intestine is connected to one end of the airtight first filter screen 14 through a diverter valve and a conduit, and the other end of the airtight first filter screen 14 is connected to one end of the first dialysis chamber 12 through a conduit, and the first dialysis The other end of the chamber 12 passes through a constant flow pump and an anaerobic dish 19 for the fermentation product of the front section of the large intestine. The junction of the shunt valve is also connected to the right end sampling tube of the rear section of the large intestine fermentation chamber 6 through an emptying delivery tube.

Fermentation area E of the rear section of the large intestine: the sampling tube at the left end of the fermentation chamber 6 of the rear section of the large intestine leads directly to the interior of the fermentation chamber 6 of the rear section of the large intestine. The fermentation chamber 6 in the rear part of the large intestine is divided into two sub-chambers, and the outside of the two sub-chambers is equipped with a constant temperature water bath circulation system, and the two sub-chambers are fixedly connected by two fixed valves. The joint of the two sub-chambers is connected with the zymolysis culture vessel 11 of the rear section of the large intestine through a flow diverter and a constant flow pump. The outlet pipe at the left end of the fermentation chamber 5 in the back section of the large intestine is connected to one end of the second airtight filter screen 15 through a diverter valve and a conduit, and the other end of the airtight second filter screen 15 is connected to one end of the second dialysis chamber 13 through a conduit, and the second dialysis chamber 13 The other end passes through a constant flow pump and the anaerobic dish 20 of the fermentation product of the rear section of the large intestine. The junction of the shunt valve is also connected to the remaining fermentation product recovery vessel 21 through an emptying delivery pipe.

How to use: Soybean protein solution is made into a certain concentration solution (5mg/ml). Add the prepared saliva, gastric juice, small intestinal juice, zymolysis medium for the first part of the large intestine, and zymolysis medium for the back part of the large intestine to the saliva dish 7, the gastric juice dish 8, the small intestine juice dish 9, and the zymolysis culture dish 10 for the first part of the large intestine. , in the fermentation culture dish 11 of the posterior segment of the large intestine. Turn on all constant temperature water bath systems and maintain a constant temperature of 37°C. Turn on all constant flow pumps and diverter valves. The speed of all constant flow pumps was controlled at 5ml/min. The soybean protein solution is added from the sample inlet 1 at the top of the digestion and fermentation system device, and the saliva is added from the saliva dish 7 to the oral digestion chamber 2 through a constant flow pump and a drainage device to perform oral digestion simulation. After 15 minutes of oral digestion, a part of the digested The final product is collected in the oral cavity digestion product dish 16 to obtain the soybean protein oral cavity digestion product, and another part of the digestion product enters the stomach digestion chamber 3 in the stomach digestion zone B along with the emptying delivery pipe. Gastric juice is added from the gastric juice dish 8 into the gastric digestion chamber 3 through a constant flow pump and a drainage device to perform gastric digestion simulation. After 5 hours of gastric digestion, a part of the digested products are collected in the gastric digest product dish 17 to obtain soybean protein gastric digest products. Another part of the digested product enters the small intestine digestion chamber 4 in the small intestine digestion zone C along with the emptying conveying pipe. The small intestine juice is added to the small intestine digestion chamber 4 through the constant flow pump and drainage device from the small intestine liquid dish 9. After the small intestine is digested for 5 hours, part of the digested products are collected in the small intestine digestion product dish 18 to obtain soybean protein small intestine digestion product, and the other part of the digestion product is Enter the fermentation chamber 5 of the front section of the large intestine in the fermentation zone D of the front section of the large intestine along with the emptying delivery tube. The fermentation medium of the front part of the large intestine is added into the fermentation chamber 5 of the front part of the large intestine from the fermentation culture dish 10 of the front part of the large intestine through a constant flow pump and a diverter. The filter screen 14 and the first dialysis chamber 12 are collected in the anaerobic dish 19 for fermentation products in the front section of the large intestine to obtain soybean protein fermentation products in the front section of the large intestine. In the zymolysis chamber 6 of the posterior segment of the large intestine. The zymolysis medium of the rear section of the large intestine is added into the fermentation room 6 of the rear section of the large intestine from the zymolysis culture dish 11 of the rear section of the large intestine through a constant flow pump and a drainage device. The airtight second filter screen 15 and the second dialysis chamber 13 are collected in the anaerobic dish 20 for the fermentation product of the rear section of the large intestine to obtain the fermentation product of the rear section of the large intestine of soybean protein. The solution product is recovered in the dish 21. The simulated digestion and fermentation process was repeated 3 times.

The oral digestion product dish 16 was placed in a boiling water bath (prepared in advance) for 5 minutes to remove the activity of salivary amylase, and then the oral digestion of soybean protein in the oral digestion product dish 16 was detected by liquid phase method (molecular weight change). The gastric digestion product dish 17 was placed in a boiling water bath (prepared in advance) for 5 minutes to remove the activity of pepsin, and then the gastric digestion (molecular weight change) of soybean protein in the gastric digestion product dish 17 was detected by liquid phase method. The small intestine digestion product dish 18 was placed in a boiling water bath (prepared in advance) for 5 minutes to remove the activity of trypsin (including pancreatic amylase and pancreatic lipase) and trypsin, and then the soybean protein in the small intestine digestion product dish 18 was detected by liquid phase method Digestion in the small intestine (molecular weight change). The anaerobic dish 19 of the fermentation product of the front section of the large intestine was placed in a boiling water bath (prepared in advance) for 30 minutes to remove the activity of the remaining flora, and the fermentation product of the fermentation product of the front section of the large intestine of soybean protein in the anaerobic dish 19 of the fermentation product of the front section of the large intestine was detected by the gas phase method. The fermentation product of the posterior large intestine anaerobic dish 20 was placed in a boiling water bath (prepared in advance) for 30 minutes to remove the activity of the remaining flora, and the fermentation of soybean protein in the anaerobic dish 20 of the fermentation product of the posterior large intestine was detected by the gas phase method solution product.

Test results: The soy protein will not be decomposed in oral simulated digestion. After gastric simulated digestion, the molecular weight was significantly reduced, indicating that the soy protein was significantly broken down in human gastric digestion. After simulated digestion in the small intestine, the molecular weight was significantly reduced, indicating that the soybean protein was largely decomposed during digestion in the human small intestine. The soybean protein can produce a certain amount of short-chain fatty acids after being fermented in the front section of the large intestine and the back section of the large intestine, and can increase the amount of acetic acid, propionic acid and butyric acid.

Claims (2)

1. A device for simulating the digestive and glycolysis system of the human body, which is characterized in that it includes a sample inlet (1), an oral cavity digestion chamber (2), a stomach digestion chamber (3), a small intestine digestion chamber (4), and a large intestine fermentation chamber ( 5), rear large intestine fermentation chamber (6), saliva dish (7), gastric juice dish (8), small intestine liquid dish (9), front large intestine fermentation culture dish (10), rear large intestine fermentation medium dish (11), first dialysis chamber (12), second dialysis chamber (13), airtight first filter screen (14), airtight second filter screen (15), mouth digestion product dish (16), stomach Digestion product dish (17), small intestine digestion product dish (18), anaerobic dish for fermentation product of the first part of the large intestine (19), anaerobic dish for fermentation product of the rear part of the large intestine (20), and recovery dish for the remaining fermentation product (21); The sample inlet (1) is connected to the sampling tube at the right end of the oral digestion chamber (2) through the connection valve and the diverter valve, and the sampling tube at the right end of the oral digestion chamber (2) leads directly to the interior of the oral digestion chamber (2); at the same time, the oral digestion chamber (2) The entire outside is equipped with a constant temperature water bath circulation system; the left end of the oral digestive chamber (2) is connected to the saliva dish (7) through a drainage device and a constant flow pump; The pump is connected to the mouth digestion product dish (16); the middle part of the outlet pipe at the left end of the mouth digestion chamber (2) is connected to the left end sample injection pipe of the stomach digestion chamber (3) through an emptying delivery pipe and a connecting valve; The sampling tube at the left end of the stomach digestion chamber (3) leads directly to the inside of the stomach digestion chamber (3); The fixed valve is fixedly connected; the junction of the two compartments is connected to the gastric juice vessel (8) through a drainage device and a constant flow pump; the outlet pipe at the right end of the gastric digestive chamber (3) is connected to a catheter through a diverter valve, and the other end of the catheter is passed through The connection valve and the constant flow pump are connected to the gastric digestion product dish (17); the connection of the diverter valve is also connected to the right end of the small intestine digestion chamber (4) through the emptying delivery pipe; The sampling tube at the right end of the small intestine digestion chamber (4) leads directly to the inside of the small intestine digestion chamber (4); the small intestine digestion chamber (4) is divided into two sub-chambers. The fixed valve is fixedly connected; at the junction of the two sub-chambers, it is connected to the small intestine liquid container (9) through a drainage device and a constant flow pump; the outlet tube at the left end of the digestive chamber of the small intestine (4) is connected to a catheter through a diverter valve, and the other end of the catheter is It is connected to the small intestine digestion product dish (18) through a connection valve and a constant flow pump; at the junction of the diverter valve, it is also connected to the left end of the large intestine fermentation chamber (5) through an emptying delivery pipe; The left end of the fermentation chamber (5) at the left end of the large intestine leads directly to the inside of the fermentation chamber (5); The two sub-chambers are fixedly connected through two fixed valves; at the junction of the two sub-chambers, it is connected to the fermentation culture dish (10) in the front section of the large intestine through a drainage device and a constant flow pump; the outlet pipe at the right end of the fermentation room (5) in the front section of the large intestine Connect one end of the airtight first filter screen (14) through a diverter valve and a conduit, and the other end of the airtight first filter screen (14) is connected to one end of the first dialysis chamber (12) through a conduit, and the first dialysis chamber (12) ) the other end is connected to the anaerobic dish (19) of fermentation product in the front section of the large intestine through a constant flow pump; at the junction of the diverter valve, it is also connected to the sampling tube at the right end of the fermentation chamber (6) in the back section of the large intestine through an emptying delivery tube; The left end of the fermentation chamber (6) at the left end of the large intestine leads directly to the interior of the fermentation chamber (6); the fermentation chamber (6) is divided into two sub-chambers, and the outside of the two sub-chambers is equipped with a constant temperature water bath circulation At the same time, the two sub-chambers are fixedly connected through two fixed valves; at the junction of the two sub-chambers, it is connected with the fermentation culture dish (11) of the rear segment of the large intestine through a drainage device and a constant flow pump; the fermentation room of the rear segment of the large intestine ( 5) The outlet pipe at the left end is connected to one end of the airtight second filter screen (15) through a diverter valve and a catheter, and the other end of the airtight second filter screen (15) is connected to one end of the second dialysis chamber (13) through a catheter, and the second dialysis The other end of the chamber (13) is connected to the anaerobic dish (20) for fermentation products of the rear section of the large intestine through a constant flow pump; at the junction of the diverter valve, it is also connected to the remaining fermentation product recovery dish (21) through an emptying delivery pipe. 2. the using method of the device of simulating human digestive fermentation system described in claim 1, it is characterized in that: Dissolve or disperse the substance to be digested in distilled water at a certain concentration to make a solution or dispersion of the substance to be tested, and prepare saliva, gastric juice, small intestinal juice, zymolysis medium in the front part of the large intestine, and zymolysis medium in the back part of the large intestine. Add them to the saliva dish (7), gastric juice dish (8), small intestine liquid dish (9), fermentation culture dish for the front part of the large intestine (10), and fermentation culture dish for the rear part of the large intestine (11); turn on all constant temperature water baths Keep the system at a constant temperature of 37°C; open all constant-flow pumps and diverter valves; control the speed of all constant-flow pumps at 4-6ml/min; put the solution or dispersion of the analyte from the uppermost sample inlet of the digestion and fermentation system device ( 1) Adding, saliva is added from the saliva dish (7) into the oral digestion chamber (2) through a constant flow pump and a drainage device to simulate oral digestion. After oral digestion for 10-15 minutes, a part of the digested products are collected in the oral digestion product Another part of the digested product enters the gastric digestion chamber (3) in the gastric digestion area (B) along with the emptying delivery tube in the vessel (16); the gastric juice is added from the gastric liquid vessel (8) to the gastric In chamber (3), simulate gastric digestion. After gastric digestion for 5-6 hours, a part of the digested product is collected in the gastric digestion product dish (17), and the other part of the digested product enters the small intestine digestion area (C) along with the emptying delivery tube. In the small intestine digestion chamber (4) in the middle; the small intestine juice is added to the small intestine digestion chamber (4) through the small intestine liquid dish (9) through a constant flow pump and a drainage device. After the small intestine is digested for 5-6 hours, a part of the digested products are collected in the small intestine In the digestion product dish (18), another part of the digestion product enters the fermentation room (5) in the fermentation zone (D) of the fermentation zone of the fermentation zone (D) of the fermentation product of the other part of the digestion product along with the emptying pipe; The base vessel (10) is added to the fermentation room (5) in the front section of the large intestine through a constant flow pump and a drainage device. After 10-12 hours of fermentation in the front section of the large intestine, a part of the products after fermentation pass through the airtight first filter screen (14) and The first dialysis chamber (12) is collected in the anaerobic dish (19) for the fermented product of the front part of the large intestine, and the other part of the fermented product enters the fermented chamber of the latter part of the large intestine in the fermented area of the latter part of the large intestine (E) In (6); the fermentation medium of the posterior large intestine is added from the fermentation culture dish of the posterior large intestine (11) to the fermentation chamber of the posterior large intestine (6) through a constant flow pump and a drainage device, and the fermentation of the posterior large intestine is 10 After -12 hours, a part of the glycolysis product was collected in the anaerobic dish (20) of the glycolysis product in the rear part of the large intestine through the airtight second filter screen (15) and the second dialysis chamber (13), and the other part of the glycolysis product was Empty the conveying tube into the remaining glycolysis product recovery dish (21); repeat the simulated digestion and zymolysis process 3 times; The oral digestion product dish (16) is placed in a boiling water bath for a few minutes to remove the activity of salivary amylase, and then the oral digestion of the test substance in the oral digestion product dish (16) is detected by the liquid phase method; the gastric digestion product dish (17) is Place in boiling water bath for a few minutes to remove the activity of pepsin, and then use liquid phase method to detect the gastric digestion of the test substance in the gastric digestion product dish (17); place the small intestine digestion product dish (18) in boiling water bath for several minutes to remove The activity of pancreatic enzymes, and then use the liquid phase method to detect the small intestinal digestion of the test substance in the small intestine digestion product dish (18); the anaerobic dish (19) of the fermentation product in the first part of the large intestine is in a boiling water bath for 30-40 minutes to remove the remaining bacteria Group activity, and use the gas phase method to detect the fermentation products of the first section of the large intestine in the anaerobic dish (19); the anaerobic dish of the fermentation product of the back section of the large intestine (20) is in a boiling water bath for 30-40 minutes to remove the remaining bacteria Group activity, and gas-phase detection of glycolysis products of the latter part of the large intestine in the anaerobic dish (20).