Equipment and process for extracting active substances of dendrobium huoshanense
阅读说明:本技术 一种霍山米斛活性物质提取设备及其工艺 (Equipment and process for extracting active substances of dendrobium huoshanense ) 是由 李晓晖 于 2019-10-23 设计创作,主要内容包括:本发明公开了一种霍山米斛活性物质提取设备,包括加热罐、一级离心机、脱蛋白罐、二级离心机和层析柱,所述加热罐内安装有隔离网,所述加热罐内开设有循环槽,所述加热罐的下方连接有一级连接管,所述一级连接管与一级离心机连接,所述一级离心机的下方设置有一级收集罐,所述一级离心机与脱蛋白罐连接,所述脱蛋白罐内开设有水浴槽,所述一级收集罐与脱蛋白罐之间连接有二级连接管,所述脱蛋白罐的上端设置有一级外接管。本发明系统化的设备能够良好的实现对米斛中多糖活性物质的提取,流水线的工作方式,有序且有效的进行活性物质的提取,进过多次的过滤,能够去除其中的蛋白质等不溶性纤维素等。(The invention discloses dendrobium huoshanense active substance extraction equipment which comprises a heating tank, a primary centrifugal machine, a deproteinizing tank, a secondary centrifugal machine and a chromatography column, wherein an isolation net is installed in the heating tank, a circulation groove is formed in the heating tank, a primary connecting pipe is connected below the heating tank and is connected with the primary centrifugal machine, a primary collecting tank is arranged below the primary centrifugal machine and is connected with the deproteinizing tank, a water bath tank is formed in the deproteinizing tank, a secondary connecting pipe is connected between the primary collecting tank and the deproteinizing tank, and a primary external connecting pipe is arranged at the upper end of the deproteinizing tank. The systematic equipment can well realize the extraction of polysaccharide active substances in the dendrobium huoshanense, the working mode of a production line, the extraction of the active substances is orderly and effectively carried out, and insoluble cellulose such as protein and the like in the dendrobium huoshanense can be removed after multiple times of filtration.)
1. The utility model provides a Huoshan huoshan equipment of drawinging, includes heating jar (1), one-level centrifuge (41), deproteinization jar (6), second grade centrifuge (12) and chromatography post (26), its characterized in that, install separation net (2) in heating jar (1), set up circulation groove (3) in heating jar (1), the below of heating jar (1) is connected with one-level connecting pipe (31), one-level connecting pipe (31) are connected with one-level centrifuge (41), the below of one-level centrifuge (41) is provided with one-level collecting tank (32), one-level centrifuge (41) and deproteinization jar (6) are connected, set up water bath (7) in deproteinization jar (6), be connected with second grade connecting pipe (4) between one-level collecting tank, the lower extreme of deproteinization jar (6) is provided with output tube (8), output tube (8) are connected with second grade centrifuge (12), the below of second grade centrifuge (12) is provided with second grade holding vessel (16), second grade holding vessel (16) are connected with chromatography column (26), be provided with tertiary connecting pipe (25) between collection chamber (17) and chromatography column (26), the below of chromatography column (26) is provided with the partial shipment device, chromatography column (26) intussuseption is filled with chromatography gel (27), install supercharging device in chromatography column (26), the upper end of chromatography column (26) is connected with input tube (29).
2. The dendrobium huoshanense active substance extraction device according to claim 1, wherein each of the primary centrifugal machine (41) and the secondary centrifugal machine (12) comprises a mounting shell (15) and a fixing frame (10), the fixing frame (10) is fixedly connected with the mounting shell (15), a rotating body (14) is rotatably mounted in the mounting shell (15), a first motor (9) is arranged in the fixing frame (10), a driving shaft (11) is fixedly connected with a driving end of the first motor (9), the first motor (9) is connected with the rotating body (14) through the driving shaft (11), an outer outlet is formed in the lower end of the mounting shell (15), an inner bearing (40) is mounted in the outer outlet, an inner outlet is formed in the lower end of the rotating body (14), a sealing column (13) is inserted in the inner outlet, a bottom plate (24) is fixedly connected with the lower end of the sealing column (13), fixed ring (39) has been cup jointed to bottom plate (24) periphery, install outer bearing (38) between fixed ring (39) and bottom plate (24), fixed ring (39) fixedly connected with thread bush (37), thread bush (37) and installation shell (15) threaded connection.
3. The dendrobe active substance extraction equipment as claimed in claim 1, wherein a plurality of collection cavities (17) are arranged in each of the secondary collection tank (16) and the primary collection tank (32), each collection cavity (17) is communicated with an output end (18), a base (19) is rotatably connected to the bottoms of the secondary collection tank (16) and the primary collection tank (32), a second motor (20) is installed in each base (19), and the driving ends of the two second motors (20) are fixedly connected with the secondary collection tank (16) and the primary collection tank (32) respectively.
4. The dendrobe active substance extracting equipment as claimed in claim 1, wherein the split charging device comprises an outer fixed box (33) and a mounting seat (34), the outer fixed box (33) is fixedly connected to the upper side of the mounting seat (34), an inner rotating box (36) is rotatably mounted in the outer fixed box (33), and a third motor (35) is mounted in the mounting seat (34).
5. The dendrobium huoshanense active substance extraction equipment according to claim 1, wherein the pressurization device comprises a pressure block (23) and a rotating plate (22), the pressure block (23) is in sliding connection with the inner wall of the chromatography column (26), the rotating plate (22) is rotatably installed in the pressure block (23), a plurality of fan-shaped openings (21) are formed in the pressure block (23), a plurality of sealing leaves corresponding to the fan-shaped openings (21) are arranged on the rotating plate (22), a moving rod (28) is fixedly connected to the upper portion of the rotating plate (22), the moving rod (28) penetrates through the pressure block (23) and the chromatography column (26), a pressure connecting pipe (30) is connected to one side of the chromatography column (26), and a limit ring is arranged in the chromatography column (26).
6. The process for extracting the active substances of dendrobium huoshanense is characterized by comprising the following steps:
s1: weighing fresh dendrobium huoshanense material; adding distilled water into an isolation net (2) of a heating tank (1) according to a material ratio of 1:3(g: ml), heating at 80 ℃, extracting filtrate once every 2h for three times, combining extracting solutions, placing the extracting solutions in an evaporator for concentration, and centrifuging by a primary centrifuge (41) at 12000rpm to remove impurities;
s2: deproteinizing the polysaccharide; introducing the extract into a deproteinization tank (6), adding a trypsin solution, carrying out enzymolysis for 5h in a constant-temperature water bath at 37 ℃, adding a 0.5% TCA solution until the optimum concentration is reached, standing overnight, centrifuging at 12000rpm of a secondary centrifuge (12) to remove protein precipitates, taking supernate, adjusting the pH of the supernate to be neutral by using NaOH, carrying out evaporation concentration, adding 95% ethanol (the ethanol is the extract: 3:16), precipitating overnight to obtain polysaccharide, collecting precipitates, dissolving the precipitates by using distilled water, repeatedly precipitating to obtain white polysaccharide precipitates, continuously centrifuging the secondary centrifuge (12), collecting the precipitates, freezing and drying, and storing in a dryer;
s3: separating by DEAE-cellulose column chromatography;dissolving the precipitate in S2, dissolving polysaccharide solution in double distilled water, filtering insoluble impurities, separating the filtered solution with chromatography column (26) by DEAE-cellulose column chromatography; eluting with double distilled water at 5 ml/min-1And collecting by sub-packaging device to obtain 5 peak values, concentrating each main peak to appropriate volume, dialyzing for desalting, and cooling for drying.
Technical Field
The invention relates to the technical field of active substance extraction, in particular to equipment and a process for extracting active substances of dendrobium huoshanense.
Background
Researches find that the polysaccharide of dendrobium huoshanense has the effects of resisting oxidation, tumors and cataract, improving the immunity of the organism and the like, and the polysaccharide is an active ingredient with higher content in dendrobium huoshanense.
The dendrobium huoshanense is used as a traditional Chinese medicinal material for a long time, but is well known and accepted by people in recent years, the dendrobium huoshanense is researched, is mostly eaten as a medicinal material or food, is eaten by branches and trunks in a use mode, and is directly used by retaining the whole dendrobium huoshanense no matter being dried or directly eaten, but the mode is an indirect use mode for users, needs to depend on consumption of own functions at a little point, and the active ingredients are utilized in a single mode, so that the gastrointestinal absorption of the human body to plants is limited, for example, cellulose in the plants can not be directly digested and absorbed by the human body, so that the absorption of the direct eating to nutrient substances is greatly reduced, and the cellulose is really a kind of polysaccharide.
Disclosure of Invention
The invention aims to solve the defects in the prior art, such as: the gastrointestinal tract of the human body has limited absorption to plants, such as cellulose in the plants, and the human body cannot directly digest and absorb the plants, so the equipment and the process for extracting the dendrobium huoshanense active substances are provided.
In order to achieve the purpose, the invention adopts the following technical scheme:
an equipment for extracting dendrobium huoshanense active substances comprises a heating tank, a primary centrifuge, a deproteinizing tank, a secondary centrifuge and a chromatography column, wherein an isolation net is installed in the heating tank, a circulation groove is formed in the heating tank, a primary connecting pipe is connected below the heating tank and is connected with the primary centrifuge, a primary collecting tank is arranged below the primary centrifuge, the primary centrifuge is connected with the deproteinizing tank, a water bath tank is formed in the deproteinizing tank, a secondary connecting pipe is connected between the primary collecting tank and the deproteinizing tank, a primary external pipe is arranged at the upper end of the deproteinizing tank, an output pipe is arranged at the lower end of the deproteinizing tank and is connected with the secondary centrifuge, a secondary collecting tank is arranged below the secondary centrifuge and is connected with the chromatography column, a tertiary connecting pipe is arranged between the collecting chamber and the chromatography column, the device comprises a chromatographic column and is characterized in that a split charging device is arranged below the chromatographic column, chromatographic gel is filled in the chromatographic column, a pressurizing device is installed in the chromatographic column, and the upper end of the chromatographic column is connected with an input pipe.
Preferably, one-level centrifuge and second grade centrifuge all include installation shell and mount, mount and installation shell fixed connection, the rotator is installed to the installation shell internal rotation, be provided with first motor in the mount, the drive end fixedly connected with drive shaft of first motor, first motor passes through the drive shaft and is connected with the rotator, the lower extreme of installation shell is provided with outer export, install the inner bearing in the export, the export in has been seted up to the lower extreme of rotator, inner export is inserted and is equipped with sealed post, the lower extreme fixedly connected with bottom plate of sealed post, solid fixed ring has been cup jointed to the bottom plate periphery, install the outer bearing between solid fixed ring and the bottom plate, gu fixed ring fixedly connected with thread bush, thread bush and installation shell threaded connection.
Preferably, all be provided with a plurality of collection chambeies in second grade collection tank and the one-level collection tank, every the collection chamber all communicates there is the output, the bottom of second grade collection tank and one-level collection tank is rotated and is connected with the base, install the second motor in the base, two the drive end of second motor respectively with second grade collection tank, one-level collection tank fixed connection.
Preferably, the partial shipment device includes external fixation case and mount pad, external fixation case fixed connection is at the upside of mount pad, rotatory case in the external fixation incasement rotation is installed, install the third motor in the mount pad.
Preferably, supercharging device includes pressure block and commentaries on classics board, the inner wall sliding connection of pressure block and chromatography post, it rotates and installs in the pressure block to change the board, a plurality of fan-shaped openings have been seted up on the pressure block, it is provided with a plurality of sealed leaves that correspond with fan-shaped opening to change the board, the top fixedly connected with carriage release lever of changeing the board, the carriage release lever passes pressure block and chromatography post, one side of chromatography post is connected with the pressure pipe, be provided with the spacing ring in the chromatography post.
A technology for extracting dendrobium huoshanense active substances comprises the following steps:
s1: weighing fresh dendrobium huoshanense material; adding distilled water into an isolation net of a heating tank according to a material ratio of 1:3(g: ml), heating at 80 ℃, extracting filtrate once every 2 hours for three times, combining the extracting solutions, placing in an evaporator for concentration, and centrifuging by a primary centrifuge at 12000rpm to remove impurities;
s2: deproteinizing the polysaccharide; introducing the extract into a deproteinization tank, adding a trypsin solution, carrying out enzymolysis for 5h in a constant-temperature water bath at 37 ℃, adding a 0.5% TCA solution until the optimum concentration is reached, standing overnight, centrifuging at 12000rpm of a secondary centrifuge to remove protein precipitates, taking supernate, regulating the pH of the supernate to be neutral by using NaOH, carrying out evaporation concentration, adding 95% ethanol (the extract: ethanol is 3:16), carrying out overnight precipitation to obtain polysaccharide, collecting precipitates, dissolving the precipitates with distilled water, repeatedly precipitating to obtain white polysaccharide precipitates, continuously centrifuging by using the secondary centrifuge to collect the precipitates, carrying out freeze drying, and storing in a dryer;
s3: separating by DEAE-cellulose column chromatography; dissolving the precipitate in S2, dissolving polysaccharide solution in double distilled water, filtering insoluble impurities, and separating the filtered solution by DEAE-cellulose column chromatography using chromatographic column; eluting with double distilled water at 5 ml/min-1And collecting by sub-packaging device to obtain 5 peak values, concentrating each main peak to appropriate volume, dialyzing for desalting, and cooling for drying.
Compared with the prior art, the invention has the beneficial effects that:
1. the extraction of polysaccharide active substances in the dendrobium huoshanense can be well realized by the aid of systematic equipment, the active substances are orderly and effectively extracted in a production line working mode, and insoluble cellulose such as protein and the like in the dendrobium huoshanense can be removed through multiple times of filtration.
2. Quantitatively controlling reactants added in the extracting solution, avoiding the loss of polysaccharide as much as possible, removing protein in the extracting solution to the maximum extent, ensuring the purity of the polysaccharide separated and extracted, and separating the polysaccharide according to the molecular mass by a chromatography.
3. According to the amplitude of free movement of different molecular masses, the chromatographic method classifies the separated polysaccharides according to the molecular masses, the polysaccharides are sequentially output from large to small according to the molecular masses, and the polysaccharides can be separated and purified while being extracted.
Drawings
FIG. 1 is a schematic structural diagram of an apparatus for extracting active substances from Dendrobium huoshanense according to the present invention;
FIG. 2 is a schematic structural diagram of a pressurizing device of an active material extraction apparatus for dendrobium huoshanense according to the present invention;
FIG. 3 is a schematic view of the structure at A in FIG. 1;
FIG. 4 is a schematic view of the structure at B in FIG. 1;
fig. 5 is a schematic structural diagram at C in fig. 1.
In the figure: 1 heating tank, 2 isolating net, 3 circulating tank, 4 second grade connecting pipe, 5 first grade external connecting pipe, 6 deproteinizing tank, 7 water bath tank, 8 output pipe, 9 first motor, 10 fixed mount, 11 driving shaft, 12 second grade centrifuge, 13 sealing column, 14 rotator, 15 mounting shell, 16 second grade collecting tank, 17 collecting cavity, 18 output end, 19 base, 20 second motor, 21 sector opening, 22 rotating plate, 23 pressure block, 24 bottom plate, 25 third grade connecting pipe, 26 chromatography column, 27 chromatography gel, 28 moving rod, 29 input pipe, 30 through pressure pipe, 31 first grade connecting pipe, 32 first grade collecting tank, 33 external fixed box, 34 mounting seat, 35 third motor, 36 internal rotating box, 37 screw sleeve, 38 external bearing,
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.
In the description of the present invention, it is to be understood that the terms "upper", "lower", "front", "rear", "left", "right", "top", "bottom", "inner", "outer", and the like, indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, are merely for convenience in describing the present invention and simplifying the description, and do not indicate or imply that the device or element being referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus, should not be construed as limiting the present invention.
Referring to fig. 1-5, an apparatus for extracting active substances from dendrobium huoshanense and a process thereof are as follows:
an active substance extraction device of dendrobium huoshanense comprises a
the primary
wherein, a plurality of collecting cavities 17 are arranged in the
S1: weighing fresh dendrobium huoshanense material; adding distilled water into an
disconnecting the
The centrifugation process is that the first motor 9 on the first-
S2: deproteinizing the polysaccharide; introducing the extract into a deproteinizing
s3: separating by DEAE-cellulose column chromatography; dissolving the precipitate in S2, dissolving the polysaccharide solution in double distilled water, and separating the dissolved solution by DEAE-cellulose column chromatography using
finally, inputting the solution with dissolved precipitates into a
when the solution passes through the
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
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