CN109738550B - Continuous purification experimental device - Google Patents
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- CN109738550B CN109738550B CN201910138582.2A CN201910138582A CN109738550B CN 109738550 B CN109738550 B CN 109738550B CN 201910138582 A CN201910138582 A CN 201910138582A CN 109738550 B CN109738550 B CN 109738550B
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Abstract
The invention relates to the field of chromatographic purification, in particular to a continuous purification experimental device. The chromatographic separation and purification device comprises a sample conveying pipeline, a plurality of groups of chromatographic separation and purification modules and sample outlet pipelines, wherein the sample conveying pipeline comprises a sample conveying main pipeline and sample conveying branches, the sample conveying ends of the chromatographic separation and purification modules are connected with the sample conveying main pipeline, the sample outlet ends of the chromatographic separation and purification modules are connected with sample outlet pipelines, control valves III are arranged on the sample outlet pipelines, one ends of the sample conveying pipelines are connected with samples to be tested, the sample conveying pipelines are respectively connected with the sample conveying main pipeline of the chromatographic separation and purification modules through the control valves I, a plurality of sample conveying branches are respectively and correspondingly arranged on each chromatographic separation and purification module, one ends of the sample conveying branches are connected with the sample conveying main pipeline of the corresponding chromatographic separation and purification module, the other ends of the sample conveying branches are connected with the sample conveying branches and the sample outlet pipelines of the other chromatographic separation and purification modules, the connecting parts of the sample conveying branches and the sample outlet pipelines are located at the inlet positions of the control valves, and the sample conveying branches are all provided with control valves II. Is suitable for research and analysis of all the current liquid chromatography separation and purification preparation processes.
Description
Technical Field
The invention relates to the field of chromatographic purification, in particular to a continuous purification experimental device.
Background
Currently, chromatographic separation and purification technology has become one of the most dominant separation and purification technologies, and has been developed from linear chromatography to nonlinear chromatography in theory from analytical experimental chromatography to preparative chromatography and mass production from the beginning of the 20 th century.
Liquid preparative chromatography is not a simple amplification of analytical chromatography, and there are many differences between them. The main factors considered in liquid phase preparative chromatography are the purity of the target product, yield, production cycle, running cost, etc. And the optimization of the preparation process is a precondition for ensuring that the equipment can achieve the expected effect. The simulation and study of the manufacturing process before production of the manufacturing type equipment is important.
The preparation process mainly comprises column structure of chromatographic column, packing, column packing method, pressure and flow rate of mobile phase in chromatographic column. The simulated chromatography preparation process can enable people to know and find chromatographic separation and purification processes and possible problems in actual production more deeply and intuitively, and further optimize the preparation process, so that development time is saved, solvents and stationary phases are selected, the preparation process is more stable and durable, cost is reduced, and equipment color marketization process is accelerated.
In the early stage of realizing large-scale preparation type chromatographic purification equipment, most of workers determine the preparation process according to experience and simple experiments, and the obtained result has larger error, long time consumption and high cost. Most of the equipment manufacturers for preparing the simulation research equipment for the preparation process in the later stage are specific experimental equipment for researching the target conforming to the self-family equipment process, the function of the equipment is relatively single, the chromatographic separation and purification process of a certain or single class of substances can only be fixedly researched, and the research on the general preparation process can not be realized.
Disclosure of Invention
The invention aims to solve the problems in the prior art, and provides a continuous purification experimental device which is suitable for research and analysis of all the existing liquid chromatography separation and purification preparation processes.
The technical scheme of the invention is as follows:
the utility model provides a continuous purification experimental apparatus, wherein, including the sample delivery pipeline, advance the pipeline, array chromatographic separation purification module and go out the appearance pipeline, advance the pipeline and advance the appearance branch road including advancing the appearance main road and advance the appearance branch road, the advance end of chromatographic separation purification module is connected with advancing the appearance main road, the play appearance end of chromatographic separation purification module is connected with the play sampling tube way, it is equipped with control valve III to go out the sampling tube way, the one end of sample delivery pipeline is connected with the sample to be surveyed, it is connected with the appearance main road of chromatographic separation purification module respectively to advance the sampling tube way through control valve I, each chromatographic separation purification module all corresponds and sets up a plurality of and advances the appearance branch road, advance the one end of appearance branch road and advance the appearance main road of corresponding chromatographic separation purification module and be connected, advance the other ends of appearance branch road and advance the appearance branch road and go out the appearance branch road and be located the entrance of control valve of appearance pipeline, advance and all be equipped with control valve II on the appearance branch road.
The chromatographic separation and purification module comprises a chromatographic column, the chromatographic column is filled with a filler, the chromatographic separation and purification module is placed in an incubator, the incubator is respectively connected with a warm air control supply system and a temperature control system, and the temperature control system is electrically connected with the warm air control supply system.
And a pipeline connected with the sample outlet end of the chromatographic separation and purification module and the corresponding sample inlet branch is connected with the comprehensive sample bottle through a control valve IV.
Preferably, the invention comprises six groups of chromatographic separation and purification modules, including chromatographic column A, chromatographic column B, chromatographic column C, chromatographic column D, chromatographic column E and chromatographic column F, wherein the sample injection branch of the chromatographic column A comprises a branch AB, a branch AC, a branch AD, a branch AE and a branch AF which are arranged in parallel, the sample injection branch of the chromatographic column B comprises a branch BA, a branch BC, a branch BD, a branch BE and a branch BF which are arranged in parallel, the sample injection branch of the chromatographic column C comprises a branch CA, a branch CB, a branch CD, a branch CE and a branch CF which are arranged in parallel, the sample injection branch of the chromatographic column D comprises a branch DA, a branch DB, a branch DC, a branch DE and a branch DF which are arranged in parallel, and the sample injection branch of the chromatographic column E comprises a branch EA, a branch EB, a branch EC, a branch ED and a branch EF which are arranged in parallel;
the branches AB, CB, DB, EB and FB are communicated with each other, the branches AC, BC, DC, EC and FC are communicated with each other, the branches AD, BD, CD, ED and FD are communicated with each other, the branches AE, BE, CE, DE and FE are communicated with each other, the branches AF, BF, CF, DF and EF are communicated with each other, and the branches BA, CA, DA, EA and FA are communicated with each other;
the sample outlet end of the chromatographic column A is communicated with the branch BA, the branch CA, the branch DA, the branch EA and the branch FA, the sample outlet end of the chromatographic column B is communicated with the branch AB, the branch DB, the branch EB and the branch FB, the sample outlet end of the chromatographic column C is communicated with the branch AC, the branch BC, the branch DC, the branch EC and the branch FC, the sample outlet end of the chromatographic column D is communicated with the branch AD, the branch BD, the branch CD, the branch ED and the branch FD, the sample outlet end of the chromatographic column E is communicated with the branch AE, the branch BE, the branch CE, the branch DE and the branch FE, and the sample outlet end of the chromatographic column F is communicated with the branch AF, the branch BF, the branch CF, the branch DF and the branch EF.
The sample outlet end of the chromatographic column A is connected with a pipeline connected with the branch BA, the sample outlet end of the chromatographic column B is connected with the branch AB, the sample outlet end of the chromatographic column C is connected with the branch AC, the sample outlet end of the chromatographic column D is connected with the branch AD, the sample outlet end of the chromatographic column E is connected with the branch AE, and the sample outlet end of the chromatographic column F is connected with the branch AF through a control valve IV respectively.
The invention has the beneficial effects that:
(1) A plurality of groups of chromatographic separation and purification modules are arranged, each group of modules are equivalent in position, the working states of the modules can be controlled at will, and sequential control is not needed;
(2) The sample and the mobile phase can be independently sampled, so that a plurality of feasibility schemes are provided for the research of the preparation process, meanwhile, the sample can be switched among any modules, an operator can randomly select and set the column structure of the chromatographic column, the packing, the column filling mode method, the pressure and the flow of the mobile phase when the mobile phase is communicated with the chromatographic column according to experimental requirements, complicated operation steps are not needed, a large amount of experimental time is saved, and accurate experimental data are obtained;
(3) Each chromatographic separation and purification module can be used for outputting samples independently or outputting samples of comprehensive samples through branches, and the specific sample outputting mode can be determined according to experimental purposes and experimental methods set by experimental staff;
(4) The intelligent degree is high, the experimental result is accurate, and meanwhile, an experimental instrument cannot be replaced due to the change of the type of an experimental sample, so that the cost is reduced;
(5) The purification effect can be monitored in real time according to the setting.
Drawings
Fig. 1 is a schematic structural view of the present invention in embodiment 1.
In the figure: 1, a sample bottle A to be tested; 2 is sampled by a sample bottle B;3, a sample feeding pump A;4, a sample feeding pump B;5 sample bottle A;6 sample bottle B;7 sample bottle C;8 sample bottles D;9 sample bottles E;10 sample bottles F;11, a comprehensive sample bottle; a warm air control supply system; 13 chromatographic column a;14 chromatographic column B; a 5-column chromatography C; a 16 chromatographic column D;17 chromatographic column E;18 chromatographic column F;19 sample injection branch AB;20 sample injection branches AC;21 sample injection branch AD;22 sample injection branch AE;23 sample injection branch AF;24 control valve IIAB; 25 control valve IIAC; 26 control valve IIAD; 27 control valve IIAE; 28 control valve IIAF; 29 sample injection branch BF;30 sample injection branch BE;31 sample injection branch BD;32 sample injection branch BC;33 sample injection branch BA;34 control valve IIBF; 35 control valve IIBE; 36 control valve IIBD; 37 control valve IIBC; 38 control valve IIBA; 39 sample injection branches CF;40 sample injection branch CE;41 sample injection branch CD;42 sample injection branch CB;43 sample injection branch CA;44 control valve II CF;45 control valve ii CE;46 control valve ii CD;47 control valve II CB;48 control valve IICA; 49 sample injection branches DF;50 sample injection branches DE;51 sample injection branch DC;52 sample injection branch DB;53 sample injection branch DA;54 control valve ii DF;55 control valve IIDE; 56 control valve IIDC; 57 control valve II DB;58 control valve IIDA; 59 sample injection branch EF;60 sample injection branches ED;61 sample injection branch EC;62 sample injection branch EB;63 sample injection branch EA;64 control valve II EF;65 control valve II ED;66 control valve II EC;67 control valve II EB;68 control valve II EA;69 sample injection branch FE;70 sample feeding branch FD;71 sample injection branch FC;72, sampling branch FB;73 sample feeding branch FA;74 control valve II FE;75 control valve II FD;76 control valve II FC;77 control valve IIFB; 78 control valve II FA;79 control valve IAL; 80 control valve iay; 81 control valve Ibl; 82 control valve iby; 83 control valve ICL; 84 control valve icy; 85 control valve idl; 86 control valve IDY; 87 control valve I EL;88 control valve IEY; 89 control valve lfl; 90 control valve ify; 91 control valve IIIA; 92 control valve IIIB; 93 control valve vc; 94 control valve IIID; 95 control valve IIIE; 96 control valve IIIF; 97 control valve IVF; 98 control valve ive; 99 control valve IVD; 100 control valve IVC; 101 control valve IVB; 102 control valve IVA.
Detailed Description
The invention is further described below with reference to the drawings and examples.
Example 1
As shown in fig. 1, the continuous purification experimental device in this embodiment includes two groups of samples to be tested and six groups of chromatographic purification modules, where the two groups of samples to be tested are respectively contained in a sample bottle A1 to be tested and a sample bottle B2 to be tested, the chromatographic purification modules include a chromatographic column a13, a chromatographic column B14, a chromatographic column C15, a chromatographic column D16, a chromatographic column E17 and a chromatographic column F18, respectively, packing is provided in the chromatographic column, and the kinds of packing in the chromatographic column are arbitrarily selected and set by an experimenter according to experimental requirements. The experimental device further comprises a sample feeding pipeline, a sample feeding pipeline and a sample discharging pipeline, and because the experimental device comprises two groups of samples to be tested, two sample feeding pipelines are correspondingly arranged, and one ends of the sample feeding pipelines are respectively communicated with the inside of the sample testing bottle A1 and the inside of the sample testing bottle B2. The sample injection pipeline comprises a sample injection main pipeline and sample injection branches, wherein the sample injection branches are connected with sample injection ends of the chromatographic columns. One end of each chromatographic column of the sample outlet pipeline is connected with the sample outlet end, and the other end is connected with the corresponding sample bottle.
The two sample feeding pipe paths are respectively connected with the sample feeding main paths of the chromatographic columns through the control valve I, and the control valve I is used for controlling whether to provide samples for the chromatographic columns and which samples are provided. The two sample feeding pipe ways are respectively connected with the sample feeding main way of the chromatographic column A through the control valve AL79 and the control valve IAY 80, are respectively connected with the sample feeding main way of the chromatographic column B through the control valve IB 81 and the control valve IB 82, are respectively connected with the sample feeding main way of the chromatographic column C through the control valve ICL 83 and the control valve ICY 84, are respectively connected with the sample feeding main way of the chromatographic column D through the control valve ICL 85 and the control valve ICY 86, are respectively connected with the sample feeding main way of the chromatographic column E through the control valve ICL 87 and the control valve ICY 88, and are respectively connected with the sample feeding main way of the chromatographic column F through the control valve ICL 89 and the control valve ICY 90.
The outlet pipe of each chromatographic column is provided with a control valve III, and the on-off of the sample outlet pipeline is controlled by the control valve III. The outlet pipe of the chromatographic column A is provided with a control valve IIIA 91, the outlet pipe of the chromatographic column B is provided with a control valve IIIB 92, the outlet pipe of the chromatographic column C is provided with a control valve IIIC 93, the outlet pipe of the chromatographic column D is provided with a control valve IIID 94, the outlet pipe of the chromatographic column E is provided with a control valve IIIE 95, and the outlet pipe of the chromatographic column F is provided with a control valve IIIF 96.
Each chromatographic column is provided with a plurality of sampling branches which are arranged in parallel, one end of each sampling branch is connected with a sampling main path of the corresponding chromatographic column, the other end of each sampling branch is connected with sampling branches and sampling pipelines of other chromatographic columns, connection of the chromatographic columns with sampling ends and/or sampling ends of other chromatographic columns is realized, and each sampling branch is provided with a control valve II which controls on-off of the sampling branch. In this embodiment, the sample injection branch of the chromatographic column a13 includes a sample injection branch AB19, a sample injection branch AC20, a sample injection branch AD21, a sample injection branch AE22 and a sample injection branch AF23 which are arranged in parallel, a control valve ii AB24 is arranged on the sample injection branch AB19, a control valve ii AC25 is arranged on the sample injection branch AC20, a control valve ii AD26 is arranged on the sample injection branch AD21, a control valve ii AE27 is arranged on the sample injection branch AE22, and a control valve ii AF28 is arranged on the sample injection branch AF 23.
The sample injection branch of the chromatographic column B14 comprises a sample injection branch BA33, a sample injection branch BC32, a sample injection branch BD31, a sample injection branch BE30 and a sample injection branch BF29 which are arranged in parallel, a control valve IIBA 38 is arranged on the sample injection branch BA33, a control valve IIBC 37 is arranged on the sample injection branch BC32, a control valve IIBD 36 is arranged on the sample injection branch BD31, a control valve IIBE 35 is arranged on the sample injection branch BE30, and a control valve IIBF 34 is arranged on the sample injection branch BF 29.
The sample injection branch of the chromatographic column C15 comprises a sample injection branch CA43, a sample injection branch CB42, a sample injection branch CD41, a sample injection branch CE40 and a sample injection branch CF39 which are arranged in parallel, wherein a control valve II CA48 is arranged on the sample injection branch CA43, a control valve II CB47 is arranged on the sample injection branch CB42, a control valve II CD46 is arranged on the sample injection branch CD41, a control valve II CE45 is arranged on the sample injection branch CE40, and a control valve II CF44 is arranged on the sample injection branch CF 39.
The sample injection branch of the chromatographic column D16 comprises a sample injection branch DA53, a sample injection branch DB52, a sample injection branch DC51, a sample injection branch DE50 and a sample injection branch DF49 which are arranged in parallel, wherein a control valve II DA58 is arranged on the sample injection branch DA53, a control valve II DB57 is arranged on the sample injection branch DB52, a control valve II DC56 is arranged on the sample injection branch DC51, a control valve II DE55 is arranged on the sample injection branch DE50, and a control valve II DF54 is arranged on the sample injection branch DF 49.
The sample injection branch of the chromatographic column E17 comprises a sample injection branch EA63, a sample injection branch EB62, a sample injection branch EC61, a sample injection branch ED60 and a sample injection branch EF59 which are arranged in parallel, wherein a control valve II EA68 is arranged on the sample injection branch EA63, a control valve II EB67 is arranged on the sample injection branch EB62, a control valve II EC66 is arranged on the sample injection branch EC61, a control valve II ED65 is arranged on the sample injection branch ED60, and a control valve II EF64 is arranged on the sample injection branch EF 59.
The sample injection branch of the chromatographic column F18 comprises a sample injection branch FA73, a sample injection branch FB72, a sample injection branch FC71, a sample injection branch FD70 and a sample injection branch FE69 which are arranged in parallel, a control valve II FA78 is arranged on the sample injection branch FA73, a control valve II FB77 is arranged on the sample injection branch FB72, a control valve II FC76 is arranged on the sample injection branch FC71, a control valve II FD75 is arranged on the sample injection branch FD70, and a control valve II FE74 is arranged on the sample injection branch FE 69.
The sample injection branch AB19, the sample injection branch CB42, the sample injection branch DB52, the sample injection branch EB62 and the sample injection branch FB72 are communicated with each other, the sample injection branch AC20, the sample injection branch BC32, the sample injection branch DC51, the sample injection branch EC61 and the sample injection branch FC71 are communicated with each other, the sample injection branch AD21, the sample injection branch BD31, the sample injection branch CD41, the sample injection branch ED60 and the sample injection branch FD70 are communicated with each other, the sample injection branch AE22, the sample injection branch BE30, the sample injection branch CE40, the sample injection branch DE50 and the sample injection branch FE69 are communicated with each other, the sample injection branch AF23, the sample injection branch BF29, the sample injection branch CF39, the sample injection branch DF49 and the sample injection branch EF59 are communicated with each other, and the sample injection branch BA33, the sample injection branch CA43, the sample injection branch DA53, the sample injection branch EA63 and the sample injection branch FA73 are communicated with each other.
At the same time, the sample outlet end of the chromatographic column A13 is communicated with a sample injection branch BA33, a sample injection branch CA43, a sample injection branch DA53, a sample injection branch EA63 and a sample injection branch FA73, the sample outlet end of the chromatographic column B14 is communicated with a sample injection branch AB19, a sample injection branch CB42, a sample injection branch DB52, a sample injection branch EB62 and a sample injection branch FB72, the sample outlet end of the chromatographic column C15 is communicated with a sample injection branch AC20, a sample injection branch BC32, a sample injection branch DC51, a sample injection branch EC61 and a sample injection branch FC71, the sample outlet end of the chromatographic column D16 is communicated with a sample injection branch AD21, a sample injection branch BD31, a sample injection branch CD41, a sample injection branch ED60 and a sample injection branch FD70, the sample outlet end of the chromatographic column E17 is communicated with a sample injection branch AE22, a sample injection branch BE30, a sample injection branch CE40, a sample injection branch DE50 and a sample injection branch FE69, and the sample outlet end of the chromatographic column F19 is communicated with a sample injection branch AF23, a sample injection branch BF29, a sample injection branch CF39 and a sample injection branch DF49 and a sample injection branch EF 59.
Through the arrangement, a group of sample injection and sample discharge management systems are independently formed by each chromatographic column, meanwhile, interoperability, randomness and equivalence are realized among each group of pipeline systems, through the control of the control valve, the working state of any control module can realize sequential flow of the tested sample among the various chromatographic columns, and also can realize non-sequential flow, namely, the tested sample can realize random flow in the whole device under the control action of the control valve.
In addition, as shown in the figure, a pipeline for connecting the sample outlet end of the chromatographic column A13 with the sample inlet branch BA33 is connected with the comprehensive sample bottle 11, and a control valve IVA 102 is arranged on the connecting pipeline; the sample outlet end of the chromatographic column B14 is connected with a pipeline connected with the sample inlet branch AB19 and the comprehensive sample bottle 11, and a control valve IVB 101 is arranged on the connecting pipeline; the sample outlet end of the chromatographic column C15 is connected with a pipeline connected with the branch AC20 and the comprehensive sample bottle 11, and a control valve IVC 100 is arranged on the connecting pipeline; the sample outlet end of the chromatographic column D16 is connected with a pipeline connected with the branch AD21 and the comprehensive sample bottle 11, and a control valve IVD 99 is arranged on the connecting pipeline; the sample outlet end of the chromatographic column E17 is connected with a pipeline connected with the branch AE22 and is connected with the comprehensive sample bottle 11, and a control valve IVA 98 is arranged on the connecting pipeline; the sample outlet end of the chromatographic column F19 is connected with a pipeline connected with the branch AF23 and the comprehensive sample bottle 11, and a control valve IVF 97 is arranged on the connecting pipeline. The valve IV is controlled to be in an off state, so that independent sample output of each chromatographic column is realized; and the valve IV is controlled to be in a connection state, so that the sample output of the comprehensive sample is realized. The specific sample-out method can be determined according to the experimental purpose and the experimental method set by the experimenter.
The device also comprises an incubator, a warm air control supply system 12 and a temperature control system, wherein six groups of chromatographic purification modules in the embodiment are all arranged in the incubator, the incubator is respectively connected with the warm air control supply system and the temperature control system, and meanwhile, the temperature control system is also connected with the warm air control supply system. The warm air control supply system comprises a box body, a plurality of self-temperature-control heating pipes, a fan blade, a connecting shaft, a motor and a heat-supply pipeline, wherein the plurality of self-temperature-control heating pipes and the fan blade are arranged in the box body, a closed space is formed in the box body, the motor is connected with the fan blade through the connecting shaft, one end of the heat-supply pipeline is communicated with the box body, and the other end of the heat-supply pipeline is connected with the incubator. The automatic temperature control heating pipe generates heat when working, the motor drives the fan blade to rotate, the whole box body is filled with the heat generated by the automatic temperature control heating pipe through the fan blade rotating piece, hot air in the box body flows, and the hot air is conveyed into the constant temperature box along the heat conveying pipe.
The temperature control system comprises a temperature sensor and a controller, wherein the temperature sensor is arranged in the incubator and is electrically connected with the controller, and the controller is connected with the self-control temperature heating tube. The temperature sensor transmits the temperature of the incubator detected in real time to the controller, and the controller controls the heating temperature of the self-temperature-control heating tube, so that the constant temperature in the incubator is ensured. The controller adopted in the implementation can be a single chip microcomputer or a control circuit, which are all in the prior art, and are not described in detail herein.
In the invention, the number of chromatographic separation and purification modules is not limited to six groups in the embodiment, and five groups, four groups and at least two groups can be selected according to actual conditions, and more groups can be selected; the sample to be measured is not limited to two kinds in the present embodiment, and more kinds may be selected.
In order to prevent the influence of the external environment on the sample from interfering with the purification purity, each chromatographic column is arranged in an incubator, and the temperature is regulated in real time by a temperature control system, so that each chromatographic column is maintained in the optimal working temperature range. Meanwhile, the constant temperature box can be internally provided with a plurality of chromatographic columns, so that the chromatographic columns are convenient to replace.
The device can realize the repeated purification of the sample, and meanwhile, the device can be switched and regulated between each chromatographic column at will, so that the precision of the result sample is improved, the purification efficiency is improved, and a large amount of working procedure time is saved.
The device provides stable and efficient experimental equipment for researching various feasibility schemes of the preparation process. The experimenter can set experimental parameters at will, select various experimental methods such as stationary phase variety at will, and the like, thus providing a great deal of possibility for the research of the preparation process. The effect of the preparation process is intuitively and accurately reflected through simulating the preparation process, the application range is wide, the purification and separation processes of chemicals, medicines and biological products can be simulated, experimental equipment does not need to be replaced because of the type of a replaced sample, and the early-stage research cost of the equipment is saved. In the following examples, the apparatus enables simulation and experimentation of purification and separation processes for chemicals, pharmaceuticals and biological products.
Example 2
The device provided by the invention is used for simulating a novel continuous separation and purification chromatographic process of L-tryptophan.
Tryptophan, also known as 2-amino-3-indolyl propionic acid, is an aromatic, heterocyclic, nonpolar alpha-amino acid. Is an important essential amino acid for nutrition, and can not be synthesized by human and animals, but can be obtained by food. The method is widely applied to industries such as medical treatment, food, feed additives, agricultural environment monitoring and the like.
By using the device to simulate continuous purification of L-tryptophan, the device is deleted in different stationary phases and process conditions, and a public welfare route which is lower in cost, higher in efficiency and more environment-friendly is developed to replace the traditional purification process.
In the embodiment, 6 groups of chromatographic separation and purification modules are adopted, one chromatographic column with the diameter of 10mm multiplied by 250mm to 5.0 mu m is arranged in each module, one group of sample injection pipelines and one group of sample discharge pipelines are arranged, and meanwhile, 2 high-pressure sample delivery pumps and a plurality of pipelines are arranged.
Because the embodiment only discusses the stationary phase and the influence of the flow rate, pressure, flow rate and temperature on the continuous chromatographic purification of L-tryptophan, 50mmoL/L potassium dihydrogen phosphate and 10% methanol solution are uniformly adopted as the mobile phase.
In this embodiment, the packing in column A is Applexion XA2041-NH4, the packing in column B is Applexion XA3114-S04, the packing in column C is Applexion XA2043, the packing in column D is Applexion XA2014-22, the packing in column E is R & H XAD1600, and the packing in column F is any combination of the above.
The simulation experiment comprises the following steps:
1. unifying flow, flow speed, column temperature and pressure, and adjusting the sample outlet sequence of the chromatographic column.
1. The flow rate is uniformly set to be 1m/min, the flow rate is 500ml/min, the column temperature is 40 ℃, the sample feeding pressure is 20mpa, and the detection wavelength is 278nm.
And (3) 50mmoL/L potassium dihydrogen phosphate, and 10% methanol solution is respectively sent into six groups of chromatographic purification modules by a sample injection pump A and a sample sending pump B, and the six groups of chromatographic purification modules simultaneously perform purification and separation actions, and samples after purification and separation are respectively collected by a sample bottle A5, a sample bottle B6, a sample bottle C7, a sample bottle D8, a sample bottle E9 and a sample bottle F10.
After 6 groups of samples are collected, pH, conductivity and L-tryptophan concentration of the samples are detected uniformly, and the samples are recorded.
2. The flow rate is uniformly set to be 1m/min, the flow rate is 500ml/min, the column temperature is 40 ℃, the sample feeding pressure is 20mpa, and the detection wavelength is 278nm.
The bottom flow liquid of the chromatographic column A enters the chromatographic column B through the feeding branch BA, and the sample bottle B6 collects the sample. The bottom flow liquid of the chromatographic column C enters the chromatographic column D through the feeding branch DC, and a sample bottle D8 collects a sample. The underflow liquid of the chromatographic column E enters the chromatographic column F through the feeding branch FE, and a sample bottle F10 collects a sample.
After the collection of 3 groups of samples is completed, the pH, conductivity and L-tryptophan concentration of the samples are detected uniformly, and the detection is recorded.
3. The flow rate is uniformly set to be 1m/min, the flow rate is 500ml/min, the column temperature is 40 ℃, the sample feeding pressure is 20mpa, and the detection wavelength is 278nm.
Sequentially discharging samples of the chromatographic columns A, B, C, D, E and F, namely, discharging bottom liquid of the chromatographic column A into the chromatographic column B through a feeding branch BA, and discharging bottom liquid of the chromatographic column A into the comprehensive sample bottle 11 through a control valve IVA 102; the bottom liquid of the chromatographic column B enters the chromatographic column C through a feeding branch CB, and the bottom liquid of the chromatographic column B enters the comprehensive sample bottle 11 through a control valve IVB 101; the bottom liquid of the chromatographic column C enters the chromatographic column B through a feeding branch DC, and the bottom liquid of the chromatographic column C enters the comprehensive sample bottle 11 through a control valve IVC 100; the bottom liquid of the chromatographic column D enters the chromatographic column E through the feeding branch EC, and the bottom liquid of the chromatographic column D enters the comprehensive sample bottle 11 through the control valve IVD 99; the bottom liquid of the chromatographic column E enters the chromatographic column F through a feeding branch FE, and the bottom liquid of the chromatographic column E enters the comprehensive sample bottle 11 through a control valve IVE 98; the bottom liquid of the chromatographic column F directly flows into the sample bottle F10, and the bottom liquid of the chromatographic column F enters the comprehensive sample bottle 11 through the control valve IVF 97.
After the collection of 2 groups of samples is completed, the pH, conductivity and L-tryptophan concentration of the samples are detected uniformly, and recorded.
4. In the next experimental step, the experimenter randomly adjusts the sample outlet sequence of the chromatographic column, so as to obtain different samples, and performs concentration detection and recording.
And secondly, according to the experimental result, obtaining an optimal scheme of the sample outlet sequence of the chromatographic column, selecting chromatographic column packing, and changing flow, flow velocity, column temperature and pressure.
1. Setting the flow rate to be 0.5m/min, the flow rate to be 250ml/min, the column temperature to be30 ℃, the sample feeding pressure to be 15mpa, and the detection wavelength to be 278nm. The packing of the chromatographic column is unchanged.
And after the sample is collected, uniformly detecting the pH, the conductivity and the concentration of L-tryptophan of the sample, and recording.
2. Setting the flow rate to be 1m/min, the flow rate to be 500ml/min, the column temperature to be 40 ℃, the sample feeding pressure to be 20mpa, and the detection wavelength to be 278nm. The packing of the chromatographic column is unchanged.
And after the sample is collected, uniformly detecting the pH, the conductivity and the concentration of L-tryptophan of the sample, and recording.
3. Setting the flow rate to be 1.5m/min, the flow rate to be 750ml/min, the column temperature to be 45 ℃, the sample feeding pressure to be 25mpa, and the detection wavelength to be 278nm. The packing of the chromatographic column is unchanged.
And after the sample is collected, uniformly detecting the pH, the conductivity and the concentration of L-tryptophan of the sample, and recording.
4. Setting the flow rate to be 2m/min, the flow rate to be 1000ml/min, the column temperature to be 50 ℃, the sample feeding pressure to be 25mpa, and the detection wavelength to be 278nm. The chromatographic column packing does not change color.
And after the sample is collected, uniformly detecting the pH, the conductivity and the concentration of L-tryptophan of the sample, and recording.
5. The flow rate is uniformly set to 2.5m/min, the flow rate is 1250ml/min, the column temperature is 55 ℃, the sample feeding pressure is 30mpa, and the detection wavelength is 278nm. The packing of the chromatographic column is unchanged.
And after the sample is collected, uniformly detecting the pH, the conductivity and the concentration of L-tryptophan of the sample, and recording.
The technological scheme with lowest cost, highest efficiency and most green environmental protection is obtained through the targeted experiments of parameters such as different flow rates, column temperatures, sample feeding pressure and the like by selecting different chromatographic column fillers.
Example 3
The device provided by the invention is used for simulating a chromatographic process for continuously separating and purifying stevioside components.
The stevioside, also called stevioside, is a mixture of diterpene glycosides containing 10 components extracted from leaves of stevia rebaudiana Bertoni of Compositae, belongs to tetracyclic diterpenoid compounds according to the division of natural phytochemistry, and is a natural sweetener, and has the characteristics of high sweetness, low calorie, safety, no toxicity and the like, and is gradually favored by people. Among 10 kinds of known stevioside, the contents of various components, taste and sweetness are different, wherein the contents of stevioside, rebaudioside a, rebaudioside C are high and account for more than 90%. Rebaudioside a has the highest sweetness about 450 times that of sucrose.
Through using this device simulation to the continuous purification of rebaudioside C, delete in different stationary phases and technological conditions, develop a set of cost lower, efficient, more green public welfare route, replace traditional purification technology.
In the embodiment, 5 groups of chromatographic separation and purification modules are adopted, one chromatographic column with the diameter of 16mm and 500mm-5.0 mu m is arranged in each module, one group of sample injection pipelines and one group of sample discharge pipelines are arranged, and meanwhile, 2 high-pressure sample delivery pumps and a plurality of pipelines are arranged.
Because this embodiment only discusses the influence of different stationary phase fillers, different flow rates, pressures, flow rates, column temperatures, etc. on the continuous chromatographic purification of rebaudioside C components in a rebaudioside extract. The mobile phase is 200ml of stevioside extract, absolute ethyl alcohol and deionized water.
In the embodiment, the filler in the chromatographic column A is macroporous adsorption resin ADS-8, the filler in the chromatographic column B is macroporous adsorption resin ADS-7, the filler in the chromatographic column C is macroporous adsorption resin ADS-17, the filler in the chromatographic column D is macroporous adsorption resin AB-8, and the filler in the chromatographic column E is macroporous adsorption resin D-06.
The simulation experiment comprises the following steps:
1. unifying flow, flow speed, column temperature and pressure, and changing the sample outlet sequence of the chromatographic column packing and the chromatographic column.
1. The flow rate is set to be 2BV/h, the flow rate is 50ml/min, the column temperature is 20 ℃, and the sample feeding pressure is 10mpa.
Filling five chromatographic columns A, B, C, D and E with the above filler, pumping ethanol into the chromatographic columns with a sample pump A at 10Mpa pressure to clean stationary phase until the liquid is no longer turbid by adding water, replacing reagent, and cleaning with deionized water until no ethanol is contained. And (3) pumping the stevioside extract into chromatographic columns by a sample pump B for adsorption, stopping adsorption when the effluent liquid and the stevioside extract are in the same phase, and recording the adsorption saturation time of each chromatographic column.
2. The flow rate is uniformly set to 2BV/h, the flow rate is 50ml/min, the column temperature is 20 ℃, the sample feeding pressure is 10mpa, and the packing in the chromatographic column is arbitrarily adjusted to be any combination of the packing according to an experimenter.
Filling five chromatographic columns A, B, C, D and E with modified filling materials, pumping ethanol into the chromatographic columns by using a sample pump A at 10Mpa pressure to clean the stationary phase until the liquid is not turbid any more by adding water, replacing the reagent, and cleaning the liquid by using deionized water until the liquid no longer contains ethanol. And (3) pumping the stevioside extract into chromatographic columns by a sample pump B for adsorption, stopping adsorption when the effluent liquid and the stevioside extract are in the same phase, and recording the adsorption saturation time of each chromatographic column.
3. The flow rate is set to be 2BV/h, the flow rate is 50ml/min, the column temperature is 20 ℃, and the sample feeding pressure is 10mpa. The packing in the chromatographic column is arbitrarily adjusted to any combination of the above-mentioned packing according to the experimenter.
Filling five chromatographic columns A, B, C, D and E with modified filling materials, pumping ethanol into the chromatographic columns by using a sample pump A at 10Mpa pressure to clean the stationary phase until the liquid is not turbid any more by adding water, replacing the reagent, and cleaning the liquid by using deionized water until the liquid no longer contains ethanol.
The sample-sending pump B pumps the stevioside extract into the chromatographic column for adsorption, and an experimenter can randomly adjust the sample-outputting sequence of the chromatographic column. The sample can be sequentially discharged from A, B, C, D and E, the sample can be sequentially discharged from E, D, C, B and A, the sample can be sequentially discharged from A, D and E, and the method is arbitrary. Stopping adsorption when the effluent liquid and stevioside are in liquid phase, and recording adsorption saturation time of each chromatographic column.
2. According to the experiment, the optimal scheme of the sample outlet sequence of the chromatographic column and the optimal filling mode of the chromatographic column packing are obtained, and the flow, the flow velocity, the column temperature and the pressure are changed.
1. The flow rate is set to be 1BV/h, the flow rate is 30ml/min, the column temperature is 10 ℃, and the sample feeding pressure is 10mpa.
And (3) pumping the stevioside extract into chromatographic columns by a sample pump B for adsorption, stopping adsorption when the effluent liquid and the stevioside extract are in the same phase, and recording the adsorption saturation time of each chromatographic column.
2. The flow rate is set to be 1.5BV/h, the flow rate is 35ml/min, the column temperature is 15 ℃, and the sample feeding pressure is 12mpa.
And (3) pumping the stevioside extract into chromatographic columns by a sample pump B for adsorption, stopping adsorption when the effluent liquid and the stevioside extract are in the same phase, and recording the adsorption saturation time of each chromatographic column.
3. The flow rate is set to be 2BV/h, the flow rate is 40ml/min, the column temperature is 20 ℃, and the sample feeding pressure is 15mpa.
And (3) pumping the stevioside extract into chromatographic columns by a sample pump B for adsorption, stopping adsorption when the effluent liquid and the stevioside extract are in the same phase, and recording the adsorption saturation time of each chromatographic column.
4. The flow rate is set to 2.5BV/h, the flow rate is 45ml/min, the column temperature is 25 ℃, and the sample feeding pressure is 20mpa.
And (3) pumping the stevioside extract into chromatographic columns by a sample pump B for adsorption, stopping adsorption when the effluent liquid and the stevioside extract are in the same phase, and recording the adsorption saturation time of each chromatographic column.
5. The flow rate is set to 3BV/h, the flow rate is 50ml/min, the column temperature is 30 ℃, and the sample feeding pressure is 30mpa.
And (3) pumping the stevioside extract into chromatographic columns by a sample pump B for adsorption, stopping adsorption when the effluent liquid and the stevioside extract are in the same phase, and recording the adsorption saturation time of each chromatographic column.
The technological scheme with lowest cost, highest efficiency, optimal green and environment protection is obtained through the targeted experiments of the parameters such as different chromatographic column packing selection modes, different flow rates, different column temperatures, different sample feeding pressures and the like.
Example 4
The iopamidol is a nonionic water-soluble iodine contrast agent, has good developing effect, small toxicity to vessel walls and nerve tissues, less adverse reaction, stable property and wide application range, and is mainly used for enhanced scanning in neuroradiology, angiography and CT examination, arthroscopy and digital subtraction angiography.
The iopamidol is difficult to separate and purify because the crude product contains more impurities with similar structures. The United states pharmacopoeia and European pharmacopoeia are both severely limited in terms of their content. At present, a macroporous adsorption resin adsorption method is used in the iopamidol production, and the method has the advantages of long production period, low separation column efficiency, low automation degree, poor process stability and unfavorable control of product quality. Meanwhile, a large amount of wastewater is generated, which is not beneficial to environmental protection.
By using the device to simulate continuous purification of lopamidol, the lopamidol is deleted in different stationary phases and process conditions, and a public welfare route which is lower in cost, higher in efficiency and more environment-friendly is developed to replace the traditional purification process.
The experiment adopts 4 groups of chromatographic separation and purification modules, wherein 250mm 4.6mm-5.0 mu m chromatographic columns are arranged in each module, one sample injection pipeline is arranged, one sample discharge pipeline is arranged, and meanwhile, 2 high-pressure sample delivery pumps are arranged, and a plurality of pipelines are arranged.
In this example, the mobile phase was selected from the group consisting of lopamidol feed solution (purity 230 g/L), ultrapure water, and industrial methanol. The stationary phase is C18-1 (13.7% carbon content), C18-2 (11.5% carbon content), C18-3 (9.3% carbon content), C18-4 (8% carbon content). The chromatographic column A is filled with C18-1, B is filled with C18-2, C is filled with C18-3, and D is filled with C18-3.
The simulation experiment comprises the following steps:
1. unifying the flow rate, the column temperature and the sample feeding pressure, and changing the sample outlet sequence of the chromatographic column.
1. The flow rate is set to 60ml/min uniformly, the column temperature is 20 ℃, and the sample feeding pressure is 20mpa.
The mobile phase A is water, the mobile phase B is methanol solution, and the injection volume of the lopamidol feed liquid is 65ml.
And (3) allowing the lopamidol feed liquid to pass through the chromatographic column A, the chromatographic column B, the chromatographic column C and the chromatographic column D simultaneously for 30min, respectively collecting samples at the sample outlet ends of the chromatographic column A, the chromatographic column B, the chromatographic column C and the chromatographic column D, and detecting and comparing the samples.
2. The flow rate is set to 60ml/min uniformly, the column temperature is 20 ℃, and the sample feeding pressure is 20mpa.
The lopamidol liquid firstly enters a chromatographic column B through a branch BA, then enters a chromatographic column C through a branch CB, then enters a chromatographic column D through a branch DC for 120min, and samples at the sample outlet end of the chromatographic column D are collected and detected for comparative analysis.
3. The flow rate is set to 60ml/min uniformly, the column temperature is 20 ℃, and the sample feeding pressure is 20mpa.
The lopamidol liquid firstly enters the chromatographic column C through the branch CD, then enters the chromatographic column B through the branch BC and then enters the chromatographic column A through the branch AB for 120min, and the sample at the sample outlet end of the chromatographic column A is collected and detected for comparative analysis.
4. The flow rate is set to 60ml/min uniformly, the column temperature is 20 ℃, and the sample feeding pressure is 20mpa.
The sequence of the lopamidol liquid through the chromatographic column is arbitrarily adjusted by an experimenter.
5. The flow rate is set to 60ml/min uniformly, the column temperature is 20 ℃, and the sample feeding pressure is 20mpa.
The sequence of the lopamidol liquid through the chromatographic column is arbitrarily adjusted by an experimenter.
6. Uniformly setting the flow rate at 60ml/min, the column temperature at 20 ℃ and the sample feeding pressure at 20mpa
The sequence of the lopamidol liquid through the chromatographic column is arbitrarily adjusted by an experimenter.
The technological scheme with lowest cost, highest efficiency and most green environmental protection is obtained through the comparison experiment of the feed liquid flowing through different stationary phases.
Claims (2)
1. The utility model provides a simulation continuous purification experimental apparatus which characterized in that: the device comprises a sample conveying pipeline, a plurality of groups of chromatographic separation and purification modules and sample outlet pipelines, wherein the sample conveying pipeline comprises a sample conveying main pipeline and sample conveying branches, the sample conveying ends of the chromatographic separation and purification modules are connected with the sample conveying main pipeline, the sample outlet ends of the chromatographic separation and purification modules are connected with sample outlet pipelines, control valves III are arranged on the sample outlet pipelines, one ends of the sample conveying pipelines are connected with samples to be tested, the sample conveying pipelines are respectively connected with the sample conveying main pipeline of the chromatographic separation and purification modules through the control valves I, a plurality of sample conveying branches are correspondingly arranged on each chromatographic separation and purification module, one ends of the sample conveying branches are connected with the sample conveying main pipeline of the corresponding chromatographic separation and purification module, the other ends of the sample conveying branches are connected with the sample conveying branches and the sample outlet pipelines of the other chromatographic separation and purification modules, the connection parts of the sample conveying branches and the sample outlet pipelines are positioned at the inlet parts of the control valves, and the sample conveying branches are all provided with control valves II;
the chromatographic separation and purification module comprises six groups, including a chromatographic column A, a chromatographic column B, a chromatographic column C, a chromatographic column D, a chromatographic column E and a chromatographic column F;
the sample injection branch of the chromatographic column A comprises a branch AB, a branch AC, a branch AD, a branch AE and a branch AF which are arranged in parallel;
the sample injection branch of the chromatographic column B comprises a branch BA, a branch BC, a branch BD, a branch BE and a branch BF which are arranged in parallel;
the sample injection branch of the chromatographic column C comprises a branch CA, a branch CB, a branch CD, a branch CE and a branch CF which are arranged in parallel;
the sample injection branch of the chromatographic column D comprises a branch DA, a branch DB, a branch DC, a branch DE and a branch DF which are arranged in parallel; the sample injection branch of the chromatographic column E comprises a branch EA, a branch EB, a branch EC, a branch ED and a branch EF which are arranged in parallel;
the sample injection branch of the chromatographic column F comprises a branch FA, a branch FB, a branch FC, a branch FD and a branch FE which are arranged in parallel;
the branches AB, CB, DB, EB and FB are communicated with each other, the branches AC, BC, DC, EC and FC are communicated with each other, the branches AD, BD, CD, ED and FD are communicated with each other, the branches AE, BE, CE, DE and FE are communicated with each other, the branches AF, BF, CF, DF and EF are communicated with each other, and the branches BA, CA, DA, EA and FA are communicated with each other;
the sample outlet end of the chromatographic column A is communicated with a branch BA, a branch CA, a branch DA, a branch EA and a branch FA, the sample outlet end of the chromatographic column B is communicated with a branch AB, a branch CB, a branch DB, a branch EB and a branch FB, the sample outlet end of the chromatographic column C is communicated with a branch AC, a branch BC, a branch DC, a branch EC and a branch FC, the sample outlet end of the chromatographic column D is communicated with a branch AD, a branch BD, a branch CD, a branch ED and a branch FD, the sample outlet end of the chromatographic column E is communicated with a branch AE, a branch BE, a branch CE, a branch DE and a branch FE, and the sample outlet end of the chromatographic column F is communicated with a branch AF, a branch BF, a branch CF, a branch DF and a branch EF;
the sample outlet end of the chromatographic column A is connected with a pipeline connected with the branch BA, the sample outlet end of the chromatographic column B is connected with the branch AB, the sample outlet end of the chromatographic column C is connected with the branch AC, the sample outlet end of the chromatographic column D is connected with the branch AD, the sample outlet end of the chromatographic column E is connected with the branch AE, and the sample outlet end of the chromatographic column F is connected with the branch AF through a control valve IV respectively.
2. The simulated continuous purification assay device of claim 1, wherein: the chromatographic separation and purification module comprises a chromatographic column, the chromatographic column is filled with a filler, the chromatographic separation and purification module is placed in an incubator, the incubator is respectively connected with a warm air control supply system and a temperature control system, and the temperature control system is electrically connected with the warm air control supply system.
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