CN106267891B - Method for rapid separation and purification of compound - Google Patents
Method for rapid separation and purification of compound Download PDFInfo
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- CN106267891B CN106267891B CN201510298605.8A CN201510298605A CN106267891B CN 106267891 B CN106267891 B CN 106267891B CN 201510298605 A CN201510298605 A CN 201510298605A CN 106267891 B CN106267891 B CN 106267891B
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- 238000000926 separation method Methods 0.000 title claims abstract description 49
- 238000000746 purification Methods 0.000 title claims abstract description 38
- 150000001875 compounds Chemical class 0.000 title claims abstract description 25
- 238000000034 method Methods 0.000 title claims abstract description 19
- 239000000945 filler Substances 0.000 claims abstract description 57
- 238000011068 loading method Methods 0.000 claims abstract description 45
- 239000002904 solvent Substances 0.000 claims abstract description 34
- 238000011049 filling Methods 0.000 claims abstract description 13
- 238000001514 detection method Methods 0.000 claims abstract description 6
- 238000011067 equilibration Methods 0.000 claims abstract description 6
- 238000004587 chromatography analysis Methods 0.000 claims description 15
- 239000007788 liquid Substances 0.000 claims description 12
- 239000002245 particle Substances 0.000 claims description 12
- 239000011148 porous material Substances 0.000 claims description 11
- 238000005086 pumping Methods 0.000 claims description 6
- 238000000605 extraction Methods 0.000 claims description 4
- 230000000903 blocking effect Effects 0.000 claims description 3
- 230000004308 accommodation Effects 0.000 claims description 2
- 239000003480 eluent Substances 0.000 description 15
- 238000012856 packing Methods 0.000 description 10
- 238000010828 elution Methods 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 230000000694 effects Effects 0.000 description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- ZOXJGFHDIHLPTG-UHFFFAOYSA-N Boron Chemical compound [B] ZOXJGFHDIHLPTG-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- CPLXHLVBOLITMK-UHFFFAOYSA-N Magnesium oxide Chemical compound [Mg]=O CPLXHLVBOLITMK-UHFFFAOYSA-N 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 229910052796 boron Inorganic materials 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000010419 fine particle Substances 0.000 description 2
- 238000011010 flushing procedure Methods 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 238000000227 grinding Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- -1 polytetrafluoroethylene Polymers 0.000 description 2
- 229920001343 polytetrafluoroethylene Polymers 0.000 description 2
- 239000004810 polytetrafluoroethylene Substances 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 1
- 239000004952 Polyamide Substances 0.000 description 1
- 239000006004 Quartz sand Substances 0.000 description 1
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000000395 magnesium oxide Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229920002647 polyamide Polymers 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
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- Treatment Of Liquids With Adsorbents In General (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Abstract
The invention provides a rapid separation and purification method of a compound, which utilizes a rapid separation and purification device to perform rapid separation and purification of the compound, and comprises the following steps: sample loading: uniformly adsorbing a sample to be separated on the filler in the sample adding groove; and (3) column loading: filling the second housing with a filler; balance: delivering solvent to the chromatographic unit through the solvent unit for equilibration; sample adding: loading the sample loading groove which is adsorbed with the sample to be separated into a chromatographic unit; eluting: eluting the sample to be separated by utilizing the solvent unit so as to obtain a target compound through separation; and (3) detection: and taking the target compound obtained by separation, and analyzing and detecting. Thereby, a rapid and efficient separation of the sample to be separated can be achieved.
Description
Technical Field
The invention belongs to the field of pharmacy or chemical synthesis, and particularly relates to a rapid separation and purification method of a compound.
Background
Column chromatography belongs to a common separation and purification means in the pharmaceutical or chemical synthesis field, and at present, it is common to separate compounds by using different types of fine fillers. However, when the number of samples to be separated is large, too small filler is easy to cause slower column passing, and too long residence time is easy to cause loss and influence efficiency.
Therefore, the separation and purification method still needs to be further improved.
Disclosure of Invention
The present invention aims to solve, at least to some extent, one of the technical problems existing in the prior art or to at least provide a useful commercial choice. To this end, an object of the present invention is to provide a method for rapid separation and purification of a compound, with which rapid and efficient separation of a sample to be separated can be achieved.
For this reason, in one aspect of the invention, a method for rapid isolation and purification of a compound is provided. According to one embodiment of the invention, the rapid separation and purification of the compound is performed using a rapid separation and purification apparatus, wherein the rapid separation and purification apparatus comprises: a solvent unit, the solvent unit comprising: a first housing defining a solvent accommodating space; the liquid outlet, the liquid outlet is located the lower extreme of first casing, chromatographic unit with solvent unit links to each other, chromatographic unit includes: a second housing defining a filler accommodating space; inlet: the inlet is connected with the liquid outlet and is positioned at the upper end of the second shell; the outlet is positioned at the lower end of the second shell, and the sample adding groove is positioned at the top of the filler accommodating space; and a receiving unit connected to the chromatography unit, the method for rapid separation and purification of the compound comprising: (1) sample loading: uniformly adsorbing a sample to be separated on the filler in the sample adding groove; (2) column loading: filling the second housing with a filler; (3) equilibration: delivering solvent to the chromatographic unit through a solvent unit for equilibration; (4) sample addition: loading the sample loading groove which is adsorbed with the sample to be separated into a chromatographic unit; (4) elution: eluting the sample to be separated by utilizing the solvent unit so as to obtain a target compound through separation; (5) detection: and taking the target compound obtained by separation, and analyzing and detecting.
Therefore, the sample to be separated is uniformly adsorbed in the filler in advance by utilizing the sample loading groove, and simultaneously, the filling column and the balancing process can be synchronously carried out by utilizing the method, so that the conventional operation of filling samples after filling the samples and balancing is avoided, the time is greatly saved, and the efficiency is improved.
According to the embodiment of the invention, the sample loading amount of the sample to be separated in the step (1) is 2-3 times of the volume of the filler in the sample loading groove. Therefore, the loss caused by excessive adsorption of the sample by the filler when the sample loading amount is too small is avoided; and when the sample loading amount is too large, the sample is overloaded, and the sample which is not adsorbed by the filler directly flows out along with the eluent, so that the separation effect is affected and the loss is caused.
According to the embodiment of the invention, the sample adding groove is a movable sample adding groove. Therefore, the sample loading groove is convenient to move out of the chromatographic unit, the sample loading step and the column loading step are synchronously carried out, and the position of the sample loading groove can be adjusted according to the amount of the filler.
According to the embodiment of the invention, the upper surface and the lower surface of the sample adding groove are provided with sample adding holes. Therefore, after the sample to be separated is adsorbed to the filler in the sample adding groove, the sample adding groove is arranged in the second shell of the chromatographic unit, and the sample adding holes on the upper surface and the lower surface of the sample adding groove are convenient for eluting the sample by the eluent, so that the effect of influencing the eluting effect can be avoided by locally flushing the filler by the eluent.
According to the embodiment of the invention, the aperture of the sample adding hole is 1/3-2/3 of the particle size of the filler. Therefore, the filler is not easy to overflow in the sample adding groove, does not block the sample adding hole, and is convenient for the eluent to flow out.
According to an embodiment of the present invention, the rapid separation and purification apparatus further comprises: and a vacuum pumping unit, wherein the vacuum pumping unit comprises: the evacuation flowing back funnel, evacuation flowing back funnel with the export with receiving element links to each other, includes: an extraction opening; and the vacuumizing piece is connected with the vacuumizing hole. Therefore, the vacuumizing unit can be utilized to realize the rapid separation and purification of the compound.
According to the embodiment of the invention, the vacuumizing liquid draining funnel is a detachable vacuumizing liquid draining funnel. Thus, the filling material can be poured out or replaced after the elution is finished.
According to an embodiment of the invention, the chromatography unit further comprises: a screen panel positioned in the filler accommodation space and below the filler for blocking the flow of filler from the outlet, comprising: at least one through hole. Thereby, the column is convenient to be installed.
According to a specific embodiment of the invention, the upper surface of the screen plate is provided with a filter membrane. Therefore, the eluting solution flowing out is prevented from bringing out the filler with extremely fine particle size, so that the filler loss is avoided, and the sample detection is prevented from being influenced.
According to the embodiment of the invention, the pore diameter of the pore on the sieve plate is 1/3-2/3 of the particle size of the filler of the chromatographic device. Thus, too small a pore size is unfavorable for the passage of a large amount of effluent, too large a pore size, and the filler is easy to block the pores.
According to an embodiment of the invention, the solvent unit further comprises a flow rate controller for controlling the outflow rate of the solvent.
According to a specific embodiment of the invention, the flow rate controller is a valve, and the flow rate controller has a flow display. Therefore, the flow rate of the eluent contained in the solvent unit is adjusted and controlled, and the flow rate of the eluent is adjusted according to the requirement, so that the parameter optimization and the data accumulation of the flow rate in the separation and purification process are facilitated, and the optimal separation and purification can be realized.
According to the embodiment of the invention, the outer wall of the hollow cylinder is provided with a longitudinal scale pattern layer. Therefore, the change of the filler height and the dosage of the eluent is convenient to observe, and the quantitative parameter screening of the separation and purification process is realized.
According to an embodiment of the invention, the second housing is a cylinder.
According to an embodiment of the invention, the second housing is made of quartz glass or high boron glass.
According to an embodiment of the invention, the inlet is a grinding port. Thus, the sealing plug with holes is matched with the liquid outlet of the solvent unit.
According to an embodiment of the invention, the first housing is a sphere.
According to an embodiment of the invention, the volume of the solvent unit may be adjusted according to experimental needs. Furthermore, the conventional 500-2000 ml can be selected for the small test experiment, so that the requirements of different amounts of samples to be separated are met.
According to the embodiment of the invention, the first flow rate controller is made of polytetrafluoroethylene material.
The separation and purification device is suitable for dry column packing or wet column packing.
The separation and purification device can be used for separating and purifying medicines, chemical products and intermediates thereof, and the filling material filled in the chromatography unit can be at least one of alumina, silica gel, polyamide, gel, magnesia, activated carbon, macroporous resin, ion exchange resin, cellulose and quartz sand. The experimenter can choose a suitable filler according to the nature of the sample to be isolated and purified.
Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention.
Drawings
Fig. 1 is a schematic structural view of a rapid separation and purification apparatus according to an embodiment of the present invention.
Detailed Description
Embodiments of the present invention are described in detail below, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to like or similar elements or elements having like or similar functions throughout. The embodiments described below by referring to the drawings are illustrative and intended to explain the present invention and should not be construed as limiting the invention.
According to one aspect of the present invention, the present invention provides a rapid separation and purification method, wherein the rapid separation and purification device is used for rapid separation and purification of the compound. The method for rapid separation and purification of the present invention will be described in detail with reference to fig. 1:
according to some embodiments of the invention, the rapid separation and purification apparatus comprises: a solvent unit 100, a chromatography unit 200, a sample addition tank 300 and a receiving unit 400. Wherein the solvent unit 100 further comprises: a first housing 120 and a liquid outlet 110, wherein the first housing defines a solvent receiving space; the liquid outlet 110 is located at the lower end of the first housing 120, the chromatographic unit 200 is connected to the solvent unit 100, and the chromatographic unit 200 further includes: a second housing 210, an inlet 220, and an outlet 230, wherein the second housing 210 defines a packing receiving space; the inlet 220 is connected to the liquid outlet 110 and is located at the upper end of the second housing 210; the outlet 230 is located at the lower end of the second housing 210. The loading well 300 is located at the top of the packing containing space. The receiving unit 400 is connected to the chromatography unit 300, and the method for rapid separation and purification of the compound comprises: (1) sample loading: uniformly adsorbing the sample to be separated on the filler in the sample adding groove 300; (2) column loading: filling the second housing 210 with a filler; (3) equilibration: feeding the solvent to the chromatography unit 200 through the solvent unit 100 for equilibration; (4) sample addition: loading the loading well 300, to which the sample to be separated is adsorbed, into the chromatography unit 200; (4) elution: eluting the sample to be separated by using the solvent unit 100 so as to obtain a target compound through separation; (5) detection: and taking the target compound obtained by separation, and analyzing and detecting.
Therefore, the sample to be separated is uniformly adsorbed in the filler in advance by utilizing the sample loading groove 300, and meanwhile, filler column loading and balancing can be synchronously performed, so that the conventional operation that the sample loading is performed after the prior filling and balancing are avoided, the time is greatly saved, and the efficiency is improved.
According to an embodiment of the present invention, the loading slot 300 is a movable loading slot, i.e., the loading slot 300 can be moved out of the chromatographic unit 200, and the loading slot 300 can be moved longitudinally within the second housing 210. Therefore, the sample loading step and the column loading step are convenient to synchronously carry out. Meanwhile, the sample adding groove 300 can be adaptively adjusted according to the filling height of the filling material in the chromatographic unit 200, and is particularly suitable for the chromatographic unit with larger caliber of the second shell 210.
According to an embodiment of the present invention, the loading well 300 has loading holes 310 on both upper and lower surfaces. Thus, after the sample to be separated is adsorbed to the filler in the loading well 300, the loading well 300 is integrally loaded into the second housing 210 of the chromatography unit 200. The loading holes 310 on the upper and lower surfaces of the loading slot 300 can facilitate the elution of the sample by the eluent, and can prevent the eluent from locally flushing up the filler, thereby affecting the elution effect.
According to the embodiment of the invention, the aperture of the sample addition hole 310 is 1/3 to 2/3 of the particle size of the filler. Thus, when the pore size of the loading well 310 is greater than 2/3 of the filler particle size, the filler is liable to overflow the loading well under the pressure of the eluent; when the pore size of the sample addition hole 310 is smaller than 1/3 of the particle size of the filler, the filler is easy to block the sample addition hole, and the elution is not easy to flow out. The filler is not easy to overflow in the sample adding groove 300 when the pore diameter is 1/3-2/3 of the particle diameter of the filler, and the sample adding hole is not blocked, so that the eluent can flow out conveniently.
According to an embodiment of the present invention, the rapid separation and purification apparatus further comprises: an evacuation unit 500, the evacuation unit 500 comprising: an evacuation drain funnel 510 and an evacuation piece 520, wherein the evacuation drain funnel 510 is connected with the outlet of the chromatography unit 200 and the receiving unit 400, and comprises: an extraction opening 511; the vacuum pumping unit 520 is connected to the pumping port 511. Thus, the evacuation unit 500 can be used to achieve rapid separation and purification of compounds.
According to an embodiment of the present invention, the evacuation drain funnel 510 is a removable evacuation drain funnel. Thus, the filling material can be poured out or replaced after the elution is finished.
According to some embodiments of the invention, the chromatography unit 200 further comprises: a screen 240, the screen 240 being located in the packing receiving space and below the packing for blocking the flow of the packing from the outlet, comprising: at least one through hole. Thereby facilitating column loading and sample effluent eluted from the chromatography unit 200 to flow out through the screen 240.
According to an embodiment of the present invention, the upper surface of the screen plate 240 is provided with a filter membrane. Therefore, the eluting solution flowing out is prevented from bringing out the filler with extremely fine particle size, so that the filler loss is avoided, and the sample detection is prevented from being influenced.
According to the embodiment of the present invention, the aperture of the through-holes on the screen plate 240 is not particularly limited, and may be selected according to different packing properties as long as the separation effect can be ensured.
According to the embodiment of the invention, the aperture of the through holes on the sieve plate 240 is 1/3-2/3 of the particle size of the filler in the chromatographic unit 200. When the pore diameter of the through holes is larger than 2/3 of the particle diameter of the packing, the packing is easy to overflow the sieve plate 240 under the pressure of the eluent; when the aperture of the through hole is smaller than 1/3 of the particle size of the filler, the filler is easy to block the sieve plate, and the effluent is not easy to flow out. The filler is not easy to overflow on the sieve plate 240 when the pore diameter is 1/3-2/3 of the particle diameter of the filler, and the sieve plate is not blocked, so that effluent is convenient to flow out.
According to an embodiment of the present invention, the solvent unit 100 further includes: a flow rate controller 130 for controlling the outflow rate of the solvent. Thereby facilitating control of the rate of solvent outflow.
According to an embodiment of the present invention, the flow rate controller 130 is a valve and has a flow display 131. Therefore, the flow rate of the eluent contained in the solvent unit 100 is adjusted and controlled, and the flow rate of the eluent is adjusted according to the requirement, so that the parameter optimization and the data accumulation of the flow rate in the separation and purification process are facilitated, and the optimal separation and purification can be realized.
According to an embodiment of the present invention, the second housing 210 has a longitudinal scale pattern layer on an outer wall thereof. Therefore, the change of the filler height and the dosage of the eluent is convenient to observe, and the quantitative parameter screening of the separation and purification process is realized.
According to an embodiment of the present invention, the second housing 210 is a cylinder.
According to some embodiments of the present invention, the second housing 210 may be made of quartz glass or high boron glass.
According to an embodiment of the present invention, the inlet is a grinding port, and is matched with the liquid outlet of the solvent unit 100 through a sealing plug with a hole.
According to an embodiment of the present invention, the first housing 120 is spherical.
According to an embodiment of the present invention, the volume of the first housing 120 may be adjusted according to experimental needs. Furthermore, the conventional 500-2000 ml can be selected for the small test experiment, so that the requirements of different amounts of samples to be separated are met.
According to an embodiment of the present invention, the flow rate controller 130 may be made of polytetrafluoroethylene.
In the description of the present invention, it should be understood that the terms "first," "second," and the like are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include one or more such feature. In the description of the present invention, the meaning of "a plurality" is two or more, unless explicitly defined otherwise.
In the description of the present specification, a description referring to terms "one embodiment," "some embodiments," "examples," "specific examples," or "some examples," etc., means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms are not necessarily directed to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, the different embodiments or examples described in this specification and the features of the different embodiments or examples may be combined and combined by those skilled in the art without contradiction.
While embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and not to be construed as limiting the invention, and that variations, modifications, alternatives and variations may be made to the above embodiments by one of ordinary skill in the art within the scope of the invention.
Claims (5)
1. A rapid separation and purification method of a compound is characterized in that the rapid separation and purification of the compound is performed by a rapid separation and purification device, wherein,
the rapid separation and purification device comprises:
a solvent unit, the solvent unit comprising:
a first housing defining a solvent accommodating space;
the liquid outlet is positioned at the lower end of the first shell,
a chromatography unit connected to the solvent unit, the chromatography unit comprising:
a second housing defining a filler accommodating space;
the inlet is connected with the liquid outlet and is positioned at the upper end of the second shell;
an outlet located at a lower end of the second housing,
the sample adding groove is positioned at the top of the filler accommodating space and is a movable sample adding groove, the upper surface and the lower surface of the sample adding groove are provided with sample adding holes, and the aperture of each sample adding hole is 1/3-2/3 of the particle size of the filler; and
a receiving unit connected with the chromatography unit,
the method for rapidly separating and purifying the compound comprises the following steps:
(1) Sample loading: uniformly adsorbing a sample to be separated on the filler in the sample adding groove;
(2) And (3) column loading: filling the second housing with a filler;
(3) Balance: delivering solvent to the chromatographic unit through a solvent unit for equilibration;
(4) Sample adding: loading the loaded sample slot into the chromatographic unit;
(5) Eluting: eluting the sample to be separated by utilizing the solvent unit so as to obtain a target compound through separation;
(6) And (3) detection: taking the target compound obtained by separation, and analyzing and detecting;
in the step (1), the sample loading amount of the sample to be separated is 2-3 times of the volume of the filler in the sample loading groove.
2. The method of claim 1, wherein the rapid separation and purification apparatus further comprises: a vacuum-pumping unit is used for pumping the air,
wherein,,
the vacuumizing unit comprises:
the evacuation flowing back funnel, evacuation flowing back funnel with the export with receiving element links to each other, includes: an extraction opening; and
and the vacuumizing piece is connected with the air extraction opening.
3. The method of claim 2, wherein the evacuation drain funnel is a removable evacuation drain funnel.
4. The method of claim 1, wherein the chromatography unit further comprises:
a screen panel positioned in the filler accommodation space and below the filler for blocking the flow of filler from the outlet, comprising: at least one through hole.
5. The method according to claim 4, wherein the pore diameter of the through-hole is 1/3 to 2/3 of the particle diameter of the filler.
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