KR101318218B1 - Microcolumn for use in chromatography for separation or detection or quantative analysis of radioactive substances and method for detecting of radioactive substances using the same - Google Patents

Abstract

The present invention relates to a microcolumn tube for chromatography used in radioisotope labeling reaction and quality control of a radioisotope labeling substance and a method for detecting radioactive substance using the same. By using the microcolumn tube of the present invention, it is possible to rapidly develop and analyze radioactive material and to accurately measure the amount of radioactive material by blocking volatilization in the air even when the radioactive material is volatile.

Description

Microcolumn tubes for separation detection or quantitative analysis of radioactive material and methods for detecting radioactive material using the same SAME}

The present invention relates to a microcolumn tube for chromatography used in radioisotope labeling reaction and quality control of a radioisotope labeling substance and a method for detecting radioactive substance using the same.

Chromatography is widely used in various industries as a method for analyzing and separating compounds, and is also useful for analyzing, separating and purifying radioactive materials. Radioactive materials, by their very nature, decay with a half-life due to radionuclides, resulting in a decrease in radiation dose over time. In particular, radionuclides used in nuclear medicine have very short half-lives, which require rapid synthesis, separation and analysis of radiotracers. In general, the chromatographic analysis method used to date is thin-layer chromatography (TLC). Thin membrane chromatography takes about 15 to 50 minutes to develop the radioactive material. This not only increases the radioactive exposure of the experimenter by lengthening the overall manufacturing time, but also is a major cause of lowering the radiochemical yield.

Japanese Patent No. 3136960 discloses that microchannels may be formed in plates used for thin-film chromatography to prevent contamination due to harmful substances and to strongly act on capillary phenomena to improve the development speed. It is not disclosed that microcolumn tubes can be used to effectively detect or analyze radioactive materials in nuclear medicine.

The present inventors completed the present invention by confirming that the chromatography method using a microcolumn tube can not only rapidly analyze radioactive material but also quantitatively measure volatile radioisotope labels.

SUMMARY OF THE INVENTION An object of the present invention is to provide a microcolumn tube for chromatography that allows for the rapid detection and separation or detection of radioactive material.

Another object of the present invention is to provide a method for detecting radioactive substances which can quickly confirm the progress of a reaction during radioisotope labeling reactions and which can be used for quality control of radioisotope labeling compounds.

In order to achieve the above object, the present invention is a microcolumn tube for chromatography for radioactive material separation detection or quantitative analysis, the injection hole for loading the radioactive material; A frit composed of porous solids present at both ends of the microcolumn tube; Solid filler in the form of particles filled between the frits; And it provides a micro-column tube, characterized by being made of a solid tube which can maintain a constant shape.

In addition, the present invention comprises the steps of loading a mixture of the radioactive material in the inlet of the microcolumn tube according to the present invention (step 1); And injecting the developing solvent into the microcolumn tube loaded with the mixed solution of the radioactive material to develop the radioactive material (step 2).

The conventional thin membrane chromatography (TLC) method, which is used for the progress of radioisotope labeling reaction and quality control of radioisotope labeling compounds, takes about 15-50 minutes to develop the compounds. If the radioactive material is a volatile material, the ratio of the actual compound is distorted. By using the microcolumn tube of the present invention, it is possible to rapidly develop and analyze radioactive material and to accurately measure the amount of radioactive material by blocking volatilization in the air even when the radioactive material is volatile.

1 is a view schematically showing the structure of a microcolumn tube of the present invention.
2 is a view briefly showing a method for detecting a radioactive material using the microcolumn tube of the present invention.
3 is a result of radio-TLC analysis using a microcolumn tube according to Table 1 of Example 2 of the present invention (10% ethyl acetate / hexane).
4 is a result of radio-TLC radio-TLC analysis using a microcolumn tube according to Table 1 of Example 2 of the present invention (10% methanol / dichloromethane).
5 is a result of radio-TLC analysis using a microcolumn tube according to Table 2 of Example 2 of the present invention (10% ethyl acetate / hexane).
6 is a result of radio-TLC analysis using a microcolumn tube according to Table 2 of Example 2 of the present invention (10% methanol / dichloromethane).
7 is a result of radio-TLC analysis using a microcolumn tube according to Table 6 of Example 3 of the present invention (50% ethyl acetate / hexane).
8 is a result of radio-TLC analysis using a microcolumn tube according to Table 7 of Example 3 of the present invention (50% ethyl acetate / hexane).
9 is a result of radio-TLC analysis using thin membrane chromatography according to Table 8 of Comparative Example 3 of the present invention (50% ethyl acetate / hexane).

Hereinafter, the present invention will be described in detail.

1 is a view schematically showing the structure of a microcolumn tube of the present invention.

According to the present invention, a microcolumn tube 100 for chromatography for radioactive material separation detection or quantitative analysis, comprising: an injection port (1) for loading radioactive material; Frits (2, 3) composed of porous solids present at both ends of the microcolumn tubes; Solid fills (4) in the form of particles filled between the frits; And it is provided with a fine column tube 100, characterized in that consisting of a solid tube 100 that can maintain a certain form.

The frit at both ends of the microcolumn tube of the present invention serves to pass only the developing solvent without the material filled therebetween, and may be used without particular limitation as long as it is a porous solid. For example, cotton or porous glass, polyethylene, polypropylene may be used, but is not limited thereto.

As the solid filler in the form of particles filled between the frits in the microcolumn tube of the present invention, any filler known to be used for column chromatography may be used. Preferably, silica gel, C4 to C18 silica gel, and aluminum oxide (Al ₂ O) are used. ₃ ), calcium carbonate, activated carbon, diatomaceous earth or cellulose powder may be used, but is not limited thereto.

The microcolumn tube of the present invention can be used for chromatography for radioactive material separation detection or quantitative analysis to quickly and accurately separate and detect radioactive material, and quantitative analysis of volatile radioactive material is also possible due to the sealed structure.

The microcolumn tube of the present invention is composed of a solid tube capable of maintaining a certain shape, and the material of the solid tube may be glass, quartz, plastic, stainless metal, but is not limited thereto. It is preferable that the inner diameter is 0.5-3.0 mm and the length is 40-200 mm. The internal diameter and length of the column are related to the analysis time and the degree of separation, and considering the rapidity and selectivity, the separation efficiency can be maximized within the numerical range.

The present invention also provides a method for detecting radioactive material using the microcolumn tube according to the present invention. Specifically, the present invention comprises the steps of loading a mixture of radioactive material in the inlet of the microcolumn tube according to the present invention (step 1); And injecting the developing solvent into the microcolumn tube loaded with the mixed solution of the radioactive material to develop the radioactive material (step 2).

2 is a view briefly showing a method for detecting a radioactive material using the microcolumn tube of the present invention. In step 1, a mixture solution of two or more radioactive materials is loaded into the inlet of the microcolumn tube according to the present invention, and in step 2, a developing solvent is injected into the microcolumn tube loaded with the mixture solution of radioactive material to expand the radioactive material. Separate the material. The developing solvent according to the present invention is not particularly limited as long as it is present as a liquid at room temperature, preferably hexane, ethyl acetate, dichloromethane, chloroform, diethyl ether, tetrahydrofuran, benzene, toluene, methanol, acetone, Acetonitrile, water and mixed solutions thereof.

In addition, in the method according to the present invention, the developing solvent is moved by using a pressure difference across the microcolumn tube, for example, by applying pressure in a portion loaded with radioactive material or by creating a vacuum in the opposite portion to move the developing solvent.

According to one embodiment of the invention, one end with the microcolumns according to the invention is connected with a rubber bulb and the other end is loaded with a mixture of radioactive material (step 1). Thereafter, while pressing the rubber bulb, the end loaded with the radioactive material may be immersed in the developing solvent, and the radioactive material may be developed while the rubber bulb is opened (step 2).

The separation detection method of the radioactive material of the present invention may further include detecting the radioactive material separated in step 2 by using a radiation detector (step 3). It is preferable to use a radiation-TLC image scanner as the radiation detector.

Hereinafter, the present invention will be described in detail with reference to examples, but these examples are only presented to more clearly understand the present invention, and are not intended to limit the scope of the present invention. It will be determined within the scope of the technical spirit of the claims.

Comparative Example  1: thin membrane Chromatography  Ready

A thin film chromatography plate (200 mm × 200 mm) coated by coating silica gel on an aluminum plate was cut to prepare a thin film chromatography plate having a length of 100 mm and a width of 15 mm ( Comparative Example 1-1 ). Further, a thin film chromatography plate (200 mm × 200 mm) coated by applying C18 silica gel to a glass plate was cut to prepare a thin film chromatography plate having a length of 100 mm and a width of 15 mm ( Comparative Example 1-2 ).

Example  One: Filling  Containing Microcolumn tube  Produce

One end of the microcolumn tube having an inner diameter of 0.8 mm and a length of 90 mm was flattened and closed using a small amount of cotton. Flash silica gel (particle size: 0.040-0.063 mm) ( Example 1-1 ), Gravity silica gel (particle size: 0.063-0.200 mm) ( Example 1-2 ), C18 Silica gel (C-18 silica gel, particle size: 0.040-0.075 ㎜) ( Example 1-3 ) Fill the microcolumn tube with the filling, chopped well for 5 hours using a shaker, and then put a small amount of cotton to flatten it. . At this time, the length of the filler filled in the microcolumn tube was 85 mm.

< Fine column tube  Comparative Experiment of Thin Film Chromatography>

The development experiment of the radioactive material using the microcolumn tube of the present invention was carried out in comparison with the development experiment by the conventional thin film chromatography. After the development experiments, each of them was evaluated for the separation effect of radioactive material through a radiation-TLC scanner. One of the positron emitting isotopes, fluorine-18 (half life 110 minutes), was used for the experimental and comparative evaluation of the present invention, but the present invention is not limited thereto and can be applied universally to all radioisotopes.

Example  2: Fine column tube  Deployment experiment

In Example 1-1 to Example 1-3, a solution in which an aqueous solution of fluorine-18 anion and a 2- (3- [ ¹⁸ F] fluoropropoxy) naphthalene compound solution was mixed was prepared and a portion thereof was taken as a microtube. One end of the prepared microcolumn tube was loaded. The other end of the microcolumn tube was connected to a glass tube with rubber parking so that the air was not counted, and then the end was connected with a rubber bulb. While pressing the rubber bulb, the tip of the microcolumn tube loaded with radioactive material was immersed in the developing solvent, and the pressed rubber bulb was released. The developing solvent developed along the microcolumn tube with the pressure generated as the rubber bulb relaxed again. Confirmed to come to the opposite end, the rubber bulb was removed and radioactive material in the microcolumn tube was detected by a radiation-TLC scanner.

The results of development experiments using the microcolumn tubes prepared in Example 1-1 are shown in Table 1 below.

Solvent 10% ethyl acetate / hexane 50% ethyl acetate / hexane 10% Methanol / Dichloromethane Deployment length (mm) 68 68 68 Deployment time (seconds) 47 47 77 Deployment speed (mm / s) 1.45 1.45 0.88 Rf value 0.54 0.89 0.93 Fluorine-18: 2- (3- [ ¹⁸ F] fluoropropoxy) naphthalene 0.435: 0.565 0.435: 0.565 0.468: 0.532

The results of development experiments using the microcolumn tubes prepared in Example 1-2 are shown in Table 2 below.

Solvent 10% ethyl acetate / hexane 50% ethyl acetate / hexane 10% Methanol / Dichloromethane Deployment length (mm) 68 68 68 Deployment time (seconds) 15 15 70 Deployment speed (mm / s) 4.53 4.53 0.97 Rf value 0.50 0.86 0.93 Fluorine-18: 2- (3- [ ¹⁸ F] fluoropropoxy) naphthalene 0.620: 0.380 0.488: 0.512 0.475: 0.525

The results of development experiments using the microcolumn tubes prepared in Examples 1-3 are shown in Table 3 below.

Solvent 50% acetonitrile / water 75% acetonitrile / water 100% acetonitrile / water Deployment length (mm) 85 85 85 Deployment time (seconds) 243 203 78 Deployment speed (mm / s) 0.35 0.42 1.09 Rf value 0.30 0.17, 0.72 0.71 Fluorine-18: 2- (3- [ ¹⁸ F] fluoropropoxy) naphthalene 0.693: 0.307 0.680: 0.320 0.597: 0.403

Comparative Example  2: thin membrane Chromatography plate  Deployment experiment

A mixed solution using an aqueous fluorine-18 anion solution and a 2- (3- [ ¹⁸ F] fluoropropoxy) naphthalene compound solution was prepared, and a portion thereof was taken as a microtube, and the droplet was applied to one end of the thin film chromatography plate prepared in Preparation Example 1. (spotting). Place this in a TLC tank containing the developing solvent, wait until the developing solvent rises to a height of 90 mm along the thin membrane chromatography, then remove the thin membrane chromatography plate from the vessel and release the radioactive TLC scanner. Detected through.

The results of development experiments using the thin film chromatography plate prepared in Comparative Example 1-1 are shown in Table 4 below.

Solvent 10% ethyl acetate / hexane 50% ethyl acetate / hexane 10% Methanol / Dichloromethane Deployment length (mm) 90 90 90 Deployment time (seconds) 900 900 1200 Deployment speed (mm / s) 0.10 0.10 0.075 Rf value 0.67 0.83 0.87 Fluorine-18: 2- (3- [ ¹⁸ F] fluoropropoxy) naphthalene 0.414: 0.586 0.318: 0.682 0.348: 0.652

The results of development experiments using the thin film chromatography plate prepared in Comparative Example 1-2 are shown in Table 5 below.

Solvent 50% acetonitrile / water 75% acetonitrile / water 100% acetonitrile / water Deployment length (mm) 90 90 90 Deployment time (seconds) 3235 1847 679 Deployment speed (mm / s) 0.028 0.049 0.13 Rf value 0.03 0.18, 0.42 0.75 (A 3- ^[18 F] Fluoro-propoxy) 2-naphthalene: fluorine -18 0.676: 0.324 0.407: 0.593 0.379: 0.621

As shown in Table 1 to Table 5, the development of the radioactive material using conventional thin membrane chromatography in the normal chromatography method using silica gel takes 15-20 minutes, whereas the microcolumn tube of the present invention The development of the used radioactive material was completed in 15 to 77 seconds depending on the particle size of each of the developing solvent and silica gel. This represents a rapid development speed of 17.1 to 60 times. In addition, in the reverse phase chromatography using C18 silica gel, the development of the radioactive material using the thin membrane chromatography takes about 11 to 53 minutes, whereas the development of the radioactive material using the microcolumn tube of the present invention is 1 to 4. It was completed in minutes. This represents about 11 to 13.2 times faster deployment speed. And, as shown in Figures 1 to 6, as a result of quantitative analysis by the radiation-TLC scanner it was confirmed that the same or similar to the conventional thin film chromatography.

< Fine column tube  membrane Chromatography plate  Comparative Experiment of Development of Volatile Isotope Labeling Compounds>

The development experiment of the volatile radioactive material using the microcolumn tube of the present invention was performed in comparison with the development experiment by the conventional thin membrane chromatography method. After the development experiments, each of them was evaluated for the separation effect of radioactive material through a radiation-TLC scanner.

Example  3: Fine column tube  Development of Volatile Isotope Labeling Compounds

A portion of the mixed solution using fluorine-18 anion aqueous solution and 5- [ ¹⁸ F] fluoro-1-pentine (volatile substance, breaking point 59.1 ± 15.0 ° C.) was taken as a microtube, and the end of the microcolumn tube prepared in Example 1 It was loaded. The other end of the microcolumn tube was connected to a glass tube with rubber parking so that air was not counted, and then the end was connected with a rubber bulb. While pressing the rubber bulb, the tip of the microcolumn tube loaded with radioactive material was immersed in the developing solvent, and the pressed rubber bulb was released. The developing solvent developed along the microcolumn tube with the pressure generated as the rubber bulb relaxed again. Confirmed to come to the opposite end, the rubber bulb was removed and the radioactive material in the microcolumn tube was detected by a radiation-TLC scanner.

Table 6 shows the results of development experiments using the microcolumn tubes prepared in Example 1-1.

Solvent 50% ethyl acetate / hexane Deployment length (mm) 70 Deployment time (seconds) 56 Deployment speed (mm / s) 1.21 Rf value 0.89 Fluorine-18: 5- [ ¹⁸ F] fluoro-1-pentine 0.499: 0.501

Table 7 shows the results of development experiments using the microcolumn tubes prepared in Example 1-2.

Solvent 50% ethyl acetate / hexane Deployment length (mm) 70 Deployment time (seconds) 14 Deployment speed (mm / s) 4.86 Rf value 0.86 Fluorine-18: 5- [ ¹⁸ F] fluoro-1-pentine 0.586: 0.414

Comparative Example  3: thin membrane Chromatography plate  Development of Volatile Isotope Labeling Compounds

A thin membrane prepared in Comparative Example 1-1 by taking a portion of a mixed solution using an aqueous fluorine-18 anion solution and 5- [ ¹⁸ F] fluoro-1-pentine (volatile substance, breaking point 59.1 ± 15.0 ° C.) as a microtube. Spotting was carried out at one end of the chromatography plate. Place this in a vessel (TLC tank) containing the developing solvent (50% ethyl acetate / hexane), wait until the developing solvent rises to a height of 90 mm along the thin membrane chromatography, and then remove the thin membrane chromatography plate from the vessel. Radioactive material was detected via a radiation-TLC scanner. The experimental results of Comparative Example 3 are shown in Table 8 below.

Solvent 50% ethyl acetate / hexane Deployment length (mm) 70 (Seconds) during deployment 900 Deployment speed (mm / s) 0.10 Rf value - Fluorine-18: 5- [ ¹⁸ F] fluoro-1-pentine 1.0: 0.0

As shown in Example 3 and Comparative Example 3, since the 5- [ ¹⁸ F] fluoro-1-pentin compound is a volatile substance having a breaking point of about 59 ° C., quantitative analysis using thin membrane chromatography is impossible (FIG. 9). Reference). However, when the microcolumn tube according to the present invention was used, it was confirmed that quantitative analysis was possible because the 5- [ ¹⁸ F] fluoro-1-pentin compound, which is a volatile substance, was trapped in the microcolumn tube. This is shown in FIGS. 7 and 8.

1: inlet
2, 3: frit
4: filling
100: fine column tube

Claims (13)

A microcolumn tube for chromatography for separation detection or quantitation of radioactive material,
An inlet for loading radioactive material;
A frit composed of porous solids present at both ends of the microcolumn tube;
Solid filler in the form of particles filled between the frits; And
Microcolumn tube, characterized in that consisting of a solid tube that can maintain a constant shape. The method of claim 1,
Microcolumn tube, characterized in that the material of the solid tube is glass, quartz, plastic or stainless metal. The method of claim 1,
A microcolumn tube, characterized in that the inner diameter of the solid tube is 0.5-3.0 mm and the length is 40-200 mm. The method of claim 1,
The filler is a silica gel, C4 to C18 silica gel, aluminum oxide (Al ₂ O ₃ ), calcium carbonate, activated carbon, diatomaceous earth, celite or cellulose powder, characterized in that the fine column tube. The method of claim 1,
The frit is a microcolumn tube, characterized in that cotton, porous glass, polyethylene or polypropylene. An inlet for loading radioactive material;
A frit composed of porous solids present at both ends of the microcolumn tube;
Solid filler in the form of particles filled between the frits; And
In the separation detection method of a radioactive material using a microcolumn tube, characterized in that the solid tube which can maintain a constant form,
Loading a mixture of radioactive material into an injection hole of the microcolumn tube (step 1); And
And developing the radioactive material by injecting the developing solvent into the microcolumn tube loaded with the mixed solution of the radioactive material (step 2). The method according to claim 6,
Detecting the radioactive material separated in step 2 using a radiation detector (step 3). The method of claim 7, wherein
And the radiation detector is a radiation-TLC image scanner. The method according to claim 6,
In step 2, the separation solvent of the radioactive material, characterized in that the developing solvent is developed using the pressure difference across the microcolumn tube. The method according to claim 6,
Load the mixture of radioactive material to the other end of the microcolumn tube of claim 1, one end of which is connected to the rubber bulb (step 1),
A method of separating and detecting a radioactive material characterized by dipping in a developing solvent while pressing a rubber bulb and developing a radioactive material in a state where the rubber bulb is opened (step 2). delete delete delete

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