CN113042111A

CN113042111A - Is used for18F-labeled miniature anion exchange adsorption column

Info

Publication number: CN113042111A
Application number: CN201911362628.5A
Authority: CN
Inventors: 李世红; 闫恒; 季琰; 余康; 王正
Original assignee: Nanjing Pet Tracer Co ltd
Current assignee: Nanjing Pet Tracer Co ltd
Priority date: 2019-12-26
Filing date: 2019-12-26
Publication date: 2021-06-29

Abstract

The invention discloses a method for¹⁸The invention discloses an F-labeled micro anion exchange adsorption column, which adopts a micro adsorption column with the anion exchange adsorbent loading capacity of 5-30 mg, and further discloses that an eluent of the micro anion exchange adsorption column comprises trace alkaline salt, K222 and an organic solvent. The micro adsorption column can simultaneously meet high-efficiency adsorption efficiency and elution efficiency, and the generated low-alkalinity reaction condition can cause high nucleophilic substitution¹⁸F marking efficiency, simple and quick, low cost and the like, and is suitable for popularization and use.

Description

Is used for18F-labeled miniature anion exchange adsorption column

Technical Field

The invention relates to the technical field of radioactive labeling, in particular to a method for labeling a polypeptide¹⁸F-labeled miniature anion exchange adsorption columns.

Background

Commercially available for adsorption and elution of radionuclides¹⁸The anion exchange adsorption column of F includes QMA strong anion exchange column, WAX weak anion exchange column, etc., and the packing of these columns is generally over 100mg, for¹⁸F has strong adsorption capacity, and only a large amount of neutral or alkalescent salt is added into the eluent to improve the elution efficiency, but excessive alkaline salt is introduced to ensure that certain nucleophilic substitution in the next step¹⁸The side reaction of the labeling reaction increases and the labeling efficiency decreases. On the other hand, if the content of the basic salt is reduced, although it is advantageous to improve the labeling efficiency, it results in¹⁸The elution efficiency of F ions from the anion exchange column decreases. Thus, commercially available for¹⁸The F-labeled anion exchange adsorption column cannot be compatible with the dual characteristics of high elution rate and high labeling rate.

Disclosure of Invention

Based on the above deficiencies of the prior art, it is an object of the present invention to provide a method for¹⁸The comprehensive efficiency of the adsorption column in three aspects of adsorption, elution and marking is superior to that of the adsorption column sold in the market¹⁸F labeled anion exchange adsorption column, and further discloses the composition of the eluent used by the adsorption column.

In order to achieve the purpose, the invention provides the following technical scheme:

the invention aims to provide a method for¹⁸The F-marked miniature anion exchange adsorption column comprises a column tube, an adsorbent, a sieve plate and joints, wherein the joints are positioned at the front end and the rear end of the column tube; the sieve plate is divided into an upper sieve plate and a lower sieve plate, the upper sieve plate is positioned behind the front end joint of the column pipe, and the lower sieve plate is positioned in front of the rear end of the column pipe; the anion exchange adsorbent is positioned between the upper sieve plate and the lower sieve plate, and the loading capacity is 5-30 mg.

Preferably, the loading amount of the adsorbent is 10-22 mg.

Preferably, the rear end joint of the column tube is¹⁸F^-The source and the leacheate are imported, and the front end joint is¹⁸F^-And the effluent outlet.

The above-mentioned¹⁸F adsorption efficiency on the column¹⁸In F dose/(adsorbent dose + effluent liquid)¹⁸Dose F).

The above-mentioned¹⁸F elution efficiency¹⁸Of dose F/adsorption¹⁸And F dose.

As a preferred scheme, the adsorbent is selected from any one of strong anion exchange adsorbent of silica gel matrix or organic polymer matrix, or weak anion exchange reverse phase adsorbent.

Further, the adsorbent is selected from a QMA strong anion exchanger or a WAX weak anion exchanger.

In a preferable scheme, the diameter of the adsorbent particles is 30-100 μm, and the preferable diameter is 35-65 μm.

As a preferable scheme, the adsorption column is subjected to KHCO before use₃、K₂CO₃Or a KOTf pretreatment.

Preferably, the adsorption column adsorbs¹⁸F^-Thereafter, the mixture is rinsed with an eluent comprising a basic salt, K222 and an organic solvent.

As a preferred embodiment, the basic salt is selected from K₂CO₃And KOTf, wherein the organic solvent is one or more selected from methanol, n-butanol, ethanol, isopropanol and acetonitrile, and preferably the organic solvent is methanol.

As a preferred embodiment, said K₂CO₃The dosage is (0.1-1) x 10^-3mmol, the dosage of KOTf is (10-100) multiplied by 10^-3mmol。

Preferably, the dosage of the K222 is (0.2-2.1) multiplied by 10^-3mmol。

The invention has the beneficial effects that:

the invention provides a method for¹⁸The invention relates to a F-marked micro anion exchange adsorption column, which can reduce the dosage of alkaline salt in eluent and simultaneously realize¹⁸F ions are eluted with high efficiency, thereby improving¹⁸F ion availability and enhancement of certain nucleophilic substitutions¹⁸Efficiency of labelling and reduction of side reactions, especially in¹⁸The F radioactive label has high field value and simple and convenient operation, and is suitable for popularization and use.

The specific implementation mode is as follows:

in order that the present disclosure may be more readily and clearly understood, the following detailed description of the present disclosure is given in conjunction with specific examples, which are included merely for purposes of illustration and description of the presently preferred modes of carrying out the disclosure, and are not intended to limit the scope of the disclosure in any way by the examples set forth herein.

The reagents used in the examples are all commercially available.

EXAMPLE 1 adsorbent column pretreatment

The method comprises the following steps:

using 250mM KHCO₃(3mL) the column was washed with air until no liquid flowed out, then 5mL of pure water was used to wash the column, and then air was used to wash the column until no liquid flowed out.

The second method comprises the following steps:

pretreatment 100mM K was used₂CO₃(3ml) the column was washed with air until no liquid flowed out, then 5ml of pure water was used to wash the column, and then air was used to wash until no liquid flowed out.

The third method comprises the following steps:

pretreatment the column was flushed with 250mM KOTf (3ml), air column flushed until no liquid flowed out, 5ml pure water column flushed, and air column flushed until no liquid flowed out.

Example 2 influence of adsorbent content on adsorption efficiency and elution efficiency, see Table 1

Adsorbent: QMA Strong anion exchanger (column 1)

Eluent: 1ml of a solution containing 0.8. mu. mol of K₂CO₃And 0.66. mu. mol K222 in methanol

TABLE 1

Serial number	Adsorbent loading/mg	Efficiency of adsorption	Efficiency of elution
				1	4	77.96％	99.89％
2	5	83.04％	99.50％
				3	10	91.55％	97.99％
4	20	97.78％	96.36％
				5	22	98.91％	93.41％
6	27	99.31％	85.02％
				7	30	99.73％	82.68％
8	50	99.80％	57.09％
				9	100	99.92％	30.21％

And (4) conclusion: the adsorption efficiency is improved along with the increase of the loading capacity of the adsorbent, and the elution efficiency is gradually reduced along with the increase of the loading capacity of the anion exchange adsorbent. When the content of the adsorbent is lower than 5mg, the adsorption efficiency is lower than 80%, and when the content of the adsorbent exceeds 30mg, the elution efficiency is lower than 80%, so that the loading capacity of the adsorbent is within 5-30 mg, the adsorption efficiency and the elution efficiency can be ensured to be higher than 80%, and the loading capacity of the adsorbent is within 10-22 mg, the adsorption efficiency and the elution efficiency can be ensured to be higher than 90%.

Example 3 Effect of eluent type on elution efficiency, see Table 2

Adsorbent: QMA Strong anion exchanger (column 1)20mg

Adsorbent: WAX Weak anion exchanger (column 2)20mg

TABLE 2

Column number	Kind of eluent	Efficiency of elution
			1	0.1μmol K₂CO₃0.21. mu. mol of K222, 0.8mL of methanol	90.1％
1	0.8μmol K₂CO₃0.66. mu. mol of K222, 1mL of methanol	96.4％
			1	10. mu. mol KOTf, 0.66. mu. mol K222, 1mL isopropanol	95.3％
1	30. mu. mol KOTf, 1.69. mu. mol K222, acetonitrile 1.5mL	84.8％
			2	1μmol K₂CO₃0.66. mu. mol of K222 and 0.8mL of n-butanol	90.7％
2	100. mu. mol KOTf, 2.1. mu. mol K222, 1mL acetonitrile	85.1％

And (4) conclusion: the alkaline salt is K₂CO₃Mu mol (0.1-1), KOTf (10-100), K222 (0.2-2.1), and organic solvent selected from methanol, isopropanol, n-butanol or acetonitrile, with an elution efficiency of more than 80%.

Example 4 Effect of eluent type on labelling efficiency, see Table 3

¹⁸F-DOPA preparation: will contain¹⁸F^-Was azeotropically dried with acetonitrile (300. mu.L). Add N, N-diBoc precursor (35mg), Cu (OTf)₂(py)₄(14mg) was dissolved in DMF (300. mu.L) and reacted at 110 ℃ for 20 min.0.1mL HI (57% H) at 130 deg.C₂O) for 10 min. The pH was then adjusted to near neutrality. Measurement by thin layer chromatography¹⁸Radiochemical labelling efficiency of the crude F-DOPA.

¹⁸F-FP-CIT preparation: will contain¹⁸F^-The eluent is evaporated to dryness at 90 ℃. The precursor with OTs group (2mg), MeCN (100. mu.L), t-amyl alcohol (500. mu.L) were added and reacted at 100 ℃ for 20 min. Measurement by thin layer chromatography¹⁸And (3) the radiochemical labeling rate of the F-FP-CIT crude product.

Each eluent pair¹⁸F-FP-CIT、¹⁸The effect of F-DOPA labeling efficiency is shown in Table 3.

TABLE 3

And (4) conclusion: the basic salt in the eluent is K₂CO₃(0.1-1) μmol, KOTf (10-100) μmol, K222 (0.2-2.1) μmol, and organic solvent selected from methanol, isopropanol, n-butanol or acetonitrile¹⁸The labeling efficiency of F reaches more than 50 percent.

Claims

1. Is used for¹⁸The F-marked micro anion exchange adsorption column comprises a column tube, an adsorbent, a sieve plate and joints, and is characterized in that the joints are positioned at the front end and the rear end of the column tube; the sieve plate is divided into an upper sieve plate and a lower sieve plate, the upper sieve plate is positioned behind the front end joint of the column pipe, and the lower sieve plate is positioned in front of the rear end of the column pipe; the anion exchange adsorbent is positioned between the upper sieve plate and the lower sieve plate, and the loading capacity is 5-30 mg.

2. The miniature anion exchange sorbent column of claim 1, wherein the loading of the sorbent is 10-22 mg.

3. The miniature anion exchange sorbent column of claim 1, wherein the sorbent is selected from any one of strong anion exchange sorbent with silica gel matrix or organic polymer matrix, or weak anion exchange reverse phase sorbent.

4. The miniature anion exchange column of claim 3 wherein the diameter of the adsorbent particles is 30-100 μm.

5. The miniature anion exchange column of claim 1, wherein said column is subjected to KHCO prior to use₃、K₂CO₃Or potassium triflate (KOTf) pretreatment.

6. The miniature anion exchange adsorbent column of any of claims 1-5, wherein the adsorbent column is adsorbing¹⁸F^-Thereafter, the mixture is rinsed with an eluent comprising a basic salt, K222 and an organic solvent.

7. The miniature anion exchange sorbent column of claim 6, wherein the basic salt is selected from the group consisting of K₂CO₃Or KOTf, wherein the organic solvent is one or more selected from methanol, n-butanol, ethanol, isopropanol and acetonitrile.

8. The miniature anion exchange sorbent column of claim 7, wherein the K is₂CO₃The dosage is (0.1-1) x 10^-3mmol, the dosage of KOTf is (10-100) multiplied by 10^-3mmol。

9. The miniature anion exchange column of claim 7, wherein the amount of K222 is (0.2-2.1) x 10^-3mmol。