HENRIKSEN, Gjermund (Schubertstraße 11, Starnberg, 82319, DE)
WEßMANN, Sarah, Hedwig (Säbenerstraße 34, München, 81547, DE)
WESTER, Hans-Jürgen, Peter (Dinkelweg 4, Ilmmünster, 85304, DE)
HENRIKSEN, Gjermund (Schubertstraße 11, Starnberg, 82319, DE)
WEßMANN, Sarah, Hedwig (Säbenerstraße 34, München, 81547, DE)
| CLAIMS 1. A method to extract out of an aqueous solution, concentrate and/or reformulate [ F]fluorides without any evaporation step, in an organic solution suitable for a subsequent radiolabeling reaction, said method comprising the following steps of : - passing said aqueous [18F]fluoride solution or solvent through a solid phase extraction column containing an anion-exchange resin so that said [18F]fluoride are trapped on said resin - rinsing the resin with an organic solvent for eliminating the residual water that may be undesirable for a subsequent chemical processing, whilst keeping the extracted anions trapped on the resin. - eluting said [18F]fluorides with an eluting solution so that to release said [18F]fluorides from said anion- exchange resin under a form which is reactive and immediately usable for a labeling reaction; - use the eluted solution for a radiolabeling reaction; characterised in that the eluting solution is a solution with a cryptate of an alkali or alkaline earth with hydroxide as counterion, containing an organic solvent suitable for the subsequent radiolabeling reaction ; 2. Method according to Claim 1, characterised in that said cryptate is formed by a suitable cryptand, such as 4,7,13,16,21,24-Hexaoxa-l,10-diazabicyclo[8.8.8]hexacosane (2,2,2-Cryptand, Kryptofix 222) and an hydroxide of an alkali or alkaline earth metal. 3. Method according to Claim 1 , characterised in that said organic solvent is selected from the group consiting of acetonitrile (MeCN) , dimethylsulfoxide (DMSO) , dimethylacetamide (DMM), dimethylformamide (DMF) , tetrahydrofuran (THF) or other solvents that are used in the methods of prior art and primary alcohols such as methanol, ethanol, n-propanol, n-butanol, amyl alcohol, n-hexyl alcohol, n-heptanol, benzyl alcohol or n-octanol, and secondary alcohols such as isopropanol, isobutanol, isoamyl alcohol or 3-pentanol, and tertiary alcohols, diols and polyols with 1 to 20 tertiary alcohol functions such as t-butanol, t-amyl alcohol, 2, 3-dimethyl- 2-butanol, 2- (trifluoromethyl) -2- propanol, 2, 8-dimethyl- 2, 8-Decanediol or 2 , 5-dimethyl-3-Octyne-2 , 5-diol. 4. Method according to Claim 1, characterised in that, prior to the elution step, a drying step comprising a flush of gas such as air, nitrogen, helium or argon, is used to purge the column and eliminate most of the remaining solvent. 5. Method according to Claim 1, characterised in that the organic solvent suitable for the subsequent radiolabeling reaction is identical to the solvent used for elution 6. Method according to Claim 1, characterised in that the eluting medium or the solid phase extraction column is heated up to enhance the elution efficiency. 7. Method according to Claim 1, characterised in that the eluted solution is diluted in a solvent suitable for the subsequent labeling reaction selected from the group consisting of acetonitrile (MeCN) , dimethylsulfoxide (DMSO), dimethylacetamide (DMAA), dimethylformamide (DMF) , tetrahydrofuran (THF) or other solvents that are used in the methods of prior art and primary alcohols such as methanol, ethanol, n-propanol, n-butanol, amyl alcohol, n-hexyl alcohol, n-heptanol, benzyl alcohol or n-octanol, and secondary alcohols such as isopropanol, isobutanol, isoamyl alcohol or 3-pentanol, and tertiary alcohols, diols and polyols with 1 to 20 tertiary alcohol functions such as t-butanol, t-amyl alcohol, 2, 3-dimethyl- 2-butanol, 2- (trifluoromethyl) -2-propanol, 2, 8-dimethyl- 2, 8-Decanediol or 2 , 5-dimethyl-3-Octyne-2 , 5-diol. 8. Method according to Claim 1, characterised in that the eluted solution is diluted with a suitable cryptate, or organic molecule, or organic salt, acting as a phase transfer catalyst, identical to or different of the cryptate used for the elution step to be added prior to labeling in the eluted [18F] fluoride solution to improve the radiolabeling yield. 9. Method according to Claim 1, characterised in that the anion exchange resin may subsequently, after the elution, be flushed with a solution of the precursor. 10. Method according to Claim 1, characterised in that the eluted solution containing the [18F] fluoride is used for the synthesis of a PEET radiotracer, without any subsequent evaporation step, the [18F] fluoride being then reactive for substitution reactions on both aliphatic and aromatic precursors. 11. Method according to Claim 9, characterised in that, in some specific labeling reactions, a suitable cryptate, inorganic salt or organic base identical to, or different to the cryptate used for the elution step is added to the eluted [18F] fluoride solution prior to labeling to improve the radiolabeling yield. |
Technical field
[0001] The present invention relates to methods for the preparation of reactive [ 18 F] fluoride in a form suitable for efficient radiolabeling without an evaporation step by the use of an anion exchange resin and alkalimetal cryptates.
Background art
[0002] [ 18 F] fluoride is produced by irradiation of water, containing H 2 18 0, with protons resulting in the reaction 18 0(p,n) 18 F. Only a minor fraction of the [ 18 0] is converted. The [ 18 F] isotope is then separated from the water and processed for production of a radiopharmaceutical agent.
[0003] In the current practice, fluoride recovery is based on the use of an anion-exchange resin. The recovery is carried out in two steps, extraction and elution: first the anions (not only fluoride) are separated from the enriched [ 18 0] water and trapped on the said resin {Coenen eta/., J. Labelled Compd. Radiopharm., 1986, vol. 23, pp. 455-467). The anions, including [ 18 F]fluoride, are then eluted into a mixture containing water, organic solvents, an activating agent or phase transfer agent or phase transfer catalyst, such as for example the complex potassium carbonate-Kryptofix 222 (K 2 C0 3 -K222) or a tetrabutylammonium salt. The [ 18 F]fluoride radiochemical recovery yield is very effective, usually exceeding 99%.
[0004] The most usual labeling method, known as nucfeophilic substitution, requires anhydrous or very low water content solutions. Thus, an evaporation step (or drying step) after [ 18 F] fluoride recovery to remove the excess water is still necessary. It usually consists in multiple azeotropic evaporations with acetonitrile or low boiling temperature organic solvent. Such evaporations require several minutes (Schlyer et al., Appl. Radiat. Isot, 1990, vol. 40, pp. 1-6).
[0005] The current trend in the automated preparation of radiopharmaceuticals for medical imaging is to develop "Lab-on-chip" devices. The aforementioned evaporation step is very difficult to implement within such a "Lab-on-chip" device. Recently, a method has been described avoiding a drying step prior to radiolabeling with [ 18 F]fluoride. This method based on the use of organic bases that allows for the removal of [ 18 F]fluoride from an anion exchange cartridge and subsequent labeling when small amounts of water are present or H-acidic compounds are added ( WO 2009/003251 Al). Aims of the invention
[0006] The current invention aims at avoiding the need for any evaporation after the elution of the anion- exchange resin.
[0007] Further the invention aim to reduce the duration of preparation, which results in an increase of the overall radiochemical yield.
[0008] A still further aim of the invention is to simplify the automated radiosynthesis equipment used for the synthesis of a radiotracer.
[0009] Further the invention aims at making the method suitable for implementation into automated "Lab-on- chip" systems .
Disclosure of the invention
[0010] The method of the invention allows the preparation of a reactive [ 18 F]fluoride solution for nucleophilic substitution reactions on both aliphatic and aromatic precursors, without any azeotropic evaporation step. The reduction of the duration of preparation results in an increase of the overall yield, and a simplification of the automated equipment needed for the synthesis of a radiotracer. In particular, the suppression of any azeotropic evaporation step facilitates the implementation of the synthesis on microfluidic devices such as "lab-one-chip" in which these evaporations are difficult to perform.
[0011] It must be noted that, according to the invention, the subsequent radiolabelling is performed using the established organic solvents, thus under identical conditions as used in the methods of prior art .
[0012] According to the invention, the eluting medium or solution is an organic solution containing at least the following species: an organic solvent suitable for a subsequent radiolabelling reaction, i.e. a polar aprotic solvent, a first compound (A) which is a cryptand and a second compound (B) that is a metal hydroxide.
[0013] The elution process is made possible by the use of hydroxide ions that can exchange with the fluoride trapped on the resin and cryptates that allows the fluoride to be solubilized in the eluting medium, and by using appropriate amounts of compound (A) and (B). The cryptate-hydoxide is thus acting as an activating agent or phase transfer agent or phase transfer catalyst allowing to remove and to elute [ 18 F]fluoride from the anion exchange resin in the absence of water.
[0014] Organic solvents suitable for radiolabeling, i.e. polar aprotic solvents, are organic solvents that have dipoles due to polar bonds, don't have H atoms that can be donated into a H-bond and in which anions are not solvated but are "naked" which allows them to be reactive as nucleophile for substitu- tion reactions. These solvents are defined by the IUPAC {IUPAC, 1994, 66, 1077 (Glossary of terms used in physical organic chemistry (IUPAC Recommendations 1994)) on page 1106) as: "solvents with a comparatively high relative permittivity (or dielectric constant), greater than ca. 15, and a sizable permanent dipole moment, that cannot donate suitably labile hydrogen atoms to form strong hydrogen bonds, e.g. dimethyl sulfoxide, ... Such solvents are usually not aprotic but protophilic (and at most weakly protogenic)...".
[0015] Chemicals suitable as compound (A) are cryptands that allow for efficient complexation of alkali or alkaline earth metals. One example for such a cryptand is as 4,7,13,16,21,24-Hexaoxa-l,10- diazabicyclo[8.8.8]hexacosane (2,2,2-Cryptand, Kryptofix 222).
[0016] According to the present invention, the eluted solution is directly usable for a radiolabeling reaction, that is no evaporation for water elimination is required. Thus, the [ 18 F] activity remains at all times in solution after the elution of the anion exchange resin, contrary to the methods of prior art where it is recovered in a "dry form" on the surface of a reactor as a result of the evaporation step.
[0017] According to the present invention, said elution step is performed by passing the eluting solution through a solid phase extraction column containing an anion-exchange resin. The [ 18 F] fluoride is released from the resin in the eluting medium as specified above and is immediately usable for efficient radiolabeling, even, depending on the reactivity of the substrate to be labeled, at room temperature.
[0018] Prior to the elution sequence, a purging sequence comprising a flush of gas such as air, nitrogen or argon is be used to purge the column and eliminate most of the remaining H 2 18 0.
[0019] After this step and prior to the elution step, the column is rinsed with an organic solvent that allows the elimination of the residua! water that may be undesirable for a subsequent chemical processing, i.e. nucleophilic substitution, whilst keeping the extracted anions trapped on the resin. This organic solvent is selected among solvents suitable for the subsequent radiolabeling reaction
[0020] The organic solvent can be selected among acetonitrile (MeCN) , dimethylsulfoxide (DMSO) , dimethylacetamide (DMAA), dimethylformamide (DMF) , tetrahydrofuran (THF) or other solvents that are used in the methods of prior art.
[0021] More specifically, according to the invention, the eluting medium is a dry solvent containing the preformed cryptate of compound (A) and the compound (B).
[0022] In some embodiments of the present invention, the solvent used for the elution step is preferably selected from the group consisting of primary alcohols such as methanol, ethanol, n- propanol, n-butanol, amyl alcohol, n-hexyl alcohol, n-heptanol, benzyl alcohol or n-octanol, and secondary alcohols such as isopropanol, isobutanol, isoamyl alcohol or 3-pentanol. [0023] In some embodiments of the present invention, the solvent used for the elution step is preferably selected from the group consisting of tertiary alcohols, diols and polyols with 1 to 20 tertiary alcohol functions such as t-butanol, t-amyl alcohol, 2, 3-dimethyl- 2-butanol, 2- (trifluoromethyl) -2-propanol, 2, 8-dimethyl- 2, 8-Decanediol or 2 , 5-dimethyl-3-Octyne-2 , 5-dioL In this context, it has been shown by CHI, D. Y. eta/ [A New Class ofSN2 Reactions Catalyzed by Protic Solvents: Facile Fluor/nation for Isotopic Labeling of Diagnostic Molecules, J. Am. Chem. Soc. , Vol.128, 50 (2006) pp.16394-16397 ; WO-A-2006/065038) 'that the addition of some tertiary alcohols to the reaction mixtures subsequently to elution, usually composed of acetonitrile with the relevant precursor, does not impact unfavorably on the subsequent nucleophilic substitution reaction ("SN2" reactions) .
[0024] In some embodiments of the present invention, the anion exchange resin may subsequently, after the elution, be flushed with a solution of the precursor.
[0025] The eluted organic solution containing the [ 18 F] fluoride can be used for the synthesis of a PET radiotracer. The [ 18 F] fluoride is then reactive, without any subsequent evaporation step, for substitution reactions on both aliphatic and aromatic precursors.
[0026] In some embodiments of the present invention, a suitable cryptate, or organic molecule or organic salt acting as a phase transfer catalyst, identical to or different of the cryptate one used for the elution step can be added prior to labelling in the eluted [ 18 F] fluoride solution to improve the radiolabeling yield.
Description of a preferred embodiment of the invention
EXAMPLES
Example 1. Trapping and release of [ 18 F]fluoride by means of anion exchange resin.
"QMA light" anion exchange cartridge (CI " form, Waters) was conditioned by flushing the resin subsequently with
1 M KOH, (2 ml), water (20 ml), 1 M K 2 C0 3 (2 ml), water (20 ml), and finally 5 ml air (conditioning QMA 1).
Alternatively, "QMA light" anion exchange cartridge (CI " form, Waters) was conditioned by flushing the resin subsequently with 1 M K 2 C0 3 (2 ml), water (20 ml), and finally 5 ml air (conditioning QMA 2). Alternatively, "QMA light" anion exchange cartridge (CI " form, Waters) was conditioned by flushing the resin subsequently with 1 M KOH, (2 ml), water (20 ml), 1 M KHC0 3 (2 ml), water (20 ml), and finally 5 ml air (conditioning QMA 3). "PSA-HCO 3 " anion exchange cartridge (HC0 3 " form, Machery-Nagel) was conditioned by applying 3 ml MeOH followed by 3 ml H 2 0 (conditioning PSA 1).
Alternatively, "PSA-HCO 3 " anion exchange cartridge (HC0 3 " form, Machery-Nagel) was conditioned by applying 0,5 mL H 2 0 (conditioning PSA 2).
Alternatively, "PSA-HCO3" anion exchange cartridge (HC0 3 ~ form, Machery-Nagel) was conditioned by applying
with 1 M KOH, (2 ml), water (20 ml), 1 M K 2 C0 3 (2 ml), water (20 ml), and finally 5 ml air (conditio ¬ ning PSA 3).
If not mentioned, commercially available cartridge amounts were used.
Aqueous [ 18 F]fluoride obtained via the 18 0(p,n) 18 F nuclear reaction was applied to the cartridge in question, the cartridge purged with 1 ml air and subsequently eluted with anhydrous 5 ml MeCN.
[ 18 F]fluoride was quantitatively retained on the cartridge during these steps.
Elution of [ 18 F]fluoride from the cartridge was evaluated with azeotropically dried solutions of
[K + 2.2.2]OH ~ of different concentrations:
[K + 2.2.2]OH ~ in acetonitrile:
solution A: 167.7 mM in KOH and 250 mM in Kryptofix 2.2.2.
500 μΙ_ of 1M KOH and 282,4 mg Kryptofix 2.2.2. were mixed, and azeotropically dried by portion wise addition of anhydrous MeCN while heating at 105 °C. Three ml of anhydrous CH 3 CN was added to the residue.
Alternatively, the residue was dissolved in three ml of anhydrous t-butanol (solution A2) or three ml of anhydrous amyl alcohol (solution A3). solution B: 334 mM in KOH and in 500 mM Kryptofix 2.2.2.
666 μΙ 1M KOH and 376 mg Kryptofix 2.2.2. were mixed and azeotropically dried by portion wise addition of anhydrous MeCN while heating at 105 °C. Two ml of anhydrous CH 3 CN was added to the residue. solution C: 83.9 mM in KOH and 125 mM in Kryptofix 2.2.2.
125 μΙ 1M KOH and 70.5 mg Kryptofix 2.2.2. were mixed and azeotropically dried by portion wise addition of anhydrous MeCN while heating at 105 °C. One and a half ml of anhydrous CH 3 CN was added to the residue.
Elution were tested using solvents of different cryptate concentrations (Figure A), different QMA conditioning procedures (Figure B), different "PSA" conditioning procedures (Figure C), different types of anion exchange cartridges (Figure D), different sorbent amounts for "QMA light" anion exchange resin (Figure E), different sorbent amounts of "PSA" anion exchange resin (Figure F), different types of anion exchange cartridge with similar sorbent amount (Figure G), different counter ions of "QMA light" cartridges (Figure H) and different counter ions of "PSA" anion exchange cartridges (Figure I), [K + 2.2.2]OH in different solvents (Figure J).
Example 2. 18 F-fluorination of precursors
18 F-labelling reactions were examined using fractions of [K + 2.2.2]OH ~ -eluate, total cartridge elution volumes of [K + 2.2.2]OH " -solution, using increasing fraction volumes of [K + 2.2.2]OH " -eluate (with measuring pH-values with increasing fraction volumes of [K + 2.2.2]OH ~ -eluate), using fractions of [K + 2.2.2]OH " -eluate from cartridges with different sorbent amounts and counter ions, using separated [K + 2.2.2]OH " -fractions from "QMA" anion exchange cartridge, using total cartridge elution volume of longer stored [K + 2.2.2]OH ~ -solution, and finally using the total cartridge elution volume of [K + 2.2.2]OH " solution in t-butanol.
18 F-fluorination of precursors using fractions of ' [K 1 2.2.2JOH -eluate
Aqueous [ 18 F]fluoride obtained via the 18 0(p,n) 18 F nuclear reaction was applied to a "QMA light" anion exchange cartridge which in advance had been conditioned by applying 1 M KOH (2 ml), water (20 ml), 1 M K 2 C0 3 (2 ml), water (20 ml), and finally 5 ml air was purged through the cartridge.
[ 18 F]fluoride was retained quantitatively on the cartridge during these steps.
Elution of [ 18 F]fluoride from the cartridge performed with an azeotropically dried solution of [K + 2.2.2]OH " in acetonitrile (167.7 mM in KOH and 250 mM in Kryptofix 2.2.2.) (solution A).
For the 18 F-fluorination, a fraction of [K + 2.2.2] 18 F solution eluted from the resin was mixed with the precursor in question, in a total volume of the reaction mixture of 250 μΙ. The concentration of KOH was 33.2 mM and the concentration of [K + 2.2.2] was 50 mM. After the desired time interval, the reaction was quenched and analyzed by means of radio thin layer chromatography. F -incorporation
Reaction efficiency,
Compound Amount Reaction time
temperature x mean ±sd [%], (mg) (min)
(° C) number of experiments #
TET 1, 7 5 85 5 90 ± 4.0 (n=2)
89.9 ± 5.0
TET 1 10 85 5
n = 2
75.3 ± 4.7
Mannose-Triflate 2 10 90 10
n = 2
82.7 ± 1.5
Mannose-Triflate 2 20 90 10
n = 2
75 ± 8
Tosyl-Fallypride 3 2.5 95 10
n = 2
3- l -Boc-5'-0- dimethoxytrityl-3'-0- 10 95 10 70.2 ± 2.0 nosyl-thymidine 4
3-/\ABoc-5'-0-
54.0 ± 4.3 dimethoxytrityl-3'-0- 20 95 10
n = 2 nosyl-thymidine 4
l-(2,3-Diacetyl-5-tosyl- (alpha- 72.1 ± 5.9
10 95 10
Darabinofuranosyl)- n = 4 2-nitroimidazole 5
# As measured by radio thin layer chromatography.
1 TET: L-Tyrosine, 0-[2-[[(4-methylphenyl)sulfonyl]oxy]ethyl]-N-(triphenylmethy l)-, 1,1-dimethylethyl ester.
2 Tosyl-fallypride. Benzamide, 2,3-dimethoxy-5-[3-[[(4-methylphenyl)sulfonyl]oxy] propyl]-N-[[l-(2- propenyl)-2-pyrrolidinyl]methyl]-, (S)-.
3 Mannose triflate. beta-D-mannopyranose, 1,3,4,6-tetraacetate 2-(trifluoromethanesulfonate).
4 3-N-Boc-5'-0-dimethoxytrityl-3'-0-nosyl-thymidine. l(2H)-Pyrimidinecarboxylic acid, 3-[2-deoxy-3-0-
[(4-nitrophenyl)sulfonyl]-5-0-(triphen
2,6-dioxo-, 1,1-dimethylethyl ester.
5 l-(2 / 3-Diacetyl-5-tosyl-(alpha-Darabinofuranosyl)-2-nitroim idazole. IH-Imidazole, l-[2,3-di-0-acetyl- 5-0-[(4-methylphenyl)sulfonyl]-alpha-D-arabino-furanosyl]-2- nitro-.
6 Tosyl-FHBG: 6H-Purin-6-one, l,9-dihydro-9-[3-[[(4-methoxyphenyl) diphenylmethoxy]methyl]- 4-[[(4- methylphenyl)sulfonyl]oxy]butyl]-2-[[(4-methoxyphenyl)diphen ylmethyl]amino]-
7 A previous attempt resulted in an efficiency of about 30%. An explanation for this result is that the TET charge used in this particular case was not fresh. 18 F-fluorination of precursors using the total cartridge elution volume of ' [K 1 2.2.2J0H -solution
Aqueous [ 18 F]fluoride obtained via the 18 0(p,n) 18 F nuclear reaction was applied to "QMA light" anion exchange cartridge in question which in advance had been conditioned by applying 1 M KOH (2 ml), water (20 ml), 1 M KHC0 3 (2 ml), water (20 ml), and finally 5 ml air through the cartridge.
Alternatively, aqueous [ 18 F]fluoride was applied to a "PSA" anion exchange cartridge which in advance had been conditioned by applying 3 mL MeOH and 3 mL H 2 0. [ 18 F]fluoride was retained quantitatively on the cartridge during these steps.
Elutions of [ 18 F]fluoride from the cartridge were performed with an azeotropically dried solution of [K + 2.2.2]OH " in acetonitrile (167.7 mM in KOH and 250 mM in Kryptofix 2.2.2.) (solution A).
For the 18 F-fluorination, cartridges were eluted with a [K + 2.2.2]OH " -volume that allows ~90 % 18 F- recovery from the sorbent amount in question. Total [K + 2.2.2] 18 F " -eluate was mixed with the precursor in question.
Volume of
[K + 2.2.2]OH ~ -eluate (pL]
105 mg QMA 480
35 mg QMA 230
18 mg PS 200
After the desired time interval, the reaction was quenched and analyzed by means of radio thin layer chromatography.
# As measured by radio thin layer chromatography.
1 Benzyl bromoacetate: Bromoacetic acid benzyl ester
2 TET: L-Tyrosine, 0-[2-[[(4-methylphenyl)sulfonyl]oxy]ethyl]-N-(triphenylmethy l)-, 1,1-dimethylethyl ester.
3 Mannose triflate. beta-D-mannopyranose, 1,3,4,6-tetraacetate 2-(trifluoromethanesulfonate). 18 F-fluo nation using the total cartridge elution volume of [K 1 2.2.2JOH -solution with increasing precursor amount
Aqueous [ 18 F]fluoride obtained via the 18 0(p,n) 18 F nuclear reaction was applied onto "QMA light" anion exchange cartridge (35 mg), preconditioned in the aforementioned manner. [ 18 F]fluoride could be retained quantitatively on the cartridge.
Elutions of [ 18 F]fluoride from the cartridge were performed with 230 μΙ of azeotropically dried [K + 2.2.2]OH ~ in acetonitrile (167.7 mM in KOH and 250 mM in Kryptofix 2.2.2.) (solution A). The entire [K + 2.2.2] 18 F ~ -eluate was mixed with the precursor.
After the desired time interval, the reaction was quenched and analyzed by means of radio thin layer chromatography.
# As measured by radio thin layer chromatography.
1 TET: L-Tyrosine, 0-[2-[[(4-methy!phenyl)sulfonyl]oxy]ethyl]-N-(triphenylmethy l)-, 1,1-dimethylethyl ester. 18 F-fluorination of precursors using increasing fraction volumes of [K' 2.2.2]OhT eluate
Aqueous [ F]fluoride obtained via the 0(p,n) F nuclear reaction was applied to 35 mg "QMA light" anion exchange cartridge in question which in advance had been conditioned by applying 1 M KOH (2 ml), water (20 ml), 1 M KHC0 3 (2 ml), water (20 ml), and finally 5 ml air was purged through the cartridge.
Alternatively, aqueous [ 18 F]fluoride was applied to a 18 mg "PSA" anion exchange cartridge which in advance had been conditioned by applying 3 mL MeOH and 3 mL H 2 0. [ 18 F]fluoride was retained quantitatively on the cartridge during these steps.
Elutions of [ 18 F]fluoride from the cartridge were performed with increasing fraction volumes of aze- otropically dried [K + 2.2.2]OH ~ in acetonitrile 167.7 mM in KOH and 250 mM in Kryptofix 2.2.2.) (solution A).
For the 18 F-fluorination, increasing volumes of [K + 2.2.2] 18 F " -solution eluted from the resins were mixed with the precursor in a total volume of the reaction mixture of 250 μΙ. With 300 pL [K + 2.2.2] 18 F ~ eluate total volume was 300 pL The concentration of KOH and [K + 2.2.2] was rising according to the added volume used for labelling. After the desired time interval, the reaction was quenched and analyzed by means of radio thin layer chromatography. p [] cooa t o% I n i r i i n ■
50 100 150 200 250 300 350
Eluted cryptate in MeCN [μΙ_]
- Eluted 18F-activity ■ Overall RCY o Incorporation
Incorporation with increasing added volume of [K + 2.2.2]OH " -solution A from 35 mg waters QMA cartridges (For detailed composition of the eluent solution see text). Labeling of FAZA- precursor ( 5 l-(2,3-Diacetyl-5-tosyl-(alpha-Darabinofuranosyl)-2-ni troimidazoler. 1H- Imidazole, l-[2,3-di-0-acetyl-5-0-[(4-methylphenyl) sulfonyl]-alpha-D-arabino- furanosyl]-2-nitro-). Reaction condition: 5 mg, 95 °C, 10 min.
Incorporation with increasing added volume of [K + 2.2.2]OH ~ -solution from 18 mg "PSA" anion exchange cartridges (Machery-Nagel) (For detailed composition of the eluent solution see text). Labeling of FAZA-precursor { s l-(2,3-Diacetyl-5-tosyl-(alpha-Darabinofuranosyl)-2- nitroimidazoler. lH-Imidazole, l-[2,3-di-0-acetyl-5-0-[(4-methylphenyl) sulfonyl]-alpha- D-arabino-furanosyl]-2-nitro-). Reaction condition: 5 mg,
95 °C, 10 min.
pH-values were measured after addition of different volumes of [K + 2.2.2.]OH ~ in H 2 0 (167.7 mM in KOH and 250 mM in Kryptofix 2.2.2.) to 35 mg QMA light anion exchange sorbent which in advance had been conditioned by applying 1 M KOH (2 ml), water (20 ml), 1 M KHC0 3 (2 ml), water (20 ml), and finally 5 ml air.
Alternatively, pH-values were measured after addition of different volumes of [K + 2.2.2.]OH " in H 2 0 (167.7 mM in KOH and 250 mM in Kryptofix 2.2.2.) to 18 mg "PSA" anion exchange sorbent which in advance had been conditioned by applying 3 mL MeOH and 3 mL H 2 0.
100 150 200 250 300 350
Volume added for pH-measurement [μί]
pH-values with increasing added volume of [K + 2,2.2]OH -solution at 35 mg waters QMA cartridges.
pH-values and incorporation with increasing added volume of [K + /2.2.2] OH " solution at 18 mg "PSA" anion exchange catrridges (Machery-Nagel). 18 F-fluorination using fractions of [K h 2.2.2]0hT-eluate from anion exchange cartridge with different sorbent amounts
Aqueous [ 18 F]fluoride obtained via the 18 0(p,n) 18 F nuclear reaction was applied to a "QMA light" anion exchange cartridge in question which in advance had been conditioned by applying 1 M KOH (2 ml), water (20 ml), 1 M K 2 C0 3 (2 ml), water (20 ml), and finally 5 ml air was purged through the cartridge. Alternatively, aqueous [ 18 F]fluoride was applied to a "PSA" anion exchange cartridge which in advance had been conditioned by applying 3 mL MeOH and 3 H 2 0. [ 18 F]fluoride was retained quantitatively on the cartridge during these steps.
Elution of [ 18 F]fluoride from the cartridge performed with an azeotropically dried solution of [K + 2.2.2]OH ~ in acetonitrile (167.7 mM in KOH and 250 mM in Kryptofix 2.2.2.) (solution A).
For the 18 F-fluorination, cartridges with different sorbent amounts were eluted with a [K + 2.2.2]OH "~ volume in question (For detailed elution volume of the different sorbent see text above). For [K + 2.2.2] 18 F solution was mixed with the precursor in a total volume of the reaction mixture of 250 μΙ. The concentration of KOH was 33.2 mM and the concentration of [K + 2.2.2] was 50 mM. After the desired time interval, the reaction was quenched and analyzed by means of radio thin layer chromatography.
# As measured by radio thin layer chromatography.
1 l-(2,3-Diacetyl-5-tosyl-(alpha-Darabinofuranosyl)-2-ni troimidazole. lH-Imidazole, l-[2,3-di-0-acetyl- 5-0-[(4-methylphenyl)sulfonyl]-alpha-D-arabino-furanosyl]-2- nitro-. 18 F-fluorination using fractions of [t 2.2.2]OrT-eluate from anion exchange cartridge with different counter ions
Aqueous [ 18 F]fluoride obtained via the 18 0(p,n) 18 F nuclear reaction was applied to a "QMA light" anion exchange cartridge in question which in advance had been conditioned by applying 1 M KOH (2 ml), water (20 ml), 1 M KHC0 3 (2 ml), water (20 ml), and finally 5 ml air was purged through the cartridge.
Alternatively, aqueous [ 18 F]fluoride was applied to a "PSA" anion exchange cartridge which in advance had been conditioned by applying 3 mL MeOH and 3 H 2 0. [ 18 F]fluoride was retained quantitatively on the cartridge during these steps.
Alternatively, aqueous [ 18 F]fluoride obtained via the 18 0(p,n) 18 F nuclear reaction were applied to" QMA light" anion exchange and "PSA" anion exchange cartridges in question which in advance had been conditioned by applying 1 M KOH (2 ml), water (20 ml), 1 M K 2 C0 3 (2 ml), water (20 ml), and finally 5 ml air was purged through the cartridges.
Elution of [ 18 F]fluoride from the cartridges was performed with an azeotropically dried solution of [K + 2.2.2]OH ~ in acetonitrile (167.7 mM in KOH and 250 mM in Kryptofix 2.2.2.) (solution A).
For the 18 F-fluorination, cartridges with different counterions were eluted with [K + 2.2.2]OH " volume in question.
(see text above). [K + 2.2.2] 18 F ~ solution was mixed with the precursor in a total volume of the reaction mixture of 250 μΙ. The concentration of KOH was 33.2 mM and the concentration of [K + 2.2.2] was 50 mM. After the desired time interval, the reaction was quenched and analyzed by means of radio thin layer chromatography.
# As measured by radio thin layer chromatography.
1 Incorporation of l-(2,3-Diacetyl-5-tosyl-(alpha-Darabinofuranosyl)-2-nitroimi dazole. lH-Imidazole, 1- [2,3-di-0-acetyl-5-0-[(4-methylphenyl)sulfonyl]-alpha-D-arab ino-furanosyl]-2-nitro-. Reaction condition: 10 mg, 95 °C, 10 min 18 F-fluorination of precursors using separated [lC 2.2.2]Ohf -fractions from "QMA" anion exchange cartridge
Aqueous [ 18 F]fluoride obtained via the 18 0(p,n) 18 F nuclear reaction was applied to 35 mg "QMA light" anion exchange cartridge which in advance had been conditioned by applying 1 M KOH (2 ml), water (20 ml), 1 M KHCO3 (2 ml), water (20 ml), and finally 5 ml air was purged through the cartridge.
[ 18 F]fluoride was retained quantitatively on the cartridge during these steps.
Elution of [ 18 F]fluoride from the cartridge performed with azeotropically dried solution of [K + 2.2.2]OH ~ in acetonitrile (167.7 rtiM in KOH and 250 mM in Kryptofix 2.2.2.) (solution A).
For the 18 F-fluorination, [K + 2.2.2] 18 F solution eluted from the resin was fractionated and collected elution aliquots mixed with the precursor in a total volume of the reaction mixture of ~250 μΙ. After the desired time interval, the reaction was quenched and analyzed by means of radio thin layer chromatography.
# As measured by radio thin layer chromatography.
1 Incorporation of l-(2,3-Diacetyl-5-tosyl-(alpha-Darabinofuranosyl)-2-nitroimi dazole. lH-Imidazole, 1- [2,3-di-0-acetyl-5-0-[(4-methylphenyl)sulfonyl]-alpha-D-arab ino-furanosyl]-2-nitro-. Reaction condition: 5 mg, 95 °C, 10 min
18 F-fluorination using total cartridge elution volume of longer stored [l 2.2.2]OH ~ -solution
Aqueous [ 18 F]fluoride obtained via the 18 0(p,n) 18 F nuclear reaction was applied to 35 mg "QMA light" anion exchange cartridge (35 mg) which in advance had been conditioned by applying 1 M KOH (2 ml), water (20 ml), 1 M KHC0 3 (2 ml), water (20 ml), and finally 5 ml air was purged through the cartridge. [ 18 F]fluoride was retained quantitatively on the cartridge during these steps.
Elution of [ 18 F]fluoride from the cartridge performed with an azeotropically dried solution of [K + 2.2.2]OH ~ in acetonitrile (167.7 mM in KOH and 250 mM in Kryptofix 2.2.2) (solution A), which was prepared at the first day of the experiments. Fractions of the solution were portioned in single vials, MeCN dried and stored in darkness. Before every experiment the residue in one vial was dissolved in the appropriate MeCN-volume evaporated before and used for elution.
For the 18 F-fluorination, [K + 2.2.2] 18 F " solution eluted from the resin was mixed with the precursor. After the desired time interval, the reaction was quenched and analyzed by means of radio thin layer chromatography. F -incorporation efficiency Charge 1 Charge 2
1 81 90
2 76
3 84 89
4 89
7 92 91
# As measured by radio thin layer chromatography
1 Benzyl bromoacetate. Bromoacetic acid benzyl ester: Reaction condition: 5 mg, 85 °C, 5 min.
18 F-fluorination of precursors using the total cartridge elution volume of [K 1 2.2.2]OH ~ solution in t- butanol
Aqueous [ 18 F]fluoride obtained via the 18 0(p,n) 18 F nuclear reaction was applied to 35 mg "QMA light" anion exchange cartridge in question which in advance had been conditioned by applying 1 M KOH (2 ml), water (20 ml), 1 M KHC0 3 (2 ml), water (20 ml), and finally 5 ml air through the cartridge.
[ 18 F]fluoride was retained quantitatively on the cartridge during these steps.
Elutions of [ 18 F]fluoride from the cartridge were performed with an azeotropically dried solution of [K + 2.2.2]Ohr in acetonitrile (167.7 mM in KOH and 250 mM in Kryptofix 2.2.2.) (solution A2).
For the 18 F-fluorination, 230 pL Total [K + 2.2.2] 18 F " -eluate was mixed with the precursor in question. After the desired time interval, the reaction was quenched and analyzed by means of radio thin layer chromatography.
1 Benzyl bromoacetate, Bromoacetic acid benzyl ester: Reaction condition: 5 mg, 85 °C, 5 min. Figure A
0 100 200 300 400 500 600
Eluted cryptate in MeCN [μΙ_]
Elution profiles for the removal of [ F]fluoride from waters QMA cartridges with solutions of different [K + 2.2.2]OH " concentration (For detailed composition of the eluent solutions see text).
Solution A (black circles): [ 18 F]fluoride activity was eluted with up to 92% efficiency from the QMA cartridge.
Solution B (white circles): [ 18 F]fluoride activity was eluted form the cartridge with up to 82% efficiency. Compared to the elution with solution A, the activity was eluted earlier, but in a comparable overall volume.
Solution C (data not shown): [ 18 F]fluoride activity could not be eluted from the cartridge even with 900 pL of solution C. Figure B
0 200 400 600 800 1000
Eluted cryptate in MeCN [μ!_]
Elution profiles for the removal of [ F]fluoride from waters QMA cartridges with solution A, but different cartridge conditioning procedures (For detailed composition of the eluent solution see text).
Conditioning QMA 1 (black circles): Flushing the resin with 1 M KOH (2 ml), water (20 ml), 1 M K 2 C0 3 (2 ml), water (20 mL), and finally 5 ml air. Subsequent trapping of 18 F-fluoride and elution with solution A.
Conditioning QMA 2 (white circles): flushing the resin with 1 M K 2 C0 3 (2 ml), water (20 mL), and finally 5 ml air. Subsequent trapping of [ 18 F]fluoride and elution with solution A.
Compared to conditioning QMA 2 conditioning with QMA 1 provides higher 18 F- recovery with a lower overall volume necessary. Figure C
0 200 400 600 800 1000
Eluted cryptate in MeCN [μΙ_]
Elution profiles for the removal of [ F]fluoride from "PSA" anionic exchange cartridges (Machery-Nagel) with solution A, but different cartridge conditioning procedures (For detailed composition of the eluent solution see text).
Conditioning PSA 1 (black circles): flushing the resin with 3 ml MeOH followed by 3 ml H 2 0. Subsequent trapping of 18 F-fluoride and elution with solution A.
Conditioning PSA 2 (white circles): flushing the resin with 0,5 mL H 2 0. Subsequent trapping of [ 18 F]fluoride and elution with solution A.
Both conditioning procedures, PSA 1 and PSA 2, leads > 95 % overall elution with similar elution volume necessary.
Figure D
400 600 800 1000
Eluted cryptate in MeCN [μΙ_]
Elution profiles for the removal of [ 8 F]fluoride from waters "QMA light" cartridges (black circles) and "PSA" anion exchange cartridge (Machery-Nagel) (white circles) with solutions A
(For detailed composition of the eluent solution see text). Conditioning of the PSA resin was carried out as described in "conditioning PSA 1"; conditioning of the QMA cartridge was carried according to "conditioning QMA Γ.
Compared to sorbent 1, [ 18 F]fluoride activity of sorbent 2 allows a higher elution volume but offers an overall elution up to >95%.
Figure E
El u ted cryptate in MeCN [μΙ_]
Elution profiles for the removal of [ F]fluoride from waters QMA cartridges (Conditioning QMA 1) with different amount of sorbent (105 mg, 50 mg, 35 mg and 25 mg) using solution A (For detailed composition of the eluent solution see text).
Black circles: 105 mg, white circles: 50 mg, black squares: 35 mg, 25 mg (data not shown).
Figure F
400 600 800 1000
Eluted cryptate in MeCN [\JL]
Elution profiles for the removal of [ 18 F]fluoride from "PSA" anion exchange cartridges (Machery-Nagel) with different amount of sorbent (6.3 mg, 15.8 mg and 6.3 mg) using solution A (For detailed composition of the eluent solution see text).
Black circles: 6.3 mg, white circles: 15.8 mg and white squares: 45 mg
Figure G
200 300 400 500 600 700 800 900
Eluted cryptate in MeCN [μ!_]
Elution profiles for the removal of [ F]fluoride from a waters QMA cartridges (black circles) and "PSA" anion exchange cartridge (Machery-Nagel) (white squares) with solution A under optimized conditions with similar sorbent amount (~45-50 mg) and geometry (For detailed composition of the eluent solution see text). Conditioning of the "PSA" resin was carried out as described in "conditioning PSA 1"; conditioning of the "QMA light" cartridge was carried according to conditioning QMA Γ (see text).
Similar overall volume necessary for similar overall elution up to 95%.
Figure H
0 50 100 150 200 250 300 350 400
Eluted cryptate in MeCN [μΙ_]
Elution profiles for the removal of [ 18 F]fluoride from waters QMA cartridges (35 mg) with solution A but different counterions (For detailed composition of the eluent solution see text).
Conditioning QMA 1 (black circles): flushing the resin with 1 M KOH (2 ml), water (20 ml), 1 M K 2 C0 3 (2 ml), water (20 mL), and finally 5 ml air. Subsequent trapping of 18 F-fluoride and elution with solution A.
Conditioning QMA 3 (white circles): flushing the resin with 1 M KOH, (2 ml), water (20 ml), 1 M KHC0 3 (2 ml), water (20 ml), and finally 5 ml air.
The F-elution form cartridges conditioned with QMA 1 started earlier but elution volumes were necessary for both cartridge types.
Figure I
150 200 250 300 350 400 450
Eluted cryptate in MeCN [μΙ_]
Elution profiles for the removal of [ 8 F]fluoride from "PSA" anion exchange cartridge (15 mg) (Machery-Nagel) with solutions of composition A (solution A), but different counteri- ons (For detailed composition of the eluent solution see text).
Conditioning PSA 1 (black circles): flushing the resin with 3 ml MeOH followed by 3 ml H 2 0. Subsequent trapping of 18 F-fluoride and elution with solution A.
Conditioning PSA 3 (white circles): flushing the resin with 1 M KOH, (2 ml), water (20 ml), 1 M K 2 C0 3 (2 ml), water (20 ml), and finally 5 ml air.
Figure J
200 300 400 500
Eluted cryptate in used solvent [μί]
Elution profiles for the removal of [ 8 F]fluoride from waters QMA cartridges (35 mg) with cryptate in t-butanol (white circles, solution A2) and cryptate in amyl alcohol (black circles, solution A3) (For detailed composition of the eluent solution see text).
Conditioning QMA 1 (black circles): flushing the resin with 1 M KOH (2 ml), water (20 ml), 1 M K 2 C0 3 (2 ml), water (20 mL), and finally 5 ml air. Subsequent trapping of 18 F-fluoride and elution with solution A.