[18F]FALLYPRIDE FOR INJECTION:MASTER BATCH RECORD
PET Radiopharmaceutical Sciences Section,Date of review: 05/17/04
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD20892
Approved by:______Initials__________ Date:______
Victor W.Pike, Ph.D
Chief, PET Radiopharmaceutical Sciences,NIMH
Batch # FAY-______Date______
Hot-cell # 5, Automated Method
1. General notes:
[18F]Fallypride for Injection is prepared by an automated method witha GE TRACERlab FXFNmodule, through nucleophilic displacement of a tosylate leaving group by no-carrier-added [18F]fluoride ion in dry acetonitrile. Endotoxin and sterility tests are performed on all samples.
2. Components:
Cyclotron material: [18F]fluoride ion productionItem / Reagent / Specifications / Manufacturer / Lot # / Quantity
1 / [18O]water / 95 atom % / Rotem Industries
Others______/ 0.25–1.5 mL
Reagents: [18F]fluoride ion-Kryptofix 2.2.2-potassium carbonatesolution
Item / Reagent /
Specifications
/ Manufacturer / Lot # / Expiry date / Quantity1 / Kryptofix 2.2.2 / Anhydrous
98% / Aldrich / 3.5−5.0 mg
2 / Potassium carbonate / Anhydrous
99.99% / Aldrich / 0.35−0.5mg
3 / Acetonitrile / Anhydrous 99.8% / Aldrich / 4– 5 mL
4 / Water / De-ionized / M; Millipore; In house / 0.5 mL
5 / Stock solution: Kryptofix 2.2.2– potassium carbonate / SOP # GP011 / In house / 80 ±20 μL
Reagents: production of [18F]Fallypride for Injection
Item /
Reagent
/Specifications
/ Manufacturer /Lot #
/ Expiry date / Quantity1 / Tosylate precursor / > 95% / ABX Advanced Biochemical Compounds / 2.5 ±0.5 mg
2 / Acetonitrile / Anhydrous 99.8% / Aldrich / 0.5–1.0 mL
3 / Acetonitrile / HPLC grade / Burdick & Jackson / ~ 500 mL per 1L of HPLC eluent
4 / HPLC water / HPLC grade / EMD Science / ~ 500 mL per 1L of HPLC eluent
5 / Fallypride preparative HPLC mobile phase / SOP # GP103 / In house / 1–4 L
6 / Fallypride analytical HPLC
mobile phase / SOP # GP103 / In house / 1–4 L
7 / Stock sodium acetate buffer
(0.25M; pH = 5.5) / SOP # GP105 / In house / 10–15 mL
8 / Triethylamine (TEA) / 99.5% / Aldrich / 0.6%TEA/L solvent (v/v)
9 / Ammonium formate / 99.995+% / Aldrich
(Cat. # 516961) / 5 mM
10 / Ethanol, absolute / USP / Warner-Graham / 100 mL
11 / Sodium Chloride / Injection, USP / APP / 9 mL
Inert components: [18F]Fallypride
Item /Component
/Specifications
/ Manufacturer /Lot #
/ Expiry date / Quantity1 / Preparative HPLC column / Luna (21.2 x 250 mm; 10 µm; 100 Å ) / Phenomenex / Serial # / 1
2 / Guard column / Luna (21.2 x 50 mm;10 µm; 100 Å) / Phenomenex / Serial # / 1
3 / Analytical HPLC column / Prodigy (4.6 x 250 mm; 5 µm) / Phenomenex / Serial # / 1
4 / C18 Sep-Pak / Plus / Waters / 1
5 / Sterile empty vial (10 or 30 mL) / Sterile, non-pyrogenic / APP / 1
6 / Filter (Millex GV; 0.22 µm; 25 mm) / Sterile, non-pyrogenic / Millipore / 1
7 / Filter (Millex GV 0.22 µm; 4 mm) / Sterile, non-pyrogenic / Millipore / 1
8 / Syringe
(1 mL size) / Luer, sterile / Norm-Ject / 1
9 / Syringe
(10 mL size) / Luer, sterile / Norm-Ject / 1
10 / Needles / 18 G x 31/2 / B-D / 3
11 / Needle / 20 G 11/2 / B-D / 1
12 / Needles / 18G11/2 / B-D / 5
13 / Activated charcoal / 4260AC / Ommifit / 4260ac / As needed
Radiosynthesis equipment
Item / Equipment / Manufacturer / Model / Serial # / Checked
1 / Synthesis apparatus / General Electric / TRACERlab FXFN / 14390
2 / HPLC pump / Beckman Coulter / System Gold 126
solvent model / 342-2187
3 / UV absorbance detector / Beckman Coulter / System Gold 166 detector / 332-2187
4 / Radioactivity detector / Bioscan / Flow-Count / 0409-316
Summary Records of [18F]Fallypride for Injection - Work Sheet
Reaction substances record
Item / Quantity / CommentsEOB (End of bombardment) / Time:
Starting radioactivity of [18O]water-[18F]fluoride ion / mCi at
Volume of [18O]water-[18F]fluoride ion / μL
Stock Kryptofix 2.2.2-potassium carbonate solution / μL
Tosyl-fallypride / mg
Acetonitrile / μL
TRACERlab FXFNPressure performance record
Item / Initial / During reaction / CommentsCompressed air pressure / kPa / kPa
Nitrogen pressure / kPa / kPa
Pressure of reactor / kPa / kPa
Radioactivity record / (mCi at______)
Item / Reagent / Radioactivity measurement
1 / Cyclotron run #
2 / Starting radioactivity / at
3 / Radioactivity in reactor of TRACERlab FXFN / at
4 / Radioactivity in final formula / at
6 / Overall yield of production
Chemist ______Initials______Date______
Batch # FAY-______Date______
3. Pre-synthesis Procedures
____1. Clean GE TRACERlab FXFN Module and Hot-Cell 5.
1.1Follow the program "CLEAN BOX" (SOP # MP101).Clean and dry the GE TRACERlab FXFN
module on the day of preparation or the day before. All the reagent vessels and transfer lines of the module should be washed with acetone and dried under nitrogen flush.
1.2Inspect hot-cell compounding/dispensing area for cleanliness. Remove extraneous materials and labels. Spray ethanol to clean this area.
1.3Inspect and set if necessary charcoal trap in the module
____2. Prepare [18F]fluoride ion-Kryptofix 2.2.2- potassium carbonatesolution
2.1Add stock solution of Kryptofix 2.2.2 and potassium carbonate (70−100 µL) to V-Vial (1 mL size).
2.2Take V-Vial to the Cyclotron Facility (CC, NIH) to collect [18F]fluoride ion in 18O-water. The fluorine-18 radioactivity should be between 100 and 250 mCi in the 18O-water (≤500 µL).
____3. Verify the integrity of the glassy carbon reaction vessel assembly (containing small magnetic bar stirrer) in the TRACERlab FXFNmodule. Follow SOP# MP202and record the data in the sheet of Summary Records of [18F]Fallypride for Injection run:
3.1. Check vacuum pressure (gauge, kPa).
3.2.Check compressed air pressure (kPa)
3.3.Check nitrogen pressure (kPa)
_____4. Prepare the preparative HPLC System
4.1Start the HPLC computer software, enter the necessary information and select the right method “Fally_Production”
4.2 Verify that mobile phase bottles are in the right position on the Beckman System Gold Delivery module.
4.2.1The solvent bottle of 0.6% TEA connects to the B1 line (marked red)
4.2.2The solvent bottle of HPLC water connects to A1 line (markedred)
The above connections should match the set-up in the method “Fally_Production”.
4.3Set acetonitrile w 0.6%TEA /HPLC water (30: 70 v/v) as initial conditions. Equilibrate the Luna preparative column with preparative mobile phase at2 mL/min.
4.4Check the pump pressure while increasing the flow rate to 9 mL/min, then reduce to 2mL/min if the pump pressure is satisfactory.
4.5.Pump the HPLC mobile phase through the preparative column at a low flow rate (1−2 mL/min) and maintain this until purification time.
_____5. Prepare at the clean bench a Sterile Filtration/Collection Unit, using sterile technique:
5.1. Attach a membrane filter unit (GV; 25 mm; 0.2 µm) to a sterile disposable needle (18 guage) and insert it completely into a sterile, empty collection vial (10 mL or 30 mL size if needed).
5.2. Attach a Chemovent needle or a combination of needle and syringe driven filter (4 mm) tothe collection vial.
5.3. Attach a prepared label for [18F]Fallypride for Injection with the current batch number and date.
_____6.Open the TRACERlab FXFNComputer Controlling System ("Admin" and "Tracerlab") and Select synthesis method: "Fallypride". Click “Synthesis-Start” to start preparation of the synthesis.
_____7. Enter all the needed information (including date, radiotracer, batch #, human preparation #) into the corresponding method of the HPLC computer software program.
_____8. Check that you have the correct UV absorbance wavelength (254 nm) and the correct settings for the gamma detector (diode/20K/2/rate).
_____9. Place the solvents and precursor solution in the corresponding module support vials:
______Support vial 1:acetonitrile (0.5 mL)
______Support vial 2:used as port (1 mL V-vial w 100 µL [18F]fluoride ion-Kryptofix 2.2.2-potassium carbonate)
______Support vial 4:dry acetonitrile (0.7−1 mL) plus tosyl-fallypride (2 ± 0.5 mg).
______Support vial 5:dry acetonitrile (0.5 mL)
______Support vial 6:dry acetonitrile (1.5 mL)
______Support vial 7:sterile saline solution (9–13.5 mL)
______Support vial 8:Ethanol for Injection (1–1.5 mL)
______Support vial 9sterile water (8 mL)
______Large 10 mL V-vial 0.5 mL sterile water with 0.6%TEA.
______Large bulb vessel:HPLC water (90 mL) and sodium acetate solution (250 mM; pH 5.5; 10 mL; see SOP # GP105).
____10. Check that the line for delivery of the complex goes through valve V2 and into the reactor vessel (glassy carbon).
____11. Place acetonitrile (0.5 mL) in a syringe (3 mL size) connected to an external line going into the "complex" vial (for recovery of [18F]fluoride ion-Kryptofix 2.2.2-potassium carbonateleft in transportation vial and its addition to the reaction vessel).
____12. Activate one C-18 Sep-Pak Plus with ethanol (10 mL) and then sterile water (10 mL) and place in its corresponding connection site in the module.
____13. Place the small double-necked round bottom flask to receive the ethanolic eluted from the Sep-Pak in its corresponding location in the module.
____14. In a lead pot place a sterile empty vial (30 mL size) fitted with a vent needle/filter and a second needle and Millev GV filter for sterilization of the final product.
____15. Place enough dry-ice-acetone mixture in the vacuum trap.
____16. Check that the nitrogen and compressed air valves close to the hot-cell are open (also check the nitrogen cylinder).
____17. Assure that you have change syringes and needles and the used de-pyrogenated reaction vessel (with magnetic stirrer), HPLC collection bulb, 10 mL V-vial, Product vial, sterile 15 mL vial etc.
____18. Get at hand a standard solution of fallypride for the validation of HPLC analysis system.
4. [18F]Fallypride synthesis and purification
_____1. Preparation of [18F]/K222/K2CO3 complex (Kryptofix 2.2.2-[18F]fluoride-potassium carbonate).
1.1. Using the dose calibrator (Biodex AtomLab 300) set at F-18, determine the amount of radioactivity that will be transferred to the clean glassy carbon reaction vessel (20 mL size). Record the data in the summary record sheet: EOB, starting radioactivity of [18F]Fluoride and volume of [18O]water.
1.2. Connect one vent needle, one needle and line from external syringe containing acetonitrile (0.5 mL) and one spinal needle and line going from the radioactive solution through V2 to reactor vial. Make sure that spinal needle is all the way down to the bottom of the radioactive solution.
_____2. Starting the preparation and transferring the [18F]/K222/K2CO3 complex.
2.1Start the TRACERrLab automated software program. Select Synthesis: "Fallypride" in method box, click “Synthesis-Start” in main menu bar, follows process instructions. Organize "Instrument screen", "Program" and "Graphics" in the computer screen.
2.2The program had already started turning the vacuum pump ON and transferring the [18F]/K222/K2CO3 complex from the V-vial (1 mL) into the reaction vessel; then MESSAGE BOX comes out. At this moment add acetonitrile (0.5 mL) from the outside syringe into the transport F-18 vial (pushing with air until the line is empty and the vial is full). Press “Enter” or click “OK” when done.
_____3. [18F]Fallypride synthesis step 1: remove water from the [18F]/K222/K2CO3 complex
3.1The reactor temperature will be increased first to 65 ºC and later to 88 ºC (T1 + 3 min) and then back to 60 ºC. At this point, the contents of V5 are added and the temperature cycle starts again: heat at 65ºC, then at 88ºC and finally at 60 ºC (drying process).
_____4. [18F]Fallypride synthesis step 2: [18F]Fallypride reaction. Check V4 and reaction temperature.
4.1Once drying is complete, V4 will open and the tosylate precursor in acetonitrile (1 mL will be added to the reaction vessel and the mixture heated at 105 ºC for 30 min. At the end of the reaction, the reaction vessel will be cooled to 35 ºC and the first part of the Program will finish.
4.2 A message will show saying "End of synthesis, Hit OK to continue to next Program". Hit CONTINUE.
_____5. Prepare Beckman HPLC system for purification.
5.1Verify that the HPLC is in Preparative and in LOAD mode and that the detectors are in the right settings.
5.2Check that method setting of Beckman HPLC instruments is correct.
5.3Set the flow rate to 9 mL/min. Wait for pressure to go up.
5.4Click “Single run” in Beckman HPLC software to start acquisition and wait the acquisition Box appears.
5.5Check that the injector is in inject position
_____6. [18F]Fallypride purification step 1: transferring the initial reaction mixture.
6.1Second program continues purification process. The reaction mixture will be transferred to the V-vial (10 mL size) containing water with 0.6%TEA (0.5 mL) for pre-dilution before HPLC injection.
6.2 Check that the injector transfer needle in V-vial (10 mL) is far above the level of the solution.
6.3 Check that valve 6 will open and the acetonitrile (1.5 mL)is added to the reaction vessel.
_____7. [18F]Fallypride purification step 2: loading and injection
7.1After transferring, a message box will appear "System now ready to load Loop. Hit OK when
completed". System will load the reaction mixture solution from T-vial (10 mL) into HPLC loop automatically.
7.2 At this moment, a new message box will appear “System now ready to inject. Hit OK when the loading is completed.”
7.3Watch the T-vial (10 mL) closely. When the loading of solution is completed, hit OK.
7.4At same time hit “OK” to start the HPLC run in Beckman software.
_____8. [18F]Fallypride purification step 3: HPLC separation and product collection.
8.1In GRAPHICS observe the UV and radioactivity traces on the data acquisition systems.
8.2When the desired peak starts to come out, click on the collect arrow. Record the Rt. as______min.
8.3The mobile phase containing the product will be added to the bulb vessel containing sterile water for dilution (90 mL) and of sodium acetate buffer (10 mL) for dilution and neutralization.
8.4At the end of collection click on the end collection arrow (the mobile phase will go into the waste reservoir).
5. [18F]Fallypride for InjectionFormulation
_____1. The program will continue and the diluted fraction will be passed through the solid phase cartridge (C-18 Sep-Pak) for concentration. This process is programmed for 6.5 min.
_____2. After passing through C-18 Sep-Pak, valve V9 will open and C-18 Sep-Pak will be washed with sterilized water.
_____3. After the water wash, V8 will open and the product will be eluted from the C-18 Sep-Pak with 1 mL ethanol into the two-necked round bottom flask.
_____4. Program will stop to allow sampling of the ethanolic solution. The message "Take QC sample and hit OK to proceed with filtration" will appear. Take sample for QC if needed.
_____5.The program can be continued hitting OK. Valve 7 will open and saline solution will be added to the round bottom flask containing [18F]Fallypride for Injection through the 0.22 µm Millex filter for filtration and sterilization.
_____6. After filtering product, the program will complete. When the message “reaction completed” comes out, remove sterile filter and measure the radioactivity of the product vial, which contains the final [18F]Fallypride for Injection.
Radioactivity of [18F]Fallypride for Injection: ______mCi at______.
_____7. Sampling for QC: The 1cc sterile polypropylene syringe is used to withdraw a 1 mL sample of the formulated [18F]Fallypride for Injection in a certified laminar flow sterile cabinet. The 1 mL sample is transferred to a sterilized depyrogenated tube labeled with the information “final [18F]FALLYPRIDE”, batch number and the date. The vial is stored in a lead pig. This aliquot will be used for the following QC tests: radioconcentration, radiochemical purity, specific radioactivity, pH, residual solvents, and bacterial endotoxins (LAL test).
Synthesis comments:
______
The approved procedure has been followed without any deviation.
Chemist ______Initials______Date______
Batch # FAY-______Date______
Document 2. [18F]Fallypride for Injection: Master Batch RecordPage 1 of 10