[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
List of SOPs for [18f]Fallypride for Injection
Name of SOP
1
Preparation of Stock Solution of Kryptofix 2.2.2* and
SOP #
SOP#GP101
Potassium Carbonate
2
Cleaning Procedure for Radiochemistry Glassware, Reactor
SOP # GP102
Vials and Magnetic Bars
3
Fallypride HPLC Mobile Phase Preparation
SOP # GP103
4
Micropore Filter Testing and Drug Product /Filter
SOP # GP104
Compatibility (Bubble Point Test).
5
Preparation of Stock Acetate Buffer
SOP # GP105
6
BOXCLEAN Program (GE Tracerlab FXFN)
SOP # MP201
7
GE Tracerlab FXFN Module. Vacuum Pressure Check
SOP # MP202
8
Production of [18F]Fallypride for Injection; Part 1: Pre-
SOP # MP203
Synthesis Procedures
9
Production of [18F]Fallypride for Injection; Part 2: Synthesis
SOP # MP204
and Purification
10
Production of [18F]Fallypride for Injection; Part 3:
SOP # MP205
Formulation
11
Standard Curve of Reference Fallypride
SOP # QA301
12
Analysis of Organic Residues by Gas Chromatography
SOP # QA304
in [18F]Fallypride for Injection
13
Analytical HPLC QC-Method
SOP # QA305
14
Sterility Test
SOP # QA306
15
P Pyrogen Test (LAL Bacterial Endotoxin Test).
SOP # QA307
Si Simplified Procedure
16
Analysis of The Purity of Fallypride and Tosyl-Fallypride by
SOP # QA308
HP HPLC
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 1 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
SOP # GP101
Preparation of Stock Solution of Kryptofix 2.2.2* and Potassium Carbonate
Approved by: _____________________________Initials________ Date:___________
Victor W Pike, Ph.D., Chief, PET Radiopharmaceutical Sciences, NIMH
Purpose: to prepare a stock solution of Kryptofix 2.2.2* and potassium carbonate
1. Procedure:
1. Using an analytical balance and a plastic screw-cap vial (20 mL size), weight out 25 mg of
potassium carbonate (Aldrich, anhydrous, 99.99%). Add deionized water (0.5 mL) and mix
to dissolve.
2. Using weighing paper, weigh 250 mg of Kryptofix 2.2.2 (anhydrous; 98%; Aldrich) and
transfer to the same vial. Add dry acetonitrile (4.5 mL) and mix to dissolve.
3. Place appropriate label showing preparation and expiry date (to be determined) and keep
the solution in the refrigerator until use (do not store in glass vial).
4. Record the data of Kryptofix 2.2.2-potassium carbonate complex in Table 1.
2. Record: Lot #__________________________
Table 1. Reagents : [18F]fluoride-Kryptofix 2.2.2-potassium carbonate complex
Item
1
2
Reagent
Specifications
Manufacturer
Kryptofix 2.2.2
Anhydrous
Aldrich Chem.
(250 mg)
98%
Co.
Potassium
Anhydrous
Aldrich Chem.
carbonate
99.99%
Co.
Anhydrous 99.8%
Aldrich Chem.
Lot #
Expiry date Quantity
(25 mg)
3
Acetonitrile
(4.5 mL)
4
Water
(0.5 mL)
Co.
De-ionized
M; Millipore, In
house
Note: 100 µL of solution contains 0.5 mg potassium carbonate and 5 mg Kryptofix 2.2.2
Chemist_____________________________Initials_________________Date________________
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 2 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
SOP # GP102
Cleaning Procedure for Radiochemistry Glassware, Reactor Vials and Magnetic Bars
Approved by: _____________________________Initials________ Date:_____________
Victor W Pike, Ph.D., Chief, PET Radiopharmaceutical Sciences, NIMH,
Purpose: to clean glassware, reactor, vials and magnetic bars that are used in the
production of [18F]Fallypride for Injection.
Procedure:
1. Rinse with acetone (HPLC grade) to remove organic residues.
2. Rinse with hexane (HPLC grade) to remove residual silicon oil.
3. Bathe the glassware in an aqueous 2% solution of Liqui-Nox (Valconox, made from dilution
of a 10% stock solution).
a) Bring solution to vigorous boiling for 5–10 min.
b) Swab material being washed with wood, cotton-tipped applicators (6 in).
4. Rinse glassware 3 times with Millipore water.
a)
Carefully spot-check each item.
b) Give final rinses with de-ionized water bottle (3 times).
c)
Leave the clean material to drip, over paper towel.
5. Place small V-vials, magnetic bars and reaction vials in beakers (Pyrex) and cover with
aluminum foil. Place material in the oven at 170−180 ºC and allow to dry overnight.
6. Keep all the material in the oven until use.
Notes:
a) Except for the aqueous Valconox that can be disposed in the sink, all other washing
solvents must be placed in their respective waste container.
b) In the Oven Log enter date and temperature IN and date and temperature OUT.
Chemist_____________________________Initials_________________Date________________
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 3 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
SOP # GP103
Fallypride HPLC Mobile Phase Preparation
Approved by: _____________________________Initials________ Date:_____________
Victor W Pike, Ph.D., Chief, PET Radiopharmaceutical Sciences, NIMH,
Purpose: to prepare mobile phase for the HPLC purification of [ 18F]fallypride and QC
analysis of [18F]Fallypride for Injection.
Procedure:
1. Preparation of HPLC solvent for the preparative column
a) Fill up a flask (1 L size) for HPLC solvent with acetonitrile (HPLC grade) containing
0.6% TEA (6 mL of TEA /L).
b) In a similar flask, prepare 1 L of HPLC water only.
2. Preparation of HPLC solvent for analytical column.
a) To prepare stock ammonium formate solution (100 mM), weigh out ammonium
formate (6.013 g) on weighing paper and then dissolve in HPLC water (1 L), mix well
and store at room temperature.
b) Put stock ammonium formate solution (50 mL; 100 mM) into a volumetric cylinder (1
L size), then dilute with HPLC water (450 mL; HPLC Grade)
c) Put acetonitrile (HPLC grade; 500 mL) into another volumetric cylinder. Mix
solutions from (a) and (b).
Chemist_____________________________Initials_________________Date________________
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 4 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
SOP # GP104
Micropore Filter Testing and Drug Product /Filter Compatibility
(Bubble Point Test).
Approved by: _____________________________Initials________ Date:____________
Victor W Pike, Ph.D., Chief, PET Radiopharmaceutical Sciences, NIMH
Purpose: Testing of Micropore filter compatibility.

Filter-Integrity test. The integrity of the Millex-MP sterile filter that was used for sterile
filtration of [18F]Fallypride for Injection is tested by the following method. The knob on the
pressure regulator that is hooked to the house compressed air gas supply is turned counter
clockwise so as to minimize outlet pressure. The filter assembly is removed from the
sterile dose vial and placed on the male luer-lock fitting of the pressure-regulated 1/8”
Teflon line. A disposable needle (1.5 inch 22 gauge) is placed on the male luer fitting of
the Millex-MP filter. A portion of this needle is submerged in a test tube (12 x 50 mm)
containing about 3 mL of HPLC grade water. The knob of the pressure regulator is slowly
turned clockwise and the pressure gauge is monitored visually. The pressure is brought to
45 p.s.i. If there is an absence of a steady stream of bubbles in the test tube when the
pressure gauge reads 45 p.s.i., then the filter passes the test. The result of the filter test is
recorded on the batch record and in the summary section of the quality control test form.
Chemist_____________________________Initials_________________Date________________
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 5 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
SOP # GP105
Preparation of Stock Acetate Buffer
Approved by: _____________________________Initials________ Date:______________
Victor W Pike, Ph.D., Chief, PET Radiopharmaceutical Sciences. NIMH
Purpose: to prepare stock acetate buffer (250 mM; pH 5.5) for use in [18F]Fallypride
for Injection prouction
A. Reagents:
Name of reagent
Manufacturer
Sodium acetate
Aldrich Chem. Co.
Quantity
Cat. # 241245, ACS reagent
Acetic acid
Aldrich Chem. Co.
Cat. # A6283, 500 mL
Water
EMD Chemicals Inc.
Cat. # WX0004-1
B. Procedure:
1.
On weighing paper, weigh out sodium acetate (14.45 g) and then dissolve this in HPLC
water (500 mL) in a bottle (500 mL size).
2.
Take acetic acid (2.15 mL) and put it into above solution.
3.
Mix well.
4.
Measured the pH of the buffer solution; the pH of this buffer should be 5.3−5.7.
Chemist_____________________________Initials_________________Date________________
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 6 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
SOP # MP201
BOXCLEAN program (GE TRACERlab FXFN)
Approved by: _____________________________Initials________ Date: _____________
Victor W Pike, Ph.D., Chief, NIMH, PET Radiopharmaceutical Sciences
Purpose: to clean the GE TracerLab FXFN module, check its functions and prepare it
for production.
1. Procedure:
1. If present, remove used Sep-Pak and re-connect line to its holder-connector.
2. Disconnect the following three lines placing the output into the waste reservoir:
a) The one coming from the Sep-Pak holder-connector into waste.
b) The one coming from the Sep-Pak holder-connector, going into the collection Vvial (10 mL size)
c) The one going from the HPLC loop waste into a V-vial (5 mL size) connected to
an external empty syringe (20 mL size), to pull diluted reaction mixture into
injector loop.
d) A fourth line, from the injector to a Blue Max (50 mL size) or to a vial (20 mL
size) (waste) remains the same.
3. Check that nitrogen and air compressed air valves are open. Check nitrogen cylinder is
open. Be sure there is a reactor vessel connected.
4. Place dry-ice in the vacuum trap.
5. Start the TracerLab automated software program.
Select Synthesis: "Boxclean", ENTER.
Follow instructions.
Add acetone as follows:
a) 1−2 mL acetone to target vial
b) 1–2 mL acetone to support vials V1 to V9 (fill small ones and half fill large ones)
c) 25 mL acetone to collection bulb vessel
d) 0.5 mL acetone to V-vial (10 mL size) (leave the spinal needle way up)
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 7 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
6. In Preparation CHECK LIST, check each item in the list, going down with repeated
ENTER. Finally, enter operator initials.
7. The screen will show the list of timed events for BOXCLEAN and the process will start
with the vacuum pup ON and the temperature increasing to 100oC.
Check that the temperature reaches 100oC and the pressure in reactor goes down.
8. At the end of the Program, the system will be flushed with nitrogen, go under vacuum to
remove most of the acetone vapors and then stop.
9. Remove the acetone that may be trapped in the vacuum trap. Dispose of it in the
appropriate container.
Copy of the BOXCLEAN Program see Attached file: Document 9: Validation Runs, attached
BOX Clean Program on the batch: FAY-032404.
Chemist_____________________________Initials_________________Date________________
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 8 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
SOP # MP202
GE TRACERlab FXFN Module. Vacuum Pressure check
Approved by: _____________________________Initials________ Date: _____________
Victor W Pike, Ph.D., Chief, NIMH, PET Radiopharmaceutical Sciences
Purpose: to check the vacuum efficiency and the leak of GE TRACERlab FXFN
module to ensure the success of production synthesis.
Procedure:
1. Open nitrogen and compressed air valves located outside hot-cell
2. Start Dell Computer-Windows 98. Enter Name and Password. After Virus Scan
Computer will open. Click twice on TRACERLab icon. Enter "Admin" and "Tracerlab"
3. Once on the Instrument SCHEME, look at the square area in the right side of the screen
that shows pressure of compressed air and nitrogen.
4. Turn on the vacuum on by click “ON” of the "Power”, which represents the vacuum on
the Instrument SCHEME. (Note: this can be turned OFF in the same way).
5. Compressed air pressure should be around _____kPa and nitrogen around______ kPa.
6. Click the mouse in V24 and V25 should open with space bar (1 = open, 0 = close), this
will set full vacuum on reactor vessel, reaching around ≤ 3 kPa. Record the lowest
pressure. _______kPa
7. Close V24 (space bar), pressure should hold (change should be ≤ 1 kPa/min).
8. If pressure is not holding find and correct the source of leak; e.g. tighten stoppers in
support vials, check valves 2 to 6, 13, 14, 20, 24; tight reactor vial, check O-ring.
9. With mouse, go to V20 (Nitrogen), open V20 with space bar (can also close with space
bar). Pressure should go up. Close with space bar and open V24. Check the pressure
again. It should go below 2 kPa again.
10. If pressure holds, go ahead with the next step.
Note. When both vacuum and pressure are open during the procedure, pressure should hold
between 21.5 to 22 kPa (V20 and V24 open).
Chemist_____________________________Initials_________________Date________________
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 9 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
SOP # MP203
Production of [18F]Fallypride for Injection; Part 1: Pre-synthesis Procedures
Approved by: _____________________________Initials________ Date: _____________
Victor W Pike, Ph.D., Chief, NIMH, PET Radiopharmaceutical Sciences
Purpose: to prepare for the synthesis of [18F]Fallypride for injection on the GE
TRACERlab FXFN module in hot-cell 5.
Procedures:
1. Check that nitrogen and compressed air valves are open.
2. Check nitrogen cylinder is open.
3. Turn on the computer with TRACERlab FXFN software and with Beckman HPLC
software.
4. Open the TRACERlab FXFN software and check the pressure of compressed air and
nitrogen pressure. The pressure of compressed air should be 400 p.s.i and nitrogen
pressure > 100 p.s.i.
5. Clean GE TRACERlab FXFN Module and hot-cell 5.
4.1
Follow the program "CLEAN BOX" (SOP # MP201). Clean and dry the GE
TRACERlab FXFN module on the day of preparation or the day before the preparation.
All the reagent vessels and transfer lines of the module to be used in the preparation
should be washed with acetone and dried under nitrogen flush before using.
4.2 Inspect hot-cell compounding/dispensing area for cleanliness. Remove extraneous
materials and labels. Spray ethanol to clean this area if necessary.
5.3 Inspect and set if necessary charcoal trap in the module
6. Prepare [18F]fluoride ion-Kryptofix 2.2.2-potassium carbonate complex solution
5.1 Add stock Kryptofix 2.2.2-potassium carbonate (70−100 µL) into V-Vial (1 mL
size).
5.2 Take V-Vial 1 to Cyclotron Facility (CC, NIH) to collect aqueous [ 18F]fluoride ion;
the activity of [18F]fluoride ion should be between 150−250 mCi water ( ≤ 500 µL).
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 10 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
6. Verify integrity of glassy reaction vessel assembly (containing small magnetic bar stirrer) in
TRACERlab FXFN Module. Follow SOP # MP202 and record the data in the sheet of
Summary Records of [18F]Fallypride for Injection run:
6.1 Vacuum pressure check (gauge, kPa).
6.2 The compress air pressure (kPa)
6.3 Nitrogen pressure (kPa)
7.
Prepare the preparative HPLC system
7.1 Start the HPLC computer software, enter the necessary information and select the
right method “ Fally_Production”
7.2 Verify that mobile phase bottles in the right position on the Beckman System Gold
Delivery module.
7.2.1
The solvent bottle of 0.6% TEA connects to B1 line (marked red)
7.2.2
The solvent bottle of HPLC water connects to A1 line (marked red)
Above connection should match the setup in the method
“Fally_Production”.
7.2.3
Set Acetonitrile w 0.6%TEA /HPLC water (30: 70 v/v) as initial
conditions. Equilibrate the Luna column (Phenomenex) with
fallypride preparative mobile phase (~ 200 mL)
7.3 Note the pump pressure while increasing the flow rate to 2 mL/min.
7.4 Pump the HPLC mobile phase through the preparative column at a low flow (1−2
mL/min) and maintain this flow until purification time.
8. Prepare in the clean bench a sterile filtration/collection Unit, using aseptic technique:
8.1. Prepare the final sterile product vial. The final sterile empty vial, product needle,
vent needle and two filters (Millex 0.22 µm) are assembled in a certified laminar
flow sterile cabinet. Attach a prepared label for [18F]fallypride with the current batch
number and date to the product vial.
8.2. Move a prepared product vial unit from the laminar flow sterile cabinet and attach it
to the product line of TracerLab FN Module.
9. Open the Tracerlab Fx Computer Controlling System ("Admin" and "Tracerlab") and Select
Synthesis method: "Fallypride". Click “Synthesis-Start” to start preparation of the
synthesis.
10. Enter all the required information (including date, radiotracer, batch #, human preparation
#) into the corresponding method of the HPLC computer software program.
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 11 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
11. Check that you have the correct UV absorbance (254 nm) and the correct settings for the
gamma 12. Place the solvents and precursor solution in the corresponding module support
vials:
________Support vial 1:
acetonitrile (0.5 mL).
________Support vial 2
[Used as a port (V-vial; 1 mL size)]: [18F]fluoride-K 2.2.2potassium carbonate solution; 100 µL).
________Support vial 4:
dry acetonitrile (1 mL) plus tosyl fallypride (2.0 ± 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 (9–14 mL)
________Support vial 8:
Ethyl alcohol for Injection (1 mL)
________Support vial 9
sterile water (8 mL)
_______ Large (10 mL size)
V-vial: 0.5 mL sterile water with 0.6%TEA.
_______ Large bulb vessel:
HPLC water (90 mL) and sodium acetate (250 mM; pH
5.5; 10 mL; see SOP # GP104).
12. Check that ‘complex solution’ delivery line goes through valve V2 and into reactor vessel
(glassy carbon).
13. Place acetonitrile (0.5 mL) in a syringe (3 mL) connected to an external line going into the
"complex" vial for recovery of [18F]fluoride-K 2.2.2-potassium carbonate solution left in
transportation vial and its addition to the reaction vessel.
14. Activate one C-18 plus Sep-Pak with ethanol (10 mL) sterile water (10 mL) and place in its
corresponding connection site in the module.
15. Place the small double-neck round bottom flask to receive the ethanolic eluate of the SepPak in its corresponding location in the Box.
16. In a lead pot, lace 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.
17. Place enough dry-ice/acetone mixture in the vacuum trap.
18. Check that the nitrogen and compressed air valves close to the hot cell are open (also
check the nitrogen cylinder).
19. Assure that syringes and needles have been changed. Use depyrogenated reaction vessel
(with magnetic stirrer), HPLC collection bulb (10 mL size) V-vial, two neck round-bottom
flask, sterile vial etc.
20. Keep the HPLC column flow at 2 mL/min.
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 12 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
Synthesis comments:
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
The approved procedure has been followed without any deviation
Chemist_____________________________Initials_________________Date________________
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 13 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
SOP # MP204
Production of [18F]Fallypride for Injection; Part 2: synthesis and purification
Approved by: _____________________________Initials________ Date: _____________
Victor W Pike, Ph.D., Chief, NIMH, PET Radiopharmaceutical Sciences
Purpose: to synthesize and purify [18F]fallypride
Procedures: [18F]Fallypride synthesis and purification
1. Preparation of [18F]fluoride ion/Kryptofix 2.2.2-potassium carbonate complex ([18O]wateracetonitrile-Kryptofix 2.2.2-[18F]fluoride ion-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 reaction vessel (20 mL
size).
Record the data: EOB, starting activity of [18F]fluoride-[18O]water and
Volume of [18F]fluoride-[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 F-18 complex.
2.1 Start the TRACERLab 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.
2.2 The program will have already started setting the vacuum pump ON and
transferring the F-18 complex from the V-vial (1 mL size) 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 F-18 complex.
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 14 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
The 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.
4.1 Once 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 100 ºC for 30 min. At
the end of the reaction, the reaction vessel will be cooled at 35 ºC and the first part of
the Program will finish.
4.2 A message will shown saying "End of synthesis, Hit OK to continue to next Program".
Hit CONTINUE.
5. Prepare Beckman HPLC system for purification.
5.1 Verify that the HPLC is in Preparative and in LOAD and that the detectors are in the
right settings.
5.2 Check that method setting of Beckman HPLC instruments is right
5.3 Set the flow rate to 9 mL/min. Wait for pressure to go up.
5.4 Click “Single run” in Beckman HPLC software to start acquisition and wait the
acquisition Box appears.
5.5 Check that the injector turn to inject position
6.
[18F]Fallypride
purification step 1: transferring the initial reaction mixture.
6.1 Second program continues purification process. The reaction mixture will be
transfered to the V- vial (10 ml size) containing sterile water (0.5 mL) for pre-dilution
before HPLC injection.
6.2 Check that the injector transfer needle in V- vial (10 mL size) is far up from 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
7.1 After 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
size) into HPLC loop automatically.
7.2 At this moment, new message box will appear “System now ready to inject. Hit OK
when the loading is completed.”
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 15 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
7.3 Watch the 10 mL T-vial closely. While the solution loading completed, hit OK.
7.4 At same time hit “OK to start the HPLC run in Beckman software
8.
[18F]Fallypride
step 3: HPLC separation and product collection.
8.1 In GRAPHICS observe the UV and and radioactivity traces on the data acquisition
systems.
8.2 When the desired peak start coming out click on the collect arrow. Record the Rt
=____________min.
8.3 The mobile phase containing the product will be added to the bulb vessel containing
sterile water (90 mL) for dilution and of sodium acetate (10 mL) for dilution and
neutralization.
8.4 At the end of collection click on the end collection arrow to stop fraction collection (the
mobile phase will go into the waste reservoir).
Synthesis comments:
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
The approved procedure has been followed without any deviation
Chemist_____________________________Initials_________________Date________________
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 16 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
SOP # MP205
Production of [18F]Fallypride for Injection, Part 3: formulation
Approved by: _____________________________Initials________ Date: _____________
Victor W Pike, Ph.D., Chief, NIMH, PET Radiopharmaceutical Sciences
S
Purpose: Formulation of [18F]Fallypride for Injection
Procedures: [18F]Fallypride formulation
1. The program will continue Fallypride Production Part 2 and the diluted fraction will be
passed through the solid phase cartridge (SPE; Waters C-18 Sep-Pak) for concentration.
This process is programmed for 6.5 min. After passing through Sep-Pak, valve V9 will open
and Sep-Pak will be washed with sterilized water.
2. After the water wash, V8 will open and the product will be eluted from the Sep-Pak with
ethanol (1mL) into the two-neck round bottom flask.
3. Program will stop to allow sampling of ethanolic solution if necessary. The message "Take
QC sample and hit OK to proceed with filtration" will appear. Take 40-100 µL of sample for
QC from the two-neck round bottom flask if needed.
4. The program can be continued hitting OK. Valve 7 will open and saline solution will be
added to the neck round bottom flask containing [18F]fallypride product.
5. The program can be continued hitting OK. The saline solution will be then transferred to
the sterile and non-pyrogenic vial (10 mL size, Flip-top- Vial – Glass) through the Millex
filter Unit (0.22 µm) for filtration and sterilization.
6. After filtering product, remove sterile filter, measure the activity of the final [18F]fallypride
product, record the number on the master batch Sheet and fill the number in the labels.
Activity of final [18F]Fallypride for Injection for _____________mCi at____________;
The volume:_____mL and the concentration:___________ mCi/mL.
The final Collection Vial and the lead shielded pot will be labeled as follows (21 CFR 361.1):
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 17 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
[18F]Fallypride for Injection
Sterile, apyrogenic solution for intravenous administration
Caution: New drug limited by Federal law to investigational use 0nly
NIMH MIB
Store at room temperature
Activity:______________ mCi
Volume: ___________
mL.
Batch #: FAY-________________
Expires 4 h after calibration
Time: _________________
Concentration: ______________
Date:_____________
In addition, one of the labels will be attached to the Radiopharmacy Form as record.
7. Sampling for QC: The 1 cc sterile polypropylene syringe may be 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 of [18F]Fallypride for Injection is transferred to sterilized
depyrogenated tube labeled “[18F]Fallypride for Injection” and the vial is dated and 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).
Standard operating procedures for these tests can be found in the quality
control and radiopharmacy forms.
8. Post-Synthesis Procedures:
8.1 HPLC and columns clean up: set the flow to 6 mL/min. and move the B pumps line into
the solvent bottle, which contains acetonitrile only. Keep the acetonitrile as 70%. Run
50-60 min and then stop running. Close the Beckman HPLC software.
8.2 Machine turn off: Turn off nitrogen and compressed air valves located outside hot-cell.
On the TRACERLab click synthesis – Reset in main menu to set all valves and vacuum
on default status, then close the software. It is ready for next Box Clean procedure
(SOP # MP201).
Synthesis comments:
______________________________________________________________________________
____________________________________________________________________
The approved procedure has been followed without any deviation
Chemist_____________________________Initials_________________Date________________
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 18 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
Diagram 1. Layout of GE TRACERlab FXFN Module
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 19 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
SOP # QA301
Standard Curve of Reference Fallypride
Approved by: _____________________________Initials________ Date: _____________
Victor W Pike, Ph.D., Chief, NIMH, PET Radiopharmaceutical Sciences
S Purpose: To set a standard curve for authentic fallypride for calculation of the mass of fallypride
in a in an analysis sample.
A) Stock solution of reference fallypride.
Weigh an accurate mass of fallypride (1−2 mg). Disolve in HPLC mobile phase (10−25
mL) in a volumetric flask for QC. Mix thoroughly for several minutes. When not in use,
store at −20 ○C in the freezer.
Concentration of solution = ~ 0.1 µg/µL
B) "Daily" dilution.
From the stock solution make a dilution 1: 100:

Example: take 100 µL of Stock solution.diluted with solvent (acetonitrile: HPLC
water 65: 35 v/v) in a volumetric flask (10 mL size)
Concentration: ~ 1−2 ng/1.0 µL
nmoles of fallypride (M.W. = 364.45) in this dilution:
1 ng/364.45 = 2.74 x 10-3 nmoles in 1.0 µL
Mix well to ensure homogeneity. When not in use, store at 4 ºC under refrigeration.
C) Procedure for obtaining standard curve
1. Open the Beckman 32 karat software and select the instrument ”Analysis System”
to start HPLC system.
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 20 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
2. Set the analytical HPLC System (Beckman Coulter) in working conditions and
keep flow for 30 min for column equilibration.
HPLC System conditions:
Column: Prodigy ODS (3) (100 μ; 4.6 x 250 mm; Phenomenex)
Mobile Phase: Acetonitrile-ammonium formate (5 mM) (50: 50 v/v)
Flow Rate: 1.0 mL/min.
UV Wavelength: 305 nm
3. Select the method ”FallyprideQ.C” in method box and check the instrument setup
on the “Instrument Setup” box, then close box.
4. Using the "daily" dilution make injections into the HPLC system varying the
injected volume, measuring the areas and ensuring that plate count and peak
shape is consistent.
5. Suggested volumes of "Daily" dilution: 10, 20, 60, 80, 100 µL.
6. Make two to four injections of each volume.
7. Perform Regression Analysis on the data, plotting area units versus µg or µmoles.
8. Using the external or internal standards, calculate the slope of calibration line (a),
Y-axis intercept of calibration line (b) and correlation coefficient (r).
9. Following is linear calibration fit of external or internal standards:
The equation for calculating the uncorrected amount is:
Y = ax+b
Chemist_____________________________Initials_________________Date________________
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 21 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
SOP # QA 304
Analysis of Organic Residues by Gas Chromatography
in [18F]Fallypride for Injection
Approved by: _____________________________Initials________ Date: _____________
Victor W Pike, Ph.D., Chief, NIMH, PET Radiopharmaceutical Sciences
Purpose: to analyze for volatile solvent residues in [18F]Fallypride for Injection
1. Responsibility 1.1 It is the responsibility of the Chief of PET Radiopharmaceutical Sciences to ensure
that personnel are trained in this procedure.
1.2 It is the responsibility of personnel to adhere to this approved SOP.
1.3 Procedure adheres to GMP/GLP guidelines.
2. Scope - Quality control testing of volatile organic solvents in NIMH produced PET
radiopharmaceuticals.
3. Reference Document(s) –
3.1 6850A GC User Information
3.2 Installation, operation and maintenance manual for hydrogen
generator (Model H2-90; Parker Balston). (Bulletin TI-H2-90C)
4. Safety Precautions
4.1 Radiation Safety – ALARA (As Low as Reasonably Acceptable)
4.2 Chemical laboratory safety
5. Materials and equipment
5.1 Agilent 6850 GC with flame ionization detector (FID)
5.2 Agilent 6850 series autosampler
5.3 J & W DBWAX column, (30 m (l) x 0.25 mm (id) x 0.25 μm (film thickness) (Alltech,
part # 122-7032)
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 22 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
5.4 Acquisition and data processing software: GC Chem Station (version: Rev. A.09.03
[1417] )
5.5 Inlet liner: split inlet glass liner with glass wool packing (Agilent part number, 5183469119251-60540)
5.6 Parker Balston H2-90 Hydrogen Generator
5.7 High purity grade (99.995 %) compressed helium compressed (Roberts Oxygen,
cat.no. R 102 F3)
5.8 In -house air purified by Parker Balston Zero Air Generator, Model 75-83NA
5.9 In -house deionized water (18 mega ohm) purified by Millipore Milli-Q j.) Autosampler
glass vial ( Agilent part no. 5182-0864) k.) Autosamper conical glass insert (Agilent
part no. 5183-2085)
5.10 Pipetman 200 L pipet (NIH stock # 6640-02-032-1955)
5.11 386 ppm internal standard aqueous solution of propionitrile prepared via 1 in 10
dilution of a 3860 ppm solution (0.5 mL propionitrile diluted to 100 mL mark with
water).
6. GC system configuration.
6.1 Injection port: split sample injection split ratio of 20:1, 250 ˚C
6.2 Carrier gas: Helium; 2 mL/min
6.3 Column temperature gradient: column temperature is initially operated at 50˚C and
held at this temperature for 1 min, and then increased to 150 ˚C at a rate of 20˚C /min.
The temperature is held at 150˚C for 0.5 min and then increased to 220˚C at a rate of
50˚C/min.
After 3 min at 220˚C the column temperature is returned to starting
temperature of 50˚C.
6.4 Detector: FID with hydrogen at 40 mL/min and air at 450 mL/min.
Helium make up
flow: 45 mL/min. Detector at 250 ˚C.
6.5 Autosampler: 10 L syringe. Sample injection volume: 1 L.
6.6 Needle/Syringe wash:
four times prior to injection of sample and two times after
injection.
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 23 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
7. Procedure before data acquisition. Ensure that the water level in the H2-90 hydrogen
generator is above the lower limit.
Otherwise, pour in 18 mega ohm water into the
reservoir until the level is just below the upper limit of water reservoir. Check the hydrogen
pressure is about 28 p.s.i. Check helium pressure is ca. 60 p.s.i. on main cylinder gauge.
Make sure that the solvents A and B (in the autosampler tray) for rinsing injection syringe
needle are filled with deionized (18 mega ohm water). Via a 200 L Pipetman pipettor,
pipette 50 L of [18F]Fallypride for Injection (test sample) into a glass insert housed in a GC
autosampler vial. Discard the radioactive pipette tip in radioactive waste. Add 50 L of the
internal standard solution of propionitrile (386 ppm) to the test sample.
autosampler vial with septum.
Cap the
Tap the bottom of the autosampler vial to remove air
bubbles. Place autosampler vial in autosampler rack and note rack position. On the GC
Control panel, use up or down arrow buttons to verify that FID is lit and the background
signal is ca. 5.
8. Data acquisition of [18F]FALLYPRIDE test sample spiked with internal standard. Double
click “Instrument 1 online” on the desktop.
Select Method and Run Control from drop
down men bar below the file menu bar. Select ISPRN.M from drop down menu box on the
right side of Method and Run Control Box. Go to sequence on the main menu and select
sequence parameter. Type in subdirectory for radiopharmaceutical that will be tested (i.e.
Fallypride). Select Sequence Table from Sequence drop down menu. Enter 1 for the
number of injections per sample and location of the injection. Enter the sample name, file
name, and injection volume (1.0 L). When the system shows a ready sign (above start
button), hit the start button. To view on line signal, go to view on the main menu bar and
select on line signal/signal window 1. After 3.5 min, the GC chromatogram and report can
be generated; this is performed by going to the desktop and opening “Instrument 1 offline”.
On the left side of the screen, scroll the drop down menu to “data analysis”. Under the file
menu, scroll to “load signal. “ Find the file name in the appropriate subdirectory on the C
drive (e.g. C:\. . . \data\Fallpride). After loading the signal, scroll down to “print preview . . .
report” that is found in the file menu. The chromatogram and report should appear on the
computer screen and this can be printed out be hitting the print button on the bottom of the
report page. Attach the chromatogram and report to the quality control form.
[18F]Fallypride
Note that
fro Injection may be released before completion of the residual solvent test
(see section
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 24 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
9. Post-run method.
Date of review: 05/17/04
Remove all samples and label radioactive samples.
Download the
method Default.M that is used to maintain the oven temperature at 150 ˚C when GC is idle.
Chemist_____________________________Initials_________________Date________________
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 25 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
SOP #QA305
Analytical HPLC QC Method
Approved by: _____________________________Initials________ Date: _____________
Victor W Pike, Ph.D., Chief, NIMH, PET Radiopharmaceutical Sciences
Purpose: to perform analytical HPLC QC on [18F]Fallypride for Injection
Procedure
1. Set the analytical HPLC system (Beckman Coulter) in working conditions and maintain flow
for 40 min for column equilibration.
HPLC system conditions:
Column: Prodigy ODS (3) (100 μm; 4.6 x 250 mm; Cat. # 00G-4244-E0;
Phenomenex)
Mobile phase: Acetonitrile-5 mM ammonium formate (50: 50 v/v)
Flow rate: 1.0 mL/min.
UV Wavelength: 305 nm
2. Download the analytical method “Fally_analysis_secondSys” or “Fally_analysis_firstSys”
from method box in Beckman software.
3. Validation of HPLC system

Inject standard fallypride (20−40 µg), which is already prepared according to SOP
# QA301, and record the exactly mass (ng) injected and retention time of fallypride
standard.

Calculate the mass of fallypride corresponding to the peak of fallypride in HPLC
chromatography, based on the standard curve of fallypride.

Compare the mass of the fallypride injected with the mass calculated, the
difference should be equal or less then 10%. (Recovery ≤ 90%)
4. HPLC system cleaning

Inject solvent (100 µL) once or twice to clean the system in 10-15 min after
standard fallypride running. Check that only the solvent front peak should shows in
the chromatography.

Inject 10% ethanol in saline (100 µL). Check that only the solvent front and saline
peaks show in the chromatography.
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 26 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
5. QC of final product: [18F]Fallypride for Injection

Measure the radioactivity of sample of final product of [18F]Fallypride for Injection
(50−150 L) before and after injection onto the HPLC system in hot-cell 5, and
record the number in QC record sheet. Calculate the activity injected (decay
correlation)

Inject the sample of [18F]Fallypride for Injection (50−150 L depending on the
radioactivity concentration of formula).

Check the retention time (Rt) of the standard and the [ 18F]fallypride for
radiochemical identity (the difference of Rt < I min)

Calculate the mass corresponding the UV area of the [18F]fallypride peak in the
chromatograph, based on the standard curve of fallypride.

Calculate the specific radioactivity according to the radioactivity injected and the
mass calculated at HPLC injection time.

Calculate the specific activity at the time of HPLC injection, EOS and EOB (decay
correction).

Calculate the mass (g) of whole formula based on the radioactivity of final
formula, which recorded on master batch Sheet Page 4, and the specific activity at
EOS.

Record the chemical purity, radiochemical purity, the concentration of the final
formula and the expired time.
6. Post-analysis HPLC system and column cleaning: to run the HPLC system for 40−50 min
with the solvent (acetonitrile-HPLC water; 70: 30 v/v) for cleaning the HPLC system and
the column. Turn off the HPLC system and software.
Chemist_____________________________Initials_________________Date________________
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 27 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
SOP # QA306
Sterility Test
Approved by: _____________________________Initials________ Date: _____________
Victor W Pike, Ph.D., Chief, NIMH, PET Radiopharmaceutical Sciences
Purpose: To test the sterility of [18F]Fallypride for Injection
Notes.
1. Routine sterility tests will be run by the Clinical Microbiology laboratory of the Department of
Laboratory Medicine at NIH, through a FDA approved procedure.
2. The radiochemistry lab will provide a sample of each formulation (in its original filtration
vial), 24 h after its preparation or when no traces of radioactivity are detected anymore (ten
or more half life periods).
3.
Before their transportation to the testing laboratory, the samples will be stored in the
refrigerator until decay.
4. A log book will be kept in the radiochemistry lab indicating the date the samples were
transported, their lot number and the date and results of the test (readings at 7 and 14 d
should be indicated).
Chemist_____________________________Initials_________________Date________________
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 28 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
SOP # QA307
Pyrogen Test (LAL Bacterial Endotoxin Test). Simplified Procedure
Approved by: _____________________________Initials________ Date: _____________
Victor W Pike, Ph.D., Chief, NIMH, PET Radiopharmaceutical Sciences
Purpose: to test the sterility of [18F]Fallypride for Injection
Materials
LAL test kit vials (e.g., Endosafe Inc., Charleston, SC or Cape Cod Associates, Falmouth MA)
Two vials 0.125 EU/mL(one for test sample, one for negative test)
One positive vial (5 EU/mL)
Sterile saline solution (from the same vial used for the final formulation)
Pyrogen-free test tubes (if not provided with kit)
Micropippets and tips (50 µL, 100 µL, 500 µL)
Tuberculin syringes (0.5 mL, 1 mL)
Incubator, 37°C
LAL Test Procedure: See “RADIOPHARMACY TEST RESULTS” form
Simplified Test Procedure ( it may not be used as a procedure of LAL test.)
a) From the final formulation in saline, take 50 µLand add to the test sample tube. To the
same LAL tube add 150 µLof saline (for a total of 200 µL).
b) To the negative test tube add 200 µL saline only.
c) To the positive test tube add also 200 µL saline.
d) Incubate all vials at 37 °C for 1 h. Remove the test vials at 20 min and in one smooth
careful motion, invert each tube (without totally disturbing the gel) and note gel
formation.
e) Continue incubation of the vials for the full 60 min. Again in one smooth motion, invert
each vial and note gel formation.
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 29 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
f)
Date of review: 05/17/04
The test sensitivity equals the concentration of the test vial multiplied by the dilution
factor. If the test sample does not gel, report the result as less than the test sensitivity.
If the test sample gels, report as greater than or equal to the test sensitivity.
Note. The results at 20 min will allow release of the formulation for injection and the 60 min
samples will confirm the results.
Chemist_____________________________Initials_________________Date______________
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 30 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
SOP #QA308
Analysis of the Purity of Fallypride and Tosyl-Fallypride by HPLC
Approved by: _____________________________Initials________ Date: _____________
Victor W Pike, Ph.D., Chief, NIMH, PET Radiopharmaceutical Sciences
Purpose: to determine the purity of Tosyl-Fallypride by HPLC
The HPLC System used in this method:
HPLC pump: System Gold 126 Solvent Model (Beckman Coulter)
UV detector: System Gold 166 Detector (Beckman Coulter)
Sample Injector: Rheodyne with 200 µLloop
The HPLC conditions used in this method:
Column: Prodigy ODS (10 µm, 4.6 X 250 mm, 100 A)
Mobile Phase: Acetonitrile-5 mM ammonium formate (50: 50 v/v) at 2 mL/min
Flow rate: 1 mL/min.
PDA: 254 nm
Procedure
1. Weigh 1−2 mg of precursor (tosyl-fallypride) in vial (20 mL size)
2. Dissolved the sample in of acetonitrile (10−20 mL). The concentration of this solution is
~ 1 µg/µL.
3. Inject 10 µLof the solution of tosyl-fallypride into HPLC and run for 30 min.
4. Use the Beckman method “Fallypride_Analysis_SecondSys” to interrogate the
Chromatograph, and get the Tosyl-fallypride purity (UV area percentage).
Chemist_____________________________Initials_________________Date__________
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 31 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
Attachment
This attachment lists the materials and equipments, which are used in the SOPs on Page 1.
Materials:
Quality grade and
Item
Name
Name of supplier
catalog #
1
4,7,13,16,21,24-Hexaoxa-1, 10-diazabicyclo
Aldrich Chem. Co.
Anhydrous, 98%
[8.8.8] hexacosane (Kryptofix 222)
1001 West St. Paul
Cat. # 29110-1G
Ave., Milwaukee, WI
53233
2
Potassium carbonate
Aldrich Chem. Co.
99.995%
1001 West St. Paul
Cat. # 367877-10G
Ave., Milwaukee, WI
53233
3
Acetonitrile (anhydrous)
Aldrich Chem. Co.
Anhydrous, 99.8%
1001 West St. Paul
Cat. # 271004-100
Ave., Milwaukee, WI
ML
53233
4
5
6
Acetonitrile
Water
Acetone
Burdick and Jackson,
HPLC Grade, ACS
B&J Brand
Cat. # 017-4
Burdick and Jackson,
HPLC Grade, ACS
B&J Brand
Cat. # 4218
Fisher Scientific
HPLC Grade, ACS
Cat #: 4218
7
Triethylamine
Aldrich Chem. Co.
HPLC Grade, ACS
1001 West St. Paul
Cat #: 471283-
Ave., Milwaukee, WI
500ML
53233
8
Sodium acetate
Aldrich Chem. Co.
99+% ACS
1001 West St. Paul
Cat. #: 241245-
Ave., Milwaukee, WI
500G
53233
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 32 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
9
10
Acetic acid
Luna C18 (10 m; 100 Å; 250 x 21.2 mm)
Date of review: 05/17/04
Malinckodt
99+% ACS
Chemicals
Cat. # V193-14
Phenomenex
Cat. # 00G-4094-
2320 W.
205th
St.
P0
Torrance, CA 905011456
11
Fallypride
ABX,
> 90%
Wilhelm-RÖnsch Str.
Cat. # 156.0025
9. D-01454
Radeberg, Germany
12
Tosyl fallypride
ABX,
> 90%
Wilhelm-RÖnsch Str.
Cat. # 155.0002
9. D- 01454
Radeberg, Germany
13
Ethyl alcohol (200 proof)
The Warner-Graham
CAS # 64-17-5
Company,
Cockeysville, MD
14
Sodium Chloride for Injection (0.9% w/v;
American
USP
USP
Pharmaceutical
CAT#:918610
Partner, Inc.
15
Sterile empty vial (10 mL)
Abbott laboratories
USP
Cat. # 5816-11
16
Sep-pak Cartridges Plus C18
Waters Corporate
Cat. # WAT020515
34 Maple Street,
Milford,
MA. 01757 USA
17
Millex filter (25 mm; 0.22 m pore size)
Millipore
Cat. # SLGVR35LS
18
Millex GV filter (25 mm; 0.22 m pore size;
Millipore
Cat. # SLGV004SL
4 mm diameter)
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 33 of 34
[18F]FALLYPRIDE FOR INJECTION: STANDARD OPERATING
PROCEDURES
PET Radiopharmaceutical Sciences Section,
Molecular Imaging Branch,
National Institute of Mental Health,
National Institutes of Health,
Bldg. 10, Rm. B3 C338,
Bethesda, MD 20892
Date of review: 05/17/04
Instruments and equipment:
Operation/Function
Manufacturer
Model
Serial #
Radiosynthesis
General Electric
TRACERlab FXFN
14390
HPLC purification
Beckman Coulter
System Gold 126
342-2187
Solvent Model
UV absorbance detection
Beckman Coulter
in HPLC purification
Radioactivity detection in
System Gold 166
332-2187
Detector
Bioscan
Flow Counter
0409-316
HPLC purification
Document 5. [18f]Fallypride for Injection: Standard Operating Procedures
Page 34 of 34