GUIDELINE FORQUALITY ASSURANCE AND QUALITY CONTROL OF
18

F-FDG (2-Deoxy-2-fluoro-D-glucose)

Forum for Nuclear Cooperation in Asia (FNCA)
Cyclotron and Positron Emission Tomography (PET) in Medicine

18 March, 20111

This guideline summarizes Quality Assurance (QA) and Quality Control (QC) of
18

F-FDG and is issued as an outcome by sub-group 2 (PET Radiopharmaceuticals)of FNCA Cyclotron and PET in Medicine.
Since a general guideline on 18F-FDG injection is already developed in eachparticipating country, it is rather difficult to develop a single guideline for all. For
that reason, we would like to entrust each country to develop a specific protocol
for each module.

2

GUIDELINE FOR QUALITY ASSURANCE AND QUALITY CONTROLOF 18F-FDG (2-Deoxy-2-fluoro-D-glucose)

CONTENTSPAGEINTRODUCTION
1.1
Scope of this document
1.2
Objectives of this guideline

5
6

QUALITY ASSURANCE (QA)
2.1
General2.2
Principles
2.3
Quality Control
2.4Standard of [18F]-FDG
2.5
Current Good Manufacturing Practice for PET Drugs

7
7
8
8
8

3

PRODUCTION OF FLUORINE-18 (18F)

10

4GENERAL REMARK ON SYNTHESIS OF [18F]-FDG

10

5

GENERAL TESTS
5.1
Gamma-ray Determination
5.1.1 Measurement of gamma-ray spectrum
5.1.2 Assay using a gamma spectrometer
5.1.3 Half-life measurement
5.2
Bacterial Endotoxin Test
5.2.1 Gel-clot technique5.2.2 Optical techniques
5.3
Sterility Test
5.3.1 Blood culture system

11
11
12
13
14
15
16
18
18

1

2

3

6

ANALYSIS METHODS FOR [18F]-FDG INJECTION
6.1
Radiochemical purity
6.2
Chemical purity and specific radioactivity
6.3
Chemical Impurities

7

QUALITY CRITERIA AND FREQUENCY

21
21
23
23

OF TEST FOR [18F]-FDG

26

8

REFERENCES

27

9TERMINOLOGY28

10

LIST OF GROUP 2 MEMBERS

29

4

1

INTRODUCTION

1. 1

Scope of this document

Positron emission tomography (PET) is a medical imaging modality involving theuse of a unique type of radiopharmaceutical drug product. The majority of PET drug
products are injected intravenously into patients for diagnostic purposes. Most PET drugs
are produced using cyclotrons at locations that are proximately to the patients to whomdrugs are administered.
Under section 501 (a) 92) (B) of the Federal Food, Drug, and Cosmetic Act (theact) (21 U.S.C. 351 (a) (2) (B)), a drug is adulterated if the methods used in, or thefacilities or controls used for, its manufacture, processing, packing, or holding do notconform to or are not operated or administered in conformity with current good
manufacturing practice to ensure that the drug meets the requirements of the act as tosafety and has the identity and strength, and meets the quality and purity characteristics,
that it purports or is represented to posses.
In the clinical application of PET radiopharmaceuticals, it is essentially important
to assure the safety of the product. Many guidelines for the production of PET
radiopharmaceuticals have been published in each country or regions such as found in the
United State Pharmacopoeia (USP), European Pharmacopoeia (EP) and JapaneseStandard. Precise procedures for the production of PET radiopharmaceuticals and their
Quality Assurance (QA)/Quality Control (QC) are described in the guidelines. There aresome minor discrepancies found among the guidelines. Each guideline does not preclude
the use of an alternative if it is equivalent or better to the method described in theguideline.
The scope of this document is to focus on the QA/QC of [18F]-FDG as a result
achieved from the FNCA 2009 Workshop on Cyclotron and Positron Emission
Tomography (PET) in Medicine held in Kuala Lumpur. As a guide to the preparation of
document, the Japanese standard “Standards for Compounds Labeled with Positron
Nuclides Approved as Established Techniques for Medical use (by the Subcommittee on
Medical Application of Cyclotron-Produced Radionuclides) (2001 revision)” and“Description of the Established Standard Techniques of Labeling Compounds with5

Positron Nuclides for use as Radiopharmaceuticals: approved by the Subcommittee on
the Medical Application of Cyclotron-Produced Radionuclides (revised in 2001)” arereferred.
1.2

Objectives of this guideline
The objectives of this guideline are:a) To provide a general knowledge and procedures to perform the QA/QC of
PET radiopharmaceutical.
b) To assure the safety of product before released for clinical application.
c) To develop a minimum QA/QC tests data required for the production of [18F]FDG PET radiopharmaceutical as a pioneer guideline to further provide otherguidelines for other types of 18F label compounds.

The minimum requirements of quality assurance and quality control for the
preparation of [18F]-FDG would be estimated and this Guideline should be read in
conjunction with all related rules and regulations enforced.

6

2

QUALITY ASSURANCE (QA)

2.1

GeneralQuality Assurance is a wide ranging concept which covers all matters which
individually or collectively influence the quality of product. It is the sum total of theorganized arrangements made with the object of ensuring that medicinal products are of
the quality required for their intended use. Quality Assurance therefore incorporates
quality control and some aspect of cGMP requirements plus other factors outside thescope of this Guideline.
2.2

Principles

There shall be a management system to oversee preparations to ensure that a PET
radiopharmaceutical of sufficient quality is prepared. It is essential to examine, approveor reject components, containers, closures, in-process materials, packaging materials,
labelling, and finished product to ensure compliance with procedures and specifications
affecting the identity, concentration, quality and purity of a PET radiopharmaceutical.
In consideration of the unique nature of PET drugs and PET drug production, thecGMP requirements for PET drug products are the following:a) Fewer required personnel with fewer organizational restrictions consistent
with the scope and complexity of operations;b) Allowance for multiple operations (or storage) in the same area as long as
space for other controls are adequate;c) Streamlined requirements for aseptic processing consistent with the nature of
the production process;d) Streamlined quality control requirements for components;e) Self-verification of significant steps in PET drug production consistent withthe scope and complexity of operations;f) Same-person oversight of production, review of batch records, anauthorization of product release consistent with the scope and complexity ofoperations;g) Specialized quality control requirements for PET drugs produced in multiple
sub-batches; and7h) Simplified labelling requirements consistent with the scope and complexity ofoperations.
These provisions reflect the unique characteristics of PET drug production, shallmake it easier for PET centers to achieve compliance with cGMP requirements.
2.3

Quality Control

PET drug product producers shall have a quality control arrangement that has the
responsibility and authority to oversee production operations to ensure that each PET
drug product meets the requirements of safety and has the identity and strength, andmeets the quality and purity characteristics that it is supposed to have according to thestandards. Each PET producer shall determine what personnel should staff the quality
control unit. At some PET centers, it may be reasonable for the same personnel to be
involved in both production and quality control.
The quality control personals shall examine and approve or reject components,
containers, closures, in-process materials, packaging materials, labelling, and finisheddosage forms to ensure compliance with procedures and specifications affecting theidentity, strength, quality, or purity of a PET drug product.
2.4

Standard of [18F]-FDG

A radiopharmaceutical that is an aqueous injection containing fluorine-18
included within the 2-deoxy-2-fluoro-D-glucose structure. This injectable can include 2deoxy-2-fluoro-D-glucose as carrier containing 90 – 110% of the fluorine-18
radioactivity labelled on the molecule at the assay day. Specific activity of this injectable
solution reaches more than 200 MBq in 1 mg of 2-deoxy-2-fluoro-D-glucose on the assay
day and time.
2.5

Current Good Manufacturing Practice for PET Drugs

Current good manufacturing practice for PET drug products is the minimum
requirements for the methods to be used in, the facilities and control used for, theproduction, quality control, holding, or distribution of PET drug products intended forhuman use. This regulation is applicable only to the manufacturer or production of PET
drug products and in this document, it refers to the [18F]-FDG product. Any human drug
8

product that does not meet the definition of a PET drug product must be manufactured inaccordance with the current good manufacturing requirements such as the Guide to Good
Manufacturing Practice For Medicinal Products in the PIC/S GMP Guide.
Current good manufacturing practice for PET drugs must address certain mattersto ensure that each drug product meets the requirements of the act that to safety and has
the identity and strength, and meets the quality and purity characteristics, that it is
supposed to have. These matters are:a) Personnel and resourcesb) Quality control systemc) Facilities and equipmentsd) Control of components, in-process materials, and finished productse) Production and process controlsf) Laboratory controlsg) Acceptance criteriah) Labelling and packaging controlsi) Complaint handling; andj) Record keeping

9

3

PRODUCTION OF FLUORINE-18 (18F)This radiopharmaceutical is prepared by using fluorine-18 obtained by irradiatingneon-20 with deuteron or oxygen-18 with proton. Due to its simplicity of managing theprocess, most cyclotron centers currently use oxygen-18 as their target material. The
manufacturing of 2-Deoxy-2-fluoro-D-glucose (18F) is then performed. After the
purification, the injection solution is prepared.
4

GENERAL REMARKS ON SYNTHESIS OF [18F]-FDG

2-Deoxy-2-fluoro-D-glucose (18F) Injection has been synthesized according to thefollowing reaction scheme:

OAc
AcO
AcO

OH
+

OTf
O

i) [K/222] F /CH3CN

OAc

Mannose triflateii) HCl or NaoH

O

HO
HO
18

F

OH

[18F]-FDG

It is then purified by passing through a series of purification column AG 50W-X8
column, AG 11A8 column, Sep-Pak C18 cartridge and Sep-Pak alumina N cartridge, andthen sterilized by passing through a 0.22 µm sterile membrane filter. Various
modifications of the fluoride ion method developed by Hamacher et al. are available toprepare [18F]-FDG, which are classified to the type of catalyst used, hydrolysis condition,
purification technique and the use of on-column method. Therefore, the impurities
contained in the product would be different and depending on the method used.
Originally, [18F]-FDG were produced by [18F]F2 method or [18F]acetyl hypofluorite
method containing a small amount of F2 gas, but [18F]fluoride methods not-adding carrier
became predominant over the original [18F]F2 method and have been used in most of the
PET facilities. Therefore, the quality control method relating to the previous methods
adding carrier were not the subject of discussion in this guideline.
[18F]-FDG have been produced in the various category PET facilities such as at
the university hospital, research centers, dispensing centers, etc. Thus the requirements
criteria for the QA/QC of the [18F]-FDG may differ from one organization to others,
which shall depend on the scale, activity and mission of each facility. In this guideline,10the sub-group 2, named as PET Radiopharmaceutical Group at the 2009 FNCA
Workshop on Cyclotron and PET In Medicine meeting agreed that the requirement
criteria for the QA/QC of [18F]-FDG is categorized into a GMP-facility and into a nonGMP facility.
5

GENERAL TESTSGeneral tests described in this Guideline should be conducted according to theGeneral tests described in “The Minimum Requirements for Radiopharmaceuticals”,“The Japanese Pharmacopoeia”, “The United State Pharmacopoeia” and “The European
Pharmacopoeia” and other authorized standards. This chapter describes only Gamma-ray
Determination, Bacterial Endotoxins Test and Sterility test which tests are not common inusual PET facilities where QA/QC should be conducted in a very short time by a small
number of staff.
5.1

Gamma-ray Determination

Gamma-ray Determination measures gamma and X-rays out of radiation rays
emitted from radionuclides. The measurement uses a gamma spectrometer which is used
for identifying radionuclides, detecting radionuclidic impurities and determining theirradioactivities.
5.1.1

Measurement of gamma-ray spectrum

This measurement identifies nuclides and detects radionuclide impurities using
gamma spectrometer through measuring and comparing gamma-ray spectrum ofradiopharmaceutical and calibration standard of the radionuclide.
Operation method
Unless otherwise specified, the operation is conducted as follows;
Take an accurately measured aliquot of a radiopharmaceutical and a calibration
standard in separate containers. Add solvent as required to achieve the concentration of 4
– 400 kBq/mL. Place the identical amount (0.1 – 10 mL) of it into containers of theidentical material and shape. Seal the containers and label as a specimen and a standard.
Make sure that the measurement devices and volumes are identical.
11Place the test specimen and the standard at a set distance from the gamma-ray
detector, and measure gamma-ray spectrum. When the standard is the same radionuclide
as that in the radiopharmaceutical, make sure that all the energy peaks of gamma-rays inthe specimen’s spectrum (“gamma-ray peak” hereafter) match the gamma-ray peaks ofthe standard, when determining radionuclides.
When using a specimen with a different radionuclide from that in theradiopharmaceutical, use the gamma-spectrum of the standard for energy calibration, andobtain the energy level of gamma-ray peaks for the specimen’s spectrum to determine theradionuclides.
5.1.2

Assay using a gamma spectrometer

This method conducts assay through calculating the counting rate of the typical
gamma-peak region emitted from the radiopharmaceutical and the standard sample.
Operation method
Unless otherwise specified, the operation is conducted as follows;
When the standard is the same radionuclide as that in the radiopharmaceutical, use
the gamma-spectrometer to measure gamma-ray spectrum and calculate the counting rateof the gamma peak region for the specimen and the standard. Use the formula below to
obtain the radioactivity (Ac) in a set aliquot of radiopharmaceutical. Radioactivity in a set
aliquot of radiopharmaceutical:
Ac = S x (A/B) x (D/D’) x G
In the above formula, A and B represent the measured value of counting rates(corrected for background region) for the same energy peak region of the specimen andthe standard D and D’ represent the dilution factors for the radiopharmaceutical andstandard respectively. S is radioactivity for a set amount of the standard. G is thegeometric correction factor on the positions of the specimen and the standard, and shouldbe equal to 1 wherever possible.
When the standard is the different radionuclide, use the gamma-spectrometer tomeasure gamma-ray spectrum, and use the energy and gamma-ray emission rate of the12radionuclide use as the standard to work out the spectrometer’s energy peak counting
efficiency (“efficiency” hereafter). Measurement conditions for a radiopharmaceuticalmust be identical to those for the standard. Use the radiopharmaceutical’s gamma-ray
spectrum obtained to calculate of the target gamma peak region (Nx). The radioactivity
for a set aliquot of the radiopharmaceutical (Ac) may be obtained with the formula below,
using the efficiency (Fx) pre-determined with the standard:
Ac = [(Nx/Fx)] x D x G
In this formula, R represents the ratio of gamma ray emission, while D represents
the dilution factor of the radiopharmaceutical. G is the geometric correction factor on thepositions of the spectrum and the standard, and should be equal to 1 wherever possible.
5.1.3

Half-life measurement

This method takes the assay using a gamma spectrometer, well-type scintillation
counter or ionization chamber.
Unless otherwise specified, the operation is conducted as follows;
Operation methoda) Using a decay curve
Take a set aliquot of the radiopharmaceutical immediately after preparation, anduse a radioactivity measuring device (well-type scintillation counter, ionization chamber,etc.) to determine chronological changes in radioactivity under the same measuring or
geometrical conditions. Plot the logarithm with time on the horizontal axis andradioactivity (or meter readings with the background corrected) on the vertical axis, andconfirm that the decay curve obtained forms a straight line. Use the straight line to workout the time required for radioactivity to fall by half.
b) Two-point determination
Take a set aliquot of the radiopharmaceutical immediately after preparation, anduse a radioactivity measuring device (well-type scintillation counter, ionization chamber,13

etc.) to determine radioactivity twice under the same measuring or geometricalconditions. The second measurement should be taken after a half-life period of theapplicable nuclide elapses. If the applicable radionuclides’s half-life period is too short or
too long to facilitate measurement, the second measurement may be conducted after a ¼
of the half life, or 3 times the half life. Use the formula T = 0.693 x t/ln (A0/A) to obtain
the half-life T, whereby A0 and A represent the radioactivity (or meter readings with thebackground corrected) at the first and second measurement respectively, and t represents
the interval between the two measurements.
When a decay curve is used to measure the half-life, confirm the half-life of thecandidate nuclide when the plotted value indicates 95 to 105% that of the target
radionuclide used. When the bracketing approach is used for measuring the half-life ofradionuclides, confirm the half-life of the nuclide used when plotted value indicates 90 to110% of the half-life of said nuclide.
5.2

Bacterial Endotoxin Test

Bacterial Endotoxin Test is designed to detect bacterial endotoxins based on the
blood-clotting reaction that occurs when bacterial endotoxins, derived from Gramnegative bacteria, activate the blood cell component LAL (Limulus amebocyte lysate)
derived from horseshoe crabs (Limulus polyphemus or Tachypleus tridentatus) or relatedorganism. Said test may be conducted with one of the following techniques; the Gel-clot
technique based on clot formation, turbidimetric technique based on turbidity changes, or
chromogenic technique based on color presentation from hydrolysis of the chromogenic
substrate. If the result is inconclusive, use the Gel-clot technique to make final judgment.
Note that gel-clotting of LAL may be triggered with a certain type of polysaccharide.
Conduct this test promptly to avoid contamination from microorganisms.
Apparatus
Wash glassware thoroughly with water, and heat it to a minimum of 250 0C in 30
minutes to inactivate endotoxins before use in the test.

14

Preparation of sample solution
Unless otherwise specified, adjust a sample solution to pH 6.0 to 8.0 with either
0.1 mol/L sodium hydroxide or 0.1 mol/L hydrochloric acid for use in the test, when andwhere applicable.
Preparation of standard endotoxin stock solution
Add endotoxin-testing water to standard endotoxin in a vial to make up a solution
with 10,000 endotoxin units (EU) in 1.0 mL, and shake well to facilitate complete
dissolution. The just prepared standard endotoxin stock solution is immediately stored at
2 – 8 0C, and should be used within 14 days of preparation.[Note 1]
Preparation of standard endotoxin solution
Rigorously shake the standard endotoxin stock solution, and place it in cold water.
Measure a designed aliquot of the stock accurately and add endotoxin-testing water toprepare a standard endotoxin solution with the endotoxin concentration specified for theradiopharmaceutical injection used in testing. Keep the solution in cold water, and use
within 90 minutes of preparation.
5.2.1

Gel-clot technique
LAL solution

LAL is freeze-dried amebocyte lysate from horseshoe crabs (Limulus polyphemus
or Tachypleus tridentatus). Add endotoxin-testing water to LAL, with constant stirring tofacilitate dissolution before storage in ice water until use. This series of procedures may
be omitted if a single-testing LAL reagent is available in a vial or ampoule.

Operational procedures
Place 0.10 mL each of LAL solution in three test-tubes (each with 10-mm innerdiameter and 75-mm length, and add 10 mL each of sample, standard endotoxin solution
or endotoxin-testing water separately to the test-tubes, respectively. In cases where the15single-testing LAL reagent is available in vials or ampoules, add the sample solution,
standard endotoxin solution or endotoxin-testing water to the vials or ampoules directly.
Place a cap on the test tubes, and mix the solution gently before standing at 37 ± 1
0
C for 60 ± 2 minutes.
Judgement
Tilt the respective test tubes containing the sample, standard endotoxin solution
and endotoxin-testing water 180 degrees gently without stirring the contents, and observe
the solutions, accordingly. The endotoxin test is considered positive if the content
agglutinates without physical distorting, but is otherwise negative.
The test is considered conformed when the test tubes treated with the sample
solution and endotoxin-testing water test negative while that treated with the standard
endotoxin solution tests positive.
If either the test tube treated with endotoxin-testing water tests positive, or thattreated with standard endotoxin solution tests negative, the test is deemed invalid.
In such cases, use new reagents or replace the equipment, and repeat the test.
Note: 1: Perform preliminary tests to confirm neither the reaction is promoted nor
inhibited by the sample solution in said test.
5.2.2

Optical techniquesa)Turbidimetric techniqueThis technique observes the changes in turbidity associated with the clotting
protein “coagulin”, which is formed in the reaction between endotoxin and LAL, andassesses the changes in light penetration. The interval it takes for the sample reaction todisplay specific turbidity from the start of the reaction is taken as the gel-clotting time(Tg). Since a calibration relationship between endotoxin concentration and Tg can bederived, the said technique can therefore quantitatively measure endotoxin.

16

LAL solution
Add endotoxin-testing water to a vial of LAL gently with constant stirring tofacilitate dissolution, and chill the preparation in ice water until use. In cases where asingle-test lyophilized LAL reagent is packed in a vial or ampoule, the procedure may beomitted.
Operational procedures
Measure three volumes of 0.10 mL each of LAL solution and place each sample
separately in a test-tube. Add 0.10-mL samples each of endotoxin-testing water, standard
endotoxin solution or sample solution to the tubes, respectively. In cases where thesingle-testing LAL reagent is available in vials/ampoules, add the sample solution,
standard endotoxin solution or endotoxin-testing water to the vials/ampoules directly.
Place a cap on the test tubes, and mix the solution gently before standing at
37±1°C. Measure the time it takes light penetration to attain and remain constant at a
certain level.
b) Chromogenic technique
The synthetic substrate technique uses a compound (e.g. Box-Leu-Gly-Arg-pNA)
to bind synthetic peptides (with an amino acid sequence similar to coagulogenhydrolysate) with chromophore p-nitroaniline (pNA). This amidase activity of theendotoxin-activated clotting enzyme promotes pNA release. This endotoxin content ismeasured using colorimetry (at 405-nm absorbance). The time required to release pNA isinversely propotional to the endotoxin content, and the logarithmic correlation between
time and endotoxin concentration shows linearity. This linear plot affords quantitative
measurements of the endotoxin content.
LAL
Mix LAL with synthetic substrate, and add 0.2 mol/L Tris-HCl buffer (pH 8.0)
before stirring gently to facilitate dissolution. Store the treated LAL solution in ice water
until use.

17

Operational procedures
Treat three 0.10 mL samples each of LAL solution with synthetic substrate inseparate test-tubes (each with 10-mm inner diameter and 75-mm length). Place mixturesin the 3 test-tubes in ice water, and add 0.10 mL each of sample solution, standard
endotoxin solution and endotoxin-testing water to the respective test-tubes. Close eachtest tube with a cap, agitate the mixture and heat at 37 0C for a fixed period of time (30minutes). Immediately transfer and chill the tubes in ice water after heating, add 0.4 mLof acetic acid solution (0.08 mol/L) to terminate the reaction. Measure the mixtures inendotoxin-testing water at 405-nm absorbance within 2 hours after terminating thereaction.
Judgement
Except for spirally administered preparation, an endotoxin concentration below
175 EU/V in non-oral preparations is considered negative, where V represents the highest
volume in mL administered in 1 hour.
The sample is considered conformed when treatment with the sample solution andendotoxin-testing water in the test-tubes yields a negative result, as well as whentreatment with standard endotoxin solution in the test-tube tests positive.
If treatment with endotoxin-testing water tests positive, or if treatment withstandard endotoxin solution tests negative, the test is deemed invalid.
Use new reagents or replace equipment to repeat the test.
5.3

Sterility Test

Sterility test may be conducted pursuant to the general tests described in theJapanese Pharmacopoeia, or in the blood culture system.
5.3.1

Blood culture system

This system examines bacterial presence in a sample through observing the flowof culture medium from the culture bottle to the growth signal reservoir, connected with athin needle, reflecting a rise in the culture bottle’s positive pressure as a result of bacterial
metabolism producing gaseous compounds.
This system consists of a blood bottle and growth signal reservoir.
18

Culture bottle
The culture bottle contains culture medium that can simultaneously grow aerobic
and anaerobic bacteria.
Measurement method
1) Volume of test solution
Although this system is capable of conducting measurement with 0.1 to 10 mL oftest solution, use at least 1 mL. Note that a test sample diluted over 100 times cannot be
used as test solution.
2) Operational methoda) Remove the bottle’s plastic cap, and use swab alcohol to thoroughly disinfectthe rubber stopper.
b) Gently agitate the sample and culture medium to mix them well.
c) Place the inoculated culture bottle in an incubator at 35 – 37 0C for
approximately 1 hour.
d) Use alcohol cotton balls to thoroughly sterilize the culture bottle’s rubber
stopper.
e) Remove the cap from the growth signal connector needle, making sure not totouch the needle itself. Insert the needle into the culture bottle’s rubber
stopper, and equalize the pressure inside the bottle (negative pressure), before
inserting the needle further until the end reaches the culture medium.
f) Press down the fixed external cylinder, and affix the growth signal reservoir tothe bottle.
g) Place the reservoir-affixed bottle (“signal unit” hereafter) in the incubator, andconduct machine shaking at least four times on Day 1. From Day 2 onwards,
conduct shaking twice a day.
h) Observed the signal unit according to the following schedule:i. Measure once each at 2 hours later and 6 hours later on Day 1, day 2(24 hours later), Day 3 (48 hours later), Day 4 (72 hours later) and Day
8 (7 days later).
ii. The measurement schedule for 2 hours later (Day 1) may be omitted.
19

iii. Conduct the following operations if the culture medium appears
cloudy at the time of observation.
i) Gently shake the bottle to mix the contents.
j) Place the nozzle of a syringe (5 mL or over) with the needle removed at theventilation hole on the cap of the growth signal reservoir, and inject air into
the signal unit. Remove the syringe to let the culture medium flow into thegrowth signal reservoir.
k) Remove the cap of the growth signal reservoir, and collect the culture medium
into a sterile syringe.
l) Repeat the operations a) – h). Add at least 5 mL of culture medium.
Judgement
Give a positive result if the culture medium is above the fixed external cylinder ofthe growth signal reservoir. Give a negative result if the culture medium does not reachthe fixed external cylinder of the growth signal reservoir even on Day 8, and if themedium does not appear cloudy. If the medium appears cloudy, conduct the operation i) –l) to continue culturing, and make a judgment on Day 8.

20

6ANALYSIS METHODS FOR [18F]-FDG INJECTION

An alternative analysis method may be used if the method is deemed to yieldhigher levels of accuracy and precision than those specified here.
6.1

Radiochemical purity

Radiochemical purity is determined by monitoring the radioactivity signal fromthe detector coupled in series with HPLC or by measuring the radioactivity distribution
with radioactivity scanners or imagers after TLC separation.
High Performance Liquid Chromatography (HPLC)[Note 1]a) Column: µBondapak Carbohydrate (3.9 mm- inner diameter x
300-mm length), Waters [Note 2].
Elution phase: CH3CN/H2O (85:15)
Flow rate: 2 mL/min
Retention time: 3.8 minutesb) Column: Asahipak NH2P-50 4E ( 4.6-mmm inner diameter x
250-mm length), Showa Denko.
Elution phase: CH3CN/ H2O (9:1)
Flow rate: 2 mL/min
Retention time: 5.9 minutesc) Column: Aminex HPX-87H (7.8-mm inner diameter x 300mm length), Bio-Rad [Note 3].
Elution phase: H2O (850C)
Flow rate: 1 mL/min
Retention time: 7.0 minutes[Note 1]: A regular analysis requires only one set of conditions. However, in cases
where the reaction criteria are required to be examined or in the event of a problem, it ispreferable to use two sets of conditions with different characteristics in the analysis. Notethat the [18F] fluoride ion is absorbed on a column and not eluted in a system adhere abuffer solution is not used as an elution phase, and [18F] fluoride ion is not included in theelution phase passing through the regular alumina column (alumina cartridge).
21

[Note 2]: Residual radioactive impurities which appear in the case of insufficienthydrolysis, can be well separated from [18F]-FDG under Condition (c), but not under
Condition (a).
[Note 3]: Bio-Rad HPX 87C column may be used in a similar manner.
Thin Layer Chromatography (TLC)a) Plate: Silica gel (soaked in 0.2 mol/L NaH2PO4, dried at theroom temperature, and heated at 110 0C for 30 minutes before use).
Elution phase: methanol/triethylamine (1000:1)
Rf value: 0.62b) Plate: Silica gel (soaked in 0.2 mol/L NaH2PO4, dried at theroom temperature, and heated at 110 0C for 30 minutes before use).
Elution phase: acetonitrile/H2O (95:5)
Rf value: 0.4c) Plate: Cellulose, Merck
Elution phase: 2-butanol/conc. NH4OH/H2O (6: 1 : 33)
Rf value: 0.67d) Plate: Silica gel (soaked in 0.2 mol/L NaH2PO4, dried at theroom temperature, and heated at 110 0C for 30 minutes before use).
Elution phase: isopropanol/H2O (4 : 1)
Rf value: 0.70e) Plate: Silica gel (soaked in 0.2 mol/L NaH2PO4, dried at theroom temperature, and heated at 110 0C for 30 minutes before use).
Elution phase: 2-butanol/CH3OH/CH3CO2H (3 : 1 : 1)
Rf value: 0.76

22

6.2

Chemical purity and specific radioactivity
High Performance Liquid Chromatography (HPLC)A high-sensitivity electrochemical detector is required to gauge the specificradioactivity of [18F]-FDG synthesized by the carrier-free fluoride ion method. Said
detector should be able to conduct high-sensitivity saccharide analysis in amperometricmode (e.g. detectors from Japan Dionex or Yokokawa Analytical System, etc.).
Quantitative limit for [18F]-FDG is 30 to 50 ppb.
However, tests for calculating specific radioactivity may be omitted for
preparations that have been manufactured by non-carrier added method and where
quantification of mass is impractical.
a) Column: Dionex CarboPac PA1 ( 4-mm inner diameter x 250-mm length),
Japan Dionex.
Elution phase: 0.1 mol/L NaOH
Post-column reagent: 0.35 mol/L NaOH
Flowrate: 1ml/L/min for both elution phaseb) Column: CarboPac PA-1(4.0 mm inner diameter x 250 mm, Dionex) +CarboPac PA-1 guard (4.0 mm inner diameter x 50 mm), 30 0C.
Elution phase: 200 mM NaOH
Flow rate: 0.3 mL/min
Detection: pulsed amperometry, electrode, gold (working) vs Ag/AgCl(reference)
Retention time: 20.0 min (FDG), 21.0 min (ClDG (2-Deoxy-2chloro-Dglucose))
6.3

Chemical Impurities
Kryptofix 222

The method using TLC is convenient to detect Kryptofix 222 during the
manufacturing of [18F]-FDG using the fluoride ion method. The detection limit is around30 ppm.[Note 1]

23

Thin Layer Chromatography (TLC)a) Plate: Silica Gel
Elution phase: CH3OH/conc. NH4OH (9 : 1)
Rf value: Kryptofix 222, 0.38;[18F]-FDG, 0.83b) Plate: Silica gel
Elution phase: triethylamine/CH3OH (0.1 : 99.9)
Rf value: [18F] fluoride ion,
Kryptofix 222, 0.38;
glucose, 0.36;[18F]-FDG, 0.62
[Note 1]: Kryptofix 222 is rarely detected during manufacturing under normal
conditions, if the positive ion-exchange resin AG50W-X8 is used for purification.
Tetrabutyl ammonium (TBA) ion
The following test is required when tetrabutylammonium bicarbonate(TBAHCO3) is used as a reaction catalyst.
To prepare the standard reagent solution, take a designed aliquot of 0.1 to 0.5 mLof 1 mmol/L standard TBA ion solution, add 3 mL of 3.75 mmol/L sodium bis (cis-1,2diethylenedithiolate) nickelate solution, 1 mL of 0.1 mmol/L acetate buffer (pH 5) andwater to make a final volume of 5 mL before standing for 20 minutes. Next, add thestandard reagent solution to a separating funnel containing 10 mL chloroform, mix
throughly, separate the lower chloroform phase, and filter with a teflon filter paper toremove water moisture. Use a spectrophotometer to measure the absorbance at 318-nm
wavelength and plot a calibration graph thereof. Measure the absorbance of a 1-mL
sample, after adjusting it with the same procedure. Based on the calibration graph, theTBA ion concentration of the material is then derived.

24

Aluminium ion
The following test is required when the alumina column (alumina cartridge) isused to purify [18F]-FDG[Note 1].
Drip a drop of [18F]-FDG solution on a test plate, and make sure it turns purple
when treated with one drop each of ammonia reagent and alizarin sulfonate solution
without turning red when acidified with acetic acid. The detection limit is around 0.65 mg(ca. 0.02 ppm).
The measurement range of commercially available ion-test paper Alumicheck(Advantek Toyo) is 2 – 100 ppm.
[Note 1]: Aluminium ion is not detected when [18F]-FDG preparation is neutral
under normal conditions.
Methanol, ethanol, acetonitrile
The type of residual solvents contained in the [18F]-FDG injection strongly
depends on the production method. Ethanol and acetonitrile are commonly contained in
the solution, but methanol and ether are in the rare case. Here, an example listed in theJapanese standard is described.
Column: HP-PLOT Q (0.52-mm inner diameter x 30 m, 40 µm, Agilent, 160 0C)
Carrier: helium. 4 mL/min
Detection: FID, 250 0C
Injection; 0.5 mL, split (2 : 1), 250 0C
Retention time: 2.4 min (methanol)
4.2 min (ethanol)
5.0 min (acetonitrile)

25

7

QUALITY CRITERIA AND FREQUENCY OF TEST FOR [18F]-FDG

The quality criteria of [18F]-FDG are slightly different among the countries. The
level of QA/QC may depend on the scale and the object of the facility, number of staff
and so on. The following criteria and frequency of the test summarised in Table 1 arecategorized in a GMP facility and In-house facility, and has been agreed and accepted by
the member of Group 2 at the FNCA Workshop on Cyclotron and Positron Emmission
Tomography (PET) in Medicine, 20 November 2007 held in Kuala Lumpur, Malaysia.
Table 1: Quality Criteria of [18F]-FDG and Frequency of Tests
No.

QC Tests

1

Appearance

2
3

pH
Radionuclidic
Identity
Radiochemical
PurityKryptofix 2.2.2
Residual
Solvents

4
5
6

7
8
9
10
11
12
13
14

26

Bacterial
Endotoxins
Sterility
Membrane Filter
Integrity
Radionuclidic
PuritySpecific
Radioactivity
ClDG
Aluminium Ion
Osmolarity

Acceptance Criteria
Clear, colorless or
slightly yellow
5.0 – 8.0
Half-life: 105 – 115 min

Frequency of Test
GMP-facility In-house
Every Batch
Every Batch
Every Batch
Every Batch

Every Batch
Every Batch> 95% (Radio-TLC)

Every Batch

Every Batch< 50 µg/mL< 0.04% acetonitrile< 0.5% alcohol< 0.5% ether
150 EU/V

Every Batch
Every Batch

Every Batch
Twice a year

Every Batch

Every Batch

Meet the Test
>50 psiEvery Batch
Every Batch

Every Batch
Not necessary>99.5% (511, 1022 keV)

Twice a year

Not necessary

No carrier added

Not necessary Not necessary< 1.0 mg/V
< 10 µg/mL-

Once a year
Not necessary
Not necessary Not necessary
Not necessary Not necessary

8

REFERENCES AND FURTHER READING
1.

2.

3.
4.
5.

6.

27

Standard for Compounds Labeled with Positron Nuclides Approved as
Established Techniques for Medical Use (by the Subcommittee on
Medical Application of Cyclotron-Produced Radionuclides) (2001
revision).
Description of the “Established Standard Techniques of Labeling
Compounds with Positron Nuclides for use as radiopharmaceuticals:
Approved by the Subcommittee on Medical Application of CyclotronProduced Radionuclides (revision in 2001).
Notice:All contents on these references are the copyright of “Japan Radioisotope
Association” unless stated. Japanese FNCA office “Nuclear Safety
Research Association (NSRA)” placed them in this FNCA Guideline withpermission from the copyright owner “Japan Radioisotope Association”.
Also, these references can be downloaded from the web of “Japan
Radioisotope Association”. URL is as follows;
http://www.jrias.or.jp/index.cfm/6,784,108,145,html
Guide to Good Manufacturing Practice For Medicinal Products in the
PIC/S GMP Guide, 2007.
Current Good Manufacturing For Positron Emission Tomography,
http://www.fda.gov.
Guidelines on the Current Good Radiopharmacy Practice (cGRPP) In the
Preparation of Radiopharmaceuticals, cGRPP-Guideline, Version 2 March
2007.
Guidelines for Good Manufacturing Practices of Radiopharmaceuticals,
IAEA, RAS/2/2009 on Quality Assurance and Good manufacturing
Practices for Radiopharmaceuticals, 2001

9

TERMINOLOGY
i.

ii.

iii.

iv.
v.
vi.
vii.

viii.
ix.
x.

xi.

xii.

28

Acceptance Criteria means numerical limit, ranges, or other criteria fortests that are used for or making a decision to accept or reject a unit, lot, orbatch of a PET drug product.
Batch means a specific quantity of PET drug intended to have uniformcharacter and quality, within specified limits, that is produced according toa single production order during the same cycle of production.
Component means any ingredient intended for use in the production of a
PET drug product, including any ingredients that may not appear in the
final PET drug product, such as precursor, reagent, and solvents.
Final release means the authoritative decision to permit the use of a batchof a PET drug in humans.
PET means positron emission tomography
PET center means a facility that is engaged in the production/applicationof a PET drug product.
PET drug means a drug that exhibits spontaneous disintegration of
unstable nuclei by the emission of positrons and is used for providing dual
photon positron emission tomographic diagnostic images.
PET drug product means a finished dosage form that contains a PET drug,
whether or not in association with one or more other ingredients.
PIC/S means the Pharmaceutical Inspection Co-operation Scheme
Production means the manufacturing, compounding, processing,packaging, labelling, reprocessing, repacking, relabeling, and testing of a
PET drug product.
Quality control means a system for maintaining the quality of PET drug
product, intermediate, analytical supplies, and other components,
including container-closure systems and in-process materials, throughprocedures, tests, analytical methods, and acceptance criteria.
Specifications means the tests, analytical procedures, and appropriate
acceptance criteria that establish the criteria to which a PET drug, PET
drug product, component, or other material used in PET drug production
must conform to be considered acceptable for its intended use.
Conformance to specifications means that a PET drug, PET drug product,when tested according to the described analytical procedures, meets thelisted acceptance criteria.

FNCA 2009 WORKSHOP ON CYCLOTRON AND PET IN MEDICINENAME OF GROUP 2: PET RADIOPHARMACEUTICAL
LIST OF MEMBERS:
Akihiro Morita, Mr.
International sales
Sumitomo Heavy Industries, Ltd.
Azahari Kasbollah, Mr.
Medical Technology Division
Malaysian Nuclear Agency
Eva Hyacinth Ocampo Torres, Ms.
PET Radiochemist
St. Luke’s Medical Center
Fan Wo, Prof.
Soochow University
Jiradanai Sarasamkan, Mr.
National Cyclotron and PET Centre
Chulaborn Hospital
Kazutoshi Suzuki, Dr
National Institute of Radiological Sciences
Nguyen Cong Duc, Mr.
PET-CT & Cyclotron Unit
Cho Ray Hospital

29

Rehir Dahalan, Dato’ Dr.
Director
Medical Technology Division
Malaysian Nuclear Agency
Suharzelim Abu Bakar, Mr.
Cyclotron and PET Centre
Hospital Putrajaya
Supaporn Sawadjoon, Ms.
National Cyclotron and PET Centre
Chulaborn Hospital
Syed Ashraf Fahlawi Wafa Syed Mohd Ghazi
Health Physics
Malaysian Nuclear Agency
Zulkifli Mohamed Hashim, Dr.
Medical Technology Division
Malaysian Nuclear Agency

30

31