10 September 2016

Lecture #4: RADIOPHARMACEUTICALS



Radionuclide Generators

A Radionuclide Generator is the major source of short–lived radionuclides used in Nuclear Medicine. In a radionuclide generator, a longer–lived radionuclide is called the parent decay while the shorter–lived radionuclide is called the daughter. The daughter can be removed periodically as it is replenished by decay of the parent.

Examples of Parent–Daughter Systems are:

1.      Germanium–68 / Gallium–68
2.      Rubidium–81 / Krypton–81
3.      Molybdenum–99 / Technetium–99

Molybdenum–99 / Technetium–99m Generator

Parts of a 99Mo/99mTc Generator

1.      Aluminum trioxide (alumina) column – this is where the parent radionuclide, 99Mo is absorbed.

2.      Porous glass – holds the alumina in place but allows saline to pass through it.



99mTc is removed or eluted from the column when a sterile, evacuated vial is in placed at the opposite end of the saline eluant. The vacuum in the collection vial causes the saline to be pulled through the alumina column, which removes the loosely bound 99mTc. Due to the different chemical properties of the parent and the daughter, 99Mo remains on the column while 99mTc is collected as the generator eluate in the form of [99mTc] sodium pertechnetate.

Types of 99Mo/99mTc Generator

1.      Wet column generator – contains a supply of sterile saline within the generator itself. To elute this type of generator, the technologist chooses the appropriate size collection vial to obtain the volume and activity concentration desired. Because no air follows the saline as it flows over the column, the column remains wet.

2.      Dry column Generator – requires that saline, provided by the generator manufacturer, is placed on the charging port, a needle connected to tubing that carries the saline to the alumina column. An evacuated vial is the place onto the needle on the collection port. The vacuum pulls the saline over the column, removing the 99mTc activity. Using an evacuated vial with a volume larger than the saline volume causes air to be pulled over the column, completely removing any saline.




Maintenance of Generators

1.      The septa of eluate and saline vials should be swabbed with alcohol

2.      Needles used to deliver saline or to collect the eluate should be replaced with new, sterile needles following each elution.

3.      Millipore filters are oftentimes present to ensure sterility of the generator

Generator yields

All 99Mo/99mTc generators contain 99Mo from the fission of Uranium–235 which has a high specific gravity. Specific gravity refers to the amount of radioactivity per unit mass of a radionuclide or radiopharmaceutical and is expressed in such units as millicuries per milligram (mCi/mg), megabecquerels per milligram (MBq/mg), curies per mole (Ci/mole) or gigabecquerels per mole (GBq/mole).

A simplified, useful rule of thumb when elution is performed only once every 24 hours is that the eluate yield will be 80% of the previous yield.

Percent of Technetium–99m activity present in generator column at selected times following Elution

                        Time since elution                          99mTc activity
                                    (hour)                                     (%99Mo activity)
                                   
                                    1                                                          9.4
                                    2                                                          18
                                    3                                                          26
                                    4                                                          32
                                    5                                                          39
                                    6                                                          44
                                    7                                                          49
                                    8                                                          54
                                    9                                                          58
                                    10                                                        61
                                    11                                                        65
                                    12                                                        68
                                    18                                                        80
                                    24                                                        87

Problems from Elution

1.      Mechanical problem such as an air leak in the tubing may prevent the saline from being drawn out of the column into collection vial.

2.      “Channeling” of the alumina column causes only a portion of the column to be exposed to saline, leaving 99mTc on that part of the column not washed with eluant.

3.      Chemical reduction of 99mTc from +7 valence state to other reduced states occurs when reducing agents are formed by the radiolysis of water. The reduced 99mTc remains bound onto the column and is not eluted.

Eluate assay and Quality Control

1.      Concentration of eluate

Concentration           =          Total 99mTc activity (mCi)
                                                     Elution volume (ml)

2.      Molybdenum–99 content

The appearance of 99Mo in generator eluate is called “molybdenum (or moly) breakthrough” which is considered a radionuclide impurity. When administered to patients, 99Mo is taken up by the parenchymal cells of the liver and delivers an unnecessary radiation dose to that organ.
The NRC states that the amount of 99Mo in 99mTc generator eluates may not exceed 0.15 µCi per 1 mCi 99mTc.

3.      Lead shield (or Moly Shield) Method

A lead container is used, which is designed to absorb Tc99m photons (140 keV) as well as permit the passage of higher energy (740, 780 keV) Mo99 photons. The generator eluate is placed in the moly shield and assayed in a dose calibrator to determine the Mo99 content. Sometimes it is necessary to apply a correction factor to the Mo99 assay. Instructions supplied with the molyshield should be consulted. To determine the amount of Mo99 contamination in the eluate, divide the Mo99 assay result by the total number if millicuries of Tc99m in the eluate measured with the dose calibrator.

            99Mo breakthrough =                      µCi 99Mo
                                                                        mCi 99mTc

4.      Mock Moly method

This method compares the unknown 99Mo to that of a cesium–137 reference standard of known activity. A well–type scintillation counter is used. The pulse–height analyzer window is set to accept counts within a range of 600 – 1000 keV which will include the principle photon of 99Mo (740, 780 keV) and 137 Cs (662 keV).

µCi 99Mo        =          net counts 99mTc eluate        x          µCi137Cs standard
                                    net counts 137Cs standard   

5.      Aluminum Ion content

Aluminum ions from the generator column appearing in the 99mTc eluate are a chemical impurity. Such impurities can affect the biodistribution of radiopharmaceutical. The established USP Al+3 concentration limit is 10 µg Al+3/ml eluate is not exceeded.

Preparation of Technetium–99m radiopharmaceuticals

1.      Technetium–99m–sodium pertechnetate eluted from a 99Mo/99mTc generator is combined with the appropriate kit reagent to form a tissue–specific compound after only a brief incubation period.

2.      Most of 99mTc–labeled compounds with the exception of 99mTc colloid, are prepared with chemically reduced 99mTc, a more active form of Technetium. For this reason, 99mTc labeling is accomplished in the presence of stannous chloride, a reducing agent. Most commercially available radiopharmaceutical preparation already has stannous chloride incorporated in the compound.
The stannous ion (Sn+2) reduces the valence state of the technetium in the pertechnetate ion from +7 to +4 or another reduced valence state. The reduced 99mTc (IV) is a more reactive species, capable of combining with a variety of compounds.

Radiopharmaceutical with specific preparation

a.      Technetium–99m–MAA is prepared with reduced 99mTc however the number of particles to be administered to the patient should be considered. The recommended number of particles is 200k to 700k particles

If too few particles are administered, the perfusion lung image may be technically unsatisfactory and may have the appearance of decreased uptake.

As 99mTc decays, the number of radioactive particles decreases, while the total number of particles in a milliliter remains constant. Therefore, as time passes, more particles must be administered to obtain the same amount of activity.

b.      Technetium–99m–sulfur colloid does not require stannous chloride for preparation. Instead, it contains an acid, sodium thiosulfate, a buffer and gelatin.

Radiopharmaceutical Quality Control

1.      99mTc is present in three different chemical forms

a.      Bound 99mTc agent – the desired form needed for examination

b.      Unbound (free) 99Tc pertechnetate – can be visualized on images as increased tracer concentration in the stomach, thyroid and salivary glands.

c.       Hydrolyzed, reduced 99mTc (HR–Tc) – is indicated through uptake in the reticuloendothelial system, especially liver.

Technetium–99m–pertechnetate and HR–Tc are radiochemical impurities
Radiochemical purity is defined as the fraction of total radioactivity present in its desired chemical form.

2.      Radiochromatography – in this method, a drop of the radiopharmaceutical to be tested is placed at a point called the origin, near one end of the paper strip. The strip is then immersed into a solvent. By capillary action, the solvent travels up the strip carrying with it the soluble components in the sample.

Insoluble components remain at the origin. As the solvent nears the top of the strip, the strip is removed, dried and cut in half. Each half is counted in a well counter or dose calibrator and the labelling or tagging efficiency, the fraction of radioactivity incorporated into the desired radiolabeled agent.

% Radiochemical impurity                        =          cpm in Part A x 100%         
                                                                        cpm in Part A + cpm part B

Labeling efficiency =          100% – [(%free 99mTc) pertechnetate + HR–99mTc]

The Rf value refers to the distance the component travels from the origin compared to the distance traveled by the solvent.

Rf value          =          distance component travels from origin
                                    distance of solvent front from origin

Unit Dose Preparation

1.      Radionuclide and chemical form of radiopharmaceutical. A radionuclide such as 99mTc may be tagged to a variety of chemical compounds so care should be taken to choose the correct radiopharmaceuticals for a given nuclear medicine procedure.

2.      Expiration time/date. The time at which the radiopharmaceutical should no longer be administered to patient to patients.

3.      Volume (in ml). The total amount of liquid in the vial

4.      Total activity. The total amount of radioactivity present in the vial at the time of assay or calibration.

5.      Concentration. The amount of activity per unit volume at the time of assay or calibration. Concentration is obtained by dividing the total activity in the vial by the total volume.

Concentration           =          Total activity
                                                Total volume

6.      Assay or calibration time/date. The time and / or date when the total activity and concentration were determined

7.      Specific gravity. The amount of activity per unit mass of a radionuclide or radiopharmaceutical. Do not confuse specific gravity with concentration. A specific gravity may not appear on all radiopharmaceutical labels.

8.      Lot number. A number assigned by the manufacturer to each batch of radiopharmaceutical. The number can be used to trace problems associated with the radiopharmaceutical.

9.      For adults, the standard dose range is already predetermined. For pediatric patients, they should receive at least 62% of the recommended adult dose or any of the following formula for computation

a.      Clark’s rule                     weight in lbs. x adult dose
150 lbs.

b.      Webster’s rule                age (yr) +1      x          adult dose
age + 7

c.       Young’s rule                   age (yr)           x          adult dose
age + 12

10.  Initial radiopharmaceutical concentration is decay – corrected for the time of administration and the volume to be administered.

Co        x          DF       =          Ct

            Co        =          initial concentration
            DF       =          decay factor
            Ct         =          concentration at time

11.  Determine the volume to be administered

Patient dose              =          volume to be administered
            Ct


            Example:

A technologist needs 3.5 mCi of Thallous chloride to perform mycordial imaging on September 6. If the concentration of the radiopharmaceutical is 2 mCi/ml on September 8, what volume should be administered to the patient?

            Co        x          DF       =          Ct

                                    Co        =          Ct/DF

                                                =          2 mCi / ml
                                                                0.634

                                                =          3.2 mCi / ml

            Patient dose              =          volume to be administered           
                        Co

                3.5 mCi                       =          1.1 ml
            3.2 mCi / ml




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