Title: Usefulness of fast protein liquid chromatography as an alternative to high performance liquid chromatography of Tc-99m-labelled human serum albumin preparations
Authors: Verbruggen, Alfons
Verbeke, Kristin * # ×
Issue Date: Jun-1996
Publisher: Pergamon-elsevier science ltd
Series Title: Journal of pharmaceutical and biomedical analysis vol:14 issue:8-10 pages:1209-1213
Abstract: Accurate analysis of the radiochemical purity of Tc-99m-labelled preparations is usually performed using an appropriate high performance liquid chromatography (HPLC) system. For the analysis of Tc-99m-HSA, the European Pharmacopoeia (Ph. fur.) prescribes an HPLC system using silica gel for size-exclusion chromatography as the stationary phase. Analysis of Tc-99m-HSA preparations with this method allows one to distinguish radiolabelled polymeric, oligomeric, dimeric and monomeric forms of albumin as well as small molecular mass impurities such as pertechnetate ((TcO4-)-Tc-99m). Due to the relatively short lifetime of this type of column, two fast protein liquid chromatography (FPLC)-type columns were evaluated for their possible usefulness for quality control of Tc-99m-HSA preparations, namely Superdex 200 HR 10/30 and Superdex 75 HR 10/30 (Pharmacia, Sweden). Both columns contain a matrix produced by the covalent bonding of dextran to highly cross-linked porous agarose beads. The fractionation range of the Superdex 75 HR 10/30 column was too low to obtain an adequate separation between albumin and its polymerized forms and this column was therefore considered unsuitable for the analysis of Tc-99m-HSA preparations. The Superdex 200 column afforded the same pattern and relative amounts of albumin and its polymerized forms as the silica column although the resolution obtained was somewhat lower. A major difference was the fact that (TcO4-)-Tc-99m, could not be eluted from the Superdex column but was eluted from the silica column, whereas other small molecular mass, negatively charged Tc-99m complexes such as Tc-99m-diethylene triamine pentaacetic acid showed an identical elution pattern on both columns. The presence of pertechnetate in Tc-99m-HSA preparations, however, can easily be checked using paper chromatography (PC) and acetone as the eluent. Since the lifetime of the Superdex column is twice that of the silica column, FPLC in combination with PC can be recommended as a money-saving and efficient analytical technique for the quality control of Tc-99m-HSA preparations.
ISSN: 0731-7085
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Radiopharmaceutical Research
Translational Research in GastroIntestinal Disorders
* (joint) first author
× corresponding author
# (joint) last author

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