TLC and HPLC in determining metabolite fractions

Separate plasma samples are collected for metabolite analysis during the PET study. Plasma proteins are precipitated or filtered to prevent clogging in the next phases. In case of small animal studies where plasma sample volumes are only few microlitres, microextraction solid-phase techniques can be used (Moein et al., 2020). A small volume of each plasma sample is applicated onto TLC disk or injected into the HPLC column. From the resulting chromatograms, the relative fractions of unchanged tracer and labelled metabolite(s) are calculated.

TLC example
Fig 1.Thin layer chromatography. © Anne Roivainen.

HPLC example
Fig 2. HPLC chromatogram. © Anne Roivainen.


See also:



References:

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Greuter HNJM Greuter, van Ophemert PLB, Luurtsema G, van Berckel BNM, Franssen EJF, Widhorst BD, Lammertsma AA. Optimizing an online SPE-HPLC method for analysis of (R)-[11C]1-(2-chlorophenyl)-N-methyl-N-(1-methylpropyl)-3-isoquinolinecarboxamide [(R)-[11C]PK11195] and its metabolites in humans. Nucl Med Biol. 2005; 32: 307-312. doi: 10.1016/j.nucmedbio.2004.12.005.

Hilton J, Yokoi F, Dannals RF, Ravert HT, Szabo Z, Wong DF. Column-switching HPLC for the analysis of plasma in PET imaging studies. Nucl Med Biol. 2000; 27: 627-630. doi: 10.1016/s0969-8051(00)00125-6.

Nics L, Hahn A, Zeilinger M, Vraka C, Ungersboeck J, Haeusler D, Hartmann S, Wagner K-H, Lanzenberger R, Wadsak W, Mitterhauser M. Quantification of the radio-metabolites of the serotonin-1A receptor radioligand [carbonyl-11C]WAY-100635 in human plasma: An HPLC-assay which enables measurement of two patients in parallel. Appl Radiat Isot. 2012; 70(12): 2730-2736. doi: 10.1016/j.apradiso.2012.08.016.

Passchier J. Fast high performance liquid chromatography in PET quality control and metabolite analysis. Q J Nucl Med Mol Imaging 2009; 53(4): 411-416. PMID: 19834451.



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Updated at: 2020-02-04
Created at: 2013-12-20
Written by: Vesa Oikonen, Richard Aarnio, Pauliina Virsu, Anne Roivainen