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@ARTICLE{Humpert:1014710,
author = {Humpert, Swen and Hoffmann, Chris and Neumaier, Felix and
Zlatopolskiy, Boris and Neumaier, Bernd},
title = {{V}alidation of analytical {HPLC} with post-column
injection as a method for rapid and precise quantification
of radiochemical yields},
journal = {Journal of chromatography / B},
volume = {1228},
issn = {1570-0232},
address = {New York, NY [u.a.]},
publisher = {Science Direct},
reportid = {FZJ-2023-03405},
pages = {123847 -},
year = {2023},
abstract = {Accurate assessment of isolated radiochemical yields (RCYs)
is a prerequisite for efficient and reliable optimization of
labeling reactions. In practice, radiochemical conversions
(RCCs) determined by HPLC analysis of crude reaction
mixtures are often used to estimate RCYs. However,
incomplete recovery of radioactivity from the stationary
phase can lead to significant inaccuracies if RCCs are
calculated based on the activity eluted from the column
(i.e. the summed integrals of all peaks). Here, we validate
a simple and practical method that overcomes problems
associated with retention of activity on the column by
determination of the total activity in the sample using
post-column injection. Post-column injections were carried
out using an additional injection valve, which was placed
between the outlet of the HPLC column and the inlet of the
detectors. 2-[18F]Fluoropyridine ([18F]FPy) and
8-cyclopentyl-3-(3-[18F]fluoropropyl)-1-propylxanthine
([18F]CPFPX) were prepared with radiochemical purities of
$>99.8\%$ and mixed with [18F]fluoride at a ratio of 1:1 to
simulate reaction mixtures obtained by radiolabeling
reactions with an RCC of $50\%.$ The samples were analyzed
on three different C18 HPLC columns using neutral and acidic
mobile phases. RCCs determined using the summed area of all
peaks in the chromatograms were compared with those
determined using post-column injection. Additionally, RCCs
determined by post-column injection were corrected for
activity losses before, during and after radiosyntheses to
afford analytical RCYs, which were compared with isolated
RCYs. Determination of RCCs based on the summed area of all
peaks gave correct results under certain chromatographic
conditions, but led to overestimation of the actual RCCs by
up to $50\%$ in other cases. In contrast, determination of
RCCs using post-column injection provided precise results in
all cases, and often significantly reduced analysis time.
Moreover, analytical RCYs calculated from RCCs determined by
post-column injection showed excellent agreement with
isolated RCYs $(<3\%$ deviation). In conclusion, HPLC
analysis using post-column injection enables reliable
determination of RCCs independent of the chromatographic
conditions and, together with a simple activity balance,
rapid and accurate prediction of isolated RCYs.},
cin = {INM-5},
ddc = {540},
cid = {I:(DE-Juel1)INM-5-20090406},
pnm = {5253 - Neuroimaging (POF4-525)},
pid = {G:(DE-HGF)POF4-5253},
typ = {PUB:(DE-HGF)16},
pubmed = {37634390},
UT = {WOS:001070859200001},
doi = {10.1016/j.jchromb.2023.123847},
url = {https://juser.fz-juelich.de/record/1014710},
}