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| 001 | 1014710 | ||
| 005 | 20231027114414.0 | ||
| 024 | 7 | _ | |a 10.1016/j.jchromb.2023.123847 |2 doi |
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| 100 | 1 | _ | |a Humpert, Swen |0 P:(DE-Juel1)132740 |b 0 |
| 245 | _ | _ | |a Validation of analytical HPLC with post-column injection as a method for rapid and precise quantification of radiochemical yields |
| 260 | _ | _ | |a New York, NY [u.a.] |c 2023 |b Science Direct |
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| 520 | _ | _ | |a 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. |
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| 700 | 1 | _ | |a Hoffmann, Chris |0 P:(DE-Juel1)180770 |b 1 |
| 700 | 1 | _ | |a Neumaier, Felix |0 P:(DE-Juel1)175142 |b 2 |
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| 700 | 1 | _ | |a Neumaier, Bernd |0 P:(DE-Juel1)166419 |b 4 |e Corresponding author |
| 773 | _ | _ | |a 10.1016/j.jchromb.2023.123847 |g Vol. 1228, p. 123847 - |0 PERI:(DE-600)1491259-4 |p 123847 - |t Journal of chromatography / B |v 1228 |y 2023 |x 1570-0232 |
| 856 | 4 | _ | |u https://juser.fz-juelich.de/record/1014710/files/Artikel%20Autorenversion.pdf |y OpenAccess |
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