A highly sensitive method for in vitro testing of fluorinated drug candidates using high-resolution continuum source molecular absorption spectrometry (HR-CS MAS).
We report here the development, optimization, and evaluation of a highly sensitive method for the determination of fluorine in biological matrices employing highresolution continuum source molecular absorption spectrometry (HR-CS MAS), suitable for pharmacological testing of fluorine-containing drug candidates. For this purpose, the most important parameters were studied in detail and subsequently optimized using a multivariate approach based on experimental design methodology. We developed a new approach employing a graphite tube lined with tantalum foil, thereby significantly enhancing sensitivity, while interferences from phosphorus monoxide (PO) molecular absorption due to the complex phosphate-rich matrix were completely eliminated. The limit of detection and the characteristic mass were 5.79 and 6.08 pg F, respectively. In order to evaluate the accuracy of the procedure, a recovery test was performed using spiked samples from three bioassays (i.e., DNA binding, protein binding, and cellular uptake) and the recovery rates ranged from 97.4 to 106.4%. The proposed method is applicable for preclinical in vitro testing of fluorinated drug molecules and thereby establishes HR-CS atomic absorption spectrometry instrumentation as a universal tool in medicinal chemistry.
Department of Pharmaceutical Chemistry, Institute of Pharmacy, Center for Molecular Biosciences Innsbruck, Center for Chemistry and Biomedicine (CCB), University of Innsbruck, Innrain 80-82, 6020, Innsbruck, Austria.
Irene
I
Würtenberger
Ronald
R
Gust
D001681:Biological Assay / Q000379:methods*; D045744:Cell Line, Tumor; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D004247:DNA / Q000737:chemistry*; D005459:Fluorides / Q000737:chemistry; D005461:Fluorine / Q000737:chemistry*; D006108:Graphite / Q000737:chemistry; D054879:Halogenation; D006358:Hot Temperature; D006801:Humans; D057230:Limit of Detection; D015999:Multivariate Analysis; D010087:Oxides / Q000737:chemistry; D010710:Phosphates / Q000737:chemistry; D010758:Phosphorus / Q000737:chemistry; D011485:Protein Binding; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013054:Spectrophotometry, Atomic; D013499:Surface Properties; D013635:Tantalum / Q000737:chemistry*
D010087:Oxides; D010710:Phosphates; D010758:Phosphorus; D005461:Fluorine; D013635:Tantalum; D006108:Graphite; D004247:DNA; D005459:Fluorides