A comparison of simple analytical methods for determination of fluoride in microlitre-volume plasma samples

The aim was to compare potential methods for fluoride analysis in microlitre-volume plasma samples containing nano-gram amounts of fluoride. Methods: A group of four laboratories analysed a set of standardised biological samples as well as plasma to determine fluoride concentration using three methods. In Phase-1, fluoride analysis was carried out using the established HMDS-diffusion method (1ml-aliquot/analysis) to obtain preliminary measurement of agreement between the laboratories. In Phase-2, the laboratories analysed the same samples using a micro-diffusion method and knownaddition technique with 200μl-aliquot/analysis. Coefficients of Variation (CVs) and intra-class correlation coefficients (ICCs) were estimated using ANOVA to evaluate the amount of variation within- and between-laboratories. Based on the results of the Phase-2 analysis, 20 human plasma samples were analysed and compared using the HMDS-diffusion method and known-addition technique in Phase-3. Results: Comparison of Phase-1 results showed no statistically significant difference among the laboratories for the overall data set. The mean between- and within-laboratory CVs and ICCs were <0.13 and ≥0.99, respectively, indicating very low variability and excellent reliability. In Phase-2, the overall results for between-laboratory variability showed a poor CV (1.16) and ICC (0.44) for the micro-diffusion method, whereas with the known-addition technique the corresponding values were 0.49 and 0.83. Phase-3 results showed no statistically significant difference in fluoride concentrations of the plasma samples measured with HMDS-diffusion method and known-addition technique, with a mean (SE) difference of 0.002(0.003) μg/ml. In conclusion, the known-addition technique could be a suitable alternative for measurement of fluoride in plasma with microlitre-volume samples.