Design of the experiment. Aggregation of untreated sample (without adding the flavonoid, AUC0) and the sample of flavonoid (AUC) are measured. If the difference of the aggregations is greater than 5% the measurement of additional two citrated bloods is performed. Otherwise, double concentration of flavonoid is taken and the first step is repeated. If the flavonoid in the analysed concentration shows statistically lower aggregation compared to the untreated sample (t-test) the analysed concentration is equal to minimal antiaggregatory concentration (MINaAC). If not analysis is performed from the beginning with double the concentration of flavonoid. This presumes that the analyzed concentration is not the first analysed (n > 1), else, the analysis is performed from beginning using the half of the concentration of flavonoid. Example of prunetin is given. Aggregation of analysed concentration of prunetin: 3.8 μM is not greater than aggregation of the untreated sample (58 U vs. 59 U). Thus additional measurement with double concentration (7.6 μM) of prunetin is performed. As the difference of aggregation is greater than 5% (ΔAUC = 17%) measurements on two additional citrated blood samples were performed. The concentration obtained is statistically different compared to the untreated sample (p = 0.033) thus minimal antiaggregatory concentration of prunetin is equal 7.6 μM. Note that red and blue lines in charts indicate two parallel measurements of the same sample. If the deviation between these two measurements is greater than 10%, quality control flag appears and measurements should be repeated.
Bojić et al. Nutrition Journal 2011 10:73 doi:10.1186/1475-2891-10-73