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Quantitation of a Dinitro Flavonoid (DNF) in Rat Plasma Using the Varian 1200L LC/MS/MS System

ring (SRM).

Results and Discussion

The ratio of the area intensities of the product ion for DNF to the product ion of the internal standard [(m/z 221)/(m/z 129)] is used to calculate the DNF concentration in unknown rat plasma samples. The calibration curve is generated from the analysis of a drug-free rat plasma matrix fortified with various amounts of DNF and a fixed amount of the internal standard. The calibration curve range for DNF is from 0.10 ng/mL to 5.00 ng/mL of rat plasma.

Figure 2 shows the excellent response and peak shape obtained at the Lower Limit of Quantitation (LLOQ) and Upper Limit of Quantitation (ULOQ). Visual examination of the calibration curve in Figure 3 shows that good linearity was obtained with the linear-log, curve-fitting model used. Additional sensitivity can easily be obtained by increasing the sample volume used for the assay.

Conclusion

The pharmacokinetic plot derived from DNF plasma levels obtained with six rats is shown in Figure 4. Visual observation of the plot reveals that at three hours post-dose there is a drop in DNF concentration, which rebounds an hour later, and from that point the concentration drops gradually in a linear fashion.

This fluctuation in concentration may be due to a phenomenon known as entero-hepatic recirculation. Enterohepatic recirculation is a process defined as a recycling of a drug through the liver by its excretion in bile, followed by reabsorption of the drug from the intestines into the portal circulation and passage back to the liver, where it is reexcreted in the bile.

Another possibility to explain the fluctuation in concentration is related to the limited solubility of DNF in aqueous media. DNF may be precipitating in the stomach and then later moving into the intestines where its absorption takes place. More studies are n
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