Background: Triflusal and Desacetyl Triflusal prevents the formation of blood clots by inhibiting
the enzyme cyclooxygenase, and reducing production of thromboxane A2 which is a stimulator
of platelet aggregation and thereby reduces the risk of strokes, heart attacks or other serious circulatory
Introduction: A rapid, sensitive, specific, accurate, isocratic reversed-phase liquid chromatography
coupled with triple quadrapole mass spectrometry method in its tandem operation mode was developed
and validated for the simultaneous quantification of Triflusal and Desacetyl Triflusal, in human plasma.
Method: Salicylic Acid and Hydrochlorothiazide are used as internal standards. The analytes were extracted
from human plasma samples by solid-phase extraction technique using a Strata-X 33 µm polymeric
sorbent. The reconstituted samples were chromatographed on an Oyster ODS3 100 x 4.6mm, 5µ
column using a mobile phase composed of acetonitrile 40:60 (v/v) - 2mM ammonium acetate solutions
(pH 3.25 ± 0.3) at a flow rate of 3.2 mL/min. The detection of precursor and product ions of analytes
and their internal standards was performed by multiple reaction monitoring mode via a turbo ion spray
interface in negative mode by MDS SCIEX API-3000.
Result: The calibration curves obtained were linear over the concentration range of 0.100 to 35.086
µg/mL for Triflusal, 1.00 to 120.06 µg/mL for Desacetyl Triflusal with a correlation coefficient r ≥
0.9952. The results of the intra- and inter-day precision and accuracy studies were well within the acceptable
limits. The intra and inter batch precision was ≤ 12.7 for Triflusal and Desacetyl Triflusal with
accuracy (%RE) between -6.6 and 6.9. Stability testing showed that all analytes were stable in plasma at
25 °C for 17.83 h, seven freeze–thaw cycles, and at 70°C for 53.65 days. The analytical method was
successfully applied to an in vivo study evaluating the pharmacokinetics of Triflusal and Desacetyl
Triflusal following single dose oral administration in 24 healthy volunteers in an open label, twoperiod,
two sequence, randomized, crossover protocol under fasting conditions.
Conclusion: Based on the 90% confidence interval of the individual ratios for Cmax and AUC0-inf, it was
concluded that the method is efficient with a very short running time of 3.2min, sufficiently sensitive
and suitable for pharmacokinetic studies.