Qualification and Application of a Liquid Chromatography-Tandem Mass Spectrometric Method for the Simultaneous Quantification of Triflusal and its Active Metabolite Desacetyl Triflusal in Human Plasma Using Solid Phase Extraction Technique

Author(s): Sathish Babu Munaga*, Rajani Kumar Valluru, Phani Bhushana Reddy Bonga, V. Sumathi Rao, Hemanth Kumar Sharma

Journal Name: Current Pharmaceutical Analysis

Volume 13 , Issue 4 , 2017

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Graphical Abstract:


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 problems.

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.

Keywords: Liquid chromatography, mass spectrometry, analysis, triflusal, desacetyl triflusal, solid phase extraction, pharmacokinetics, bioavailability.

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Article Details

Year: 2017
Published on: 13 June, 2017
Page: [384 - 397]
Pages: 14
DOI: 10.2174/1573412913666161222143118
Price: $65

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