生物等效性和生物利用度杂志

抽象的

Determination of Fingolimod in Whole Blood by LC-MS/MS: Application to Bioequivalence Study

Tawfiq Arafate, Ahmed Abu-Awwad, Basil Arafat, Asaad Shahin, Hadil Alotaibi, Mona Bustami

A new bioanalytical liquid chromatography tandem mass spectrometric method has developed and validated for a clinical determination of fingolimod hydrochloride in whole blood. The method has applied in bioequivalence study following parallel open-label design, randomized single dose, and two treatments by healthy adult male subjects under fasting conditions. Fingolimod with labeled Internal Standard (IS; Fingolimod-D4) was extracted from human blood by liquid-liquid extraction and separated chromatographically from their matrix by Fortis UniverSil Cyano column. The established method was validated over calibration dynamic range of 12-1200 pg/ml for spiked blood. In single period, each subject of 42 healthy volunteer was received a single dose of 0.5 mg fingolimod hydrochloride capsule of either the test or reference product. The pharmacokinetic parameters of C_max, AUC_0–t and T_max were calculated for fingolimod in blood of test product and compared with reference product. Fingolimod and IS have detected by a positive Multiple Reaction Monitoring (MRM) scan mode at mass filter of m/z 308.4 → 255.3 for fingolimod and 312.4 → 259.3 for IS. All validation results were within the acceptance criteria according to European guideline for bioanalytical methods. The current findings of AUC_0-72, C_max and T_max for Gilenya^® were bio-comparable to corresponding parameters derived from test product. The validated method was succefully applied in bioequivalence study to investigate figolimod 0.5 mg capsule from human whole blood following parallel periods design.