Determination of Dydrogesterone in Human Plasma by Tandem Mass Spectrometry: Application to Therapeutic Drug Monitoring of Dydrogesterone in Gynecological Disorders

Abstract

A sensitive and specific tandem mass spectrometric (MS–MS) method was developed and validated for the determination of dydrogesterone (Duphaston®), an orally active synthetic progestogen, in human plasma. Multiple reaction monitoring (MRM) scans at m/z 313.1 > 105.5 (dydrogesterone) and m/z 393 > 147 (dexamethasone, internal standard) were selected to determine dydrogesterone by the internal standard method. Linear correlations (r: ∼0.99 ± 0.05) of the calibration curves were established over the concentration range 10–60 ng mL−1 with a lower limit of quantification (LLQ) of 10 ng mL−1 (RSD% 14.9 and %DEVs −10.5 to +15.6). Solid-phase extraction (SPE) technique was used for extraction of dydrogesterone and internal standard from patient plasma samples using Oasis® Max C18 cartridges. Ion suppression studies indicated negligible effects of plasma matrix on the mass ions detection of dydrogesterone and IS, when measured in MRM mode. Validation data showed that RSD% values were <22.0%, whereas %DEV values were in the range of −20.2 to +13.3 for intra- and inter-day precision and accuracy, respectively. Analytical recoveries of dydrogesterone from supplemented plasma samples with the drug were in the range of 100.7–112%, indicating the efficiency of the SPE for separation of dydrogesterone from human plasma. Stability studies conducted at −20 °C, showed that dydrogesterone was stable in plasma as indicated from the measured degradation kinetic parameters. The developed method was applied for monitoring plasma levels of dydrogesterone in 25 patients treated with Duphaston® tablets at a dose of 10 mg three times daily. Mean plasma concentration of 16.1 ± 3.5 ng mL−1 of dydrogesterone were measured at the steady state. The data suggest the utility of tandem mass method in therapeutic drug monitoring of plasma levels of dydrogesterone in gynecological disorders treated with Duphaston® tablets.