Development and validation of ultra high performance liquid chromatography–mass spectrometry method for LBH589 in mouse plasma and tissues

An ultra high performance liquid chromatography tandem mass spectrometry method (UHPLC-MS/MS) was developed and validated for the quantitation of LBH589, a novel histone deacetylase inhibitor (HDACi), in mouse plasma and tissues (liver, spleen, kidney and lung). Tobramycin was employed as the internal standard. Separation was performed on an Acquity UPLC™ BEH column, with a mobile phase consisting of 10% water (with 0.1% of trifluoroacetic acid) and 90% methanol (with 0.1% trifluoroacetic acid). LBH589 and tobramycin were determined using an electrospray ionization (ESI) interface. Detection was performed on electrospray positive ionization mass spectrometry by multiple reaction monitoring of the transitions of LBH589 at m/z 349.42 → 157.95 and of tobramycin at 468.2 → 163. Calibration curves for the UHPLC method (0.0025–1 μg/mL for plasma and tissue homogenates, equivalent to 0.0357–14.2857 μg/g for tissue samples) showed a linear range of detector responses (r > 0.998). Intra-batch and inter-batch precision expressed as coefficient of variation (CV) ranged from 0.92 to 8.40%. Accuracy expressed as bias, ranged from −2.41 to 2.62%. The lower limit of quantitation (LLOQ) was 0.0025 μg/mL for both plasma and tissue homogenate samples, equivalent to 0.0357 μg/g tissue. This method was successfully applied to quantify LBH589 in plasma and tissue samples obtained after the intraperitoneal administration of a single dose of 20 mg/kg of LBH589 in BALB/c mice.

► The method is specific, accurate and reproducible for biological analysis of LBH589. ► Liquid–liquid extraction allowed acceptable recovery and matrix effect values. ► The method is valuable for the determination of the pharmacokinetic behavior of LBH589 in mice.