Mitoxantrone Quantification by HPLC-MS/MS in Caco-2 Culture Media

Mitoxantrone is a marker substrate of breast cancer resistance protein (BCRP). BCRP involved in number pharmacokinetic drug-drug interactions. The transporter’s possible saturability makes it advisable to use low concentrations mitoxantrone for vitro studies. Consequently, quantification requires method with high sensitivity. aim the study was develop and validate procedure Caco-2 culture media by HPLC-MS/MS. Materials methods . authors used an Ultimate 3000 HPLC system TSQ Fortis triple quadrupole mass spectrometer Thermo Fisher Scientific Selectra C18 column (4.6×100 mm, 5 μm, 100 Å) United Chemical Technologies. elution ran gradient mode mobile phase 1% formic acid solution methanol. Experimental parameters were as follows: eluent flow rate, 0.3 mL/min; separation temperature, 35 °C; injection volume, μL; ana lysis time, 10 min; approximate retention 5.51 min. sample preparation precipitation from medium methanol, followed centrifugation at 13,000 g detection performed using electrospray ionisation positive ion mode. Detection voltage, 3700 V; sheath gas 50 L/min; auxiliary sweep 1 ion-transfer tube 300 evaporator 350 °C. set transitions m/z 455 88.2 358.1, collision energy these amounting 25 V 18 V, respectively. source fragmentation 0, CID pressure 2 mTorr. Results analytical showed selectivity, sensitivity (limit detection, nmol/L; lower limit quantification, nmol/L), accuracy, precision, linearity concentration range 50–1000 nmol/L. observed no carryover or matrix effects. A simulation real-life storage conditions demonstrated stability samples. Thus, enables preclinical evaluation medicinal product effects on functional activity BCRP, based assessing transcellular transport presence test product. Conclusion developed validated