Development and Validation of an LC-MS/MS Assay for the Quantitation of MO-OH-Nap Tropolone in Mouse Plasma: Application to In Vitro and In Vivo Pharmacokinetic Studies

• Published in: Molecules (Basel, Switzerland) Switzerland), 2024-09, Vol.29 (18), p.4424
• Main Authors: Aldhafiri, Wafaa N, Chhonker, Yashpal S, Ahmed, Nusrat, Singh, Sandeep K, Haney, Staci L, Ford, James B, Holstein, Sarah A, Murry, Daryl J
• Format: Article
• Language: English
• Subjects: Accuracy; Acids; Animals; biodistribution; Blood proteins; Bone cancer; Chromatography; Chromatography, Liquid - methods; Cytotoxicity; in vitro metabolism; LC-MS/MS; Liquid Chromatography-Mass Spectrometry; Melanoma; Metabolism; Metabolites; Mice; Multiple myeloma; Optimization; Pharmacokinetics; Plasma; plasma protein binding; Protein binding; Proteins; Quality control; Reproducibility of Results; Retention; Tandem Mass Spectrometry - methods; tropolone; Tropolone - analogs & derivatives; Tropolone - blood; Tropolone - chemistry; Tropolone - pharmacokinetics
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules29184424

A rapid, selective, and sensitive liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantitation of MO-OH-Nap tropolone (MO-OH-Nap) in mouse plasma. MO-OH-Nap is an α-substituted tropolone with anti-proliferative properties in various cancer cell lines. Detection and separation of analytes was achieved on an ACE Excel C18 (1.7 µm, 100 × 2.1 mm, MAC-MOD Analytical, Chadds Ford, PA, USA) column with mobile phase consisting of 0.05% trifluoroacetic acid in water (mobile phase A) and 0.05% trifluoroacetic acid in acetonitrile (mobile phase B), with an isocratic elution of 15:85% (A:B) at a total flow rate of 0.25 mL/min. The LC-MS/MS system was operated at unit resolution in multiple reaction monitoring (MRM) mode, using precursor ion > product ion combination of 249.10 > 202.15 / for MO-OH-Nap and 305.10 > 215.05 / for the internal standard (IS), BA-SM-OM. The MS/MS response was linear over a concentration range of 1 to 500 ng/mL with a correlation coefficient (r ) of ≥0.987. The within- and between-batch precision (%RSD) and accuracy (%Bias) were within acceptable limits. The validated method was successfully applied to determine MO-OH-Nap metabolic stability, plasma protein binding, and bio-distribution studies of MO-OH-Nap in CD-1 mice.