Can Celecoxib Assay in Preclinical Studies Be Improved?

Abstract

Celecoxib, a cyclooxygenase-2 inhibitor (COX-2), is attracting considerable interest owing to its potential anticancer activity. The repurposing strategy of this drug, however, requires preclinical assessment involving the use of increasingly improved analytical methods. In this work, a rapid, accurate, precise, and sensitive reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed for the quantification of celecoxib in five mouse matrices (plasma, brain, spleen, liver, and kidney). Chromatographic separation was achieved within 8 min on a reversed-phase C18 column at 35 °C using a mixture of acetonitrile and 2% (v/v) acetic acid (50:50) as mobile phase, at a flow rate of 0.6 mL/min. Celecoxib and curcumin, as the internal standard, were analyzed at 425 nm and 250 nm, respectively. Linearity was observed (r2 ≥ 0.996) in the concentration ranges selected for celecoxib. Overall precision was below 14.9%, and accuracy was between −14.9% and 13.2%. The acceptance criteria specified in FDA and EMA guidelines were met. Celecoxib was reproducibly recovered (≥84%) and showed stability in all biological matrices at room temperature for 24 h. The method was then effectively applied for the quantification of celecoxib to understand in vivo biodistribution following its intraperitoneal administration in mice.