Enantiomeric Separation of Dioxopromethazine and its Stereoselective Pharmacokinetics in Rats by HPLC-MS/MS.

Abstract

Dioxopromethazine (DPZ) is a popular phenothiazine antihistamine that is widely used as a racemic drug in clinical to cure respiratory illness. In our work, a reliable, specific, and rapid enantioselective HPLC-MS/MS method has been established and fully validated for the quantification of R- and S-DPZ in rat plasma. After plasma alkalization (with 1M Na2CO3), DPZ enantiomers and diphenhydramine (IS) were extracted using ethyl acetate. Completely separation of R- and S-DPZ (Rs=2.8) within 12min was implemented on Chiralpak AGP column (100×4.0mm i.d., 5μm) employing ammonium acetate (10mM; pH 4.5) - methanol (90:10, v/v) as mobile phase. Themultiple reaction monitoring (MRM) mode was used for the detection of DPZ enantiomers and IS. The transitions of m/z 317.2→86.1 and 256.2→167.1 werechosen for monitoring DPZ enantiomers and IS, respectively. Good linearity (r2 > 0.995) was achieved for each DPZ enantiomer over the linear ranges of 1.00 - 80.00ng/mL, with the lower limit of quantitation (LLOQ) of 1.00ng/mL. The intra-day and inter-day precisions (RSDs,%) were below 12.3%, and accuracies (REs,%) were in the scope of-10.5% to 6.6%, which were within the admissible criteria. The validated bioanalytical approach was applied to the stereoselective pharmacokinetic (PK) research of DPZ in rat plasma for the first time. It was found that significant differences (p < 0.05) exist between the main PK parameters of R- and S-DPZ, indicating the pharmacokinetic behaviors of DPZ enantiomers in rats were stereoselective. The chiral inversion of the enantiomers did not occur during the assay.