Simultaneous determination of carboprost methylate and its active metabolite carboprost in dog plasma by liquid chromatography–tandem mass spectrometry with positive/negative ion-switching electrospray ionization and its application to a pharmacokinetic s

• First simultaneously quantitate carboprost methylate and carboprost in dog plasma.
• An effective approach to select esterase inhibitors to stabilize carboprost methylate in dog blood was developed.
• Indomethacin was added to plasma to inhibit prostaglandins synthesis after sampling.
• Limit of detection (LOD) values were 10 and 20 pg/mL for carboprost and carboprost methylate.
• The short running time meets the requirement of high throughput analysis.
• This study reveals the pharmacokinetic profile of carboprost methylate in beagle dogs.

A liquid chromatography–tandem mass spectrometric (LC–MS/MS) method using positive/negative electrospray ionization (ESI) switching for the simultaneous quantitation of carboprost methylate and carboprost in dog plasma has been developed and validated. After screening, the esterase inhibitor, dichlorvos was added to the whole blood at a ratio of 1:99 (v/v) to stabilize carboprost methylate during blood collection, sample storage and LLE. Indomethacin was added to plasma to inhibit prostaglandins synthesis after sampling. After liquid–liquid extraction of 500 μL plasma with ethyl ether-dichloromethane (75:25, v/v), analytes and internal standard (IS), alprostadil-d4, were chromatographed on a CAPCELL PAK Phenyl column (150 × 2.0 mm, 5 μm) using acetonitrile-5 mM ammonium acetate as mobile phase. Carboprost methylate was detected by positive ion electrospray ionization followed by multiple reaction monitoring (MRM) of the transition at m/z 400.5 → 329.3; the carboprost and IS were detected by negative ion electrospray ionization followed by MRM of the transitions at m/z 367.2 → 323.2, and 357.1 → 321.2, respectively. The method was linear for both analytes in the concentration range 0.05–30 ng/mL with intra- and inter-day precisions (as relative standard deviation) of ≤6.75% and accuracy (as relative error) of ≤7.21% and limit of detection (LOD) values were 10 and 20 pg/mL, respectively. The method was successfully applied to a pharmacokinetic study of the analytes in beagle dogs after intravaginal administration of a suppository containing 0.5 mg carboprost methylate.