Background and Aims The present study aimed to develop a simple and sensitive method for quantitative determination of monocrotaline (MCT) in mouse blood employing ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI/MS/MS) using rhynchophylline as an internal standard. Methods Proteins present in the blood samples were precipitated using acetonitrile. MCT was separated using a 1.7-μm ethylene bridged hybrid (BEH) C18 column (2.1 mm × 50 mm) with a gradient elution program and a constant flow rate of 0.4 mL/min. The LC mobile phase consisted of 10 mmol/L ammonium acetate (containing 0.1% formic acid) and acetonitrile. The total elution time was 4.0 min. The analytes were detected on a UPLC-ESI mass spectrometer in multiple reaction monitoring (MRM) mode and quantified. Results The new method for the determination of MCT has a satisfactory linear detection range of 1-2000 ng/mL and excellent linearity (r = 0.9971). The lower limit of quantification (LLOQ) of MCT is 1.0 ng/mL. Intra- and interassay precisions of MCT were ≤13% with an accuracy from 96.2% to 106.6%. The average recovery of the new method was >75.0%, and matrix effects were between 89.0% and 94.3%. Based on the pharmacokinetics data, the bioavailability of MCT in mice was 88.3% after oral administration. Conclusions The results suggest that the newly standardized method for quantitative determination of MCT in whole blood is fast, reliable, specific, sensitive, and suitable for pharmacokinetic studies of MCT after intravenous or intragastric administration.

The purpose of this study was to develop an ultra-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS) method to determine armepavine in mouse blood. Nuciferine was used as internal standard. Chromatographic separation was performed on a UPLC BEH (2.1 × 50 mm, 1.7 μm) column with a gradient elution of acetonitrile and 10 mmol/L ammonium acetate solution (containing 0.1% formic acid). The quantitative analysis was conducted in multiple reaction monitoring mode with m/z 314.1 → 106.9 for armepavine and m/z 296.2 → 265.1 for nuciferine. Calibration curves were linear (r > 0.995) over the concentration range 1-1000 ng/mL in mouse blood with a lowest limit of quantitation of 1 ng/mL. The intra- and inter-day precisions of armepavine in mouse were < 13.5 and 10.8%, respectively. The accuracy ranged between 86.8 and 103.3%. Meanwhile, the average recovery was >70.7% and the matrix effect was within the range 109.5-113.7%. All of the obtained data confirmed the satisfactory sensitivity and selectivity of the developed method which was then successfully applied to evaluate the pharmacokinetic behavior of armepavine in mouse for the first time. The bioavailability of armepavine in mouse was calculated to be 11.3%.

In this work, a selective and sensitive ultra-performance liquid chromatography tandem mass spectrometry method was established and validated for determination of corypalmine in mouse blood after oral or intravenous administration. A UPLC BEH C column was used to separate corypalmine and berberrubine (internal standard) at 40°C. The mobile phase was composed of acetonitrile and 10 mmol/L ammonium acetate (containing 0.1% formic acid) at a flow rate of 0.4 mL/min, and the total run time was 4.0 min. Electrospray ionization in positive ion mode was applied; target fragment ions m/z 342.2 → 178.0 for corypalmine and m/z 322.1 → 307.0 for berberrubine were identified with multiple reaction monitoring mode. The linear range was 1-1000 ng/mL (r > 0.995) and the lower limit of quantification for corypalmine in plasma was 1.0 ng/mL. The intra- and inter-day precisions were both <14%. The range of accuracy in this method was 97.5-109.0%. Mean recovery was >69.6%, and the matrix effect was 96.8-107.6%. Based on its high sensitivity, specificity and reliability, this method was successfully applied to study the pharmacokinetic parameters of corypalmine in mouse by oral and intravenous administration, and finally, the bioavailability of corypalmine was identified at 4.6%.