Simultaneous determination of acetaminophen and dihydrocodeine in human plasma by UPLC–MS/MS: Its pharmacokinetic application

Abstract An ultra performance liquid chromatography tandem mass spectrometry (UPLC–MS/MS) method was developed and validated to determine acetaminophen (AAP) and dihydrocodeine (DHC) in human plasma simultaneously. Plasma samples were prepared using protein precipitation with acetonitrile, the two analytes and the internal standard midazolam were separated on an Acquity UPLC BEH C18 column and mass spectrometric analysis was performed using a QTrap5500 mass spectrometer coupled with an electro-spray ionization (ESI) source in the positive ion mode. The MRM transitions of m / z 151.2 → 110.0 and m / z 302.3 → 199.2 were used to quantify for AAP and DHC, respectively. The linearity of this method was found to be within the concentration range of 50–10000 ng/mL for AAP, and 1–100 ng/mL for DHC in human plasma, respectively. The lower limit of quantification (LLOQ) was 50 ng/mL and 1 ng/mL for AAP and DHC in human plasma, respectively. The relative standard deviations (RSD) of intra and inter precision were less than 10% for both AAP and DHC. The analysis time of per sample was 1.0 min. The developed and validated method was successfully applied to a pharmacokinetic study of AAP (500 mg) with DHC (20 mg) capsule in Chinese healthy volunteers ( N  = 20).