A sensitive LC–MS/MS method for simultaneous determination of amygdalin and paeoniflorin in human plasma and its application

• A novel LC–MS/MS method for simultaneous determination of AD and PF in human plasma.
• Ammonium acetate was used to stabilize and promote the formation of the [M+NH4]+.
• The LLOQs of 0.6 ng/mL for AD and PF were achieved, which were the lowest reported so far.
• Pharmacokinetics of the analytes in human was reported for the first time.

A simple and sensitive HPLC–MS/MS method was developed and fully validated for the simultaneous determination of amygdalin (AD) and paeoniflorin (PF) in human plasma. For both analytes, the method exhibited high sensitivity (LLOQs of 0.6 ng/mL) by selecting the ammonium adduct ions ([M+NH4]+) as the precursor ions and good linearity over the concentration range of 0.6–2000 ng/mL with the correlation coefficients > 0.9972. The intra- and inter-day precision was lower than 10% in relation to relative standard deviation, while accuracy was within ±2.3% in terms of relative error for both analytes. The developed method was successfully applied to a pilot pharmacokinetic study of AD and PF in healthy volunteers after intravenous infusion administration of Huoxue-Tongluo lyophilized powder for injection.

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