Compensation of Matrix Effects in a Standard Addition Method for Metformin in Postmortem Blood Using Liquid Chromatography–Electrospray–Tandem Mass Spectrometry

This work describes a procedure to evaluate matrix effects in a combined dilution and standard addition method (SAM) using liquid chromatography–electrospray–tandem mass spectrometry. The method was validated and applied to an analysis of metformin in postmortem blood samples. The analytical method included protein precipitation with methanol, followed by liquid chromatographic separation of metformin on Gemini NX-C18 reversed-phase column using a gradient consisting of methanol and ammonium acetate at pH 3.2. The mass spectrometric analysis was performed with a quadrupole-linear ion trap mass spectrometer equipped with a turbo ion spray interface in a positive ion mode using selected reaction monitoring. Quantitation was performed based on an SAM. Validation for metformin revealed a practical limit of quantification of 0.1 mg/L, a linear range from 0.1 to 3.0 mg/L, average precision 10%, accuracy (bias) 9% and reproducibility 10%. Combined matrix effects were evaluated by k-values (slopes) of calibration plots, postextraction addition approach and a comparison of within- and between-sample precision (relative standard deviation). It was demonstrated that the method contained matrix effects which were fully compensated for using dilution and the SAM.