LAUSR

The paper presents a method for the determination of methadone, EDDP, and EMDP in postmortem biological materials using liquid‐liquid extraction with ethyl acetate (pH9) and UHPLC‐MS/MS technique. Methadone‐d9 and EDDP‐d3 were used as the internal standards. The method validation results for blood and urine were as follows: linearity: 0.5–1000 ng/ml; R2 > 0.9993 for methadone, EDDP and R2 > 0.9944 for EMDP. Intra‐ and inter‐day precision: 0.1%–7.5% and 0.3%–8.6%, respectively; intra‐ and inter‐day accuracy: −11.8% to 13.9% and −9.3 to 14.8%, respectively; recovery: 91.5%–123.0%; matrix effect: 83.5%–123.9%. This study also describes 18 postmortem cases, where methadone concentrations ranged 2.3–1180 ng/ml in blood (n = 17), from 11.0 to >10,000 ng/ml in urine (n = 13) and 135.2–409.0 in vitreous humor (VH, n = 3). EDDP concentrations ranged from not detectable to 180 ng/mL in blood, from 42.4 to >10,000 ng/ml in urine and 18.3–36.5 in VH. EMDP concentrations were found in four cases in blood from below LLOQ to 1.8 ng/ml and in seven cases in urine, ranged 2.1–243.0 ng/ml. EMDP was not detected in VH samples. The EDDP/methadone ratios and blood/urine ratios for methadone and EDDP in EMDP‐positive and negative cases were performed. The paper presents mass spectra of other methadone metabolites, than EDDP and EMDP (ring hydroxylated methadone, ring hydroxylated EDDP, ring hydroxylated EMDP, methadol, and DDP). Simultaneous determination of methadone and its metabolites in order to unequivocally interpret the results of toxicological tests seems to be useful in cases related to prescription/illicit use of methadone.