We developed and validated an UHPLC-MS/MS method for the quantification of phosphatidylethanol (PEth) in dried blood spots (DBS). Sample preparation was a liquid extraction and chromatographic separation was performed in an Acquity C8 column. The mobile phases were 4 mM ammonium acetate in water (A) and acetonitrile (B) at 24:76 (v/v). The total analytical run time was 6 minutes, with retention time of 3.36 min for PEth and 4.13 min for phosphatidylpropanol. The method was linear from 10 to 3000 ng mL-1, specific, with, precise and accurate. The analyte was stable in DBS stored from -20 to 45 °C for 21 days. Matrix effect was compensated with the internal standard (-5.21% to + 6.09%). The method was applied in the evaluation of alcohol consumption in DBS from 25 chemical dependents. The PEth concentrations ranged from 14.5 to 2380.8 ng mL-1, with significant correlation with the self-report alcohol consumption AUDIT scores (r=0.41).

We developed a UPLC-MS/MS method for the quantification of the ethanol metabolites ethyl-glucuronide (EtG) and ethyl-sulfate (EtS) in blood samples. Sample preparation was a protein precipitation with acetonitrile 0.1% formic acid containing internal standards (IS) 0.5 µg mL-1 EtG-D5 and EtS-D5. Chromatographic separation was performed in a fluoro-phenyl column, mobile phases were 0.1% formic acid in water (A) and acetonitrile (B) in gradient elution from 5% B to 70% B, with 10 min run time. Ionization was performed with electrospray source at 550 °C in negative mode, quantification mass transitions of m/z EtG 221- 75, EtS 125- 80.1, EtS-D5 130- 79.9, and EtG-D5 226- 75.1. The method was linear from 0.1 to 18 µg mL-1 for EtG and 0.02 to 6 µg mL-1 for EtS, specific, with no interfering peaks during the blank samples analyses. The method was precise and accurate, the matrix effect was compensated with the IS (-12% a + 6.1%). EtG and EtS were analyzed in 28 blood samples from trauma patients with positive alcoholaemia. The concentrations of EtG and EtS ranged from 0.25 to 14.11 µg mL-1 and 0.13 to 5.53 µg mL-1, with high correlation with ethanol concentrations, r=0.739 and r=0.817 (p <0,001), respectively.

A highly sensitive ultra-performance liquid chromatography tandem mass spectrometry method for the determination of Docetaxel and metabolites (M1+M3) in human plasma samples was developed and validated. The method employs a liquid-liquid extraction and a reversed phase separation Acquity® C18 column BEH C18 (150 x 2.1 mm, 1.7 µm). Mobile phase was a mixture of formic acid 0.1% (eluent A) and acetonitrile plus 0.1% formic acid (eluent B) in isocratic mode. Total analytical run time was 7 min. The interval of method linearity interval was 50 to 3000 ng mL-1 for docetaxel and 5.0 to 300 ng mL-1, for M1+M3 metabolites. Intra-assay and inter-assay precision was coefficient of variation 2.5-15.7% and 5.9-12.6%, respectively and accuracy was 89.0-114.0%. The method was successfully applied in the measurement of docetaxel plasma levels and area under the curve calculation in 31 cancer patients, demonstrating adequate analytical performance for the clinical application in the therapeutic drug monitoring and pharmacokinetics studies.