Simultaneous Screening and Quantification of Basic, Neutral and Acidic Drugs in Blood Using UPLC-QTOF-MS

Abstract

An analytical method using ultra performance liquid chromatography (UPLC) quadrupole time-of-flight mass spectrometry (QTOF-MS) was developed and validated for the targeted toxicological screening and quantification of commonly used pharmaceuticals and drugs of abuse in postmortem blood using 100 µL sample. It screens for more than 185 drugs and metabolites and quantifies more than 90 drugs. The selected compounds include classes of pharmaceuticals and drugs of abuse such as: antidepressants, antipsychotics, analgesics (including narcotic analgesics), anti-inflammatory drugs, benzodiazepines, beta-blockers, amphetamines, new psychoactive substances (NPS), cocaine and metabolites. Compounds were extracted into acetonitrile using a salting-out assisted liquid-liquid extraction (SALLE) procedure. The extracts were analyzed using a Waters ACQUITY UPLC coupled with a XEVO QTOF mass spectrometer. Separation of the analytes was achieved by gradient elution using Waters ACQUITY HSS C18 column (2.1 mm x 150 mm, 1.8 μm). The mass spectrometer was operated in both positive and negative electrospray ionization modes. The high-resolution mass spectrometry (HRMS) data was acquired using a patented Waters MSE acquisition mode which collected low and high energy spectra alternatively during the same acquisition. Positive identification of target analytes was based on accurate mass measurements of the molecular ion, product ion, peak area ratio and retention times. Calibration curves were linear over the concentration range 0.05-2 mg/L for basic and neutral analytes and 0.1-6 mg/L for acidic analytes with the correlation coefficients (r2) > 0.96 for most analytes. The limits of detection (LOD) were between 0.001-0.05 mg/L for all analytes. Good recoveries were achieved ranging from 80% to 100% for most analytes using the SALLE method. The method was validated for sensitivity, selectivity, accuracy, precision, stability, carryover and matrix effects. The developed method was tested on a number of authentic forensic samples producing consistent results that correlated with results obtained from other validated methods.

Polydopamine coated hypodermic needles as a microextraction device for the determination of tricyclic antidepressants in oral fluid by direct infusion MS/MS

“…Table 2 compares the sensitivity of the proposed method with other alternatives reported for the determination of tricyclic antidepressants in biouids, including saliva, urine, plasma and blood. 1,6,7,9,12,16,[49][50][51][52][53] Our method provides a wider linear range, allowing the determination of the target compounds at the therapeutic level. In the oral uid applications, the sensitivity of the new method is intermediate and only surpassed by the method based on high-resolution MS, although the latter provided a narrower linearity range.…”

Medikamente und Fahrsicherheit

“…Mit hochauflösender Massenspektrometrie kann das Spektrum potenziell erfassbarer Substanzen nach einfacher Probenvorbereitung, z. B. einer Proteinfällung, erheblich erweitert werden [19,20]. Literatur…”

Retrospective analysis for valproate screening targets with liquid chromatography–high resolution mass spectrometry with positive electrospray ionization: An omics‐based approach

“…13 Valproate (valproic acid; VPA) is a fatty acid and frequently used anti-epileptic drug, and is therefore generally included in STA in forensic toxicology. 14,15,[17][18][19] Metabolites of VPA have been used for confirmation, 14,17,18 and adducts of VPA with common mobile phase additives enable use of real mass transitions with increased sensitivity. 16 Analysis of VPA using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and LC-HRMS is often performed in negative electrospray ionization mode (ESI − ) with pseudo-multiplereaction monitoring (MRM) transition of mass-to-charge (m/z) 143 → 143, corresponding to the [M-H] − precursor.…”

“…16 Analysis of VPA using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and LC-HRMS is often performed in negative electrospray ionization mode (ESI − ) with pseudo-multiplereaction monitoring (MRM) transition of mass-to-charge (m/z) 143 → 143, corresponding to the [M-H] − precursor. 14,15,[17][18][19] Metabolites of VPA have been used for confirmation, 14,17,18 and adducts of VPA with common mobile phase additives enable use of real mass transitions with increased sensitivity. 20 However, LC-HRMS screening in toxicology is often performed solely using positive electrospray ionization (ESI + ).…”