Academic Journal

Prediction and Validation of Phase II Glucuronide Conjugates in Urine Using Combined Non-Targeted and Targeted LC–HRMS/MS Workflows and Their Validation for over 200 Drugs

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Τίτλος: Prediction and Validation of Phase II Glucuronide Conjugates in Urine Using Combined Non-Targeted and Targeted LC–HRMS/MS Workflows and Their Validation for over 200 Drugs
Συγγραφείς: Camila Bardy, Luis Manuel Menéndez-Quintanal, Gemma Montalvo, Carmen García-Ruiz, Begoña Bravo Serrano, Jose Manuel Matey
Πηγή: Analytica ; Volume 7 ; Issue 1 ; Pages: 18
Στοιχεία εκδότη: Multidisciplinary Digital Publishing Institute
Έτος έκδοσης: 2026
Συλλογή: MDPI Open Access Publishing
Θεματικοί όροι: high-resolution mass spectrometry (HRMS), orbitrap, metabolism, spectral libraries, exact mass list, compound discoverer, Structured Query Language (SQL), glucuronide, neutral loss, predictive
Περιγραφή: High-resolution mass spectrometry (HRMS) enables non-targeted detection of drugs and metabolites in complex matrices. Phase II metabolites—especially glucuronides—are often the only detectable biomarkers in late or postmortem samples but are underrepresented in commercial libraries. This work pursued the prediction of phase II-glucuronide conjugates in diluted urine samples by non-targeted/targeted LC-HRMS workflows. A simply “dilute-and-shoot” qualitative UHPLC-HRMS/MS method (Q Exactive HF, ddMS2) was integrated with Compound Discoverer® software for data processing. The workflow incorporated predictive strategies such as exact mass suspect lists, Structured Query Language (SQL)-based filters, compound-class and diagnostic neutral-loss rules (including the characteristic loss of 176.0321 Da for glucuronides) and MS/MS confirmation using both in-house and public spectral libraries. An additional part of the application’s performance assessment involved its validation for diluted urine sample. A qualitative validated method for more than two hundred drugs in urine samples was performed, including the method’s selectivity/specificity, limit of identification, matrix effects, and potential carryover. Most analytes fulfilled the qualitative acceptance criteria, with more than 60% successfully identified at a concentration of at least 2.5 ng/mL. Matrix effects were within acceptable limits for most compounds, and no severe ion suppression was observed. A non-targeted workflow was applied to real forensic samples (n = 16), allowing a reduction of approximately 66,800 detected features to 225 glucuronide candidates, while a targeted workflow based on exact mass lists yielded 31 high-confidence identifications. Characteristic neutral losses and diagnostic fragment ions led to the tentative identification of some glucuronide phase II metabolites such as mirtazapine–glucuronide, morphine-6–glucuronide, and glucuronide conjugates of benzodiazepines and synthetic opioids. In conclusion, the integration of biotransformation ...
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Γλώσσα: English
Relation: https://dx.doi.org/10.3390/analytica7010018
DOI: 10.3390/analytica7010018
Διαθεσιμότητα: https://doi.org/10.3390/analytica7010018
Rights: https://creativecommons.org/licenses/by/4.0/
Αριθμός Καταχώρησης: edsbas.B7C3A94C
Βάση Δεδομένων: BASE