Page 71 - Clinical-Medical Solution for Clinical Chemistry

Quantitative Multi Target Screening (MTS) using liquid
chromatography-tandem mass spectrometry with MS/MS library
based identi cation for forensic toxicology

LC-MS/MS method set up for simultaneous full scan and MRM data acquisition with polarity switching
Type Event Polarity Name | m/z Time (0-13mins)
MRM 5 + Target | 7-aminonitrazepam 252.10>121.10
Product Ion Scan 6 + > CE: -10, 30.00-1000.00
Product Ion Scan 7 + > CE: -35, 30.00-1000.00
Product Ion Scan 8 + > CE: -50, 30.00-1000.00
MRM 9 + Target | 7-aminoclonazepam 286.05>121.10
Product Ion Scan 10 + > CE: -10, 30.00-1000.00
Product Ion Scan 11 + > CE: -35, 30.00-1000.00
Product Ion Scan 12 + > CE: -50, 30.00-1000.00
MRM 13 + Target | 3-Hydroxybromazepam 322.00>287.00
Product Ion Scan 14 + > CE: -10, 30.00-1000.00
Product Ion Scan 15 + > CE: -35, 30.00-1000.00

Spectral Library >1200 compounds

Each library spectrum was acquired by authentic standard and peak area measured to enable reference ion-ratio
ow injection at collision energies 10-60V. Compounds calculation. RT analysis included internal standard
that ionised ef ciently with more than one adduct state compounds for relative RT calculation. Compound
were saved resulting in 1476 Library entries from 1207 information was populated including: CAS number,
compounds (1278 positive mode, 229 negative mode). formula, synonyms, compound class/properties,
Spectral Library information was registered for CE 10, 35 ChemSpider URL and ID number, mol le, InChI and
and 55V. Optimised MRM transitions were determined InChIKey.
for all compounds with chromatographic retention time

Toxicological Screening

Compounds were spiked into whole blood, prepared in were prepared, the rst measuring benzodiazepines (36
triplicate at a concentration range 1-1000 µg/L compounds), the second measuring antiepileptics,
(calibration curves typically ranged 5-500 µg/L). Quality antipsychotics, barbiturates and cannabinoids (35
control samples were prepared (5x) at three compounds).
concentrations (20, 100, 500 µg/L). Two MTS methods

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