A data-reduction algorithm for broad-spectrum LC/MS screening of psychoactive substances in urine Middleware compound identification algorithm Performance Danijela Konfortea, Difei Suna, Jan Palatyb a)LifeLabs Medical Laboratories, Toronto ON b) LifeLabs Medical Laboratories, Burnaby BC [email protected] Reporting LC/MS ~90 min to report 90 samples Situation of which ~94% are positive for at least one compound. Broad-spectrum LC/MS drug screen for ~600 urine sample per day from addiction patients. results with Goals  Automated data review using MS Excel middleware exception-based  Simple, exception-based, semi-quantitative report format

 Graded identification scheme  Use known metabolites to strengthen ID confidence middleware adds  Built-in controls for carryover, crosstalk, %hydrolysis, adulteration, unusual compounds “Found” = common Exceptioninvariably proven as a true Found

Substitution / pipetting error Method speed, consistency and Report used by Operator for reviewing results Flagging rate extremely low new process control Only 1 case to date traced to pipetting error. Cross-talk Carryover (CO) Hydrolysis HIAA Adulteration* F1 F3 tools.

Compound list

Acetylcodeine Desalkylflurazepam Lorazepam Noroxycodone Acetylfentanyl Acetylnorfentanyl Desipramine Lurasidone Oxycodone Oxymorphone Acetylmorphine Morphine Desmethylclobazam MBDB Paroxetine Alfentanil Dextromethorphan Dextrorphan MDEA PCP Aminoclonazepam DiMeMethcathinone MDMA MDA Pentazocine *) Data from our BC lab using virtually identical method. Aminoflunitrazepam Diphenhydramine MDPV Pentedrone_NE Aminonitrazepam Doxepin Desmethyldoxepin MeCathinone PMA Amoxapine Duloxetine Meperidine Normeperidine Pregabalin Hydrolysis Aripiprazole Dehydroaripiprazole Ephedrine Mephedrone Promethazine Asenapine Ethylone Methadone EDDP Pseudoephedrine 900 consecutive samples: 37 (4.1%) flagged Benzoylecgonine F1 Methamphetamine Amphetamine PVP Summary Bromazepam F3 Methaqualone Norquetiapine for decreased hydrolysis of Morphine-Glc-D3, Brompheniramine Fentanyl Norfentanyl Methedrone Methedrone_NPE Quetiapine HO-Quetiapine Buprenorphine Norbuprenorphine Fluoxetine Norfluoxetine Methedrone_NPE Risperidone HO-Risperidone Fast and accurate analysis of > 300, 000 urine samples since 2016 typically due to high levels of analyte. Bupropion HO-Bupropion Fluphenazine Methoxetamine Ritalinic_acid Buspirone Gabapentin Methylone Sertraline Butorphanol Haloperidol Methylphenidate Ritalinic_acid Sufentanil User-friendly – requires no previous toxicology or LCMS knowledge/experience Cross-talk & Carryover Butylone HIAA Mirtazapine N-Desmethylmirtazapine Tapentadol Desmethyltapentadol Carbamazepine Carbamazepine_EPX HO-Alprazolam Mitragynine Temazepam Oxazepam Robust QA – automation of error detection that cannot be done any other way Carfentanil Norcarfentanil HO-Midazolam Morphine-D3 TFMPP Carisoprodol Meprobamate HO-Triazolam Nalbuphine THCA Chlorpheniramine Hydrocodone Norhydrocodone Nalorphine Tilidine Nortilidine Flexible – user-defined rules; continual improvement since original implementation Chlorpromazine Hydromorphone Naloxone Tramadol O-Desmethyltramadol Citalopram Desmethylcitalopram Imipramine Naltrexone Naltrexol Trazodone mCPP Client Education Tool – drug-specific interpretive comments added to patient reports Clomipramine N-Desmethylclomipramine JWH-018_4-OH Nordiazepam Oxazepam Trimipramine Desmethyltrimipramine Clozapine N-Desmethylclozapine JWH-019_5-OH Norpropoxyphene UR-144_5-OH Cocaethylene Benzoylecgonine JWH-073_3-OH Nortriptyline Venlafaxine O-Desmethylvenlafaxine Codeine Codeine-6Glc Dehydronorketamine Olanzapine N-Desmethylolanzapine XLR-11_4-OH Cyclobenzaprine N-Desmethylcyclobenzaprine Ketamine Norketamine Zolpidem Zolpidem-PCA Levamisole Benzoylecgonine Zopiclone Desmethylzopiclone Lidocaine Acknowledgements Blue = metabolite Pink = manual review (due to peak shape or propensity for RT shift) We thank Dawn-Marie Murphy and Maria Cataluna from LifeLabs (ON) for assistance at the lab bench and staff from Operations and Quality and Regulatory Affairs for help with implementation.