This application note describes the extraction of 96 licit and illicit drugs of abuse from urine prior to UPLC-MS/MS analysis using EVOLUTE® HYDRO CX 96-well plates. EVOLUTE® HYDRO CX plates offer an efficient way to perform hydrolysis in the well of the extraction plate. This method provides high analyte recovery, reduced extraction time due to the elimination of a sample transfer step as well as the elimination of the column conditioning and equilibration steps, and a reduced risk for sample carryover or cross-contamination due to the elimination of the sample transfer step.

6-acetyl codeine 6-acetylmorphine (MAM) 7-Aminoclonazepam 7-Aminoflunitrazepam 96-well plate Alprazolam Amitriptyline Amphetamine Amphetamines Analgesics Application Notes Aripiprazole Atomoxetine Barbiturates Benzodiazepines Benzoylecgonine Buprenorphine Bupropion Buspirone Butalbital Carbamate hypnotic Carbamate muscle relaxant Carbamates Carbazepine Carisoprodol Chlordiazepoxide Chlorpheniramine Chlorpromazine Clinical Clomipramine Clonazepam Clonidine Clozapine Cocaethylene Cocaine Cocaines Codeine Cyclobenzaprine Desalkylflurazepam Designer stimulants Dextromethorphan Diazepam Dihydrocodeine (DHC) Drugs of Abuse Duloxetine EDDP English Ethyl Sulfate (EtS) Extrahera Fentanyl Fluoxetine Forensic Gabapentin Haloperidol Hydrocodone Hydrolysed urine Hydromorphone Hydroxyalprazolam Hydroxybupropion Hydroxymidazolam Hydroxytriazolam Imipramine Ketamine LC-MS/MS Lamotrigine Levetiracetam Lidocaine Lorazepam MDA MDEA MDMA Meperidine Meprobamate Methadone Methamphetamine Methaqualone Methcathinone Midazolam Morphine N-des-tapentadol N-desmethylclomipramine N-desmethyltapentadol Naloxone Norbuprenorphine Nordiazepam Norfentanyl Norhydrocodone Norketamine Normeperidine Noroxymorphone Norpropoxyphene Nortryptiline Opiates Opioid Opioid agonist Oxazepam Oxcarbazepine Oxycodone Oxymorphone Pain Management Pentobarbital Pharmaceuticals Phencyclidine (PCP) Phenobarbital Pregabalin Quetiapine Ritalinic acid Secobarbital Stimulants THC-COOH Tapentadol Temazepam Topiramate Tramadol Triazolam Trimipramine Urine Venlafaxine Zolpidem Zolpidem-phenyl-4-COOH mCPP z-drugs

Analysis of drug panels in urine samples can be challenging, and the trend towards larger panels including multiple drug classes compounds the issues faced during method development. This white paper examines a number of aspects of sample preparation, and their impact on the success of subsequent LC-MS/MS analysis of broad urine panels. Section 1 examines the applicability of various sample preparation techniques: supported liquid extraction, reverse phase SPE and mixed-mode SPE, to the various classes of drugs extracted. In addition, hydrolysis approaches: enzyme type and protocol used (time, temperature), are compared. Mixed-mode reverse phase/cation exchange SPE is widely used for extraction of basic drug classes from urine, but the inclusion of drugs and metabolites that exhibit ‘non-typical’ functionality within urine panels can be problematic. Section 2 examines the impact of various parameters (interference wash strength, elution solvent composition) on analyte retention, elution and extract cleanliness with particular focus on zwitterionic (gabapentin, pregabalin) and non-ionic (carisoprodol, meprobamate) drugs.

11-nor- Δ9-carboxy tetrahydrocannabinol (THC-COOH) 6-acetylmorphine (MAM) 7-Aminoclonazepam 96-well plate Alprazolam Amitriptyline Amphetamine Amphetamines Anaesthetic Anticonvulsant Application Notes Barbiturates Benzodiazepines Benzoylecgonine Buprenorphine Butalbital Cannabinoid Carbamate hypnotic Carbamate muscle relaxant Carbamates Carisoprodol Chlordiazepoxide Clinical Clonazepam Cocaine Cocaines Codeine Diazepam Dihydrocodeine (DHC) EDDP English Fentanyl Forensic Gabapentin Hallucinogen Hydrocodone Hydrolysed urine Hydromorphone Ketamine LC-MS/MS Lorazepam MDMA Meperidine Meprobamate Methadone Methamphetamine Morphine N-des-tapentadol Naloxone Norbuprenorphine Nordiazepam Norfentanyl Norhydrocodone Norketamine Normeperidine Noroxymorphone Norpropoxyphene Nortryptiline O-desmethyltramadol Opiates Opioid Opioid agonist Oxazepam Oxycodone Oxymorphone Pentobarbital Phencyclidine Phencyclidine (PCP) Phenobarbital Pregabalin Product Notes Ritalinic acid Secobarbital Stimulants Sympathomimetic amine THC metabolites THC-COOH Tapentadol Temazepam Tramadol Tricyclic Antidepressant White Papers Zolpidem Zolpidem-phenyl-4-COOH z-drugs α-OH alprazolam

This poster evaluates the performance of a novel 96-well filter plate (ISOLUTE PLD+ Protein and Phospholipid Removal Plate) for the simultaneous removal of proteins and phospholipids from serum samples prior to LC-MS/MS analysis. ASMS2014

96-well plate Acidic Amitriptyline Analgesics Atenolol Basic Bretylium Clinical English Fluoxetine Forensic Ketoprofen LC-MS/MS Metoprolol Mianserin NSAIDs Naltrexone Pharma Plasma Posters Procainamide Quinidine Ranitidine Salbutamol Serum Sulindac Tricyclic Antidepressant β-blockers

This poster presents the ISOLUTE PLD+ Protein and Phospholipid removal plate, highlighting its ease of use and excellent performance with respect to sample clean up, analyte recovery and elimination of back pressure build up in the UPLC system. IMSC 2014

96-well plate Acetaminophen Acidic Amitriptyline Atenolol Basic Bretylium Brompheniramine Clinical English Fluoxetine Forensic Ketoprofen LC-MS/MS Metoprolol Mianserin Naltrexone Neutral Pharma Pharmaceuticals Plasma Posters Procainamide Quinidine Ranitidine Salbutamol Sulindac p-toluamide

EVOLUTE® EXPRESS SPE columns and plates combine sorbent wettability with optimized SPE components, allowing better flow consistency and in many cases eliminating the need for SPE column conditioning; thus simplifying and reducing extraction processing. This poster compares the performance of various EVOLUTE EXPRESS column chemistries using 10-500 mg bed weights, in order to investigate whether sorbent conditioning and equilibration steps are truly required. ASMS 2015

Acidic Amitriptyline Atenolol Benzenesulfonic acid Bretylium Clinical Column Diethyldithiophosphate (DEDTP) Dimethylthiophosphate (DMTP) English Extrahera Forensic Hexanesulfonic acid Ketoprofen LC-MS/MS Metoprolol Mianserin Organophosphate Pesticide Over the counter pharmaceuticals Pain Management Pharma Pharmaceuticals Posters Prednisolone Procainamide Quaternary Amines Quinidine Salbutamol Sulindac Tricyclic Antidepressant Urine β-blockers

Most drugs are excreted in urine as glucuronide conjugates. Hydrolysis using a beta-glucuronidase enzyme to convert the metabolites to their “free” form for analysis increases sensitivity. Red abalone (Kura Biotech), abalone (Campbell Scientific), and recombinant (IMCSzyme) beta-glucuronidase enzymes were evaluated to determine which provided the most complete hydrolysis of glucuronide metabolites without effecting the overall recovery of non-conjugated compounds. EVOLUTE EXPRESS CX 96-well plates were used to extract hydrolysed urine samples, and the impact of th enzymes was compared. MSACL 2017, Palm Springs SOFT 2017, Boca Raton

6-acetylmorphine (MAM) 7-Aminoclonazepam 96-well plate Alprazolam Amitriptyline Amphetamine Amphetamines Analgesics Barbiturates Benzodiazepines Benzoylecgonine Butalbarbital Carbamates Carisoprodol Chlordiazepoxide Clinical Clonazepam Cocaine Cocaines Codeine Diazepam Dihydrocodeine (DHC) Drugs of Abuse EDDP English Fentanyl Forensic Gabapentin Hydrocodone Hydrolysed urine Hydromorphone Ketamine LC-MS/MS Lorazepam MDMA Meperidine Meprobamate Methadone Methamphetamine Morphine N-des-tapentadol Naloxone Norbuprenorphine Nordiazepam Norfentanyl Norhydrocodone Norketamine Normeperidine Nortryptiline Opiates Over the counter pharmaceuticals Oxazepam Oxycodone Oxymorphone Pain Management Pharmaceuticals Phencyclidine (PCP) Phenobarbital Posters Pregabalin Ritalinic acid Secobarbital Stimulants THC metabolites THC-COOH Tapentadol Zolpidem Zolpidem-phenyl-4-COOH z-drugs α-OH alprazolam

This application note describes the use of the ISOLUTE® PLD+ Protein and Phospholipid Removal Plate for clean-up of a range of acidic, basic and neutral drugs from plasma. ISOLUTE® PLD+ Protein and Phospholipid Removal Plates are suitable for clean-up of a range of analytes with widely differing functionality and polarity characteristics from plasma, giving high recoveries, good reproducibility and excellent extract cleanliness.

96-well plate Acetaminophen Acidic Amitriptyline Application Notes Atenolol Basic Bretylium Brompheniramine Clinical English Fluoxetine Forensic Ketoprofen LC-MS/MS Metoprolol Mianserin NSAIDs Naltrexone Neutral Pharma Plasma Procainamide Quinidine Ranitidine Salbutamol Sulindac Tricyclic Antidepressant p-toluamide β-blockers

This Application Note describes the extraction of a variety of basic drugs from plasma using EVOLUTE® CX mixed-mode resin-based SPE. The analyte suite includes basic drugs with wide ranging pKa and logP values. Basic Drugs, Biotage, Ion Exchange, EVOLUTE, Pharmaceuticals

Amitriptyline Application Notes Atenolol Basic Clinical Fluoxetine LC-MS/MS Metoprolol Mianserin Naltrexone Plasma Procainamide Quinidine Ranitidine Salbutamol

This application note describes the fast extraction and quantitation of 10 different antidepressant drugs in urine using ISOLUTE SLE+ 1 mL columns.

Amitriptyline Clinical Clomipramine Column Desipramine Doxepin English Forensic Imipramine Nortryptiline Paroxetine Sertraline Tricyclic Antidepressant Trimipramine Urine

This poster compares manual SPE (EVOLUTE EXPRESS CX) processing using the Biotage Pressure+ 96 with automated processing using Biotage Extrahera, using a suite of 98 drugs of abuse analytes in whole blood . Parameters such as the total time for extraction of 96 samples, and consistency of analyte recovery and RSD are explored. SOFT 2019

96-well plate Amitriptyline Anticonvulsant Antipsychotics Barbiturates Benzodiazepines Cannabinoid Clinical English Forensic Hallucinogen Ketamine LC-MS/MS Posters Stimulants Whole blood

This application note describes an automated extraction procedure from Tricyclic Antidepressants from Whole Blood Using ISOLUTE HCX Columns and the RapidTrace Workstation. TCAD, TCA, Tricyclic antidepressants, ISOLUTE,

Amitriptyline Application Notes Clinical Column Desipramine Imipramine Nortryptiline RapidTrace Tricyclic Antidepressant Whole blood

The typical range of drugs screened using this generic method are Cocaine, D3-Cocaine, Benzoylegonine-TMS, D3-Benzoylegonine-TMS, Amphetamine, D5-Amphetamine, Methamphetamine, Codeine, Morphine, Amitriptyline HCL, Nortriptyline HCL, Imipramine HCL, Desipramine HCL. These drugs represent a panel of the most significantly abused drugs as recommended by The National Institute on Drug Abuse (NIDA). drugs of abuse, drugs testing, amphetamines, BZE, Cocaine, Codeine, Ion exchange,

Amitriptyline Amphetamine Application Notes Benzoylecgonine Biological fluids Butylone Codeine Column Desipramine Forensic Imipramine Methamphetamine Morphine Nortryptiline Plasma RapidTrace Tricyclic Antidepressant Urine

This poster evaluates 3 different sample preparation approaches (ISOLUTE SLE+, EVOLUTE EXPRESS ABN, EVOLUTE EXPRESS CX) for extraction of large multi-drug urine panels. Each approach is assessed in terms of suitability for extraction of analytes with different different properties (pka, LogP etc). ASMS 2017

11-nor- Δ9-carboxy tetrahydrocannabinol (THC-COOH) 6-acetylmorphine (MAM) 7-Aminoclonazepam 96-well plate Alprazolam Amitriptyline Amphetamine Amphetamines Analgesics Barbiturates Benzodiazepines Benzoylecgonine Buprenorphine Butalbital Cannabinoid Carbamate hypnotic Carbamate muscle relaxant Carisoprodol Chlordiazepoxide Clinical Clonazepam Cocaine Cocaines Codeine Diazepam Dihydrocodeine (DHC) Drugs of Abuse EDDP English Fentanyl Forensic Gabapentin Hydrocodone Hydrolysed urine Hydromorphone Ketamine LC-MS/MS Lorazepam MDMA Meperidine Meprobamate Methadone Methamphetamine Morphine N-des-tapentadol Naloxone Norbuprenorphine Nordiazepam Norfentanyl Norhydrocodone Norketamine Normeperidine Nortryptiline Opiates Opioid Oxazepam Oxycodone Oxymorphone Pain Management Phencyclidine Phenobarbital Posters Pregabalin Ritalinic acid Secobarbital Tapentadol Temazepam Tramadol Urine Zolpidem Zolpidem-phenyl-4-COOH z-drugs α-OH alprazolam