The ability to extract a broad range of different drugs from a biological matrix allows for the expedited analysis of a patient sample using LC-MS/MS. Typically small molecules are extracted from matrices like urine based on their polarities. A fast and reliable sample preparation method that could be implemented to extract drugs of different polarities from urine could be used as a screening tool to quickly identify the presence of illicit drugs in patient samples using LC-MS-MS. This poster demonstrates the utility of supported liquid extraction for the extraction of over 30 different acidic, basic and neutral drugs in urine prior to LC-MS/MS. MSACL 2016

11-hydroxy- Δ9-tetrahydrocannabinol (THC-OH) 11-nor- Δ9-carboxy tetrahydrocannabinol (THC-COOH) 6-acetyl codeine 96-well plate Acidic Alprazolam Amphetamine Barbiturates Basic Benzodiazepines Benzoylecgonine Buprenorphine Butalbital Clinical Clonazepam Codeine Dextromethorphan Diazepam Drugs of Abuse EDDP English Fentanyl Flunitrazepam Forensic Hydrocodone Hydrolysed urine Hydromorphone LC-MS/MS MDEA Morphine Naltrexone Neutral Nitrazepam Norbuprenorphine Norcodeine Norfentanyl Opiates Oxazepam Oxycodone Oxymorphone Pain Management Pentazocine Pentobarbital Phenobarbital Posters Propoxyphene Secobarbital THC THC metabolites THC-COOH Temazepam Δ9-tetrahydrocannabinol (THC)

This application note describes the extraction of acidic, basic, and neutral drugs from urine for screening purposes using ISOLUTE® SLE+ supported liquid extraction plates prior to LC-MS/MS analysis.

11-hydroxy- Δ9-tetrahydrocannabinol (THC-OH) 11-nor- Δ9-carboxy tetrahydrocannabinol (THC-COOH) 6-acetyl codeine 96-well plate Acidic Alprazolam Amphetamine Application Notes Barbiturates Basic Benzodiazepines Benzoylecgonine Buprenorphine Butalbital Clinical Clonazepam Codeine Dextromethorphan Diazepam Drugs of Abuse EDDP English Fentanyl Flunitrazepam Forensic Hydrocodone Hydrolysed urine Hydromorphone LC-MS/MS MDEA Naltrexone Neutral Nitrazepam Norbuprenorphine Norcodeine Norfentanyl Normeperidine Opiates Oxazepam Oxycodone Oxymorphone Pain Management Pentazocine Pentobarbital Phenobarbital Propoxyphene Secobarbital THC metabolites Temazepam Urine Δ9-tetrahydrocannabinol (THC)

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

Over the past decade, the need for non-invasive drug screening that that precludes sample adulteration has become attractive. As a result, detection using oral fluid devices for Drugs of Abuse (DOA) has come to the vanguard of the scientific community. The use of Supported Liquid Extraction (ISOLUTE® SLE+) prior to LC/MS or GC/MS can improve sample cleanliness without forfeiting sample detection within a diverse panel of DOAs. Here, we demonstrate the effects of altering elution solvent polarity and pH for sample pretreatment upon the simultaneous recovery of 34 compounds comprised of opioids, benzodiazepines, and stimulants to directly measure the effects of the oral fluid buffer, OraSure™, upon extraction and signal intensity at presumed LOQs. ASMS 2017

6-acetylmorphine (MAM) 7-Aminoclonazepam Alprazolam Amphetamine Benzodiazepines Benzoylecgonine Buprenorphine Clinical Clonazepam Cocaine Cocaines Codeine Column Cotinine Diazepam EDDP English Forensic Hydrocodone Hydromorphone LC-MS/MS Lorazepam MDA MDEA MDMA Meperidine Methadone Methamphetamine Midazolam Morphine Norbuprenorphine Nordiazepam Opiates Opioid Oral fluid Oxazepam Phencyclidine Phencyclidine (PCP) Posters Stimulants Temazepam Triazolam α-OH Triazolam α-OH alprazolam

This poster shows an extraction protocol for 12 common drugs of abuse (DOA) to be detected in breast milk using mixed-mode polymeric cation exchange solid phase extraction (SPE). Using the combination of reliable automation (on Biotage Extrahera) and SPE sample preparation techniques, a method was developed demonstrating the precision, accuracy, linearity, and sensitivity necessary for a robust quantitative workflow. MSACL NA 2019

96-well plate Amphetamine Amphetamines Barbiturates Benzodiazepines Benzoylecgonine Breast milk Caffeine Clinical Cocaines Cotinine Drugs of Abuse English Forensic LC-MS/MS Methadone Methamphetamine Morphine Opiates Opioid Oxazepam Oxycodone Phencyclidine (PCP) Posters Secobarbital THC metabolites THC-COOH

This poster compares two different procedures for quantification of drugs of abuse from postmortem brain tissue.

7-Aminoflunitrazepam Amphetamine Benzoylecgonine Brain Cocaine Codeine Column Diazepam Drugs of Abuse English Forensic GC-MS LC-MS/MS Methadone Morphine Posters

Evaporative crosstalk is well-to-well cross contamination during extract evaporation after SLE or SPE extraction in a 96 well plate. It is often observed with volatile analytes like methamphetamine and amphetamine, but it can also occur with other drugs and metabolites when urine samples have very high analyte concentrations. ASMS, 2020.

Amphetamine Benzoylecgonine Drugs of Abuse Forensic Hydromorphone MDA MDEA MDMA Methamphetamine Morphine Posters Urine

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

Miniaturisation within various fields of analytical chemistry is not a new phenomenon. Early phase small animal drug trials have largely been the driver due to limited sample sizes available. However, this trend is gaining popularity in forensic/clinical toxicology with respect to alleviating patient discomfort, particularly in paediatrics. Increased LC-MS/MS sensitivity has reignited this focus area. This poster will evaluate a novel low-volume 96-well solid phase extraction (SPE) format and potential application to forensic and clinical toxicology. ASMS, 2020.

6-MAM Alprazolam Amphetamine Buprenorphine Clinical Clonazepam Cocaine Codeine Diazepam Drugs of Abuse EDDP Estazolam Fentanyl Flunitrazepam Flurazepam Forensic Hydrocodone Hydromorphone Ketamine Lorazepam Lysergic acid diethylamide (LSD) MDA MDEA MDMA Mephedrone Methadone Methamphetamine Midazolam Mikro plate Morphine Nitrazepam Norbuprenorphine Nordiazepam Norfentanyl Norketamine Oxazepam Oxycodone Phencyclidine (PCP) Plasma Posters Temazepam Triazolam Zolpidem Zopiclone

In postmortem cases, where drugs or pesticides have been used for their poisonous properties, traditional matrices such as urine and whole blood may be inappropriate for qualitative and quantitative analysis. As the site of metabolism for most drugs and toxins, the liver may provide more insight to cause of death than other bodily fluids. This poster describes the use of ISOLUTE SLE+ supported liquid extraction columns to extract a range of drug and pesticide classes form homogenised liver using a simple, streamlined workflow. SOFT 2016

Alprazolam Amphetamine Atrazine Bead Ruptor Bendiocarb Biphenthrin Butabarbital Chlorothalonil Clinical Clonazepam Cocaine Column Diazepam Drugs of Abuse English Estazolam Forensic GC-MS Ketamine Liver MDMA Malathion Methadone Methamphetamine Midazolam Nitrazepam Pesticides Phenobarbital Posters Propanil Secobarbital THC Triazolam

This poster demonstrates the extraction of a range of drugs of abuse from oral fluid, collected with common collection devices, prior to UPLC-MS/MS analysis. The target analyte list includes benzodiazepines, z drugs, amphetamines, cathinones, opiates, cocaine, buprenorphine, PCP, THC-COOH, fentanyl and ketamine. SOFT 2014

11-nor- Δ9-carboxy tetrahydrocannabinol (THC-COOH) 2-OH-Et-Flurazepam 6-acetylmorphine (MAM) 7-Aminoclonazepam 7-Aminoflunitrazepam Alprazolam Amphetamine Amphetamines Benzodiazepines Benzoylecgonine Bromazepam Buprenorphine Cathinones Clonazepam Cocaine Codeine Column Diazepam Dihydrocodeine (DHC) EDDP English Estazolam Fentanyl Flunitrazepam Flurazepam Forensic Hydrocodone Hydromorphone Ketamine LC-MS/MS Lorazepam MDA MDEA MDMA Mephedrone Methadone Methamphetamine Midazolam Morphine Nitrazepam Norbuprenorphine Nordiazepam Norfentanyl Norketamine Opiates Opiates Oral fluid Oxazepam Oxycodone Oxymorphone Phencyclidine (PCP) Posters Saliva Temazepam Triazolam Zaleplone Zolpidem Zopiclone z-drugs α-OH Triazolam α-OH alprazolam

This poster demonstrates the extraction of a range of drugs of abuse from oral fluid collection devices using supported liquid extraction suitable for UPLC-MS/MS analysis. Unlike some sample preparation techniques, SLE allows for the simultaneous extraction of cross-functional analytes in a single extraction protocol without forfeiting extract cleanliness. The target analyte list includes benzodiazepines, z drugs, amphetamines, cathinones, opiates, cocaine, buprenorphine, PCP, THC-COOH, fentanyl and ketamine. TIAFT 2015

2-OH-Et-Flurazepam 6-acetylmorphine (MAM) 7-Aminoclonazepam 7-Aminoflunitrazepam Alprazolam Amphetamine Amphetamines Benzodiazepines Benzoylecgonine Bromazepam Buprenorphine Cathinones Clonazepam Cocaine Cocaines Codeine Column Diazepam Dihydrocodeine (DHC) Drugs of Abuse EDDP English Estazolam Fentanyl Flunitrazepam Flurazepam Forensic Hydrocodone Hydromorphone LC-MS/MS Lorazepam MDA MDMA Mephedrone Methadone Midazolam Morphine Norbuprenorphine Nordiazepam Norfentanyl Norketamine Opiates Oral fluid Oxazepam Oxycodone Oxymorphone Pain Management Phencyclidine (PCP) Posters Temazepam Triazolam Zaleplone Zolpidem Zopiclone α-OH Triazolam α-OH alprazolam

This poster describes the extraction of a range of drugs of abuse (including barbiturates, THC and metabolites, benzodiazepines, z drugs, amphetamines,cathinones, opiates, cocaine, buprenorphine, PCP, fentanyl and ketamine) from oral fluid using supported liquid extraction (ISOLUTE SLE+) columns prior to GC-MS and LC-MS/MS analysis. ASMS 2014

11-nor- Δ9-carboxy tetrahydrocannabinol (THC-COOH) 6-acetylmorphine (MAM) 7-Aminoclonazepam 7-Aminoflunitrazepam Alprazolam Amphetamine Amphetamines Barbiturates Benzodiazepines Benzoylecgonine Bromazepam Buprenorphine Clonazepam Cocaine Codeine Column Designer stimulants Diazepam Dihydrocodeine (DHC) EDDP English Estazolam Fentanyl Flunitrazepam Flurazepam Forensic GC-MS Hydrocodone Hydromorphone Hydroxyalprazolam Hydroxytriazolam Ketamine LC-MS/MS Lorazepam MDA MDEA MDMA Mephedrone Methadone Methamphetamine Midazolam Morphine Nitrazepam Norbuprenorphine Nordiazepam Norfentanyl Norketamine Oral fluid Oxazepam Oxycodone Oxymorphone Phencyclidine (PCP) Posters THC metabolites Temazepam Triazolam Zaleplone Zolpidem Zopiclone

This poster demonstrates a fast, reliable protocol to extract multiple drug of abuse panels from whole blood using a common supported liquid extraction methodology. This benefits laboratory workflow where multiple assays are run each day, saving both worker hours and consumable costs. SOFT 2015 ASMS 2016 TIAFT 2016

2-Hydroxyethylflurazepam 2C-B 5-APB 6-APB 6-acetylmorphine (MAM) 7-Aminoclonazepam 7-Aminoflunitrazepam Alprazolam Amobarbital Amphetamine Anhydroecgonine Methyl Ester AEME BZP Benzoylecgonine Bromazepam Butabarbital Butalbarbital Butylone Clinical Clonazepam Cocaethylene Cocaine Cocaines Codeine Column Diazepam Dihydrocodeine (DHC) Drugs of Abuse Ecgonine Methyl Ester EME English Estazolam Ethylone Flunitrazepam Flurazepam Forensic GC-MS Hexobarbital Hydrocodone Hydromorphone Lorazepam MDA MDEA MDMA MDPV Mephedrone Methamphetamine Methcathinone Methedrone Methylone Midazolam Morphine Naphyrone Nitrazepam Nordiazepam Oxazepam Oxycodone Oxymorphone Pentobarbital Phenobarbital Posters Secobarbital TFMPP Temazepam Triazolam Whole blood mCPP pMMA α-OH Triazolam α-OH alprazolam

This poster compares the performance of manual processing to a novel automated sample preparation system prior to GC/MS or LC-MS/MS analysis in forensic toxicology applications. Emphasis is placed on the potential for 96-well cross contamination and strategies for its elimination. TIAFT 2016 SOFT 2016

2-Hydroxyethylflurazepam 2-OH-Et-Flurazepam 6-acetylmorphine (MAM) 7-Aminoclonazepam 7-Aminoflunitrazepam 96-well plate Alprazolam Amphetamine Amphetamines Benzodiazepines Bromazepam Buprenorphine Clinical Clonazepam Codeine Column Diazepam Dihydrocodeine (DHC) Drugs of Abuse EDDP English Estazolam Extrahera Fentanyl Flunitrazepam Flurazepam Forensic GC-MS Hydrocodone Hydromorphone Ketamine LC-MS/MS Lorazepam MDA MDEA MDMA Mephedrone Methadone Methamphetamine Midazolam Morphine Nitrazepam Norbuprenorphine Nordiazepam Norfentanyl Norketamine Opiates Opiates Oxazepam Oxycodone Oxymorphone Phencyclidine (PCP) Posters Temazepam Triazolam Urine Whole blood Zaleplone Zolpidem Zopiclone z-drugs α-OH Triazolam α-OH alprazolam

Amphetamine, methamphetamine and Ecstasy continue to be widely abused in many parts of the world. Urine analysis is the most popular approach to determining drug intake. This poster examines various sample preparation approaches in the analysis of amphetamines prior to gas chromatography-mass spectrometry. Comparisons were performed between silica-based and polymer-based SPE (ISOLUTE HCX and EVOLUTE EXPRESS CX) as well as supported liquid extraction (SLE, ISOLUTE SLE+). TIAFT 2019

Amphetamine Amphetamines Clinical English Forensic MDA MDEA MDMA Methamphetamine Posters Urine

This poster describes an improved workflow for the analysis of drugs of abuse from hair. TIAFT 2018, Ghent, Belgium

2-OH-Et-Flurazepam 6-acetylmorphine (MAM) 7-Aminoclonazepam 7-Aminoflunitrazepam 96-well plate Alprazolam Amphetamine Amphetamines Benzodiazepines Benzoylecgonine Bromazepam Buprenorphine Clonazepam Cocaine Codeine Column Diazepam Dihydrocodeine (DHC) EDDP English Estazolam Extrahera Fentanyl Flunitrazepam Flurazepam Forensic Hair Hydrocodone Hydromorphone Ketamine LC-MS/MS Lorazepam Lysera Lysergic acid diethylamide (LSD) MDA MDEA MDMA Methadone Methamphetamine Midazolam Morphine Nitrazepam Norbuprenorphine Nordiazepam Norfentanyl Norketamine Opiates Opioid Oxazepam Oxycodone Oxymorphone PCP Posters Temazepam Triazolam Zaleplone Zolpidem Zopiclone z-drugs α-OH Triazolam α-OH alprazolam

Hair analysis is growing in popularity due to the non-invasive nature of the sample collection. Although not used as routinely as other matrices such as blood or urine it does have advantages in that the matrix can indicate prolonged drug exposure. NCFM 2020, Reykjavik, Iceland

2-OH-Et-Flurazepam 6-MAM 7-amino-flunitrazepam 96-well plate Alprazolam Amphetamine Bromazepam Buprenorphine Cannabinoid Clonazepam Cocaine Codeine Diazepam Drugs of Abuse EDDP Estazolam Fentanyl Flunitrazepam Flurazepam Forensic Hair Hydrocodone Hydromorphone Ketamine Lorazepam Lysera Lysergic acid diethylamide (LSD) MDA MDEA MDMA Mephedrone Methadone Methamphetamine Midazolam Morphine Nitrazepam Norbuprenorphine Nordiazepam Norfentanyl Norketamine Oxazepam Oxycodone Oxymorphone PCP Posters THC THC-COOH Temazepam Triazolam UPLC-MS/MS Zaleplone

Sample preparation for hair analysis is often lengthy involving multiple manual labor steps from cutting, washing, homogenization/pulverization, digestion, sample extraction and analysis. This poster aims to demonstrate a streamlined sample preparation workflow for hair analysis. The workflow is evaluated using a suite of drugs of abuse, including THC and metabolites. MSACL NA 2019

6-MAM 7-Aminoclonazepam 7-Aminoflunitrazepam 96-well plate Amphetamine Amphetamines Benzodiazepines Bromazepam Clinical Clonazepam Codeine Drugs of Abuse EDDP English Flunitrazepam Flurazepam Forensic Hair Hydromorphone LC-MS/MS Lysera MDA MDEA MDMA Mephedrone Methadone Methamphetamine Midazolam Morphine Nitrazepam Opiates Opioid Oxycodone Oxymorphone Posters THC THC metabolites THC-COOH

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