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)

The low-level detection required for cannabis use combined with the complexity of hair testing makes for a challenging application. This poster aims to demonstrate simplified sample preparation workflow for low level analysis of THC and metabolites from hair, using Biotage Lysera and ISOLUTE SLE+ columns for sample preparation. TIAFT 2019 SOFT 2019

Cannabidiol Cannabinoid Clinical Column English Forensic Hair LC-MS/MS Lysera Posters THC THC metabolites THC-COOH

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)

The method described in this application note achieves high recoveries of THC and an extended suite of common metabolites in oral fluid from Quantisal (Immunalysis) oral fluid collection devices. This application note describes effective and efficient ISOLUTE SLE+ protocols optimized for sample loading volumes of either 300 μL or 800 μL. The simple sample preparation procedure delivers clean extracts and analyte recoveries greater than 64% with RSDs of <10% for all analytes.

11-hydroxy- Δ9-tetrahydrocannabinol (THC-OH) 11-nor- Δ9-carboxy tetrahydrocannabinol (THC-COOH) 11-nor-9-carboxy- Δ9-tetrahydrocannabivarin (THC-V-COOH) 11-nor-9-carboxy- Δ9tetrahydrocannabinol glucuronide (THC-COOH-g 11-nor-9-carboxy-Δ9-THC glucuronide Application Notes Cannabidiol Cannabigerol Column Drugs of Abuse English Forensic LC-MS/MS Oral fluid Saliva THC THC metabolites THC-COOH Δ9-tetrahydrocannabinol (THC) Δ9-tetrahydrocannabivarin (THCV)

This application note describes the fully automated extraction of carboxy-THC from urine, following base hydrolysis prior to GC/MS analysis. The method was automated using Biotage® ExtraheraTM, configured for use with EVOLUTE EXPRESS ABN columns. The method delivers clean extracts and analyte recovery of 90% with RSD lower than 10% for carboxy-THC and its deuterated internal standard. Using the Biotage® ExtraheraTM, 24 samples were extracted in approximately 38 minutes. The limit of quantitation of 2 ng/mL is below the SAMHSA/ EWDTS confirmation cut off of 15 ng/mL for workplace testing applications.

11-nor- Δ9-carboxy tetrahydrocannabinol (THC-COOH) Application Notes Column English Extrahera Forensic GC-MS Hydrolysed urine THC metabolites THC-COOH

This application note describes the fully automated extraction of carboxy-THC from urine, following base hydrolysis prior to GC/MS analysis. The method was automated using Biotage® ExtraheraTM, configured for use with ISOLUTE SLE+ columns. The simple sample preparation procedure delivers clean extracts and analyte recoveries greater than 80% with RSDs lower than 10% for carboxy-THC and its deuterated internal standard. Using Biotage Extrahera, 24 samples are extracted in approximately 35 minutes. Limit of quantitation is below the SAMHSA/EWDTS confirmation cut off of 15 ng/mL for workplace testing applications.

11-nor- Δ9-carboxy tetrahydrocannabinol (THC-COOH) Application Notes Column English Forensic GC-MS Hydrolysed urine THC metabolites THC-COOH

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

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 discusses the potential for a single extraction protocol for various drugs of abuse classes in whole blood prior to UPLC-MS/MS analysis. SOFT 2017, Boca Raton also presented at SFTA, Marseille, France, 2018

Amphetamines Benzodiazepines Cannabinoid Cathinones Clinical Cocaines Column Drugs of Abuse English Extrahera Forensic LC-MS/MS Opiates Posters THC metabolites Whole blood z-drugs

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

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

This application note describes the extraction of 11-nor-9-carboxy-THC from a urine matrix, prior to GC/MS analysis. Carboxy-THC is the primary metabolite of THC, a key indicator of illicit marijuana usage. In urine, ~80% of the carboxy-THC metabolite is present in the form of its glucuronide metabolite. Therefore, to effectively quantify the THC-COOH, urine is hydrolyzed before extraction. This application note describes optimized extraction of urine samples prepared by either enzymatic or base hydrolysis.

11-nor- Δ9-carboxy tetrahydrocannabinol (THC-COOH) Application Notes Column Drugs of Abuse English Forensic GC-MS Hydrolysed urine THC metabolites THC-COOH Urine

This application note describes the extraction, using ISOLUTE SLE+ supported liquid extraction columns, of 49 drugs of abuse from whole blood, prior to UPLC-MS/MS analysis. High, reproducible drug recoveries are achieved, with sub ng/mL LOQs for most analytes. The method is easily automated using Biotage® Extrahera.

11-nor- Δ9-carboxy tetrahydrocannabinol (THC-COOH) 2-Hydroxyethylflurazepam 6-acetylmorphine (MAM) 7-Aminoclonazepam 7-Aminoflunitrazepam Alprazolam Amphetamine Benzodiazepines Benzoylecgonine Bromazepam Buprenorphine Clonazepam Cocaine Cocaines Codeine Column Diazepam Dihydrocodeine (DHC) Drugs of Abuse EDDP English Estazolam Extrahera Fentanyl Flunitrazepam Flurazepam Forensic Hydrocodone Hydromorphone Ketamine LC-MS/MS Lorazepam Lysergic acid diethylamide (LSD) MDA MDEA MDMA Mephedrone Methadone Methamphetamine Midazolam Morphine Nitrazepam Norbuprenorphine Nordiazepam Norfentanyl Norketamine Opiates Oxazepam Oxycodone Oxymorphone Pain Management Phencyclidine (PCP) THC metabolites THC-COOH Temazepam Triazolam Whole blood Zaleplone Zolpidem Zopiclone z-drugs α-OH Triazolam α-OH alprazolam

This application note describes a simple but effective method for the extraction of THC and metabolites from whole blood using ISOLUTE SLE+ Supported Liquid Extraction 96-well plates.

11-hydroxy- Δ9-tetrahydrocannabinol (THC-OH) 11-nor- Δ9-carboxy tetrahydrocannabinol (THC-COOH) 96-well plate Application Notes Drugs of Abuse English Forensic LC-MS/MS THC THC metabolites THC-COOH Whole blood Δ9-tetrahydrocannabinol (THC)

This application note describes a procedure for sample pre-treatment and extraction of THC and metabolites from human hair, using Biotage® Lysera for matrix pulverization of the sample prior to clean up using ISOLUTE® SLE+ supported liquid extraction. Manual processing protocols were developed using the Biotage® PRESSURE+ 96 (plate format) or 48 (column format) positive pressure manifolds. For automated processing, protocols were developed using Biotage® ExtraheraTM.

11-hydroxy- Δ9-tetrahydrocannabinol (THC-OH) 11-nor- Δ9-carboxy tetrahydrocannabinol (THC-COOH) 96-well plate Application Notes Bead Ruptor Cannabidiol Cannabinol Column English Extrahera Forensic Hair LC-MS/MS Lysera THC THC metabolites THC-COOH Δ9-tetrahydrocannabinol (THC)

This application note describes a procedure for sample pre-treatment and extraction of THC and its metabolites from human nail samples, using the Biotage® Lysera for matrix pulverisation, and pre concentration of the sample prior to clean up using ISOLUTE® SLE+ supported liquid extraction.

11-hydroxy- Δ9-tetrahydrocannabinol (THC-OH) 11-nor- Δ9-carboxy tetrahydrocannabinol (THC-COOH) 96-well plate Application Notes Cannabidiol Cannabinol Column Drugs of Abuse English Forensic LC-MS/MS Lysera Nail (human) THC THC metabolites THC-COOH Δ9-tetrahydrocannabinol (THC)

This application note describes the simultaneous extraction of THC and its major metabolites, including 11-nor-9-carboxy-Δ9-THC glucuronide, from urine using supported liquid extraction (ISOLUTE® SLE+ in both plate and column formats) prior to analysis by LC-MS/MS.

11-hydroxy- Δ9-tetrahydrocannabinol (THC-OH) 11-nor- Δ9-carboxy tetrahydrocannabinol (THC-COOH) 11-nor-9-carboxy-Δ9-THC glucuronide 96-well plate Application Notes Cannabidiol Cannabinol Column Drugs of Abuse English Forensic LC-MS/MS THC metabolites Urine Δ9-tetrahydrocannabinol (THC)

This application note describes the extraction, using ISOLUTE SLE+ supported liquid extraction columns, of THC, THCA and Carboxy-THC from oral fluid matrix collected using the NeoSal device, prior to GC/MS analysis.

11-hydroxy- Δ9-tetrahydrocannabinol (THC-OH) 11-nor- Δ9-carboxy tetrahydrocannabinol (THC-COOH) Application Notes Column Drugs of Abuse English Forensic GC-MS Oral fluid Saliva THC THC metabolites THC-COOH Δ9-tetrahydrocannabinol (THC)

This application note describes the extraction of Δ9-THC, 11-hydroxy- Δ9-THC and 11-nor-9-carboxy-THC from whole blood matrix, prior to GC/MS analysis. An extremely simple sample pre-treatment and extraction method is employed to extract the parent THC and main metabolites from complex whole blood matrix, delivering high, reproducible analyte recoveries.

11-hydroxy- Δ9-tetrahydrocannabinol (THC-OH) 11-nor- Δ9-carboxy tetrahydrocannabinol (THC-COOH) Application Notes Column Drugs of Abuse English Forensic GC-MS THC THC metabolites THC-COOH Whole blood Δ9-tetrahydrocannabinol (THC)