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

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

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 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

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 method describes the use of ISOLUTE SLE+ supported liquid extraction 1 mL sample volume columns to extract a range of barbiturates from human urine. The analysis of these analytes was carried out by GC-MS. This simplified and efficient extraction method has significant analyte recoveries ranging from 103-108% with LOQs of 10 ng/mL and RSDs <10%. Barbs, Barbiturates, SLE, SLE+, ISOLUTE, GCMS, DoA, Forensic, Work Place Drug Testing, Supported Liquid Extraction, GC

Amobarbital Application Notes Barbiturates Butabarbital Butalbarbital GC-MS Phenobarbital Secobarbital Urine

This application note describes the extraction of barbiturates from oral fluid matrix collected using the Intercept Oral Fluid Drug Test Kit(Orasure Technologies), prior to GC/MS analysis. The ISOLUTE SLE+ protocol is optimized for 400 μL and 1 mL sample capacity formats. The simple sample preparation procedure delivers clean extracts and analyte recoveries greater than 90% with RSDs lower than 7% for all analytes.

Amobarbital Application Notes Barbiturates Butabarbital Butalbarbital Column Drugs of Abuse Forensic Hexobarbital LC-MS/MS Oral fluid Pentobarbital Phenobarbital Saliva Secobarbital

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

Amobarbital Application Notes Barbiturates Butabarbital Butalbarbital Column Drugs of Abuse English Forensic GC-MS Hexobarbital Oral fluid Pentobarbital Phenobarbital Saliva Secobarbital

This application note describes the extraction of Butalbarbital, Butabarbital, Amobarbital, Pentobarbital,Secobarbital, Hexobarbital and Phenobarbital from oral fluid matrix collected using the Oral-Eze® collection devices, prior to GC/MS analysis.

Amobarbital Application Notes Barbiturates Butabarbital Butalbarbital Column English Forensic GC-MS Hexobarbital Oral fluid Pentobarbital Phenobarbital Saliva Secobarbital

This application note describes the extraction of barbiturates from oral fluid matrix collected using Quantisal™ collection devices (Immunalysis), prior to GC/MS analysis. The ISOLUTE SLE+ protocol is optimized for 400 μL and 1 mL sample capacity formats. The simple sample preparation procedure delivers clean extracts and analyte recoveries greater than 88% with RSDs lower than 6% for all analytes.

96-well plate Amobarbital Application Notes Butabarbital Butalbarbital Forensic GC-MS Hexobarbital Oral fluid Pentobarbital Phenobarbital Saliva Secobarbital

This poster describes the extraction of barbiturates from oral fluid collected using common collection devices using ISOLUTE SLE+ Supported Liquid Extraction columns, optimised for GC-MS analysis. SOFT 2014

Amobarbital Barbiturates Butabarbital Butalbarbital Column English Forensic GC-MS Hexobarbital Oral fluid Pentobarbital Phenobarbital Posters Secobarbital

The following method was developed for the extraction of barbiturate drugs from urine, including: butabarbital, amobarbital, cyclobarbital and phenobarbital. Typical recoveries for these drugs are > 85%. ISOLUTE, BIOTAGE, HCX, DOA, Nervous System

Amobarbital Application Notes Barbiturates Butabarbital Clinical Cyclobarbital GC-MS Phenobarbital Urine

This application note describes the extraction of barbiturate compounds from whole blood, prior to GC/MS analysis. This protocol also allows the simultaneous extraction of various other drugs of abuse classes: amphetamines, benzodiazepines, cocaine and opiates. ISOLUTE® SLE+ columns with 1 mL sample capacity are used to extract whole blood samples following a straightforward sample dilution. No protein precipitation or other pre-treatment is required prior to sample loading. The sample preparation procedure delivers clean extracts, good recoveries and RSD values and LLOQs from 20 ng/mL (analyte dependant).

Amobarbital Application Notes Barbiturates Butabarbital Butalbarbital Clinical Column English GC-MS Hexobarbital Pentobarbital Phenobarbital Secobarbital Whole blood

This application note describes the extraction of a broad range of analytes from liver tissue matrix prior to GC/MS analysis using ISOLUTE® SLE+ supported liquid extraction columns. A protocol has been developed that allows the simultaneous extraction of various drugs of abuse classes: amphetamines, barbiturates, benzodiazepines, cocaine, ketamine, THC. In addition to these drug panels, simultaneous extraction of carbamate, organochlorine, organophosphate, pyrethroid, and triazine pesticide classes is demonstrated.

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

Testing for drugs of abuse in oral fluids can strongly benefit the criminal justice field as a less invasive and cost-effective approach for drug detection when compared to blood or urine sampling. Oral fluid analysis has facilitated laboratory analysis for many drugs of abuse and is a constantly evolving analysis procedure which benefits from increasingly sensitive methods of detection. The Laser Diode Thermal Desorption (LDTD) source combined with Mass spectrometry is presented as a new screening tool for drug analysis in Oral Fluids. Analysis speed of LDTD provides accurate results in seconds in combination with exceptional specificity of MS instruments make a powerful platform for the screening of different drugs of abuse and new emerging drugs. Different extraction procedures are available; however those methods depend on specific drug conditions: basic or acid drugs / hydrophilic or hydrophobic. A new extraction approach, Supported Liquid Extraction (SLE+) is evaluated as generic extraction procedure. MSACL 2014

96-well plate Amphetamine Benzoylecgonine Butalbital Cocaine Codeine Drugs of Abuse English Fentanyl Forensic LDTD-MS/MS MDA MDEA MDMA Methadone Methamphetamine Oral fluid Phenobarbital Posters Secobarbital

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

This poster describes a simple but effective method for screening a range of pesticides and drugs of abuse from liver samples. Liver tissue is homogenized using the Biotage® Lysera, and the subsequent extract is cleaned up using ISOLUTE SLE+ supported liquid extraction columns. ASMS 2017

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