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

Most drugs, both licit and illicit, are excreted in urine as glucuronide conjugates. Hydrolysis using beta-glucuronidase converts the glucuronidated metabolites back to their “free” or non-conjugated form, increasing sensitivity for LC-MS/MS analysis. Hydrolysis using red abalone, abalone, and recombinant beta-glucuronidase enzymes was evaluated using an in-well hydrolysis plate to determine which provided the most efficient hydrolysis of glucuronide metabolites without affecting overall recovery of nonconjugated compounds. A glucuronide control containing 8 glucuronide compounds was used to determine the extent of hydrolysis that occurred. A non-conjugated control containing 96 licit and illicit drugs of abuse was evaluated to determine if signal suppression occurred as a result of enzyme hydrolysis. ASMS 2017

96-well plate Clinical Codeine Drugs of Abuse English Forensic Hydrolysed urine Hydromorphone LC-MS/MS Lorazepam Morphine Norbuprenorphine Oxazepam Oxymorphone Pain Management Posters Urine

This poster presents a method for detection of approximately 100 drugs and metabolites in urine using mixed-mode polymeric cation exchange solid phase extraction (SPE). Four different glucuronidase enzymes and several incubation times and temperatures were evaluated to determine optimum hydrolysis conditions for seven commonly detected glucuronide metabolites. Samples were subjected to offline hydrolysis and SPE (EVOLUTE® EXPRESS CX, Biotage, Charlotte, NC) and results compared to an SPE plate that incorporates in-well hydrolysis (EVOLUTE HYDRO CX, Biotage). MSACL 2018, Palm Springs, CA

96-well plate Clinical Drugs of Abuse English Forensic Hydrolysed urine LC-MS/MS Pain Management Posters Urine

Whether you need targeted methods for analytes such as Vitamin D metabolites in serum, or methods suitable for extraction of a wide panel of drugs and metabolites from urine, sample preparation before analysis is essential. This compendium highlights a selection of clinical sample preparation applications from Biotage.

Alcohol metabolite Application Notes Biomarker Brochures Catecholamines Clinical English Pain Management Peptides Product Notes Steroid Hormones Steroid metabolite Thyroid Hormone Tobacco Specific Nitrosamines (TSNAs) Vitamin D Vitamins Water Soluble Vitamins

This poster demonstrates that a large urine panel, comprised of 43 DOAs, from multiple drug classes, can be simultaneously screened by mixed-mode cation exchange SPE (using EVOLUTE EXPRESS CX 96 well plates) despite their disparate intermolecular traits, by thoughtfully selecting appropriate organic wash and elution conditions that simultaneously enable sample isolation and detection along with minimizing sample matrix effects. The extraction method is automated using the Biotage® Extrahera™ Automated sample Preparation Platform. MSACL 2017, Palm Springs SOFT 2017, Boca Raton

96-well plate Benzodiazepines Carbamates Carisoprodol Clinical Drugs of Abuse English Extrahera Forensic Gabapentin Hydrolysed urine LC-MS/MS Meprobamate Opiates Pain Management Posters Pregabalin Stimulants Urine

Buprenorphine and Norbuprenorphine are typically problematic for analysis due to analyte stability issues during sample preparation. This poster will demonstrate two fast and robust methods for the extraction of buprenorphine and norbuprenorphine in urine (using EVOLUTE EXPRESS CX), oral fluid (using EVOLUTE EXPRESS CX) and whole blood (using ISOLUTE SLE+). MSACL 2016

96-well plate Buprenorphine Clinical Column English Forensic LC-MS/MS Norbuprenorphine Oral fluid Pain Management Posters Saliva Synthetic urine Urine Whole blood

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

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

Product Note for EVOLUTE® HYDRO Solid Phase Extraction Plates with In-Well Hydrolysis Capability

96-well plate Alcohol metabolite Amphetamines Anaesthetic Analgesics Barbiturates Benzodiazepines Carbamate hypnotic Carbamate muscle relaxant Carbamates Clinical Designer stimulants Drugs of Abuse English Forensic GC-MS Hydrolysed urine LC-MS/MS NSAIDs Opiates Opioid Opioid agonist Pain Management Pharmaceuticals Product Notes Stimulants Urine z-drugs

The use of schedule I drugs for patient pain management therapy warrants constant monitoring of therapeutic levels in the patient. Screening patient urine samples for the free drugs is complicated by the metabolism process which converts the free drug to the -glucuronide form. Patient urine samples can be enzymatically hydrolyzed and extracted to detect the drugs using Supported Liquid Extraction (ISOLUTE SLE+) which offers an efficient alternative to traditional liquid-liquid extraction (LLE) for bioanalytical sample preparation. LOQs for recovered drugs ranged from 10 ng/mL to 0.5 ng/mL with recoveries above 80% and RSDs <10% SLE, Supported Liquid Extraction, Pain Mangement, Hydrolysis, Urine, Buprenorphine, Oxycodone, Temazepam,

Alprazolam Application Notes Benzodiazepines Buprenorphine Clinical Clonazepam Codeine Diazepam Drugs of Abuse English Fentanyl Flunitrazepam Hydromorphone LC-MS/MS Meperidine Methadone Methamphetamine Morphine Naloxone Naltrexone Nitrazepam Oxycodone Pain Management Temazepam Urine

This application note describes the extraction of 47 drugs of abuse from oral fluid matrix after sampling via Quantisal collection devices prior to analysis by UPLC-MS/MS. Optimised protocols for 200 uL and 500 uL sample volumes are included.

2-OH-Et-Flurazepam 6-acetylmorphine (MAM) 7-Aminoclonazepam 7-Aminoflunitrazepam Amphetamine Amphetamines Application Notes Benzodiazepines Benzoylecgonine Bromazepam Buprenorphine Cathinones Clinical Clonazepam Cocaine Codeine Column Diazepam Dihydrocodeine (DHC) Drugs of Abuse 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 Opiates Oral fluid Oxazepam Oxycodone Oxymorphone Pain Management Phencyclidine Phencyclidine (PCP) Saliva Temazepam Triazolam Zaleplone Zolpidem Zopiclone α-OH Triazolam α-OH alprazolam

This application note describes the extraction of 47 drugs of abuse and metabolites from oral fluid matrix after sampling via Oral-Eze collection devices, prior to UPLC-MS/MS analysis using ISOLUTE SLE+ supported liquid extraction columns. Estimated LOQs of between 0.005 and 0.75 ng/mL (analyte dependant) are achieved,.

2-OH-Et-Flurazepam 6-acetylmorphine (MAM) 7-Aminoclonazepam 7-Aminoflunitrazepam Alprazolam Amphetamine Amphetamines Application Notes Basic Benzodiazepines Benzoylecgonine Bromazepam Buprenorphine Clinical Clonazepam Cocaine Cocaines Codeine Column Diazepam Dihydrocodeine (DHC) Drugs of Abuse 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 Oral fluid Over the counter pharmaceuticals Oxazepam Oxycodone Oxymorphone Pain Management Pharmaceuticals Phencyclidine (PCP) Saliva Temazepam Triazolam Zaleplone Zolpidem Zopiclone α-OH Triazolam α-OH alprazolam

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 and robust mixed mode cation exchange solid phase extraction protocol using EVOLUTE® EXPRESS CX 96 well plates for the extraction of buprenorphine and norbuprenorphine from oral fluid. Excellent extract cleanliness allows quantitation of the analytes across the range 0.1 ng/mL to 100 ng/mL using LC-MS/MS.

96-well plate Application Notes Buprenorphine Clinical English Forensic LC-MS/MS Norbuprenorphine Oral fluid Pain Management Saliva

This application note describes a simple and robust mixed mode cation exchange solid phase extraction protocol using EVOLUTE® EXPRESS CX 96 well plates for the extraction of buprenorphine and norbuprenorphine from urine. Excellent extract cleanliness allows low level quantitation (0.1 ng/mL) of the analytes using LC-MS/MS.

96-well plate Application Notes Buprenorphine Clinical English Forensic Norbuprenorphine Pain Management Urine

This application note describes the extraction of buprenorphine and norbuprenorphine from whole blood using ISOLUTE® SLE+ Supported Liquid Extraction prior to LC-MS/MS analysis.

96-well plate Application Notes Buprenorphine Clinical English Forensic LC-MS/MS Norbuprenorphine Pain Management Whole blood

The screening of patient urine samples for basic drugs is typically performed following a lengthy sample preparation methodology that requires a time consuming dry down step. A novel elution protocol coupled to the EVOLUTE EXPRESS advanced plate technology affords a fast sample preparation solution that follows a LOAD-WASH-ELUTE-ANALYZE strategy. Basic drugs are fortified into urine and extracted in 3 easy steps using EVOLUTE EXPRESS CX solid phase extraction in a 96-well plate format.

6-acetyl codeine 6-acetylmorphine (MAM) 96-well plate Alprazolam Application Notes Basic Buprenorphine Clinical Clonazepam Codeine Diazepam Drugs of Abuse EDDP English Fentanyl Flunitrazepam Forensic Hydrocodone Hydromorphone LC-MS/MS Morphine Naltrexone Nitrazepam Normeperidine Oxazepam Oxycodone Oxymorphone Pain Management Temazepam Urine