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

When analyzing human urine for drugs of abuse, one of the most common tests is for the cocaine metabolite Benzoylecgonine (BZE). A contract laboratory has automated this labor intensive procedure using the RapidTrace.

Application Notes Benzoylecgonine Column Drugs of Abuse Forensic GC-MS RapidTrace Urine

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

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 Zolpidem Zolpidem-phenyl-4-COOH z-drugs α-OH alprazolam

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

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 a Supported Liquid Extraction (SLE) protocol for the extraction of various drugs of abuse from hydrolyzed urine prior to LC-MS/MS analysis. drugs of abuse, sle, multiclass

2-Hydroxyethylflurazepam 7-Aminoclonazepam 96-well plate Alprazolam Amphetamine Amphetamines Anhydroecgonine methyl ester Application Notes Benzodiazepines Benzoylecgonine Bromazepam Buprenorphine Clonazepam Cocaethylene Cocaine Cocaines Codeine Column Diazepam Ecgonine methyl ester Estazolam Fentanyl Flunitrazepam Hydrolysed urine Hydroxyalprazolam Hydroxytriazolam Ketamine Methamphetamine Midazolam Morphine Nitrazepam Norbuprenorphine Norcocaine Nordiazepam Norfentanyl Norketamine Opiates Opioid Oxazepam Temazepam Triazolam Zaleplone Zolpidem Zopiclone z-drugs

This application note describes a Supported Liquid Extraction (SLE) protocol for the extraction of various drugs of abuse from non- hydrolyzed urine prior to LC-MS/MS analysis. drugs of abuse, sle, multiclass

2-Hydroxyethylflurazepam 6-acetylmorphine (MAM) 7-Aminoclonazepam 7-Aminoflunitrazepam Alprazolam Amphetamine Amphetamines Application Notes Benzodiazepines Benzoylecgonine Bromazepam Buprenorphine Clonazepam Cocaine Cocaines Codeine Diazepam Dihydrocodeine (DHC) EDDP English Estazolam Fentanyl Flunitrazepam Flurazepam Forensic Hydrocodone Hydromorphone Hydroxyalprazolam Hydroxytriazolam Ketamine LC-MS/MS MDA MDEA MDMA Methadone Methamphetamine Midazolam Morphine Nitrazepam Norbuprenorphine Nordiazepam Norfentanyl Norketamine Opiates Opioid Oxazepam Oxymorphone RapidTrace TECAN Temazepam Triazolam Urine Zaleplone Zolpidem Zopiclone z-drugs

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 collected using the Intercept Oral Fluid Drug Test Kit (Orasure Technologies), prior to UPLC-MS/MS analysis. The sample preparation is optimised to minimise matrix effects due to the buffers used in the collection device. Estimated LOQs range from 0.1-1 ng/mL for the various analytes.

2-Hydroxyethylflurazepam 2-OH-Et-Flurazepam 6-acetylmorphine (MAM) 7-Aminoclonazepam 7-Aminoflunitrazepam Alprazolam Amphetamine Amphetamines Application Notes Basic drugs 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 Oxazepam Oxycodone Oxymorphone Pharmaceuticals 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

The method addresses the extraction of all three classes of drugs of abuse from urine using non-polar and cation exchange retention mechanisms on the ISOLUTE HCX mixed mode column. This protocol was optimized to eliminate the use of dichloromethane in the elution of the acidic / neutral drug fraction.

Acidic drugs Amphetamine Application Notes Basic drugs Benzodiazepines Benzoylecgonine Cocaines Codeine Column Diazepam Forensic Morphine Naphyrone Norcocaine Nordiazepam Opiates Oxazepam THC-COOH Temazepam Urine

This application note describes effective and efficient ISOLUTE SLE+ protocols optimized for sample (hydrolysed urine) volumes of either 200 μL or 1 mL. Procedures for high throughput (96-well plate) and column format are included. The simple sample preparation procedure delivers clean extracts and analyte recoveries of 82-100% with RSDs of <5% for all analytes. Sub ng/mL LOQs can be achieved.

Anhydroecgonine Methyl Ester AEME Application Notes Benzoylecgonine Butylone Cocaines Column Ecgonine Methyl Ester EME Forensic Hydrolysed urine LC-MS/MS Norcocaine