This poster describes a strong cation exchange SPE protocol for extraction of Vitamin B3 (Niacin) and related metabolites from serum. The ISOLUTE SCX-3 96 well plate format demonstrated as a viable option for serum measurements over a relevant concentration range in clinical diagnostics. It is anticipated that strategies presented in this poster would be of broad-based interest to clinical laboratories focused on combining parent/metabolite assays into a single analysis. MSACL 2014

96-well plate B Vitamins Clinical English LC-MS/MS Niacin (nicotinic acid) Niacinamide Nicotinuric acid Posters Serum Vitamin B3 Vitamins

This application note describes the extraction of Vitamin B3 (Niacin, nicotinic acid) and its key metabolites from serum using a strong cation exchange SPE retention mechanism. High reproducible analyte recoveries are achieved.

96-well plate Application Notes B Vitamins Clinical English LC-MS/MS Niacin (nicotinic acid) Niacinamide Nicotinuric acid Serum Vitamin B3 Vitamins

This application note describes the extraction of nine phthalate metabolites from human urine using ISOLUTE® ENV+ solid phase extraction columns.

Application Notes Biomarker Clinical Column English LC-MS/MS Mono(2-ethyl-5-carboxypentyl) phthalate (MECPP) Mono(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP) Mono(2-ethylhexyl) phthalate (MEHP) Monobenzyl phthalate (MBzP) Monobutyl phthalate (MBP) Monoethyl phthalate (MEP) Monohexyl phthalate (MHxP) Monoisononyl phthalate (MiNP) Monomethyl phthalate (MMP) Phthalate Metabolites Phthalates Urine

Pain management therapy warrants constant monitoring of therapeutic levels of prescribed drug levels in patient urine samples. The number of samples being submitted for analysis has increased dramatically in the last 10 years with improvements in high throughput automated screening capabilities. Patient samples analysis is complicated by the need for an effective sample preparation methodology that can extract target analytes from complex matrices with good efficiency. Further complicating the process is the need to enzymatically hydrolyse the glucoronidated metabolites prior to extraction from the urine matrix. A fully automated sample preparation process using a TECAN Freedom EVO® 100 was designed to incorporate both the enzymatic hydrolysis and subsequent sample preparation assay as one continuous workflow. Supported Liquid Extraction (ISOLUTE SLE+) which offers an efficient alternative to traditional liquid-liquid extraction (LLE) and solid phase extraction (SPE) techniques was used to extract a suite of pain management drugs from spiked urine samples. A recovery and quantitation assay was run on the TECAN Freedom EVO® 100 using mock patient samples to demonstrate utility of automation process. MSACL, Pain Management, Biotage, SPE, SLE, LLE, Supported Liquid Extraction, Drugs, MSACL, San Diego, 2013

6-acetylmorphine (MAM) 96-well plate Benzodiazepines Clinical Clonazepam Codeine Diazepam Dihydrocodeine (DHC) Drugs of Abuse Flunitrazepam Hydrocodone Hydrolysed urine Hydromorphone LC-MS/MS Methadone Methamphetamine Morphine Nitrazepam Opiates Oxycodone Oxymorphone Posters Urine

This application note describes the extraction of ethyl glucuronide and ethyl sulfate from human urine using ISOLUTE® NH2 solid phase extraction columns. The method has been applied real patient samples that had been previously analyzed with a validated referee method. The results of the orthogonal measurements agreed, to provide similar diagnostic values.

Alcohol metabolite Application Notes Clinical Column Ethyl Sulfate (EtS) Ethyl glucuronide (EtG) Forensic LC-MS/MS Urine

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 poster describes a reduced workflow solid phase extraction method for extraction of vitamin B7 (biotin) from human serum. Using EVOLUTE EXPRESS AX 96-well plates, the traditional sorbent conditioning and equilibration steps are not required, meaning the sample can be applied directly to the dry plate. Post extraction evaporation is also eliminated. ASMS 2014

96-well plate Biotin Clinical English LC-MS/MS Posters Serum Vitamin B7 Vitamins

This report details a “load, wash, elute” weak cation exchange solid phase extraction procedure amenable to both plasma and urine samples. The extracts are subsequently injected into an LC-MS/MS system. The preliminary sample preparation method was developed at the Biotage US Applications lab (Charlotte, NC).The method was then transferred to Ionics (Bolton, ON, Canada) to facilitate the nmole/L measurements of the selected biomarkers by laminar flow tandem mass spectrometry. The SPE-LC-ESIMS/ MS method parameters were first optimized using pooled mixed gender plasma. A set of patient samples (n=32) was later supplied by the Mayo Clinic (Rochester, MN) that had previously been analyzed by a validated referee method. The population represented measured values across a range of clinical relevance. AACC 2014

96-well plate Catecholamines Clinical English LC-MS/MS Metanephrine Normetanephrine Plasma Posters Urine

This poster describes the use of ISOLUTE ENV+ columns to extract important phthalate metabolites from human urine. MSACL Europe 2014

Clinical Column LC-MS/MS Mono(2-ethyl-5-carboxypentyl) phthalate (MECPP) Mono(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP) Mono(2-ethylhexyl) phthalate (MEHP) Monobenzyl phthalate (MBzP) Monobutyl phthalate (MBP) Monoethyl phthalate (MEP) Monohexyl phthalate (MHxP) Monoisononyl phthalate (MiNP) Monomethyl phthalate (MMP) Phthalate Metabolites Phthalates Posters Urine

Cortisol measurement in hair samples scan be a useful indicator of chronic stress. This poster compares a new LC-MS/MS method with an existing immunoassay based method. The Biotage Bead Ruptor is used effectively for hair pre-treatment. MSACL 2016

Bead Ruptor Clinical Corticosteroids Cortisol English Hair LC-MS/MS Posters

The use of LC/MS analysis in the clinical lab has increased exponentially over the last 10 years. Modern mass spectrometers are extremely sensitive allowing low level detection of many target analytes. However, this sensitivity can come at a price, in that levels of contamination not previously detected with less sensitive instruments can now have larger impact on analysis. The complexity of common matrices such as plasma/serum and urine while presenting different challenges can have a marked influence on method performance. As a result sample preparation is an extremely important part of the process in order to provide robust methods. This seminar focuses on some of the method development challenges our lab faced when looking at two clinical assays: Endogenous steroid hormone extraction from serum, and catecholamine extraction from plasma and urine. Particular emphasis is placed on the various sample preparation options we looked at for the extraction of these analytes. During optimization of the extraction process we investigated analyte recovery, co-extractable matrix components, HPLC column degradation, calibration curve performance and limits of quantitation. MSACL US 2018 Breakfast Seminar

96-well plate Catecholamines Clinical English LC-MS/MS Plasma Serum Steroid Hormones Steroid metabolite Urine

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 poster describes an automated procedure for isolation and clean up of 15 F2t-isoprostane (8-iso Prostaglandin 2a)(a lipid marker of oxidative stress) from BSA.

15 F2t-isoprostane 8-iso Prostaglandin2a BSA Biological fluids Biomarker Clinical Column English GC-MS Lipids Posters RapidTrace

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

A GC-MSMS method of simultaneously measuring legacy/emerging POPs PCBs, OCPs and PBDEs) in serum was developed and applied to this study of exposure of mothers to harmful environmental contaminants.

BHC Clinical DDE DDT English Environmental GC-MS HCB Organochlorine pesticides (OCP) Organochlorines Oxychlordane PBDE PBDEs PCB PCBs Persistant organic pollutants (POP) Posters RapidTrace Serum Trans-nonachlor

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 a sample preparation technique calls for removal of compounds using SPE, it is standard practice to determine what concentrations will produce an acceptable linear range for the assay. Traditional linear regression is often performed manually using pipettes to generate an external standard curve that unknown samples are measured against.

Application Notes Clinical English Polysorbate 20 SmartPrep

This guide describes simple practical strategies, hints and tips designed to mitigate or avoid cross-well contamination or ‘cross talk’ in high throughput 96-well based sample preparation techniques.

96-well plate Brochures Clinical English Forensic Pharma

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