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
The potential harm caused by synthetic cannabinoids is a significant concern since exposures and reports of human fatalities due to these substances is widespread . Detection of these relatively new type designer drugs in blood is essential in forensic toxicology, other scientific and medical fields. Cannabinoids, SLE, SLE+, Supported Liquid Extraction,
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)
Vitamin D deficiency can result in various health issues such as osteoporosis, liver and kidney problems and is associated with increased risk of cancers and multiple sclerosis. From this standpoint vitamin D analysis has extremely important clinical relevance. This poster presents a novel method for the extraction of 25 hydroxyvitamin D demonstrating acceptable validation parameters from serum calibrators and DEQAS supplied samples using ISOLUTE SLE+.
This poster describes a supported liquid extraction method for extraction of a class of novel psychoactive substances (NBOMes) with analysis using laser diode thermal desorption (LDTD) MS/MS. Samples are oral fluid collected using Salivettes. TAMS 2014
The objective was to develop a GC-MS assay for the determination of free benzodiazepines using Supported Liquid Extraction (SLE). The SLE extraction mechanism is very efficient, delivering higher analyte recoveries and cleaner extracts than equivalent LLE methods. ISOLUTE, SLE, SLE+, Supported Liquid Extraction, Benzodiazepines, Forensic, Drugs, DOA, Drugs of Abuse, SOFT,
4-Hydroxynordiazepam 7-Aminoclonazepam 7-Aminoflunitrazepam 7-Aminonitrazepam Benzodiazepines Bromazepam Column Demethylflunitrazepam Flurazepam Forensic GC-MS Hydroxyalprazolam Hydroxymidazolam Hydroxytriazolam Midazolam Nitrazepam Posters 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)
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)
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 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
This application note describes the extraction of a range of SPICE drugs and metabolites in urine which are typically screened in forensic toxicology panels using ISOLUTE® SLE+ in a 96-well plate format. Both manual (Biotage Pressure+ 96) and automated (TECAN Freedom EVO® 100) processing conditions are described.
96-well plate Application Notes English Forensic JWH 018-N-(4-hydroxypentyl) JWH-018 JWH-018 JWH-018-(5-hydroxypentyl) JWH-018-N-5-(pentanoic acid) JWH-073 JWH-073-N-(3-hydroxybutyl) JWH-200 JWH-250 JWH-250-N-(5-hydroxypentyl) LC-MS/MS Synthetic Cannabinoids (SPICE) TECAN UR-144 UR-144-(5-hydroxypentyl) UR-144-(pentanoic acid) UR144-(5-chloropentyl) Urine XLR-11
This poster compares the use of supported liquid extraction (ISOLUTE® SLE+) and a novel protein and phospholipid depletion plate (ISOLUTE® PLD+), for the extraction of 25-hydroxy vitamin D. The manual extraction protocols were transferred to an SPE automation platform (Biotage® Extrahera)and method performance versus manual processing compared. MSACL EU 2015
This poster summarizes various sample preparation strategies for the extraction of MMA from serum without the necessity for derivatization, prior to LC-MS/MS analysis. A range of sample preparation techniques of varying complexity were evaluated: protein precipitation, phospholipid depletion, supported liquid extraction and solid phase extraction using both silica and polymer-based mixed-mode anion exchange chemistries. Method performance was evaluated for evaporative effects, assay recovery, ion suppression and phospholipid removal. MSACL 2016
At the Forensic Medicine Lab at Toho University, researchers use ISOLUTE® SLE+ columns from Biotage. When dealing with samples that easily form emulsions like urine or blood, it allows researchers to use the established liquid-liquid extraction technique, saving significant amount of time on analysis. We spoke with the Head of the Forensic Medicine Lab Professor Masaru Terada.
This poster describes the development and validation of a method for supported liquid extraction of serum cortisol, with analysis by UPLC-MS/MS. The aim of this study was to develop a candidate reference method that could then be used to underpin the UK NEQAS Cortisol scheme. MSACL EU 2014
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
Endogenous phospholipids present in biological fluids are a major problem in LCMS/ MS analysis as they are often very difficult to remove during sample preparation. When phospholipids are not removed, they retain very strongly on reversed phase analytical columns. If high organic (end of run) washes are not incorporated into the LC methods these matrix components may elute in subsequent analyses causing regions of suppression/enhancement leading to inaccurate quantitation. This poster evaluates the use of polymer-based solid phase extraction (SPE) sorbents, incorporating hydrophobic and various mixedmode retention mechanisms to address the problems associated with phospholipid removal. Phospholipid, EVOLUTE, STRATA X, OASIS, WATERS, AX, WAX, CX, WCX, ABN, ASMS 2011
Endogenous phospholipids (outline structure shown in Figure 1.) present in biological fluids are a major problem in LC-MS/MS analysis. Due to their strong retention characteristics in reversed phase chromatography phospholipids tend not to elute as discrete peaks and are often very difficult to separate from analytes of interest. This co-elution often leads to areas of suppression or enhancement in the chromatogram which in turn can cause quantitation issues. Supported liquid extraction (SLE) is an analogous technique to traditional liquidliquid extraction. This poster compares phospholipid removal using a wide variety of solvent combinations, pH control and polar extraction solvents on supported liquid extraction plates ASMS 2011
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