Part No: P052Issued year: 2013File size: 0.48mbFile type: pdf
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
Part No: P144Issued year: 2016File size: 0.6mbFile type: pdf
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.
Part No: PPS443Issued year: 2017File size: 2.31mbFile type: pdf
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.
Part No: P112Issued year: 2014File size: 1.4mbFile type: pdf
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.
Part No: P132Issued year: 2015File size: 1.55mbFile type: pdf
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.
Part No: P087Issued year: 2014File size: 0.94mbFile type: pdf
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.
Part No: P138Issued year: 2015File size: 0.82mbFile type: pdf
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
Part No: P151Issued year: 2016File size: 0.96mbFile type: pdf
This poster compares the performance of manual processing to a novel automated sample preparation system prior to GC/MS or LC-MS/MS analysis in forensic toxicology applications. Emphasis is placed on the potential for 96-well cross contamination and strategies for its elimination.
Part No: P156Issued year: 2017File size: 0.23mbFile type: pdf
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
Part No: AN764Issued year: 2012File size: 0.52mbFile type: pdf
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,
Part No: AN832Issued year: 2014File size: 1.78mbFile type: pdf
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.
Part No: AN836Issued year: 2015File size: 1.81mbFile type: pdf
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.
Part No: AN837Issued year: 2015File size: 2.27mbFile type: pdf
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,.
Part No: AN875Issued year: 2017File size: 3.36mbFile type: pdf
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.
Part No: AN770Issued year: 2012File size: 0.26mbFile type: pdf
This application note demonstrates an effective and efficient suported liquid extraction protocol for the clean up and concentration of a range of forensically significant opiates and their metabolites
opiates, sle, urine, GC-MS, forensic, drugs of abuse, DOA, UCT, agilent
Part No: AN790Issued year: 2013File size: 1.75mbFile type: pdf
This application note describes the extraction of different drugs in hemolyzed whole blood, which are typically screened for forensic toxicology panels, using ISOLUTE® SLE+ supported liquid extraction columns in the 1 mL and 2 mL sample capacity formats.
illicit drugs, pain management, supported liquid extraction, LC-MS/MS, blood, post mortem blood, hemolysed whole blood,
Part No: AN788Issued year: 2013File size: 1.57mbFile type: pdf
This application note describes the extraction of a range of illicit drugs, which are typically screened in forensic toxicology panels, from oral fluid using ISOLUTE® SLE+ supported liquid extraction columns.
oral fluid, Quantisal, Immunalysis, Intercept, Orasure, supported liquid extraction
Part No: AN741Issued year: 2012File size: 0.6mbFile type: pdf
This method describes the use of ISOLUTE SLE+ supported liquid extraction plates (96-well) and 1 mL sample volume columns to extract a range of opiates including opiate treatment analytes (methadone and its metabolite EDDP)) from human urine. This simplified and efficient extraction method has significant analyte recoveries ranging from 70-102% with LODs as low as 500 pg/mL.
Opiates, SLE, SLE+, Supported Liquid Extraction, Urine, Oasis, Waters, SPE, Solid Phase Extraction, Sample Prep, quick, easy, DOA, Drugs of Abuse
Part No: AN866Issued year: 2016File size: 1.4mbFile type: pdf
This application note describes the extraction, using ISOLUTE SLE+ supported liquid extraction columns, of opiate compounds from oral fluid matrix collected using the NeoSal device, prior to GC/MS analysis.