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: P044Issued year: 2012File size: 0.89mbFile type: pdf
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,
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: P157Issued year: 2017File size: 0.8mbFile type: pdf
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
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: 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: P050Issued year: 2013File size: 0.46mbFile type: pdf
The use of schedule I drugs for patient pain management therapy warrants constant monitoring of therapeutic levels in patient urine samples. Screening patient urine samples is further complicated by the metabolic process which converts the free drug to the - glucuronide form. The target drugs in patient urine samples can be enzymatically hydrolysed and extracted using Supported Liquid Extraction (ISOLUTE SLE+) which offers an efficient alternative to traditional liquid-liquid extraction (LLE) and solid phase extraction (SPE) techniques typically used for this type of clinical sample preparation.
San Diego, MSACL, 2013
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: IST1044AIssued year: 2011File size: 0.11mbFile type: pdf
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.
UCT CleanScreen DAU, Varian Bond Elut Certify, ISOLUTE, Drugs of Abuse, Neutral, Basic, Acidic,
Part No: AN731Issued year: 2010File size: 0.09mbFile type: pdf
This procedure is recommended for the extraction of extensive panel of significant therapeutic and illicit drugs of abuse from whole blood and urine using mixed-mode cation exchange SPE columns. ISOLUTE HCX is a silica based mixed-mode sorbent that combines non-polar and strong cation exchange retention mechanisms for extraction of basic drugs from biological fluids.
Ion exchange, Drugs of Abuse, DOA, Clinical,
Part No: AN751Issued year: 2011File size: 0.36mbFile type: pdf
Benzodiazepines are psychoactive drugs used to treat various conditions including anxiety, insomnia, seizures and alcohol withdrawal. Long-term use can cause physical dependence and withdrawal upon cessation of use, due to this their application as medication has to be monitored and measured. This application note details an optimized extraction protocol for the use of ISOLUTE SLE+ supported liquid extraction plates (96-well) and 1 mL sample volume columns to extract a range of benzodiazepines from human urine. This simplified and efficient extraction method has analyte recoveries from 85% with LOQs as low as 100 pg/mL.
SLE+, SUPPORTED LIQUID EXTRACTION, BENZOS, BENZODIAZEPINES, URINE,