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: AN886Issued year: 2017File size: 1mbFile type: pdf
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.
Part No: P164Issued year: 2017File size: 0.69mbFile type: pdf
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.
Part No: P163Issued year: 2017File size: 0.58mbFile type: pdf
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.
Part No: AN918Issued year: 2019File size: 1.33mbFile type: pdf
This application note describes the fully-automated extraction of opiates from acid-hydrolyzed urine and non-hydrolyzed urine prior to GC/MS analysis. The methods were automated using Biotage® Extrahera™ configured for use with ISOLUTE® SLE+ supported liquid extraction columns.
Using the Biotage® Extrahera™, 24 hydrolyzed samples were extracted in under 31 minutes and 24 non-hydrolyzed samples were extracted in under 36 minutes. The limits of quantitation meet or exceed the sensitivity requirements set by SAMHSA and EWDTS for workplace testing applications.
Part No: PPS443.V.1Issued year: 2019File size: 2.98mbFile 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: P194Issued year: 2019File size: 1.08mbFile type: pdf
Sample preparation for hair analysis is often lengthy involving multiple manual labor steps from cutting, washing, homogenization/pulverization, digestion, sample extraction and analysis. This poster aims to demonstrate a streamlined sample
preparation workflow for hair analysis.
The workflow is evaluated using a suite of drugs of abuse, including THC and metabolites.
MSACL NA 2019
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
SOFT 2017, Boca Raton
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: AN768Issued year: 2013File size: 1.86mbFile type: pdf
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