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: 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: 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: 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: P153Issued year: 2016File size: 0.52mbFile type: pdf
In postmortem cases, where drugs or pesticides have been used for
their poisonous properties, traditional matrices such as urine and
whole blood may be inappropriate for qualitative and quantitative
analysis. As the site of metabolism for most drugs and toxins, the
liver may provide more insight to cause of death than other bodily
This poster describes the use of ISOLUTE SLE+ supported liquid extraction columns to extract a range of drug and pesticide classes form homogenised liver using a simple, streamlined workflow.
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: AN769Issued year: 2013File size: 1.2mbFile type: pdf
This application note describes a Supported Liquid Extraction (SLE) protocol for the extraction of various drugs of abuse from non- hydrolyzed urine prior to LC-MS/MS analysis.
drugs of abuse, sle, multiclass
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: AN864Issued year: 2016File size: 2.43mbFile type: pdf
This application note describes the extraction of a broad range of analytes from liver tissue matrix prior to GC/MS analysis using ISOLUTE® SLE+ supported liquid extraction columns.
A protocol has been developed that allows the simultaneous
extraction of various drugs of abuse classes: amphetamines, barbiturates, benzodiazepines, cocaine, ketamine, THC.
In addition to these drug panels, simultaneous extraction of carbamate, organochlorine, organophosphate, pyrethroid, and triazine pesticide classes is demonstrated.