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: AN818.v1Issued year: 2014File size: 1.54mbFile type: pdf
This application note describes the extraction of ethyl glucuronide and ethyl sulfate from human urine using ISOLUTE® NH2 solid phase extraction columns. The method has been applied real patient samples that had been previously analyzed with a validated referee method. The results of the orthogonal measurements agreed, to provide similar diagnostic values.
Part No: P091Issued year: 2014File size: 0.38mbFile type: pdf
This report details a “load, wash, elute” weak cation exchange solid phase extraction procedure amenable to both plasma and urine samples. The extracts are subsequently injected into an LC-MS/MS system. The preliminary sample preparation method was developed at the Biotage US Applications lab (Charlotte, NC).The method was then transferred to Ionics (Bolton, ON, Canada) to facilitate the nmole/L measurements of the selected biomarkers
by laminar flow tandem mass spectrometry. The SPE-LC-ESIMS/ MS method parameters were first optimized using pooled mixed gender plasma. A set of patient samples (n=32) was later supplied by the Mayo Clinic (Rochester, MN) that had previously been analyzed by a validated referee method. The population represented measured values across a range of clinical relevance.
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: 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: AN793Issued year: 2013File size: 2.04mbFile type: pdf
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.
Part No: RP-DS-04Issued year: 2010File size: 0.19mbFile type: pdf
When analyzing human urine for drugs of abuse, one of the most common tests is for the cocaine metabolite Benzoylecgonine (BZE). A contract laboratory has automated this labor intensive procedure using the RapidTrace.
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: P143Issued year: 2016File size: 0.27mbFile type: pdf
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+).
Part No: PN43Issued year: 2011File size: 0.27mbFile type: pdf
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
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: P045Issued year: 2012File size: 0.32mbFile type: pdf
This poster will demonstrate SPE sample preparation strategies for the simultaneous extraction of both ethyl glucuronide and ethyl sulphate from urine, allowing cleaner extraction protocols compared to traditional dilute and shoot approaches.
EtG, EtS, Urine, Poster, SOFT, Forensic, UPLC, LC-MS/MS, EVOLUTE, AX, WAX
Part No: P127Issued year: 2015File size: 0.17mbFile type: pdf
EVOLUTE® EXPRESS SPE columns and plates combine sorbent wettability with optimized SPE components, allowing better flow consistency and in many cases eliminating the need for SPE column conditioning; thus simplifying and reducing extraction processing. This poster compares the performance of various EVOLUTE EXPRESS column chemistries using 10-500 mg bed weights, in order to investigate whether sorbent conditioning and equilibration steps are truly required.
Part No: AN777Issued year: 2013File size: 0.86mbFile type: pdf
Cortisol is a steroid hormone that in urine can be used to diagnose hyper or hypo cortisol diseases such as Cushing’s Syndrome. This methodology has been designed to give an effective and efficient supported liquid extraction protocol for the clean-up and concentration of urinary cortisol levels. Analyte recoveries achieved using this method ranged from 99-101% with RSDs below 5% for all analytes.
Cortisol, Steroid hormone, Cushing's Syndrome, Cushing's Disease, Clinical, SLE, Supported Liquid Extraction, Stress,