Part No: MSACL US 2018 Breakfast SeminarIssued year: 2018File size: 3.24mbFile type: pdf
The use of LC/MS analysis in the clinical lab has increased exponentially over the last 10 years. Modern mass spectrometers are extremely sensitive allowing low level detection of many target analytes. However, this sensitivity can come at a price, in that levels of contamination not previously detected with less sensitive instruments can now have larger impact on analysis. The complexity of common matrices such as plasma/serum and urine while presenting different challenges can have a marked influence on method performance.
As a result sample preparation is an extremely important part of the process in order to provide robust methods. This seminar focuses on some of the method development challenges our lab faced when looking at two clinical assays:
Endogenous steroid hormone extraction from serum, and catecholamine extraction from plasma and urine.
Particular emphasis is placed on the various sample preparation options we looked at for the extraction of these analytes. During optimization of the extraction process we investigated analyte recovery, co-extractable matrix components, HPLC column degradation, calibration curve performance and limits of quantitation.
MSACL US 2018 Breakfast Seminar
Part No: P175Issued year: 2018File size: 2.14mbFile type: pdf
This poster presents a method for detection of approximately 100 drugs and metabolites in urine using mixed-mode polymeric cation exchange solid phase extraction (SPE).
Four different glucuronidase enzymes and several incubation times and temperatures were evaluated to determine optimum hydrolysis conditions for seven commonly detected glucuronide metabolites. Samples were subjected to offline hydrolysis and SPE (EVOLUTE® EXPRESS CX, Biotage, Charlotte, NC) and results compared to an SPE plate that incorporates in-well hydrolysis (EVOLUTE HYDRO CX, Biotage).
MSACL 2018, Palm Springs, CA
Part No: P155Issued year: 2017File size: 0.61mbFile type: pdf
This poster discusses the impact of optimization of various parts of the method development process to maximize assay performance while delivering a combined assay for the analysis of urinary catecholamines and metanephrines. The extraction method using EVOLUTE EXPRESS WCX 96-well plates is successfully automated using the Biotage® Extrahera™ Automated Sample Preparation Platform.
MSACL 2017, Palm Springs
ASMS 2017, Indianapolis
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
SOFT 2017, Boca Raton
Part No: AN871Issued year: 2017File size: 3.46mbFile type: pdf
This application note describes a mixed-mode weak cation exchange SPE protocol for the extraction of three catecholamines (epinephrine, norepinephrine and dopamine) and three metanephrine metabolites (metanephrine, normetanephrine and 3-methoxytyramine) from urine prior to LC-MS/MS detection.
Included in this application note are the detailed settings for implementing the method on the Biotage® Extrahera(TM) automation system.
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: P165Issued year: 2017File size: 0.32mbFile type: pdf
This poster evaluates 3 different sample preparation approaches (ISOLUTE SLE+, EVOLUTE EXPRESS ABN, EVOLUTE EXPRESS CX) for extraction of large multi-drug urine panels.
Each approach is assessed in terms of suitability for extraction of analytes with different different properties (pka, LogP etc).
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: 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: 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: AN841Issued year: 2015File size: 0.82mbFile type: pdf
This application note describes the extraction of 11-nor-9-carboxy-THC from a urine matrix, prior to GC/MS analysis.
Carboxy-THC is the primary metabolite of THC, a key indicator of illicit marijuana usage. In urine, ~80% of the carboxy-THC metabolite is present in the form of its glucuronide metabolite. Therefore, to effectively quantify the THC-COOH, urine is hydrolyzed before extraction. This application note describes optimized extraction of urine samples prepared by either enzymatic or base hydrolysis.
Part No: AN843Issued year: 2015File size: 0.86mbFile type: pdf
Ethylestrenol and stanozolol are anabolic steroids which can be used to increase muscle mass and enhance performance. These drugs have been linked to instances of doping in race horses.
This application note describes a Supported Liquid Extraction (SLE) protocol for the extraction of ethylestrenol and stanozolol from horse urine prior to LC-MS/MS analysis.
The method achieves high reproducible analyte recoveries from both gelding and filly urine. Protocols for 48-well, 96-well plate and column formats are described.
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: AN846Issued year: 2015File size: 1.04mbFile type: pdf
This application note describes a simple and robust mixed mode cation exchange solid phase extraction protocol using EVOLUTE® EXPRESS CX 96 well plates for the extraction of buprenorphine and norbuprenorphine from urine. Excellent extract cleanliness allows low level quantitation (0.1 ng/mL) of the analytes using LC-MS/MS.
Part No: AN809Issued year: 2014File size: 1.3mbFile type: pdf
This application note describes the simultaneous extraction of THC and its major metabolites, including 11-nor-9-carboxy-Δ9-THC glucuronide, from urine using supported liquid extraction (ISOLUTE® SLE+ in both plate and column formats) prior to analysis by LC-MS/MS.