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: 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: P137Issued year: 2015File size: 0.37mbFile type: pdf
This poster examines various factors in the development and optimisation of a SPE-LC-MS/MS method for extraction of catecholamines from plasma.
The final method utilizes EVOLUTE EXPRESS WCX plates in either standard SPE or fast Load-wash-elute modes, and incorporates a series of wash steps to optimise analytical sensitivity. Low elution volumes mean no evaporation step is required, and LLOQ is 20mg/mL or lower for each of the analytes.
Part No: P137v2Issued year: 2016File size: 0.39mbFile type: pdf
This poster investigates various parts of the SPE method development process to develop a highly sensitive LC-MS/MS method for analysis of catecholamines (epinephrine, norepinephrine, dopamine).
A number of steps are optimized to improve sensitivity of the
analytes. This includes reducing the buffer strength in the mobile
phase, reducing the buffer strength of washes, careful pH control
throughout the extraction, avoiding an evaporation step and
incorporating IPA in the reconstitution solvent.
Part No: PPS362Issued year: 2014File size: 1.23mbFile type: pdf
This product sheet compares automated sample preparation using the Biotage®Extrahera™ to an equivalent manual method utilizing a vacuum manifold. A selection of beta blocker drugs were extracted from pooled
stripped plasma using a supported liquid extraction procedure.
Part No: PPS366Issued year: 2014File size: 1.76mbFile type: pdf
Automated sample preparation using the Biotage®Extrahera™ was compared to an equivalent manual method utilizing a vacuum manifold. Analytes were extracted from pooled stripped plasma using a supported liquid extraction procedure.
Part No: PPS361Issued year: 2014File size: 1.28mbFile type: pdf
This document compares automated sample preparation using the Biotage®
Extrahera™ to an equivalent manual method utilizing a vacuum manifold. A selection of non-steroidal anti-inflammatory drugs (NSAIDS) were extracted from pooled stripped plasma using a supported liquid extraction procedure.
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: P088Issued year: 2014File size: 1.06mbFile type: pdf
This poster evaluates the performance of a novel 96-well filter plate (ISOLUTE PLD+ Protein and Phospholipid Removal Plate) for
the simultaneous removal of proteins and phospholipids from serum samples prior to LC-MS/MS analysis.
Part No: P104Issued year: 2014File size: 0.77mbFile type: pdf
This poster presents the ISOLUTE PLD+ Protein and Phospholipid removal plate, highlighting its ease of use and excellent performance with respect to sample clean up, analyte recovery and elimination of back pressure build up in the UPLC system.
Part No: P026Issued year: 2008File size: 0.36mbFile type: pdf
Matrix components, particularly salts, proteins and
phospholipids, can lead to ion suppression resulting in
inaccurate quantitation and reduced detection limits.
Resin-based mixed-mode cation exchange SPE sorbents
are widely used for the extraction of basic compounds
from biological fluids. The dual retention mechanism
allows a two stage interference wash protocol, which
results in extremely clean extracts.
This poster will investigate the performance of
EVOLUTE™ CX for the extraction of a wide range of basic
drugs from plasma, showing high analyte recovery and
advanced extract cleanliness. The analyte suite was
selected to cover a variety of basic analytes with wide
ranging polarities (LogP values). Extract cleanliness
experiments were performed showing overall ion
suppression, then individual matrix components examined
in terms of protein and phospholipid removal.
Part No: AN830Issued year: 2014File size: 1.44mbFile type: pdf
This application note describes the use of the ISOLUTE® PLD+ Protein and Phospholipid Removal Plate for clean-up of a range of acidic, basic and neutral drugs from plasma. ISOLUTE® PLD+ Protein and Phospholipid Removal Plates are suitable for clean-up of a range of analytes with widely differing functionality and polarity characteristics from plasma, giving high recoveries, good reproducibility and excellent extract cleanliness.
Part No: AN889Issued year: 2018File size: 1.03mbFile type: pdf
This application note describes a solid phase extraction protocol for the extraction of three catecholamine metabolites (vanillylmandelic acid, homovanillic acid and 5-hydroxyindoleacetic acid) from plasma prior to LC-MS/MS detection. The method described in this application achieves high reproducible recoveries and excellent linearity (>0.99) in the range 10-200 ng/mL.
Part No: AN888Issued year: 2017File size: 0.91mbFile type: pdf
This application note describes a supported liquid extraction protocol for the extraction of three acidic catecholamine metabolites (vanillylmandelic acid, homovanillic acid and 5-hydroxyindoleacetic acid) from plasma prior to LC-MS/MS detection. The method described in this application note achieves high reproducible recoveries (>80%, RSD < 5%) for these analytes in plasma with linearity > 0.995 in the range
Part No: AN702Issued year: 2011File size: 0.11mbFile type: pdf
This Application Note describes the extraction of a variety of basic drugs from plasma using EVOLUTE® CX mixed-mode resin-based SPE. The analyte suite includes basic drugs with wide ranging pKa and logP values.
Basic Drugs, Biotage, Ion Exchange, EVOLUTE, Pharmaceuticals
Part No: P170Issued year: 2017File size: 0.58mbFile type: pdf
Catecholamine metabolites vanillylmandelic acid (VMA), homovanillic acid (HVA) and 5-hydroxyindoleacetic acid (5-HIAA) are biomarkers for neuroblastoma and catecholaminesecreting tumors.
This poster describes optimization of the method development process to maximize analyte sensitivity in the extraction and quantitation of these metabolites from plasma. Parameters investigated include MRM transitions, chromatography and sample preparation protocols.
MSACL EU 2017
Part No: P176Issued year: 2018File size: 0.72mbFile type: pdf
Catecholamine metabolites vanillylmandelic acid (VMA), homovanillic acid (HVA) and 5-hydroxyindoleacetic acid (5 HIAA) are biomarkers for neuroblastoma and catecholamine-secreting tumors.
This poster presents optimization of the method development process to maximize analyte sensitivity in the extraction and quantitation of these metabolites from plasma. Sample preparation protocols using solid phase extraction (EVOLUTE EXPRESS ABN and AX) and supported liquid extraction (ISOLUTE SLE+) are presented.
MSACL NA 2018 Palm Springs, CA
Part No: AN711Issued year: 2011File size: 0.11mbFile type: pdf
The following methods have been developed and optimized for the extraction of chloramphenicol from a variety of sample matrixes including milk, plasma, honey, urine, and shrimp/prawns for subsequent LC-MS/MS analysis. The methods are highly reproducible and offer low limits of detection.
Part No: AN602Issued year: 2011File size: 0.23mbFile type: pdf
This Application Note highlights the use of ISOLUTE® SLE+ supported-liquid extraction plates for the extraction of corticosteroids from human plasma, specifically Triamcinolone, Prednisolone, Hydrocortisone, Prednisone, Betamethasone, Cortisone, Dexamethasone, Flumethasone, Corticosterone, Beclomethasone, Triamcinolone, Fluocinolone, Budesonide 1, Budesonide 2 and 5-Pregnen-3β−ol-20-one.