Part No: P070Issued year: 2014File size: 0.55mbFile type: pdf
This poster describes a strong cation exchange SPE protocol for extraction of Vitamin B3 (Niacin) and related metabolites from serum.
The ISOLUTE SCX-3 96 well plate format demonstrated as a viable option for serum measurements over a relevant concentration range in clinical diagnostics.
It is anticipated that strategies presented in this poster would be of broad-based interest to clinical laboratories focused on combining parent/metabolite assays into a single analysis.
Part No: AN814Issued year: 2014File size: 1.19mbFile type: pdf
This application note describes the extraction of Vitamin B3 (Niacin, nicotinic acid) and its key metabolites from serum using a strong cation exchange SPE retention mechanism. High reproducible analyte recoveries are achieved.
Part No: P081Issued year: 2014File size: 0.42mbFile type: pdf
This poster describes a reduced workflow solid phase extraction method for extraction of vitamin B7 (biotin) from human serum. Using EVOLUTE EXPRESS AX 96-well plates, the traditional sorbent conditioning and equilibration steps are not required, meaning the sample can be applied directly to the dry plate. Post extraction evaporation is also eliminated.
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: P136Issued year: 2015File size: 0.6mbFile type: pdf
This poster compares the use of supported liquid extraction (ISOLUTE® SLE+) and a novel protein and phospholipid depletion plate (ISOLUTE® PLD+), for the extraction of 25-hydroxy vitamin D. The manual extraction protocols were transferred to an SPE automation platform (Biotage® Extrahera)and method performance versus manual processing compared.
MSACL EU 2015
Part No: P142Issued year: 2016File size: 0.57mbFile type: pdf
This poster summarizes various sample preparation strategies for the
extraction of MMA from serum without the necessity for derivatization, prior to LC-MS/MS analysis. A range of sample preparation techniques of varying complexity were evaluated: protein precipitation, phospholipid depletion, supported liquid extraction and solid phase extraction using both silica and polymer-based mixed-mode anion exchange chemistries.
Method performance was evaluated for evaporative effects, assay recovery, ion suppression and phospholipid removal.
Part No: AN933Issued year: 2020File size: 2.25mbFile type: pdf
This application note describes the extraction of a panel of estrogenic hormones from human serum using EVOLUTE® EXPRESS ABN solid phase extraction plates prior to LC/MS analysis. The simple sample preparation procedure delivers clean extracts and analyte recoveries greater than 90% with RSDs lower than 5% for all analytes. Linearity of greater than 0.99 is achieved for Estrone (E1) and Estradiol (E2) from 5–2000 pg/mL, and 50–20000 pg/mL for Estriol (E3). No derivatization is required, and detection limits are enhanced using a fluorinated mobile phase.
Part No: P216.V.2Issued year: 2020File size: 2.46mbFile type: pdf
Monitoring estrogenic compounds such as Estrone (E1), Estradiol
(E2), and Estriol (E3) in human serum is essential in clinical research and diagnostics. Detection in men, post-menopausal women and children requires very low limits of detection (LODs) which can be challenging.
Part No: Issued year: 2014File size: 0.12mbFile type: pdf
This poster describes the development and validation of a method for supported liquid extraction of serum cortisol, with analysis by UPLC-MS/MS. The aim of this study was to develop a candidate reference method that could then be used to underpin the UK NEQAS Cortisol scheme.
MSACL EU 2014
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: P126Issued year: 2015File size: 0.48mbFile type: pdf
This poster demonstrates the use of a novel protein and phospholipid depletion plate, for the extraction of 25-hydroxy vitamin D from serum. The extraction protocol was ultimately transferred to an SPE automation platform and method performance versus manual processing was compared.
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: P064Issued year: 2014File size: 0.47mbFile type: pdf
This poster presents a simple method for the extraction and subsequent detection of both the traditional hydroxy metabolites and the biologically active dihydroxy metabolites in serum. High analyte recoveries and low ion suppression were demonstrated, allowing limits of quantitation at low pg/mL levels for the di-OH metabolites and < 1 ng/mL levels for the OH metabolites.
Part No: P120Issued year: 2015File size: 0.63mbFile type: pdf
This poster describes a comparison of 25-hydroxy Vitamin D Extraction using Supported Liquid Extraction and Phospholipid Depletion Plate Technology using Manual and Automated Sample Preparation prior to LC/MS Analysis. It was presented at MSACL 2015, San Diego
Part No: AN842.V.1Issued year: 2016File size: 2.19mbFile type: pdf
ISOLUTE® PLD+ Protein and Phospholipid Removal plates offer a substantial improvement in extract cleanliness compared to traditional protein precipitation techniques for bioanalytical sample preparation.
This application note describes a simple, effective ISOLUTE® PLD+ protocol for the extraction of 25-hydroxy vitamin D from serum, demonstrating high, reproducible analyte recoveries with low protein and phospholipid content in the extracts.
Included in this application note are the detailed settings for implementing the method on the Biotage Extrahera automation system.
Part No: AN939Issued year: 2020File size: 1.32mbFile type: pdf
This application note describes the extraction of steroid hormones from human serum using Biotage® Mikro ABN microelution plates prior to LC/MS-MS analysis. The simple sample preparation procedure delivers clean extracts and analyte recoveries mostly greater than 70% with RSDs lower than 5% for most analytes. Linearity of greater than 0.99 is achieved for all analytes from 1–1000 pg/mL. An elution volume of 30 μL is used, which is simply diluted prior to analysis, avoiding a time consuming evaporation step.
Part No: AN940Issued year: 2020File size: 1.28mbFile type: pdf
This application note describes the extraction of steroid hormones from human serum using Biotage® Mikro ABN microelution plates prior to LC/MS-MS analysis. The simple sample preparation procedure delivers clean extracts and analyte recoveries mostly greater than 90% with RSDs lower than 5% for most analytes. Linearity of greater than 0.99 is achieved for all analytes from 1–1000 pg/mL.
Note: if DHEAS is required, see application note AN939