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: 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: P144Issued year: 2016File size: 0.6mbFile type: pdf
The ability to extract a broad range of different drugs from a biological matrix allows for the expedited analysis of a patient sample using LC-MS/MS. Typically small molecules are extracted from matrices like urine based on their polarities. A fast and reliable sample preparation method that could be implemented to extract drugs of different polarities from urine could be used as a screening tool to quickly identify the presence of illicit drugs in patient samples using LC-MS-MS.
This poster demonstrates the utility of supported liquid extraction for the extraction of over 30 different acidic, basic and neutral drugs in urine prior to LC-MS/MS.
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: 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: Issued year: 2016File size: 0.94mbFile type: pdf
Cortisol measurement in hair samples scan be a useful indicator of chronic stress. This poster compares a new LC-MS/MS method with an existing immunoassay based method.
The Biotage Bead Ruptor is used effectively for hair pre-treatment.
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: 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: 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: 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: AN094-HORIssued year: 2014File size: 1.41mbFile type: pdf
When a sample preparation technique calls for removal of compounds using SPE, it is standard practice to determine what concentrations will produce an acceptable linear range for the assay. Traditional linear regression is often performed manually using pipettes to generate an external standard curve that unknown samples are measured against.
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