Development of a high-throughput mass spectrometry based analytical method to support anin vitroOATP1B1 inhibition screening assay

2016 ◽  
Vol 30 (15) ◽  
pp. 1787-1796 ◽  
Author(s):  
Andrew D. Wagner ◽  
Lisa Elkin ◽  
Kathy Mosure ◽  
Lizbeth Gallagher ◽  
Lindsey K. Stavola ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Throughput ◽  
Analytical Method ◽  
Screening Assay

scholarly journals Application of an innovative high-throughput liquid chromatography-tandem mass spectrometry method for simultaneous analysis of 18 hazardous drugs to rule out accidental acute chemotherapy exposures in health care workers

2019 ◽  
Vol 26 (4) ◽  
pp. 794-802 ◽  
Author(s):  
Pan Shu ◽  
Ting Zhao ◽  
Bo Wen ◽  
Kari Mendelsohn-Victor ◽  
Duxin Sun ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Health Care ◽  
Tandem Mass Spectrometry ◽  
High Throughput ◽  
Health Care Workers ◽  
Analytical Method ◽  
Tandem Mass ◽  
Care Workers ◽  
Hazardous Drugs ◽  
Rule Out

Objectives Despite safe handling guidelines published by several groups, health care worker exposure to hazardous drugs continues to occur due to suboptimal engineering controls and low use of protective equipment. Simple, multi-target and specific analytical methods are needed so that acute exposures to these drugs in the workplace can be assessed rapidly. Our aim was to develop an analytical method for simultaneous detection and quantification of widely used cancer drugs to rule out accidental acute chemotherapy exposures in health care workers. Methods We examined the feasibility of alternate high-performance liquid chromatographic-tandem mass spectrometry methods to simultaneously detect eighteen chemotherapy analytes in plasma and urine. The linear concentration ranges tested during assay development were 0.1–50 ng/mL. After development of a multi-analyte assay protocol, plasma samples (n = 743) from a multi-center cluster-randomized clinical trial (n = 12 sites) of an hazardous drug educational intervention were assayed. Confirmatory assays were performed based on the individual acute-spill case-histories. Results An innovative HPLC-multiple reaction monitoring-information dependent acquisition-enhanced production ion (MRM-IDA-EPI) analytical method was developed to simultaneously detect: cytarabine, gemcitabine, dacarbazine, methotrexate, topotecan, mitomycin, pemetrexed, irinotecan, doxorubicin, vincristine, vinblastine, ifosamide, cyclophosphamide, vinorelbine, bendamustine, etoposide, docetaxel, and paclitaxel. The retention times ranged from 4 min to 13 min for the analytical run. The limit of detection (MRM-IDA-EPI) and limit of quantitation (MRM) was 0.25 ng/mL and 0.1 ng/mL, respectively for most analytes. No detectable plasma concentrations were measured at baseline, post-intervention and in cases of documented acute spills. Use of a secondary tandem mass spectrometry approach was able to successfully rule out false positive results. Conclusions Development of a sensitive high-throughput multi-analyte cancer chemotherapy assay is feasible using an MRM-IDA-EPI method. This method can be used to rapidly rule out systemic exposure to accidental acute chemotherapy spills in health care workers.

scholarly journals Optimization of a Higher Throughput Microsomal Stability Screening Assay for Profiling Drug Discovery Candidates

2003 ◽  
Vol 8 (4) ◽  
pp. 453-462 ◽  
Author(s):  
Li Di ◽  
Edward H. Kerns ◽  
Yan Hong ◽  
Teresa A. Kleintop ◽  
Oliver J. Mc Connell ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Drug Discovery ◽  
High Throughput ◽  
High Throughput Screening ◽  
Parent Compound ◽  
Metabolic Stability ◽  
Screening Assay ◽  
Drug Candidates ◽  
Increasing Demand ◽  
The Stability

Metabolic stability plays an important role in the success of drug candidates. First-pass metabolism is one of the major causes of poor oral bioavailability and short half-life. Traditionally, metabolic stability was evaluated at a later stage of drug discovery and required laborious manual manipulations. With the advance of high-throughput screening, combinatorial chemistry, and early profiling of drug-like properties, automated and rapid stability assays are needed to meet the increasing demand of throughput, speed, and reproducibility at earlier stages of drug discovery. The authors describe optimization of a simple, robust, high-throughput microsomal stability assay developed in a 96-well format. The assay consists of 2 automated components: robotic sample preparation for incubation and cleanup and rapid liquid chromatography/mass spectrometry/mass spectrometry (LC/MS/MS) analysis to determine percent remaining of the parent compound. The reagent solutions and procedural steps were optimized for automation. Variables affecting assay results were investigated. The variability introduced by microsome preparations from different sources (various vendors and batches) was studied and indicates the need for careful control. Quality control and normalization of the stability results are critical when applying the screening data, generated at different times or research sites, to discovery projects.

Simple high-throughput analytical method using ultra-performance liquid chromatography coupled with tandem mass spectrometry to quantify total 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol in urine

2015 ◽  
Vol 53 (8) ◽  
Author(s):  
Jun-Young Yang ◽  
Hyun-Kyong Ahn ◽  
Si-Won Lee ◽  
You-Jung Han ◽  
Young-Jun Oh ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
High Throughput ◽  
Tobacco Smoke ◽  
Analytical Method ◽  
Ultra Performance Liquid Chromatography ◽  
Urinary Concentration ◽  
Tandem Mass ◽  
Biomarker Of Exposure

AbstractSince the urinary concentration of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) is a reliable biomarker of exposure to tobacco smoke, we developed a relatively simple high-throughput chromatographic method to quantify total urinary NNAL concentrations in the general population.The high-throughput analytical method was developed using ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) to identify and quantify total urinary NNAL concentrations in 10 non-smokers and 15 otherwise healthy smokers.Loss of nitric oxide atAn UPLC-MS/MS analytical method to quantify total urinary NNAL concentrations in smokers that does not require sample derivatization is presented herein. The method could be useful in clarifying the toxicities associated with human exposure to cigarette smoking. However, quantification might be adversely affected by co-eluting interfering compounds or selective ion suppression or enhancement as a result of having only one ion transition to monitor NNAL and NNAL-methyl-

Development of a high-throughput screening assay for stearoyl-CoA desaturase using rat liver microsomes, deuterium labeled stearoyl-CoA and mass spectrometry

2008 ◽  
Vol 627 (1) ◽  
pp. 105-111 ◽  
Author(s):  
Patricia Soulard ◽  
Meg McLaughlin ◽  
Jessica Stevens ◽  
Brendan Connolly ◽  
Rocco Coli ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Rat Liver ◽  
High Throughput ◽  
High Throughput Screening ◽  
Liver Microsomes ◽  
Rat Liver Microsomes ◽  
Screening Assay ◽  
High Throughput Screening Assay ◽  
Stearoyl Coa Desaturase

Development of a High-Throughput Screening Assay to Identify Inhibitors of the Major M17-Leucyl Aminopeptidase from Trypanosoma cruzi Using RapidFire Mass Spectrometry

2020 ◽  
Vol 25 (9) ◽  
pp. 1064-1071
Author(s):  
Maikel Izquierdo ◽  
De Lin ◽  
Sandra O’Neill ◽  
Martin Zoltner ◽  
Lauren Webster ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Trypanosoma Cruzi ◽  
High Throughput ◽  
High Throughput Screening ◽  
Drug Targets ◽  
Signal To Noise Ratio ◽  
Screening Method ◽  
Cellular Functions ◽  
Screening Assay ◽  
High Throughput Screening Assay

Leucyl aminopeptidases (LAPs) are involved in multiple cellular functions, which, in the case of infectious diseases, includes participation in the pathogen-host cell interface and pathogenesis. Thus, LAPs are considered good candidate drug targets, and the major M17-LAP from Trypanosoma cruzi (LAPTc) in particular is a promising target for Chagas disease. To exploit LAPTc as a potential target, it is essential to develop potent and selective inhibitors. To achieve this, we report a high-throughput screening method for LAPTc. Two methods were developed and optimized: a Leu-7-amido-4-methylcoumarin–based fluorogenic assay and a RapidFire mass spectrometry (RapidFire MS)–based assay using the LSTVIVR peptide as substrate. Compared with a fluorescence assay, the major advantages of the RapidFire MS assay are a greater signal-to-noise ratio as well as decreased consumption of enzyme. RapidFire MS was validated with the broad-spectrum LAP inhibitors bestatin (IC50 = 0.35 μM) and arphamenine A (IC50 = 15.75 μM). We suggest that RapidFire MS is highly suitable for screening for specific LAPTc inhibitors.

High-performance mass spectrometry as a drug discovery tool: a high-throughput screening assay to identify RNA-binding ligands

2001 ◽  
Author(s):  
Kristin A. Sannes-Lowery ◽  
Jared J. Drader ◽  
Richard H. Griffey ◽  
Steven A. Hofstadler
Keyword(s):  
Mass Spectrometry ◽  
Drug Discovery ◽  
High Throughput ◽  
High Throughput Screening ◽  
High Performance ◽  
Rna Binding ◽  
Screening Assay ◽  
High Throughput Screening Assay

scholarly journals High-Throughput Screening Assay for Sphingosine Kinase Inhibitors in Whole Blood Using RapidFire® Mass Spectrometry

2011 ◽  
Vol 16 (2) ◽  
pp. 272-277 ◽  
Author(s):  
Maureen K. Highkin ◽  
Matthew P. Yates ◽  
Olga V. Nemirovskiy ◽  
William A. Lamarr ◽  
Grace E. Munie ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Throughput ◽  
Whole Blood ◽  
High Throughput Screening ◽  
Kinase Inhibitors ◽  
Screening Assay ◽  
High Throughput Screening Assay ◽  
Mass Spectrometry Technology ◽  
Sphingosine 1 Phosphate ◽  
Mass Spectrometry System

To facilitate discovery of compounds modulating sphingosine-1-phosphate (S1P) signaling, the authors used high-throughput mass spectrometry technology to measure S1P formation in human whole blood. Since blood contains endogenous sphingosine (SPH) and S1P, mass spectrometry was chosen to detect the conversion of an exogenously added 17-carbon-long variant of sphingosine, C17SPH, into C17S1P. The authors developed procedures to achieve homogeneous mixing of whole blood in 384-well plates and for a method requiring minimal manipulations to extract S1P from blood in 96- and 384-well plates prior to analyses using the RapidFire® mass spectrometry system.

Mass Spectrometry Imaging of Neurotransmitters

2020 ◽  
Author(s):  
Katherine A. Stumpo
Keyword(s):  
Mass Spectrometry ◽  
High Throughput ◽  
Analytical Method ◽  
Biological Sample ◽  
Mass Spectrometry Imaging ◽  
Broad Applicability ◽  
Chemical Background ◽  
Target Molecules ◽  
Small Molecule Detection ◽  
New Strategies

Mass spectrometry imaging (MSI) is a powerful analytical method for the simultaneous analysis of hundreds of compounds within a biological sample. Despite the broad applicability of this technique, there is a critical need for advancements in methods for small molecule detection. Some molecular classes of small molecules are more difficult than others to ionize, e.g., neurotransmitters (NTs). The chemical structure of NTs (i.e., primary, secondary, and tertiary amines) affects ionization and has been a noted difficulty in the literature. In order to achieve detection of NTs using MSI, strategies must focus on either changing the chemistry of target molecules to aid in detection or focus on new methods of ionization. Additionally, even with new strategies, the issues of delocalization, chemical background noise, and ability to achieve high throughput (HTP) must be considered. This chapter will explore previous and up-and-coming techniques for maximizing the detection of NTs.

High‐throughput screening assay for the quantification of Cer d18:1/16:0, d18:1/24:0, d18:1/24:1, d18:1/18:0, d18:1/14:0, d18:1/20:0, and d18:1/22:0 in HepG2 cells using RapidFire mass spectrometry

2020 ◽  
Vol 34 (5) ◽  
Author(s):  
Sreekanth Dittakavi ◽  
Lavanya Mahadevan ◽  
Devaraj V. Chandrashekar ◽  
Ravi Kanth Bhamidipati ◽  
Juluri Suresh ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Throughput ◽  
Hepg2 Cells ◽  
High Throughput Screening ◽  
Screening Assay ◽  
High Throughput Screening Assay
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