LC–MS

Using a liquid chromatography–mass spectrometry (LC–MS) method in conjunction with two complementary types of chromatographic retention modes - reversed phase and aqueous normal phase - various compounds present in mesquite flour extracts were identified. Because of the diverse types of chemical constituents found in such natural product extracts, a single chromatographic mode may not be sufficient for a comprehensive characterization. However, the combination of reversed-phase and aqueous normal phase LC can encompass a wide range of analyte polarity. This characterization of the composition of mesquite flour could be used in future studies to elucidate the beneficial health effects of its consumption.

Part IV of this series takes a closer look at clustering. Clustering can be very useful at observing your data when the sample dimensionality is large. This is a barbarian term meaning that diversity among your samples may be wide. In that case, the space reduction provided by principal component analysis (PCA) is not always convincing, because the simplification provided by a single two-dimensional plot erases too much information. Clustering allows you to preserve more information.

Tips and tricks to help you recognize and solve the most common SPE problems.

How to optimize the key variables in LC–MS analysis

Here we propose an exemplary workflow for the analysis of phenolic extracts (i.e. wine) enabling confident differential analysis using high performance liquid chromatography in combination with low-field drift tube ion mobility quadrupole time-of-flight mass spectrometry (HPLC×IMS-QTOFMS). In this workflow, single-field collisional cross section values from low-field drift-tube IMS using nitrogen as drift gas (DTCCSN2) are readily extracted in addition to a retention time and a high resolution mass spectrum for each compound. “Alternating frames” experiments utilizing post-drift tube fragmentation also allow drift time-aligned MS/MS spectra to be obtained. Molecular feature extraction was highly repeatable with average precision values of 0.28% for retention time, 0.18% for drift time, and 1.5 ppm m/z determined for 233 molecular features found in all six technical replicates. The improved selectivity of this strategy increases confidence in intersample molecular feature alignment (i.e. compound identity confirmation), including the resolution of co-eluting isomeric compounds.

The power of nontargeted metabolite profiling is illustrated in a study focused on the determination of molecular markers in malting barley that are predictive of desirable malting quality for brewing applications. The metabolite extraction, detection, and analysis methods are high throughput and reproducible, and therefore, this approach represents a practical addition to the plant breeder’s molecular toolbox.

This study of a selected group of PPCP contaminants in eastern North Carolina demonstrates the advantages and disadvantages of LC–MS and GC–MS as well as SPE and liquid–liquid extraction.

An LC–MS method for simultaneous quantification of buprenorphine and three metabolites: norbuprenorphine, buprenorphine glucuronide, and norbuprenorphine glucuronide.

Validation of this rapid bioanalytical method for the determination of moxidectin in cattle hair demonstrated that the method is accurate, reliable, and reproducible.

High-resolution mass spectrometry (MS) is not readily compatible with chromatographic techniques using high-salt mobile phases. This study presents the development and use of 2D-LC–MS via an on-line desalting step for the analysis of monoclonal antibodies and antibody–drug conjugates.

Measuring trypsin inhibitors in legumes is important to feed processors, who are concerned with providing high-quality products for animal feed. In this study, a rapid, accurate, and precise method for the quantification of trypsin inhibitor activity is evaluated.

The development of various analytical MS methods to investigate the chemical composition of liquids used in electronic cigarettes and characterize their quality is presented in this study.

A review of the central role LC–MS plays in characterizing mAbs and biosimilars, including highlights of top-down and middle-up approaches to measure the intact masses of mAbs and their subunits, the current state of peptide mapping techniques, and glycoprofiling methods.

The last decade has seen a series of advances in the field of liquid chromatography that have resulted in improvements for many clinical diagnostic services. These innovations have included the expansion of superficially porous particle columns, new or improved stationary phase options, and “user-friendly” multiple-channel HPLC instrument options that allow sequential analysis-a boon for low and moderate throughput laboratories with limited hardware. As a result, diagnostic services are able to offer faster turn-around-times and measure analytes in patient types and disease states that were previously problematic. This article presents examples of the impact these innovations have had in a number of hospital settings.

Adequate detection of trans-resveratrol in wine is complicated by two factors: relatively low levels and interferences from matrix components. Here, we present two useful approaches to overcoming these issues depending on the instrumentation available. For HPLC–UV analyses, matrix peaks can be removed by microextraction using packed sorbent while simultaneously concentrating the trans-resveratrol peak by a factor of two. For LC–MS, the extracted ion chromatogram for the [M + H]+ analyte ion can be used to obtain specificity without prior extraction procedures.

This article presents a step-by-step DoE optimization strategy for an electrospray ionization source with the aim to quantitatively analyze 18 compounds in a metabolomic study of human urine.

This article provides useful tips for smooth validation of multi-analyte LC–MS-MS methods and summarizes important validation outcomes for 295 analytes, including more than 200 mycotoxins.

Non-targeted metabolite profiling by ultrahigh-performance liquid chromatography coupled with mass spectrometry (UHPLC–MS) is a powerful technique to investigate the influence of genetic and environmental influence on metabolic phenotype in plants. The approach offers an unbiased and in-depth analysis that can reveal molecular markers of desirable phenotypic traits which can be complementary to genetic markers in plant breeding efforts. Here, the power of non-targeted metabolite profiling is illustrated in a study focused on the determination of molecular markers in malting barley that are predictive of desirable malting quality for brewing applications.

Experiments presented here demonstrate the suitability of LC–SAI-MS for the detection and quantification of pharmaceuticals, with limits of detection in the low parts-per-trillion range. A comparison of LC–ESI-MS to LC–SAI-MS also yielded favorable results for SAI.

Enzyme immunoassay (EIA) is a conventional drug screening technique, but it can be limited by cross-reactivity that can lead to high false positive rates.

We recently spoke to Gary Duncan and Wendy Russell of the Rowett Institute of Nutrition & Health in Aberdeen, Scotland, about the significance of phytochemical bioavailability to human health and the important role of liquid chromatography linked to tandem mass spectrometry (LC–MS-MS) in their research.

Click here to view the complete E-Separation Solutions newsletter from February 20, 2014.

Secondary parameters in the interface and mass analyzer can have a major impact on sensitivity and reproducibility. Here, we examine how and when to consider optimizing these parameters through a study of the working principles of LC–MS analysis.

We assessed a simple method based on recovery for the detection of matrix effects and two alternative methods for the rectification of matrix effects in LC–MS: standard addition and the coeluting internal standard method.

Redox reactions are integral parts of many cellular processes. Thus, they are extensively studied in vitro and in vivo. Electrochemistry (EC) represents a pure instrumental approach to characterize direct and indirect effects of redox reactions on bioorganic molecules.