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This instalment of Tips & Tricks shows the influence of analytical parameters on gel permeation chromatography/size-exclusion chromatography (GPC/SEC)–light scattering results when artificially wrong parameters reflecting typical experimental errors are used.

Matrix-assisted laser desorption–ionization time-of-flight mass spectrometry (MALDI-TOF-MS) is actually one of the most versatile tools for fingerprinting mixtures.

Traditionally mass detection instruments have been for the mass spec experts and not played a major role in the majority of chromatography labs. With the advent of smaller, more accessible mass detectors, the potential of mass data is coming more and more within the reach of the chromatographer. With this webcast we look to see how the landscape of the chromatography lab is changing and how the value of mass data can be realized by the chromatographer.

Click the title above to open The Column July 08, 2016 Europe & Asia issue, Volume 12, Number 12, in an interactive PDF format.

Click the title above to open The Column July 08, 2016 North American issue, Volume 12, Number 12, in an interactive PDF format.

How to optimize the key variables in GC analysis–GC columns and detectors

Microflow LC–MS-MS has seen a surge of attention, development, and popularity among research scientists and bioanalysts over the last few years. The potential of this technology to provide better sensitivity, less solvent waste, near-zero dead volume, and high through-put are a big part of this renewed interest. However, microflow LC techniques are hardly a new idea. More than 40 years ago, in 1974, a group at Nagoya University in Japan first developed a microcolumn liquid chromatography system, elements of which can be found in today’s commercial products. With the advances in technology over the last several years, development and implementation of this technique have been kicked into high gear. In this article, we discuss the history of microflow LC–MS-MS, the current state of the art, and where the future might lead for this rapidly growing technology.

Incognito debates the pros and cons of onshore support.

The winners of the 2016 Thermo Scientific Tandem Mass Tag (TMT) Research Award were celebrated at a ceremony held at the 64th American Society of Mass Spectrometry (ASMS) Conference on Mass Spectrometry and Allied Topics in San Antonio, Texas, USA.

Postnova Analytics has opened a dedicated office to enhance local support for its UK customers.

The quantitative performance of the latest generation of high-resolution instruments is comparable to that of a triple quadrupole MS, even though different scanning modes are used. Higher-resolution instrumentation also allows flexibility concerning compound identification because the experiment can be set up for targeted quantitation, screening, or both. In an Orbitrap-based instrument, the parallel reaction monitoring (PRM) mode performs most closely to a triple quadrupole mass analyzer using selected reaction monitoring (SRM) mode. This study looks at the performance of an Orbitrap-based LC–MS method for EPA Method 539.

A preview of the upcoming 31st International Symposium on Chromatography (ISC 2016), which will be held 28 August–1 September at University College Cork, Ireland.

The use of antibodies in “bottom-up” LC–MS workflows to determine low abundant biological active proteins in complex human samples has increased in recent years: immuno-capture analysis combines the workflow of conventional immunological assays with LC–MS analysis. This paper describes typical challenges, such as cross reactivity and the mass spectrometer’s dynamic range, as well as the advantages of isoform differentiation and multiplexing. Additionally, some experimental formats of immuno-capture bottom-up LC–MS analysis of biological active proteins in complex human samples will be discussed.

Ultrahigh-pressure liquid chromatography (UHPLC) instruments from different manufacturers and instruments with different configurations can produce significant variations in chromatographic separation. The variety in instrument configuration increases the complexity of the method development process, which now requires a more thorough evaluation of the effect of instrument variations on the method. The studies presented here determined the typical interinstrument variations in dwell volume, extracolumn dispersion, and mixing efficiency as measured by mobile-phase compositional accuracy. Additionally, the dwell volume and extracolumn dispersion were independently and systematically varied to evaluate the resulting impact on resolution for a small-molecule test mixture during gradient elution. To account for these interinstrument variations, dwell volume and wash-out volume method translation and adjustment techniques were evaluated.

A reliable autosampler is one key requirement for unattended operation of a liquid chromatograph.

While gas chromatographers may take their septa for granted, in fact these small and seemingly unremarkable polymer disks keep air out of the carrier-gas stream when used in an inlet and keep samples intact and uncontaminated when used in sample vials. Choosing the wrong septa can compromise method accuracy and repeatability as well as reduce column life in extreme cases. This instalment addresses septa for inlets and sample vials.

In recent years industry has been moving to columns with smaller and smaller inner diameters - moving from 4.6 mm and 3.0 mm i.d. columns to 2.1 mm, 1.0 mm, and even smaller. While small inner diameter columns have some clear advantages, they also bring challenges. Reduction of extracolumn volumes must be given greater consideration by both customers and manufacturers. This article focuses on the sources of band broadening within high performance liquid chromatography (HPLC) columns with an emphasis on eddy dispersion. The physical mechanisms of dispersion are discussed and a review of the current literature as it pertains to small inner diameter columns is presented.

Glycosylation of monoclonal antibody (mAb) therapeutics is widely recognized by the regulators and the industry as a critical quality attribute (CQA). Hence, it is necessary that glycosylation is measured and adequately controlled during production. This instalment reviews the various process parameters and raw material attributes that affect glycosylation, as well as the different analytical tools that are used for characterization, with greater emphasis on the chromatographic methods of analysis. Key recent advancements that have occurred in the past five years are also discussed briefly. While significant progress has been made in the monitoring of glycosylation, its real time control has yet to be demonstrated.

The 10th annual International Conference on Packed Column SFC (pcSFC) will be held 5–7 October 2016 at the Melia’ Vienna hotel in Vienna, Austria.

This work will demonstrate a simple methodology using automated solid-phase extraction (SPE) and HPLC coupled with mass spectrometric detection. This work will demonstrate a simple methodology using automated solid-phase extraction (SPE) and HPLC coupled with mass spectrometric detection. This work will demonstrate a simple methodology using automated solid-phase extraction (SPE) and HPLC coupled with mass spectrometric detection.

The Diablo 5000A RTGA-MS allows the process stream to be seen in real-time and provides quantitative data with reliable mass information. The result is visualization.

Click the title above to open the LCGC Europe July 2016 regular issue, Vol 29, No 7, in an interactive PDF format.