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Quality and consistency in reagents is critical to successful drug discovery and development. When targeting a particular protein of interest, in vitro experiments should be performed with proteins of biological properties similar to those for in vivo tests. It is important that molecularity, purity, shape, and degree of heterogeneity remain the same when any alterations are made to the model protein or the formulation buffer. Multi-angle light scattering (MALS) combined with size-exclusion chromatography (SEC-MALS) is a very useful technique to monitor the solution properties of the protein as changes to reagents are made.

In this application note, an antibody–drug conjugate (ADC) was analyzed using a TSKgel® Butyl-NPR column, the least hydrophobic of the TSKgel HIC columns. Both unconjugated and drug conjugated Trastuzumab samples were successfully separated with baseline resolution. The baseline resolution enabled an easy integration and quantification of different drug pay loads in ADC characterization.

Projects in drug discovery and safety constantly aim at development of novel and safer drugs, therapeutics, and diagnostics. During active pharmaceutical ingredient (API) development, drug stereoisomerism is recognized as an issue having clinical and regulatory implications. Enantiomers have essentially identical physical and chemical properties, while potentially showing large differences in toxicity.

Chromatographic techniques with mass spectrometric detection are important enablers in modern drug discovery. With the development of robust instrumentation and implementation of user-friendly software (or software packages), non-expert users can now walk up to easily accessible advanced chromatographic systems and perform experiments at their own convenience. Although remarkable improvements in robustness and ease-of-use have happened since the introduction of the first high performance liquid chromatography–mass spectrometry (HPLC–MS) systems, the instrument performance still needs to be qualified and monitored to ensure consistent high-quality results. This article will demonstrate how a simple test mixture of carefully selected compounds can facilitate both the development of generic ultrahigh-pressure liquid chromatography–mass spectrometry (UHPLC–MS) methods and automated performance monitoring of multiple instruments located in separate laboratories and buildings.

Interest in chromatography using hydrophilic interaction liquid chromatography (HILIC) has continued to build in recent years. Adoption of the technique has been slowed by experiences of poor reproducibility. In particular, reequilibration times in HILIC have been reported as being exceptionally long as compared to reversed-phase chromatography. In this study, reequilibration times in HILIC for both aqueous–organic gradients and buffer gradients are systematically explored. The results not only promise to improve method development practices, but also provide insight into HILIC retention mechanisms across mechanistically differing polar stationary phases.

A brief preview of this year’s ASMS conference, taking place June 4–8, 2017, in Indianapolis, Indiana.

Click the title above to open the LCGC Asia Pacific March 2017 regular issue, Vol 20, No 1, in an interactive PDF format.

Click the title above to open the LCGC Europe March 2017 regular issue, Vol 30, No 3, in an interactive PDF format.

Click the title above to open the March 2017 issue of Current Trends in Mass Spectrometry, Volume 15, Number 1, in an interactive PDF format.

The 2017 Chromatography Forum of the Delaware Valley Dal Nogare Award will be presented Monday morning at Pittcon 2017 to Professor Andras Guttman from the Csaba Horváth Laboratory of Bioseparation Sciences at the University of Debrecen in Hungary.

Wyatt TechnologyReversed-phase chromatography represents one of the most popular applications of high performance liquid chromatography (HPLC), with particular importance for protein characterization. Because elution depends on the hydrophobicity of the sample, it is generally impossible to identify the separated products on the basis of their elution time (volume). Frequently, each eluted fraction must be isolated further and analyzed with other techniques to gain some understanding of the molecular behaviour of the protein.

Students who have an internship on their resume, and are seeking jobs in a particular sector, are doing so with an informed opinion. While an internship comes in many forms, that real-world experience has provided a clear touchstone of understanding of what it would be like to work in a given sector.

The Satinder Ahuja Award for Young Investigator in Separation Sciences has been presented to Omar K. Farha, a research professor of chemistry at Northwestern University, Distinguished Adjunct Professor at King Abdulaziz University, and president of NuMat Technologies.

The 2017 Chromatography Forum of the Delaware Valley Dal Nogare Award has been presented to András Guttman, head of the Horváth Csaba Laboratory of Bioseparation Sciences at the University of Debrecen in Hungary, at Pittcon 2017, held this year in Chicago, Illinois, USA.

The 2017 Pittsburgh Analytical Chemistry Award has been presented to Janusz Pawliszyn of the University of Waterloo, Canada.

Few fruits are as notorious as the Durian fruit. Native to Southeast Asia but fast becoming available around the globe, these fruits hold the unsavoury title of “the world’s smelliest fruit”. The compounds responsible for such a complex and at times difficult to stomach aroma have intrigued scientists for decades, but until recently the data relating to those chemicals was inconsistent.

The implementation of the FSMA (Food Safety Modernization Act) and the continued globalization of the food supply have increased the need for improved analytical methods for quality and safety testing. Food safety and quality requirements are increasingly stringent, as are consumer expectations. In addition to new methods, the industry continues to look for improvements in laboratory productivity and new ways to reduce costs. These trends extend to dietary supplements and natural products. This article discusses the current state of efforts to modernize US Pharmacopeia (USP) compendial methods to take advantage of new chromatography technology, including the use of “allowable adjustments”.

A preview of RAFA 2017, which will be held 7–10 November 2017 in Prague, Czech Republic.

Luigi Mondello spoke to The Column about his group’s research into characterizing pigments in bell peppers using offline multidimensional supercritical fluid chromatography (SFC) combined with reversed-phase liquid chromatography (LC).

Click the title above to open The Column February 21, 2017 Europe & Asia issue, Volume 13, Number 3, in an interactive PDF format.

Click the title above to open The Column February 21, 2017 North American issue, Volume 13, Number 3, in an interactive PDF format.

The 2017 Guiochon Fellowship will be awarded to Dwight Stoll, an Associate Professor and Co-Chair of Chemistry at Gustavus Adolphus College in St. Peter, Minnesota, USA.

Delivering samples to the analytical column in “clean” mobile phase is important for robust methods and high-quality results.

Raw data is a term that is used in both good manufacturing practice (GMP) and good laboratory practice (GLP) laboratories but it can create misunderstanding. What exactly does raw data mean and what electronic records are within the scope of the term?

The carotenoid test allows one to build a simple classification map of stationary phases used in reversed‑phase liquid chromatography, on the basis of the shape recognition (plotted on the x axis), the polar surface activity (plotted on the y axis), and the phase hydrophobicity (related by the bubble size).

Gel permeation chromatography/size-exclusion chromatography (GPC/SEC) is used to determine molar mass averages and the complete molar mass distribution with just one injection. This is possible because GPC/SEC is a fractionating technique. The fractionation power allows the higher molar mass fraction to be characterized and more about low molar mass compounds such as oligomers, additives, or residual educts to be learned.