Different Stationary Phase Selectivities and Morphologies for Intact Protein Separations

The central dogma of biology proposed that one gene encodes for one protein. We now know that this does not reflect reality. The human body has approximately 20,000 protein-encoding genes; each of these genes can encode more than one protein. Proteins expressed from a single gene can vary in terms of their post-translational modifications, which often regulate their function within the body. Understanding the proteins within our bodies is a key step in understanding the cause, and perhaps the solution, to disease. This is one of the application areas of proteomics, which is defined as the study of all proteins expressed within an organism at a given point in time. The human proteome is incredibly complex. The complexity of biological samples requires a combination of technologies to achieve high resolution and high sensitivity analysis. Despite the significant advances in mass spectrometry, separation techniques are still essential in this field. Liquid chromatography is an indispensable tool by which low-abundant proteins in complex samples can be enriched and separated. However, advances in chromatography are not as readily adapted in proteomics compared to advances in mass spectrometry. Biologists in this field still favour reversed-phase chromatography with fully porous particles. The purpose of this review is to highlight alternative selectivities and stationary phase morphologies that show potential for application in top-down proteomics; the study of intact proteins

Surface Acoustic Wave Nebulisation Mass Spectrometry for the Fast and Highly Sensitive Characterisation of Synthetic Dyes in Textile Samples

Surface acoustic wave nebulisation (SAWN) mass spectrometry (MS) is a method to generate gaseous ions compatible with direct MS of minute samples at femtomole sensitivity. To perform SAWN, acoustic waves are propagated through a LiNbO3 sampling chip, and are conducted to the liquid sample, which ultimately leads to the generation of a fine mist containing droplets of nanometre to micrometre diameter. Through fission and evaporation, the droplets undergo a phase change from liquid to gaseous analyte ions in a non-destructive manner. We have developed SAWN technology for the characterisation of organic colourants in textiles. It generates electrospray-ionisation-like ions in a non-destructive manner during ionisation, as can be observed by the unmodified chemical structure. The sample size is decreased by tenfold to 1000-fold when compared with currently used liquid chromatography-MS methods, with equal or better sensitivity. This work underscores SAWN-MS as an ideal tool for molecular analysis of art objects as it is non-destructive, is rapid, involves minimally invasive sampling and is more sensitive than current MS-based methods

Size distributions of droplets produced by ultrasonic nebulizers

Abstract In many applications where small, similar-sized droplets are needed, ultrasonic nebulizers are employed. Little is known about the mechanism of nebulization, for example about what determines the median droplet size. Even less understood, is the droplet size distribution, which is often simply fitted with a log-normal distribution or assumed to be very narrow. We perform the first systematic study of droplet size distributions for different nebulizer technologies, showing that these distributions can be very well fitted with distributions found for sprays, where the size distribution is completely determined by the corrugation of ligaments and the distribution of ligament sizes. In our case, breakup is believed to be due to pinch-off of Faraday instabilities. The droplet size distribution is then set by the distribution of wavelengths of the standing capillary waves and the roughness of the pinch-off ligaments. We show that different nebulizer technologies produce different size distributions, which we relate to (variation in) wavelengths of the waves that contribute to the droplet formation. We further show that the median droplet size scales with the capillary wavelength, with a proportionality constant that depends only slightly on the type of nebulizer, despite order-of-magnitude differences in other parameters