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A new ultrafast ultrasonic-based method for shotgun proteomics as well as label-free protein quantification in urine samples is developed. The method first separates the urine proteins using nitrocellulose-based membranes and then proteins are in-membrane digested using trypsin. The enzymatic digestion process is accelerated from overnight to four minutes using a sonoreactor ultrasonic device. Overall,...
We report on the new microplate horn ultrasonic device as a powerful tool to speed proteomics workflows with unparalleled throughput. 96 complex proteomes were digested at the same time in 4min. Variables such as ultrasonication time, ultrasonication amplitude, and protein to enzyme ratio were optimized. The "classic" method relying on overnight protein digestion (12h) and the sonoreactor-based...
In this study, laser-induced breakdown spectroscopy (LIBS) was combined with chemometric strategies (PCA, Principal Component Analysis) and scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM–EDS) to investigate the metal composition of a printed circuit board (PCB) sample from a mobile phone. Scanning electron microscopy–EDS was used for two main reasons: it was possible at...
The use of chemical protein depletion in conjunction with gold-based nanoparticles for fast matrix assisted laser desoption ionization time of flight mass spectrometry-based human serum profiling was assessed. The following variables influencing the process were optimized: (i) amount of nanoparticles, (ii) sample pH, (iii) amount of protein and (iv) incubation time. pH was found the most important...
The variables affecting the direct matrix assisted laser desorption ionization mass spectrometry-based analysis of wine for classification purposes have been studied. The type of matrix, the number of bottles of wine, the number of technical replicates and the number of spots used for the sample analysis have been carefully assessed to obtain the best classification possible. Ten different algorithms...
The decision peptide-driven tool implements a software application for assisting the user in a protocol for accurate protein quantification based on the following steps: (1) protein separation through gel electrophoresis; (2) in-gel protein digestion; (3) direct and inverse 18 O-labeling and (4) matrix assisted laser desorption ionization time of flight mass spectrometry, MALDI analysis. The...
Several ultrasound-based platforms for DNA sample preparation were evaluated in terms of effective fragmentation of DNA (plasmid and genomic DNA)—ultrasonic probe, sonoreactor, ultrasonic bath and the newest Vialtweeter device. The sonoreactor showed the best efficiency of DNA fragmentation while simultaneously assuring no cross-contamination of samples, and was considered the best ultrasonic tool...
We studied in this work the performance of the new ultrasonic multiprobe in terms of throughput, handling and robustness. The study was conducted using the multiprobe to speed two different proteomics workflows. The “classic” method relaying on overnight protein digestion (12h), was used as the standard procedure. This work clearly shows the importance of testing variables such as ultrasonic amplitude...
The estimation of the time since death known as postmortem interval (PMI) is a main issue in the field of forensic science and legal medicine. In this work it is proposed a sequential injection system for the determination of hypoxanthine and potassium in the same sample of vitreous humor since the concentrations of both parameters change with PMI and the vitreous humor has been regarded as the ideal...
An efficient methodology for the determination of carbohydrate content in soils, employing acid hydrolysis and subsequent capillary electrophoresis analysis (CE), is here described. Polysaccharides present in soil samples were hydrolyzed, at 100°C during 4h, to their monosaccharide form, by addition of 2moldm −3 trifluoroacetic acid (TFA) directly to soil. The resulting monosaccharides were...
An easy to use and low time consuming capillary electrophoresis (CE) method was developed and applied to the simultaneous determination of six antibiotics (ampicillin, amoxicillin, cloxacillin, penicillin, tetracycline and chloramphenicol) in spiked milk samples. Samples of milk were cleaned up by solid-phase extraction (with a C 18 cartridge) after protein precipitation. Analysis was performed...
Milk samples can be efficiently digested using a focused microwave oven, however the conventional procedure of addition of concentrated acids to the liquid sample leads to digestates with elevated acidity and residual carbon concentrations. In this work a focused microwave oven was applied for acid digestion of bovine milk samples using a conventional and an alternative procedure based on gradual...
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