Food and feed laboratories share several similarities when facing the implementation of liquid-chromatographic analysis. Using the experience acquired over the years, through application chemistry in food and feed research, selected analytes of relevance for both areas were discussed. This review focused on the common obstacles and peculiarities that each analyte offers (during the sample treatment or the chromatographic separation) throughout the implementation of said methods. A brief description of the techniques which we considered to be more pertinent, commonly used to assay such analytes is provided, including approaches using commonly available detectors (especially in starter labs) as well as mass detection. This manuscript consists of three sections: feed analysis (as the start of the food chain); food destined for human consumption determinations (the end of the food chain); and finally, assays shared by either matrices or laboratories. Analytes discussed consist of both those considered undesirable substances, contaminants, additives, and those related to nutritional quality. Our review is comprised of the examination of polyphenols, capsaicinoids, theobromine and caffeine, cholesterol, mycotoxins, antibiotics, amino acids, triphenylmethane dyes, nitrates/nitrites, ethanol soluble carbohydrates/sugars, organic acids, carotenoids, hydro and liposoluble vitamins. All analytes are currently assayed in our laboratories.

Liquid Chromatography Analysis of Common Nutritional Components, in Feed and Food.

A natural occurrence

Composite edible coatings from commercial pectin, corn flour and beetroot powder minimize post-harvest decay, reduces ripening and improves sensory liking of tomatoes.

Structural features, antioxidant and acetylcholinesterase inhibitory activities of polysaccharides from stem of Physalis alkekengi L.

Functional characterization of plant-based protein to determine its quality for food applications

Quantification of sugars and organic acids in tomato fruits

Glutamic and aspartic acid fulfil numerous functions in organisms. They are proteinogenic amino acids, they function as neurotransmitters, and glutamic acid links the citrate cycle with amino acid metabolism. In addition, glutamic acid is a precursor for many bioactive molecules like γ-aminobutyric acid (GABA). In tomatoes, glutamic acid accumulates in ripening fruits. Here we present a simple and rapid method for quantification of glutamate and aspartate in tomatoes. A cleared extract is prepared and 2-aminoadipic acid added as internal standard. Subsequently, the amino acids are derivatised with 2,4-dinitro-1-fluorobenzene under alkaline conditions. The derivatives are separated by ultra-high performance liquid chromatography using a phenyl-hexyl column and 50 mM N-methylmorpholine/acetate buffer pH 7.4 containing 12% acetonitrile as eluent and detected by UV absorption at 363 nm. The whole analysis time including separation and column equilibration takes less than 2.8 min with a flow rate of 1 mL/min and less than 1.6 min with a flow rate of 2 mL/min, making this method suitable for high-throughput applications. The method shows excellent reproducibility with intra- and inter-day SDs of approximately 4% for both aspartic and glutamic acid. Using this method we show that the glutamate/aspartate ratio changes significantly during fruit ripening.

/pdf/quantification-of-glutamate-and-aspartate-by-ultra-high-1gm3oho4u3.pdf

Quantification of Glutamate and Aspartate by Ultra-High Performance Liquid Chromatography

Hydroponic cultivation of vegetables avoids problems with soil-borne plant pathogens and may allow higher yield. In arid climates and particularly on islands, high concentrations of sodium chloride can be present in the groundwater. For instance, in many sites of Malta, the groundwater contains more than 10 mM sodium chloride. Here we investigated the effects of sodium chloride at levels typically found in Malta on yield, physiology and fruit quality of tomato, the economically most important vegetable. We selected cherry tomatoes since their production is attractive due to their high marketing value. While the yield declined at higher salinity levels tested (17 and 34 mM), the quality increased significantly as indicated by higher total soluble solids and fructose and glucose levels. The type of substrate—coco peat, perlite or Rockwool—had only minor effects. Although the concentration of citric acid and malic acid remained unaffected, the pH dropped by approximately 0.1 unit and the titratable acidity increased slightly. This might be explained by a high uptake of chloride but a lower increase of the sodium content and a reduced potassium level in the fruits, shifting the equilibrium of the organic acids more to their protonated forms. Proline increased significantly, while the level of glutamic acid, which is crucial for the taste, remained unchanged. Our results show that cherry tomatoes can be cultivated in nutrient solutions prepared with salt-containing groundwater, as found in Malta. The yield declined to some extent but the quality of the produced fruits was higher compared to cultivation in salt-free media. 

/pdf/the-effect-of-salinity-on-fruit-quality-and-yield-of-cherry-1bdgdk0o.pdf

The Effect of Salinity on Fruit Quality and Yield of Cherry Tomatoes

: Hydroponic cultivation of vegetables avoids problems with soil-borne plant pathogens and may allow higher yield. In arid climates and particularly on islands, high concentrations of sodium chloride can be present in the groundwater. For instance, in many sites of Malta, the groundwater contains more than 10 mM sodium chloride. Here we investigated the effects of sodium chloride at levels typically found in Malta on yield, physiology and fruit quality of tomato, the economically most important vegetable. We selected cherry tomatoes since their production is attractive due to their high marketing value. While the yield declined at higher salinity levels tested (17 and 34 mM), the quality increased signiﬁcantly as indicated by higher total soluble solids and fructose and glucose levels. The type of substrate—coco peat, perlite or Rockwool—had only minor effects. Although the concentration of citric acid and malic acid remained unaffected, the pH dropped by approximately 0.1 unit and the titratable acidity increased slightly. This might be explained by a high uptake of chloride but a lower increase of the sodium content and a reduced potassium level in the fruits, shifting the equilibrium of the organic acids more to their protonated forms. Proline increased signiﬁcantly, while the level of glutamic acid, which is crucial for the taste, remained unchanged. Our results show that cherry tomatoes can be cultivated in nutrient solutions prepared with salt-containing groundwater, as found in Malta. The yield declined to some extent but the quality of the produced fruits was higher compared to cultivation in salt-free media.

Plants are an indispensable cornerstone of sustainable global food supply. While immense progress has been made in decoding the genomes of crops in recent decades, the composition of their proteomes, the entirety of all expressed proteins of a species, is virtually unknown. In contrast to the model plant Arabidopsis thaliana, proteomic analyses of crop plants have often been hindered by the presence of extreme concentrations of secondary metabolites such as pigments, phenolic compounds, lipids, carbohydrates or terpenes. As a consequence, crop proteomic experiments have, thus far, required individually optimized protein extraction protocols to obtain samples of acceptable quality for downstream analysis by liquid chromatography tandem mass spectrometry (LC-MS/MS). In this article, we present a universal protein extraction protocol originally developed for gel-based experiments and combined it with an automated single-pot solid-phase-enhanced sample preparation (SP3) protocol on a liquid handling robot to prepare high-quality samples for proteomic analysis of crop plants. We also report an automated offline peptide separation protocol and optimized micro-LC-MS/MS conditions that enables the identification and quantification of ~10,000 proteins from plant tissue within 6 h of instrument time. We illustrate the utility of the workflow by analyzing the proteomes of mature tomato fruits to an unprecedented depth. The data demonstrate the robustness of the approach which we propose for use in upcoming large-scale projects that aim to map crop tissue proteomes.

/pdf/getting-ready-for-large-scale-proteomics-in-crop-plants-1uj0lnwr.pdf

Carlos Agius

Papers

Quantification of sugars and organic acids in tomato fruits

Quantification of Glutamate and Aspartate by Ultra-High Performance Liquid Chromatography

The Effect of Salinity on Fruit Quality and Yield of Cherry Tomatoes

The Effect of Salinity on Fruit Quality and Yield of Cherry Tomatoes

Getting Ready for Large-Scale Proteomics in Crop Plants