Rapid quantification of disaccharide isomers by derivatization in combination with ion mobility spectrometry in beer and milk.

The subtle structural variations among carbohydrate isomers pose significant challenges for their identification and quantification. Here, we propose a strategy for rapid identification and quantification of isomeric disaccharides via derivatization with 4-(3-methyl-5-oxo-pyrazolin-1-yl) benzoic acid (CPMP) and analysing by ion mobility spectrometry (IMS). After derivatization, the ionization efficiency of disaccharides was significantly improved. The disaccharide isomers were distinguished by determining the different ion mobilities of CPMP-labelled disaccharides. Among them, [M + 2CPMP + H]+ was separated with a resolution of 1.484, almost achieving baseline separation. Subsequently, [M + CPMP + Na]+ was used for the relative quantification of lactose and maltose, showing a good linear relationship with R2 > 0.990. Finally, the method was successfully applied to the identification of lactose and maltose in beer and milk. The method is fast, accurate and effective for the identification of disaccharide isomers in complex samples.

A rapid liquid chromatography-electrospray ionization-ion mobility spectrometry method for monitoring nine representative metabolites in the seedlings of cucumber and wheat.

A fast and convenient high-performance liquid chromatography-electrospray ionization-ion mobility spectrometry method was developed to determine nine representative metabolites in the seedlings of cucumber and wheat. The analytical conditions were obtained by optimizing the parameters of high-performance liquid chromatography and ion mobility spectrometry. Briefly, acetonitrile-0.1% formic acid solution was selected as the mobile phase for gradient elution at a flow velocity of 0.4 mL/min. Under negative electrospray ionization mode, spray voltage of ion mobility spectrometry was 4.5 kV, and drift tube temperature was set at 90°C. The metabolites from seedling leaves were extracted using 80% acetonitrile as the solvent at 4°C for 12 h. Results showed that under soilless culture conditions, the contents of maltose, citric acid, and p-hydroxybenzoic acid in the seedlings of cucumber and wheat were reduced by low concentration of itaconic acid, succinic acid, and citric acid. Importantly, this analytical approach demonstrated high sensitivity, good linear response, and high selectivity. The lowest limit of detection was 0.004 µg for p-hydroxybenzoic acid. Overall, this high-performance liquid chromatography-electrospray ionization-ion mobility spectrometry method is sensitive and efficient for rapid separation and identification of plant metabolites.

New application of a traditional method: colorimetric sensor array for reducing sugars based on the in-situ formation of core-shell gold nanorod-coated silver nanoparticles by the traditional Tollens reaction.

An effective and robust colorimetric sensor array for simultaneous detection and discrimination of five reducing sugars (i.e., glyceraldehyde (Gly), fructose (Fru), glucose (Glu), maltose (Mal), and ribose (Rib)) has been proposed. In the sensor array, two negatively charged polydielectrics (sodium polystyrenesulfonate (NaPSS) and sodium polymethacrylate (NaPMAA)), which served as the sensing elements, were individually absorbed on the surface of the cetyltrimethylammonium bromide (CTAB)-coated gold nanorods (AuNR) with positive charges through electrostatic action, forming the designed sensor units (NaPSS-AuNR and NaPMAA-AuNR). In the presence of Tollens reagent (Ag(NH3)2OH), Ag+ was absorbed on the surface of negatively charged NaPSS-AuNR and NaPMAA-AuNRs. When confronted with differential reducing sugars, different reducing sugars exhibited differential levels of deoxidizing abilities toward Ag+, thus Ag+ was reduced to diverse amounts of silver nanoparticles (AgNPs) in situ to form core-shell AuNR@AgNP by the traditional Tollens reaction method, leading to distinct colorimetric response patterns (value of AS/AL (the ratio of absorbance at 360 nm to that at 760 nm in Ag+-NaPMAA-AuNR, and the ratio of absorbance at 360 nm to that at 740 nm in Ag+-NaPSS-AuNR)). These response patterns are characteristic for each reducing sugar, and can be quantitatively distinguished by linear discriminant analysis (LDA) at concentrations as low as 10 nM with relative standard deviation (RSD) of 4.11% (n = 3). The practicability of this sensor array has been validated by recognition of reducing sugars in serum and urine samples. A colorimetric sensor array for reducing sugar discrimination based on the reduction of Ag+ and in situ formation of AuNR@AgNP.

A temperature-mediated two-step saccharification process enhances maltose yield from high-concentration maltodextrin solutions.

BACKGROUND: Designing a high-concentration (50%, w/w) maltodextrin saccharification process is a green method to increase the productivity of maltose syrup. RESULTS: In this study, a temperature-mediated two-step process using ß-amylase and pullulanase was investigated as a strategy to improve the efficiency of saccharification. During the saccharification process, both pullulanase addition time and temperature adjustment greatly impacted the final maltose yield. These results indicated that an appropriate ß-amylolysis in the first stage (the first 8 h) was required to facilitate saccharification process, with the maltose yield of 8.46% greater than that of the single step saccharification. Molecular structure analysis further demonstrated that a relatively low temperature (50 °C), as compared with a normal temperature (60 °C), in the first stage resulted in a greater number of chains polymerized by at least seven glucose units and a less heterogeneity system within the residual substrate. The molecular structure of the residual substrate might be beneficial for the subsequent cooperation between ß-amylase and pullulanase in the following 40 h (second stage). CONCLUSION: Over a 48 h saccharification, the temperature-mediated two-step process dramatically increased the conversion rate of maltodextrin and yielded significantly more maltose and less byproduct, as compared with a constant-temperature process. The two-step saccharification process therefore offered an efficient and green strategy for maltose syrup production in industry. © 2020 Society of Chemical Industry.

Fast and Robust Quantification of Detergent Micellization Thermodynamics from Isothermal Titration Calorimetry.

Detergents are widely used in modern in vitro biochemistry and biophysics, in particular to aid the characterization of integral membrane proteins. An important characteristic of these chemicals in aqueous solutions is the concentration above which their molecular monomers self-associate to form micelles, termed the critical micellar concentration (CMC). Micelles are supramolecular assemblies arranged with the hydrophobic portions oriented inward and the hydrophilic head groups positioned outward to interact with the aqueous solvent. Knowledge of the CMC is not only of practical relevance but also of theoretical interest because it provides thermodynamic insights. Isothermal titration calorimetry (ITC) is a powerful method to determine CMCs, as it furnishes additional information on the enthalpy and entropy of micellization. Here we describe our extension of previous methods to determine CMCs and other thermodynamic parameters from ITC demicellization curves. The new algorithm, incorporated into the stand-alone software package D/STAIN, analyzes ITC demicellization curves by taking advantage of state-of-the-art thermogram-integration techniques and automatically providing rigorous confidence intervals on the refined parameters. As a demonstration of the software's capabilities, we undertook ITC experiments to determine the respective CMCs of n-octyl ß-d-glucopyranoside (OG), n-dodecyl ß-d-maltopyranoside (DDM), and lauryldimethylamine N-oxide (LDAO). Motivated by the fact that in vitro membrane protein studies often require additives such as precipitants (e.g., polyethylene glycol (PEG)), we also carried out ITC demicellization studies in the presence of PEG3350, finding in all cases that PEG had significant effects on the thermodynamics of detergent micellization.

Short communication: Characterization of an atypical maltose-negative Staphylococcus aureus through the use of phenotypic and molecular techniques.

The most clinically relevant staphylococci in veterinary medicine are those that are coagulase-positive, namely Staphylococcus aureus. During microbiological udder health monitoring (2009-2018), a new S. aureus strain (coagulase-positive and maltose-negative) was discovered as an emerging udder pathogen during routine examinations of South African dairy herds. This study challenged the conventional microbiological diagnosis of staphylococci by comparing its results to those of the MALDI-TOF mass spectrometry and 16S rRNA sequencing. Both of these tests confirmed that the maltose-negative staphylococcus (MNS), identified as Staphylococcus pseudintermedius by conventional microbiology, was S. aureus ST2992. Multi locus sequence typing was performed on 3 of the MNS isolates and indicated that these isolates were of single origin. These strains tested positive for both MALA and MALR genes (control: S. aureus ATCC 25923). Although the α-glucosidase gene was present, it was not expressed phenotypically. The latter may be attributed to the abnormal stop codon identified in the MALA gene sequence of S. aureus ST2992 (GenBank accession number, MN531305). The newly identified MNS has a field behavior different to that of maltose-positive S. aureus, and more similar to the low virulence of non-aureus staphylococci.

Salivary α-Amylase Activity and Starch-Related Sweet Taste Perception in Humans.

Starch-related sweet taste perception plays an important role as a part of the dietary nutrient sensing mechanisms in the oral cavity. However, the release of sugars from starchy foods eliciting sweetness has been less studied in humans than in laboratory rodents. Thus, 28 respondents were recruited and evaluated for their starch-related sweet taste perception, salivary alpha-amylase (sAA) activity, oral release of reducing sugars, and salivary leptin. The results demonstrated that a 2-min oral mastication of starchy chewing gum produced an oral concentration of maltose above the sweet taste threshold and revealed that the total amount of maltose equivalent reducing sugars produced was positively correlated with the sAA activity. In addition, respondents who consistently identified the starch-related sweet taste in two sessions (test and retest) generated a higher maltose equivalent reducing sugar concentration compared to respondents who could not detect starch-related sweet taste at all (51.52 ± 2.85 and 29.96 ± 15.58 mM, respectively). In our study, salivary leptin levels were not correlated with starch-related sweet taste perception. The data contribute to the overall understanding of oral nutrient sensing and potentially to the control of food intake in humans. The results provide insight on how starchy foods without added glucose can elicit variable sweet taste perception in humans after mastication as a result of the maltose generated. The data contribute to the overall understanding of oral sensing of simple and complex carbohydrates in humans.

The use of of inulin, maltitol and lecithin as fat replacers and plasticizers in a model reduced-fat mozzarella cheese-like product.

BACKGROUND: Mono-, di- and oligosaccharides, polyhydric alcohols and lipids are three main types of plasticizers used to process food materials. In the present study, inulin, maltitol and lecithin were selected as representative oligosaccharide, polyhydric alcohol and lipid fat replacers, respectively. Their effects on the physicochemical properties of reduced-fat mozzarella cheese were evaluated. RESULTS: Lecithin reduced the hardness and increased the degree of free oil released. Inulin and lecithin decreased the hydrophobic interaction of reduced-fat cheese. Maltitol improved the elasticity of the reduced-fat cheese and increased the hydrophobic interaction within the casein matrix. Maltitol-added cheese had a lower glass transition temperature (Tg ) than the other cheeses. Maltitol significantly improved the stretchability of the reduced-fat cheese. CONCLUSION: The results obtained in the present study suggest that maltitol is an effective fat replacer in reduced-fat mozzarella cheese and might enhance the cheese's functional properties. The Tg of cheese was related to the water and fat content, fat replacer addition and cross-linking degree of casein. The relationship between Tg and the physicochemical properties of cheese will be studied in further research. © 2019 Society of Chemical Industry.

Effects of maltose and lysine treatment on coffee aroma by flash gas chromatography electronic nose and gas chromatography-mass spectrometry.

BACKGROUND: Arabica coffee is a sub-tropical agricultural product in China. Coffee undergoes a series of thermal reactions to form abundant volatile profiles after roasting, so it loses a lot of reducing sugars and amino acids. Adding carbonyl compounds with amino acids before roasting could ensure the nutrition and flavour of coffee. The technology is versatile for the development of coffee roasting process. This investigation evaluates the effects of combining maltose and lysine (Lys) to modify coffee aroma and the possibly related mechanisms. Arabica coffee was pretreated with a series of solvent ratios of maltose and Lys with an identical concentration (0.25 mol L-1 ) before microwave heating. RESULTS: It was found that the combination of maltose and Lys significantly (P ≤ 0.05) influenced quality indices of coffee (pH and browning degree). Ninety-six aromatic volatiles have been isolated and identified. Twelve volatile profiles revealed the relationship between fragrance difference and compound content in coffee. Moreover, coffee aroma was modified by a large number of volatiles with different chemical classes and character. CONCLUSION: Thus, our results suggest that the combination of reagents changed overall aroma quality through a series of complex thermal reactions, especially the ratio of Lys/maltose over 2:1. © 2017 Society of Chemical Industry.

[Method of Quantitative Analysis by HPLC and Confirmation by LC-MS/MS of Erythritol, Maltitol, Lactitol and Trehalose in Foods].

A quantitative determination method of erythritol, maltitol, lactitol and trehalose in foods by HPLC, and confirmation method by LC-MS/MS were developed. HPLC analysis was performed on a separation column packed with amino group-binding polymer with acetonitrile-water (80 : 20) as the mobile phase. The column was operated at room temperature, and the three sugar alcohols and trehalose were quantified. LC-MS/MS confirmation was performed on an amino group-bound column with acetonitrile-ammonium acetate solution as the mobile phase, with detection in the SRM mode. At low sample dilution ratios, the analysis may be affected by matrix derived from the sample, but this can be suppressed by 1,000-fold or greater dilution. Recoveries of the three sugar alcohols and trehalose spiked into food samples, such as tea, jelly, tablets (ramune candy), and chocolate, exceeded 90% (CV≦6.1%) in HPLC and 94% (CV≦4.8%) in LC-MS/MS.

Sucrose replacement in high ratio white layer cakes.

BACKGROUND: Several commercially available alternative sweeteners have potential in reducing the caloric content of baked products. Sugar alcohols and natural sweeteners have similar bulk as sucrose and can replace sucrose directly. High intensity sweeteners have high potency but light weight so bulking agents are often added. This study determined alternative sweeteners and combinations of alternative sweetener and a bulking agent that produced good quality white layer cakes. RESULTS: Cakes made with maltitol were acceptable but erythritol and fructose produced undesirable cakes. Maltodextrin and polydextrose were acceptable bulking agents, producing cakes that were similar to control cakes. The flavor of cakes sweetened with sucralose was acceptable but those with stevia had a disagreeable metallic aftertaste. Cakes made with sucralose plus maltodextrin were preferred over those containing sucralose plus polydextrose. Consumer acceptance of flavor, texture and overall liking of cakes containing maltitol was similar to sucrose and both were preferred over cakes containing maltodextrin plus sucralose. CONCLUSION: Replacing sucrose with maltitol in white layer cakes reduced the caloric content by 16% with no loss in quality. © 2016 Society of Chemical Industry.

Encapsulation of FRET-based glucose and maltose biosensors to develop functionalized silica nanoparticles.

Silicate nanoparticles with immobilized FRET-based biosensors were developed for the detection of glucose and maltose. Immobilization of the protein biosensor in the nanoparticle was achieved through specific interaction between the hexa-histidine tag of the protein and a calcium-silicate complex of the silica matrix. Encapsulation of the biosensors preserved the affinity for the respective sugar. Compared to the free biosensors, encapsulation had a stabilizing effect on the biosensor towards chemical and thermal denaturation. The demonstrated immobilization strategy for specific sensing proteins paves the way towards the development of protein-inorganic nanostructures for application in metabolite analyses.

Phenylboronic acid modified solid-phase extraction column: Preparation, characterization, and application to the analysis of amino acids in sepia capsule by removing the maltose.

Maltose, a common auxiliary material of pharmaceutical preparation, may disturb the analysis of total amino acids in sepia capsule by aldolization. Therefore, it is necessary to remove the maltose through a convenient method. In this work, a phenylboronic acid modified solid-phase extraction column has been synthesized and used to remove the maltose. The materials were synthesized by one step "thiol-ene" reaction and the parameters of the column such as absorption capacity, recovery, and absorption specificity have been investigated. The results showed the column (0.5 cm of length × 0.5 cm of inner diameter) can absorb 4.6 mg maltose with a linear absorption and absorption specificity. Then this technique was applied in the quantification of amino acids in sepia capsule. After the optimization of the method, four kinds of amino acids, which were the most abundant, were quantified by high-performance liquid chromatography with diode array detection. The amounts of the four kinds of amino acids are 1.5∼2 times more than that without the treatment of solid-phase extraction column, which almost overcomes the influence of the maltose. All the results indicate that the phenylboronic acid modified solid-phase extraction column can successfully help to accurately quantify the total amino acids in sepia capsule.

A portable FRET analyzer for rapid detection of sugar content.

Fluorescence resonance energy transfer (FRET) is widely used as a core process in biometric sensors to detect small molecules such as sugars, calcium ions, or amino acids. However, FRET based biosensors with innate weak signal intensity require the use of expensive, high-sensitive equipment. In the present study, these shortcomings were overcome with the fabrication of a sensitive, inexpensive, and portable analyzer which provides quantitative detection of small molecules in a liquid sample. The usability of the developed analyzer was successfully tested by measuring sucrose and maltose contents in commercially available beverage samples, with better performance than the conventional monochromator-type spectrofluorometer. It is anticipated that miniaturization of the equipment and improving the FRET based biosensors will contribute to the practical use of this hand-held analyzer in conditions where high-end equipment is not available.

Study of the physicochemical parameters and spontaneous fermentation during the traditional production of yakupa, an indigenous beverage produced by Brazilian Amerindians.

Yakupa is a traditional non-alcoholic cassava beverage produced by Brazilian Amerindians. In this work the microbial dynamics and metabolites involved in yakupa fermentation were investigated by PCR-denaturing gradient gel electrophoresis and chromatography analysis, respectively. The lactic acid bacteria (LAB) population was higher than yeast in the beginning of fermentation (5 log CFU mL(-1) and 3 log CFU mL(-1), respectively) and after 36 h both population increased reaching 7 log CFU mL(-1), remaining constant until 60 h. Culture dependent and independent methods in combination identified the bacteria Lactobacillus fermentum, L. plantarum, Weissela cibaria and W. confusa, and yeasts Saccharomyces cerevisiae and Pichia kudriavzevii. Maltose (41.2 g L(-1)), ethanol (6.5 g L(-1)) and lactic acid (7.8 g L(-1)) were the most abundant compounds identified by high performance liquid chromatography. Aldehydes, acids, alcohols and esters were identified by gas chromatography flame ionization detection. By the metabolites and PCA analysis we may assign the beverage's flavor to the microbial metabolism. Heterolactic LAB and S. cerevisiae dominated the yakupa fermentation, being responsible for the organoleptic characteristics of the final product. This is the first time that the microbial dynamics and physicochemical parameters were investigated in the yakupa beverage and it may contribute to the future selection of starter cultures to perform yakupa fermentations.

Distinguishing Ontario ginseng landraces and ginseng species using NMR-based metabolomics.

The use of (1)H-NMR-based metabolomics to distinguish and identify unique markers of five Ontario ginseng (Panax quinquefolius L.) landraces and two ginseng species (P. quinquefolius and P. ginseng) was evaluated. Three landraces (2, 3, and 5) were distinguished from one another in the principal component analysis (PCA) scores plot. Further analysis was conducted and specific discriminating metabolites from the PCA loadings were determined. Landraces 3 and 5 were distinguishable on the basis of a decreased NMR intensity in the methyl ginsenoside region, indicating decreased overall ginsenoside levels. In addition, landrace 5 was separated by an increased amount of sucrose relative to the rest of the landraces. Landrace 2 was separated from the rest of the landraces by the increased level of ginsenoside R(b1). The Ontario P. quinquefolius was also compared with Asian P. ginseng by PCA, and clear separation between the two groups was detected in the PCA scores plot. The PCA loadings plot and a t-test NMR difference plot were able to identify an increased level of maltose and a decreased level of sucrose in the Asian ginseng compared with the Ontario ginseng. An overall decrease of ginsenoside content, especially ginsenoside R(b1), was also detected in the Asian ginseng's metabolic profile. This study demonstrates the potential of NMR-based metabolomics as a powerful high-throughput technique in distinguishing various closely related ginseng landraces and its ability to identify metabolic differences from Ontario and Asian ginseng. The results from this study will allow better understanding for quality assessment, species authentication, and the potential for developing a fully automated method for quality control.

The use of sensory attributes, sugar content, instrumental data and consumer acceptability in selection of sweet potato varieties.

BACKGROUND: As eating quality is important for adoption of new varieties, nine orange-fleshed and three cream-fleshed sweet potato varieties were assessed for sensory characteristics, dry mass and free sugar content, instrumental texture and colour and consumer acceptability (n = 216) in a peri-urban South African setting. RESULTS: Cream-fleshed varieties were higher in yellow-green colour and sweet potato-like flavour and lower in graininess. Orange-fleshed varieties were higher in pumpkin-like flavour, orange colour, discolouration and sucrose content. Partial least squares regression analysis showed that the most accepted varieties (Impilo, Excel, Resisto, 2001_5_2, Serolane, W-119 and Monate) were associated with sweet flavour, dry mass and maltose content, while the least accepted varieties (Beauregard, Khano and 1999_1_7) were associated with wateriness. Pearson correlation analysis highlighted correlations of sensory attributes yellow and orange with instrumental colour measurements (colour a* and colour b*), instrumental firmness with sensory firmness, dry mass with sensory wateriness, and maltose content with sensory sweet and sweet potato-like flavour. The varieties were clustered into three groups. Consumer acceptability for eating quality correlated with maltose content, dry mass and sweet flavour. CONCLUSION: Chemical and instrumental measurements were identified to evaluate key attributes and will be useful in the intermediate phases of sweet potato varietal development.

Gas chromatography-mass spectrometry analysis of 13C labeling in sugars for metabolic flux analysis.

Metabolic flux analysis, using 13C labeled substrates, has become a powerful methodology for quantifying intracellular fluxes. Most often, analysis is restricted to nuclear magnetic resonance or mass spectrometry measurement of 13C label incorporation into protein amino acids. However, amino acid isotopomer distribution insufficiently covers the entire network of central metabolism, especially in plant cells with highly compartmented metabolism, and analysis of other metabolites is required. Analysis of label in saccharides provides complementary data to better define fluxes around hexose, pentose, and triose phosphate pools. Here, we propose a gas chromatography-mass spectrometry (GC-MS) method to analyze 13C labeling in glucose and fructose moieties of sucrose, free glucose, fructose, maltose, inositol, and starch. Our results show that saccharide labeling for isotopomer quantification is better analyzed by chemical ionization than by electron ionization. The structure of the generated fragments was simulated and validated using labeled standards. The method is illustrated by analysis of saccharides extracted from developing rapeseed (Brassica napus L.) embryos. It is shown that glucose 6-phosphate isomerase and plastidial glucose 6-phosphate transport reactions are not at equilibrium, and light is shed on the pathways leading to fructose, maltose, and inositol synthesis.

Discrimination of neutral oligosaccharides through a nanopore.

The detection of oligosaccharides at the single-molecule level was investigated using a protein nanopore device. Neutral oligosaccharides of various molecular weights were translocated through a single α-hemolysin nanopore and their nano-transit recorded at the single-molecule level. The translocation of maltose and dextran oligosaccharides featured by 1→4 and 1→6 glycosidic bonds respectively was studied in an attempt to discriminate oligosaccharides according to their polymerization degree and glycosidic linkages. Oligosaccharides were translocated through a free diffusion regime indicating that they adopted an extended conformation during their translocation in the nanopore. The dwell time increased with molecular mass, suggesting the usefulness of nanopore as a molecular sizing device.