A rapid and inexpensive microplate assay for the enzymatic determination of glucose, fructose, sucrose, <scp>L</scp>‐malate and citrate in tomato (<i>Lycopersicon esculentum</i>) extracts and in orange juice

Velterop, Joyce S.; Vos, Femke

doi:10.1002/pca.598

Cited by 71 publications

(44 citation statements)

References 9 publications

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“…Starch remaining in the pellet after polar compounds extraction was converted to glucose using amyloglucosidase (Moing et al, 1994) and the resulting glucose was analyzed enzymatically following NADH production at 340 nm (Kunst et al, 1984) with adaptation to a microplate spectrophotometer (Dynatech, St Cloud, France) (Velterop and Vos, 2001). …”

Section: Starch Analysismentioning

confidence: 99%

Quantitative metabolic profiles of tomato flesh and seeds during fruit development: complementary analysis with ANN and PCA

et al. 2007

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Tomato, an essential crop in terms of economic importance and nutritional quality, is also used as a model species for all fleshy fruits and for genomics of Solanaceae. Tomato fruit quality at harvest is a direct function of its metabolite content, which in turn is a result of many physiological changes during fruit development. The aim of the work reported here was to develop a global approach to characterize changes in metabolic profiles in two interdependent tissues from the same tomato fruits. Absolute quantification data of compounds in flesh and seeds from 8 days to 45 days post anthesis (DPA) were obtained through untargeted (proton nuclear magnetic resonance, 1 H-NMR) and targeted metabolic profiling (liquid chromatography with diode array detection (LC-DAD) or gas chromatography with flame ionization detection (GC-FID)). These data were analyzed with chemometric approaches. Kohonen self organizing maps (SOM) analysis of these data allowed us to combine multivariate (distribution of samples on Kohonen SOMs) and univariate information (component plane representation of metabolites) in a single analysis. This strategy confirmed published data and brought new insights on tomato flesh and seed composition, thus demonstrating its potential in metabolomics. The compositional changes were related to physiological processes occurring in each tissue. They pointed to (i) some parallel changes at early stages in relation to cell division and transitory storage of carbon, (ii) metabolites participating in the fleshy trait and (iii) metabolites involved in the specific developmental patterns of the seeds.

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Section: Starch Analysismentioning

confidence: 99%

Quantitative metabolic profiles of tomato flesh and seeds during fruit development: complementary analysis with ANN and PCA

et al. 2007

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“…An enzymatic method was chosen to determine the sucrose concentration, and the method was able to provide a high degree of specificity and sensitivity. 16 A series of enzymatic reactions began with the hydrolysis of sucrose by b-fructosidase (invertase) into D-glucose and D-fructose at pH 4.6. Then, hexokinase (HKase) catalyzes the phosphorylation of D-glucose by adenosine-5-triphosphate (ATP), producing adenosine-5-diphosphate (ADP) and D-glucose-6-phosphate (G-6-P).…”

Section: Discussionmentioning

confidence: 99%

“…15 It is nontoxic to humans and can be enzymatically detected at low levels. 16 Sucrose is hydrophilic and has a low molecular weight (342 Da), compared to the rather large molecule (about 10 6 Da) for endotoxins. Therefore, sucrose has the potential to be a new and stable tracer.…”

Section: Introductionmentioning

confidence: 99%

Sucrose as a stable tracer for quantifying endodontic leakage

Cheung

Deng

et al. 2012

Journal of Dental Sciences

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Background/purpose: Coronal or apical leakage is a major cause of endodontic treatment failure. To evaluate sucrose as a tracer for leakage testing for various filling materials involved in endodontic treatment. Materials and methods: The stability of sucrose and glucose was examined by immersing 11 common filling materials (1.5 Â 3 mm blocks, cured for 1 week at 37 C in 100% relative humidity; n Z 10 for each) in a 10mM solution of either sucrose or glucose. The concentration of the solution was measured after 1 week, 2 weeks, 4 weeks, and 8 weeks of immersion and compared. Then, the two tracers were used to test the sealing ability of zinc oxideeeugenol cement (IRM) and amalgam root-end fillings. Each material (n Z 40 for each) was equally divided into two subgroups, these were evaluated with either glucose or sucrose as the tracer substance, and the amount of leakage was determined after 24 h, and 1 week, 2 weeks, 3 weeks, 4 weeks, 6 weeks, and 8 weeks. Results: Sucrose was stable with all materials at all time points. The concentration of glucose had significantly diminished after 1 week of immersion with mineral trioxide aggregate (MTA) or Sealapex (P < 0.001), but not with other filling materials (P > 0.05). Leakage results were similar (P > 0.05) when glucose and sucrose were used as the tracer substance, respectively. Amalgam leaked significantly less than IRM after 3 (sucrose test) and 4 weeks (glucose test) (P < 0.05).Conclusion: Sucrose appears to be stable in the presence of various endodontic materials, and can be used as a stable tracer substance for detecting endodontic microleakage.

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“…Glucose, fructose and sucrose assays were adapted from Jones et al (1977). Citric and malic acid assays were modified from Velterop and Vos (2001). Modifications are described in Basson et al (2010).…”

Section: Metabolite Extraction and Quantificationmentioning

confidence: 99%

Upregulation of pyrophosphate: fructose 6-phosphate 1-phosphotransferase (PFP) activity in strawberry

et al. 2010

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Pyrophosphate: fructose 6-phosphate 1-phosphotransferase (PFP) is a cytosolic enzyme catalyzing the first committed step in glycolysis by reversibly phosphorylating fructose-6-phosphate to fructose-1,6-bisphosphate. The position of PFP in glycolytic and gluconeogenic metabolism, as well as activity patterns in ripening strawberry, suggest that the enzyme may influence carbohydrate allocation to sugars and organic acids. Fructose-2,6-bisphosphate activates and tightly regulates PFP activity in plants and has hampered attempts to increase PFP activity through overexpression. Heterologous expression of a homodimeric isoform from Giardia lamblia, not regulated by fructose-2,6-bisphosphate, was therefore employed to ensure in vivo increases in PFP activity. The coding sequence was placed into a constitutive expression cassette under control of the cauliflower mosaic virus 35S promoter and introduced into strawberry by Agrobacterium tumefaciens-mediated transformation. Heterologous expression of PFP resulted in an up to eightfold increase in total activity in ripe berries collected over two consecutive growing seasons. Total sugar and organic acid content of transgenic berries harvested during the first season were not affected when compared to the wild type, however, fructose content increased at the expense of sucrose. In the second season, total sugar content and composition remained unchanged while the citrate content increased slightly. Considering that PFP catalyses a reversible reaction, PFP activity appears to shift between gluconeogenic and glycolytic metabolism, depending on the metabolic status of the cell.

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A rapid and inexpensive microplate assay for the enzymatic determination of glucose, fructose, sucrose, L‐malate and citrate in tomato (Lycopersicon esculentum) extracts and in orange juice

Cited by 71 publications

References 9 publications

Quantitative metabolic profiles of tomato flesh and seeds during fruit development: complementary analysis with ANN and PCA

Quantitative metabolic profiles of tomato flesh and seeds during fruit development: complementary analysis with ANN and PCA

Sucrose as a stable tracer for quantifying endodontic leakage

Upregulation of pyrophosphate: fructose 6-phosphate 1-phosphotransferase (PFP) activity in strawberry

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