Determination of Metabolites by 1H NMR and GC: Analysis for Organic Osmolytes in Crude Tissue Extracts

Fan, Teresa W.‐M.; Colmer, Timothy D.; Lane, Andrew N.; Higashi, Richard M.

doi:10.1006/abio.1993.1486

Cited by 85 publications

(57 citation statements)

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“…Based on published chemical shifts (Fan et al, 1986;Fan et al, 1993;Behar et al, 1983, http://www.bmrb.wisc.edu) and two-dimensional Cosy spectra run on perchloric extracts, the following chemical shifts were assigned to NMR spectra for metabolite comparisons. Alanine was assigned 1.47 ppm, proline 2.1 ppm, and carbohydrates (primarily sucrose and glucose), due to overlap of peaks, were assigned to the area between 3.8 and 4.2 ppm (Fig.…”

Section: Changes In Leaf Compatible Solute Concentrationsmentioning

confidence: 99%

Physiological effects of short-term salinity changes on Ruppia maritima

2003

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Changes in Ruppia maritima L. leaf-tissue osmolality, compatible solute synthesis in leaf tissues, and maximum effective quantum yield in response to short-term changes in salinity were investigated. Plants cultured at 20‰ S were exposed to 0‰ S, 10‰ S (half-ambient), 20‰ S (ambient), and 40‰ S (twice-ambient) salinities. Total and non-vacuolar leaf osmolality for cultured plants significantly decreased (total: from 1464 ± 266 to 712 ± 210 mmol kg −1 ; non-vacuolar: from 880 ± 108 to 257 ± 80 mmol kg −1 ) or increased (total: from 1464 to 2532 ± 673 mmol kg −1 , non-vacuolar from 880 to 1168 ± 15 mmol kg −1 ), within 1 min of exposure to 0 and 40‰ S, respectively. After the initial rapid change in leaf osmolality, values were relatively constant for the first 180 min of exposure. Osmolality then changed again over the period from 1 to 2 days post-treatment with values again increasing (40‰ S: total = 3152 ± 335 mmol kg −1 , non-vacuolar = 1967 ± 103 mmol kg −1 ) or decreasing (0‰ S: total = 357 ± 46 mmol kg −1 , non-vacuolar = 74 ± 32 mmol kg −1 ) with salinity. Soluble and total carbohydrates in leaf tissues responded differently to changing salinity. Total carbohydrates decreased by 65%, while soluble levels increased by 34%, in high salinity. Total and soluble proline levels increased (63 and 18%, respectively), and decreased (−36 and −20% for 10‰ S; −72 and −32% for 0‰ S, respectively), with salinity. These results suggest that both proline and soluble carbohydrates act as compatible solutes. Maximum quantum yields (F v /F m ) were measured over a 48 h period in response to changes in medium salinity and inorganic carbon (ambient and ∼2.0 mM bicarbonate-equilibrium treatments). F v /F m exhibited significant variation in response to salinity, bicarbonate-level and time as main effects, with significant interactions. Quantum yields were lowest in the 0 and 40‰ S treatments; the 10 and 20‰ S treatments had significantly higher quantum yields. These short-term responses indicated that both increases and reductions of external ion concentrations are initially stressful for R. maritima, but that it can physiologically adjust after several days.

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Section: Changes In Leaf Compatible Solute Concentrationsmentioning

confidence: 99%

Physiological effects of short-term salinity changes on Ruppia maritima

2003

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“…The concentrations of at least 36 metabolites, including Gly betaine, SUC, Glc, amino acids, and organic acids, in perchloric acid extracts of leaf blade tissue were determined using the combined 'H NMR spectroscopy and GLC procedure described by Fan et al (1993).…”

Section: Perchloric Acid Extraction Of Metabolitesmentioning

confidence: 99%

“…Low-molecular-weight, water-soluble solutes were extracted twice from 100-mg samples of leaf tissue with 3 mL of 5% (w/v) perchloric acid at 4°C as described by Fan et al (1993). Aliquots of the extracts were put into GC vials and lyophilized for subsequent GLC analysis.…”

Section: Perchloric Acid Extraction Of Metabolitesmentioning

confidence: 99%

Differential Solute Regulation in Leaf Blades of Various Ages in Salt-Sensitive Wheat and a Salt-Tolerant Wheat x Lophopyrum elongatum (Host) A. Love Amphiploid

1995

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“…The area of peak is proportional to the number of nuclei responsible for the peaks and NMR determination of peak area is a conventional and validated method for metabolic profiling. 16,17 Peaks were assigned by comparing the observed chemical shifts with reference spectra using the databases in the Human Metabolome Database (HMDB) and using Chenomx NMR Suite 8.2 (Chenomx, Edmonton, Alberta, Canada) and most of the chemical shift values were already reported in the previously results.…”

Section: Methodsmentioning

confidence: 99%

Assessment of Metabolism of the Eastern Oyster and Eastern Elliptio Using NMR Spectroscopy

Lee¹

2016

J. Braz. Chem. Soc.

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Metabolites are known to characterize the functional responses of a cell. Therefore, a quantitative measurement of metabolite profiles can provide insight into the underlying effect of genetic or environmental actions on cell metabolism. It follows then that the study of the metabolite profiles of bivalve mollusks, such as oysters, can be particularly worthwhile in assessing changes in physiology, pharmacology and toxicology that result from their adaptation to changing environments. We have determined the metabolic profiles of three different organ groups in freshwater mussel and oysters by using 1 H (proton) and 13 C (carbon) NMR (nuclear magnetic resonance) spectroscopy following the infusion of 13 C glucose or 13 C glycine, respectively. The result shows infused glucose formed glycogen by glycogen synthesis, alanine by glycolysis, and glutamate and aspartate through the Krebs cycle and glycine formed serine by the glycine cleavage system in oysters. Decrease in adenosine triphosphate (ATP), glucogen, putrescine and ornithine were observed in the fasting state freshwater mussel. Our result opens the possibility that organ specific metabolic fingerprints can be established to interpret functional adaptations to environmental and nutritional challenges using NMR spectroscopy.

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Determination of Metabolites by 1H NMR and GC: Analysis for Organic Osmolytes in Crude Tissue Extracts

Cited by 85 publications

References 0 publications

Physiological effects of short-term salinity changes on Ruppia maritima

Physiological effects of short-term salinity changes on Ruppia maritima

Differential Solute Regulation in Leaf Blades of Various Ages in Salt-Sensitive Wheat and a Salt-Tolerant Wheat x Lophopyrum elongatum (Host) A. Love Amphiploid

Assessment of Metabolism of the Eastern Oyster and Eastern Elliptio Using NMR Spectroscopy

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