Measurement of the 13C/12C ratio of soil-plant individual sugars by gas chromatography/combustion/isotope-ratio mass spectrometry of silylated derivatives.
Carbohydrate is an important pool in the terrestrial carbon cycle. The potential offered by natural and artificial 13C-labelling techniques should therefore be applied to the investigation of the dynamics of individual sugars in soils. For this reason, we evaluated the method of 13C sugar analysis by gas chromatography/combustion/isotope-ratio mass spectrometry (GC/C/IRMS) after hydrolysis and direct trimethylsilylation. Trimethylsilylation involved the addition of several carbon atoms per sugar. These atoms have to be taken into account in the estimation of the carbon isotope ratio. The analysis of standard and natural pentoses and hexoses of known 13C enrichments revealed that the number of analysed added carbon atoms was less than expected from stoichiometry. This was attributed to incomplete derivatization and/or incomplete oxidation of methylsilyl carbon before IRMS. Using a calibration of the number of analysed added carbon atoms, the isotope excess of enriched samples could be determined with a relative error close to 5%. Concerning the determination of natural abundances by GC/C/IRMS, we could measure the delta 13C of standard C3- and C4-derived sugars with an accuracy of +/-1.5 per thousand using the previous calibration. We were able to apply this technique to plant-soil systems labelled by pulse-chase of 13CO2, revealing the nature and dynamics of sugars in the plant rhizosphere.
Derrien
Delphine
D
Laboratoire d'Ecologie Microbienne de la Rhizosphère (LEMIR), UMR 163 CNRS/CEA, CEA/DSV/DEVM, Cadarache, 13108 Saint-Paul-lez-Durance, France.
Balesdent
Jérôme
J
Marol
Christine
C
Santaella
Catherine
C
D000465:Algorithms; D002138:Calibration; D002241:Carbohydrates / Q000737:chemistry; D002247:Carbon Isotopes / Q000032:analysis; D002250:Carbon Radioisotopes / Q000032:analysis; D008401:Gas Chromatography-Mass Spectrometry; D006868:Hydrolysis; D007202:Indicators and Reagents; D010944:Plants / Q000737:chemistry*; D011134:Polysaccharides / Q000737:chemistry; D012987:Soil / Q000032:analysis*; D014297:Trimethylsilyl Compounds / Q000737:chemistry*; D014908:Triticum / Q000737:chemistry
D002241:Carbohydrates; D002247:Carbon Isotopes; D002250:Carbon Radioisotopes; D007202:Indicators and Reagents; D011134:Polysaccharides; D012987:Soil; D014297:Trimethylsilyl Compounds