Analytical Procedures for the Sequential Extraction of ¹⁴C‐Labeled Constituents from Leaves, Bark and Wood of Cottonwood Plants

Abstract

Abstract A procedure for the extraction, separation, and measurement of photosynthetically fixed 14 C in up to 8 chemical fractions (CHCl), amino acids, organic acids, sugars, protein, starch, hemicellulose, and residue) from small samples (1 to 100 mg) of cottonwood ( Populus deltoides Bartr.) leaf material is described. The different chemical fractions are extracted in a sequence of chemical, ion exchange, and enzymatic steps. The 14 C‐activity in these major fractions is then determined with liquid scintillation spectrometry. These major fractions ( e.g. , sugars, amino acids, organic acids) can be further separated into their individual chemical components by standard thin‐layer or gas‐ chromatographic methods for quantitative analysis if specific activities are desired. The major advantage of the procedure is that many chemical fractions can be sequentially separated with good reproducibility from a small amount of plant material without transfer of the material from the original homogenizer or centrifuge tube.