Adaptation of a simple method to determine the total terpenoid content in needles of coniferous trees

Abstract

In ecological research, quantitative methods are often used to measure the total content of metabolites groups (i.e., phenols, carbohydrates). Until recently, there has been no simple and effective method to determine the total terpenoid content with satisfactory repeatability and sensitivity. The procedure proposed by Ghorai et al. (2012) requires the use of fresh plant material. That may be problematic when experimental units are located far from a laboratory. Our goals were to optimize the procedure, and to find the threshold of misestimation using the procedure adjusted to work with dried material. Needles of Pinus sylvestris were used to test the effect of changes in drying, grinding, storage, and extraction on determined total terpenoid content. All applied changes in material storage and grinding decreased the quantity of the terpenoids in needles. Only air-dried and ball-milled material produced similar results to those obtained with fresh material – can be recommended if the fresh material unattainable. Air-dried material may be stored for up to three months, but it resulted in greater variation and then greater sample size is needed. Lower sample mass and solvent volume have no impact on accuracy. Shorter extraction time, oven-drying or microwaving leads to unreliability of measurements.