Determination of organic acids in seven wheat varieties by capillary gas chromatography

Fresh wheat tops were extracted with acidic 90% ethanol, and the ethanol was evaporated and a portion of the aqueous residue loaded onto DEAE-Sephadex. Organic acids were eluted with pyridinium formate and then lyophilized and the dried residue was derivatized with 1% trimethylchlorosilane in bis(trimethylsilyl)trifluoroacetamide. The acids were then quantitatively determined using capillary gas chromatography and identified using capillary gas chromatography-mass spectrometry. The acidic ethanol extraction of fresh plant tissue was quantitative for all acids except citric while losses in the remaining procedures were controlled by using an internal standard. The ion exchange chromatography made the greatest contribution to experimental error, imposing a minimum loading requirement of 0.1 mumol of each acid for adequate precision. Organic acid profiles were determined for seven wheat cultivars (Triticum aestivum cv Carazinho, Teal, Lance, Warigal, Isis, Maringa, and BH1146) grown on gravel in solution culture for 30 days. Profiles were simple, consisting of only malic, aconitic, and citric acids, with levels of each acid for all varieties falling within the range 2-5 mumol/g fresh tissue. Storage of samples led to a large increase in sampling error and increased the amount of extractable citric acid.