| Summary: | Sugar and organic acid contents are major factors for tomato fruit flavour and are important breeding traits. Here we provide an improved protocol for accurate quantification of the main sugars, glucose and fructose, and the organic acids, citric acid and malic acid, present in tomato. The tomato extract is spiked with lactose and tricarballylic acid as internal standards and loaded onto a NH2 solid phase extraction (SPE) column. The sugars appear in the flow-through and are subsequently analysed by HPLC using a Nucleodur NH2 column and a refractive index detector. The organic acids bind to the SPE column and are eluted with 400 mM phosphoric acid. For analysis, the organic acids are separated by HPLC using a Nucleodur C18ec column and detected by UV absorption at 210 nm. The method shows excellent inter-day and intra-day reproducibility for glucose, fructose and citric acid with standard deviations of 1–5%. Quantification of citric acid by HPLC and GC–MS showed perfect agreement with a deviation of less than 3%. • Simple method for quantification of glucose, fructose, citric acid and malic acid in tomato. • Efficient removal of interfering compounds by solid phase extraction. • High intra and inter-day reproducibility. Method name: Quantification of sugars and organic acids by HPLC and GC–MS, Keywords: Citric acid, Fructose, GC–MS, Glucose, HPLC, Malic acid, Solid phase extraction
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