Development of a high-performance liquid chromatography method for simultaneous quantification of sixteen polyphenols and application to tomato

Abstract

The main purpose of the current work was to develop a new method to evaluate and quantify sixteen polyphenol compounds from tomato fruit using high-performance liquid chromatography (HPLC). The separation of 16 polyphenols from tomato fruit was achieved in < 60 min by using a Waters Symmetry C18 column (250 × 4.6 mm i. d, 5 µm particle sizes) with a gradient system of ultrapure water (1 % acetic acid) and 100 % methanol, a temperature of 30 °C, an injection volume of 10 μL and a flow rate of 1.1 mL/min, respectively. The analytical characteristics of evaluation method provide sufficient sensitivity for all tomato polyphenols compounds within normal range 0.1–20 μg·mL−1 (R2≥0.999) with 0.069–0.365 μg·mL−1 LOD, and 0.171–1.106 μg·mL−1 LOQ, with good system suitability (<2 % RSD of retention time, peak area, and tailing factor, 6,000–1,336,000 N, and >1.5 peak resolution), <10 % RSD of precision, stability, repeatability, and robustness, and 99.2 - 105.0 % of recovery. The applicability of this method was demonstrated by the determination of polyphenols in nine cultivars of tomatoes. The results showed that ‘184′ possessed the highest content of total polyphenols (1249.53 μg·g−1 DW) followed by ‘Disease resistance 184′ (1064.93 μg·g−1 DW). The main polyphenol components were rutin, quercetin, gallic acid, chlorogenic acid, 2,5-dihydroxy benzoic acid, caffeic acid and benzoic acid in tomato fruits. In conclusion, this novel HPLC method is useful and acceptable to analyze polyphenols in tomato fruit.