Development and validation of a fast, high-throughput UPLC-DAD Method for simultaneous quantification of 38 polyphenols in natural waste streams

Efficient analytical methods are necessary to determine the polyphenol content and antioxidant properties in natural waste streams. To avoid long analysis times, excessive solvent consumption, and the cost-intensive nature of MS detectors, the prospect of formulating a fast and accurate UPLC-DAD method, applicable within analytical laboratories, becomes particularly compelling. This poster describes the development and validation of a fast, efficient, and high-throughput analytical method for the simultaneous analysis of 38 polyphenols, based on reversed-phase ultra-performance liquid chromatography (RP-UPLC) coupled to a photodiode array detection (DAD) system. An Infinitylab Poroshell 120 Å SB-C18-column was employed and in optimal conditions, 38 polyphenols were separated in less than 21 minutes. The developed and optimized UPLC-DAD method was validated according to the ICH guideline with respect to the following parameters: linearity, precision (including inter- and intraday precision), limit of detection limit (LOD), limit of quantification (LOQ), accuracy and robustness. The developed method showed good linearity (R2 > 0.999) for all 38 polyphenols. Polyphenols LOD and LOQ values ranged between 0.0074 – 0.1179 mg/L and 0.0225 – 0.3572 mg/L, respectively. The accuracy of polyphenols recovery, determined for three concentrations levels, ranged between 95.0 and 104% and both inter- and intraday precision showed a variation coefficient lower than 5%. Ultimately, the developed UPLC-DAD method was applied to extracts obtained through ultrasonic-assisted extraction with ethanol-water mixtures from natural waste streams, specially applewood, mandarin peels, pine needles, onion skins, root peels, and rosemary, confirming effective identification and quantification of the polyphenols.