A fast and sensitive isocratic high performance liquid chromatography method for determination of guaraná (Paullinia cupana) flavan-3-ols

Abstract Most methods for quantification of catechins in guarana and other food matrices rely on lengthy gradients to resolve all peaks, and the analysis time for one sample is around 45–135 min. The present method is a fast, sensitive and simple HPLC-UV method, with a 16-minute isocratic run that deliver the high throughput needed to process a large number of samples without compromising the analyte stability. This method is suitable for the determination of catechins, procyanidins (up to degree of polymerisation 2) and caffeine in guarana extracts, in vitro digesta, and Caco-2 cell permeates. The higher sensitivity was achieved by detection at 210 nm, after checking for the absence of interfering substances in the matrices. The method was validated and LODs of 0.019, 0.030, 0.028, 0.030, and 0.043 nmol/mL were achieved for catechin, epicatechin, procyanidin B1, procyanidin B2 and caffeine, respectively. Standard recovery at three different concentrations were within 97–109%. Intra- and inter-day variabilities (RSD) were both under 4%. • This is an isocratic HPLC-UV method for the quantification of flavan-3-ols from guarana (Paullinia cupana), which is 3–8 times faster and >80 times sensitive than previously published methods using UV or diode array detectors. • It is cost-effective as it uses the widely available and affordable UV detector and consumes less solvent due the short analysis time. • With low LODs, fast sample preparation and the short analytical run, this method can be used for the quantification of flavan-3-ols and caffeine in guarana extracts as well as guarana in vitro digesta and its Caco-2 permeates.