Internal Validation of Rapid and Performance Method for Patulin Determination in Apple Cider by High-Performance Liquid Chromatography

An analytical method was developed and validated for separation, detection and quantification of patulin in apple cider by high performan celiquid chromatography (HPLC-DAD). Extraction and clean-up of patulin from apple cider are achieved on AFFINIMIP (R) SPE PATULIN cartridge (3mL/100mg). Patulin was separated on a C18 Hypersil GOLD column (150 mm x 4 mm, 5 mu m) coupled to a Hypersil Gold guard column (10 x 4.0 mm, 5 mu m) using mobile phase consisting of water:acetonitrile (95:5, v/v) and detected at 276 nm. The method has a good sensitivity (LOD = 2.63 mu g/mL and LOQ = 8.45 mu g/mL) and a good precision (RDS (r) = 0.32...0.91% for injection repeatability; RSD (r) = 1.05....2.98% for analysis repeatability intra-day; RSD (R) = 1.35....3.24% for intermediate precision; RSD (R) = 1.64....3.86% for intra-laboratory reproducibility). The method was applied by analyzing samples of apple cider purchased from commerce. In case of 80% of the analyzed samples, patulin was not detected, and for the rest of them concentration of patulin was < LOQ.