Determination of Patulin in Apple Products using HPLC with Photodiode Array Detector and Ultra Performance Liquid Chromatography with Electrospray-Tandem Mass Spectrometry

Abstract

The mycotoxin, patulin (4-hydroxy-4H-furo[3,2c] pyran-2[6H]-one), is produced by a number of fungi common to fruit- and vegetable-based products, most notably, apples. A rapid, simple, and economical method was described for determination of patulin in apple products. The sample was extracted with ethyl acetate and then the extract was cleaned up by extraction with a sodium carbonate solution. Patulin was determined by reverse-phase liquid chromatography using a C₁₈ column and a photodiode array detector and ultra performance liquid chromatography with electrospray-tandem mass spectrometry. Patulin was separated by an HPLC/DAD method on 125 × 4 mm I.D Purospher Star RP-18 endcapped column (Merck) with 5 µm C₁₈ stacionary phase and it was detected at 275 nm. The mobile phase used was acetonitrile-water containing 1% acetonitrile (5/95 v/v) and the flow-rate was 0.5 mL/min. Ultra performance liquid chromatography (UPLC) was performed on an Acquity UPLC BEH C₁₈ (100 mm × 2.1 mm) column. The mobile phase was acetonitrile -0,01 mol/L amonium acetate (95/5 v/v) for UPLC/MS/MS analysis and the flow-rate was 0.3 mL/min. The limits of detection for patulin were found to be 1 µg/kg for the HPLC/DAD method and 10 µg/kg for the UPLC/MS/MS method.

Keywords:

• Apple products
• Mycotoxin
• Patulin
• UPLC/MS/MS