Patulin Background. Background. Structure. Regulations

Transcription

1 Patulin Background Background Produced by molds such as Aspergillus and Penicillium First investigated for use as an antibiotic until found to be toxic Most commonly found on bruised apples, but may also occur on pears, grapes, peaches, and apricots and juices made from these fruits Structure g/mol Not stable at high ph Interference analytes such as 5-(Hydroxymethyl) furfural (HMF) can make the analysis more challenging Regulations In the US, the FDA maintains a limit of 50 μg/kg EU limits 50 μg/kg for apple juice and apple products 25 μg/kg for solid apple products 10 μg/kg for apple products intended for children 1

2 Patulin in Recent News Ricker Hill Farms in Maine was issued a warning February 15, 2018 Patulin test results were 125 ppb and FDA suggested that a route cause analysis be performed and improvements to the Hazard Analysis and Critical Control Point (HACCP) plan be made Company issued a voluntary recall of two cider brands Hood River Juice Company in Oregon was issued a warning letter from the FDA on April 25, 2018 after an inspection in the fall of 2017 Patulin test results were > 50 ppb Damaged fruit was found on conveyer lines after inspection and sorting Apples were stored outside without temperature control The company blended juices with high levels of patulin with juices that had low levels of patulin Response from company: We consider this FDA warning letter a chance to improve upon our process and food safety program. Continuous improvement is incorporated into our Food Safety Policy. 2

3 Spiked Apple Juice/No clean-up uv HALO 90 Å Biphenyl, 2.7 µm, 2.1 x 100 mm Part Number: HAL µl; 0.5 ml/min; 30 C A= 0.1% Acetic Acid in Water B= 0.1% Acetic Acid in Acetonitrile 5-95% B in 10 min HMF Patulin Black = apple juice no clean-up Blue = apple juice spiked with HMF and patulin min 3

4 Faster Method HALO 90 Å Biphenyl, 2.7 µm, 2.1 x 100 mm Part Number: HAL µl; 0.6 ml/min; 40 C A= 0.1% Acetic Acid in Water B= 0.1% Acetic Acid in Acetonitrile 5-90% B in 2.6 min uv HM F Patuli n min 4

5 Complex Sample Matrices HALO 90 Å Biphenyl, 2.7 µm, 2.1 x 100 mm Part Number: HAL µl; 0.6 ml/min; 40 C A= 0.1% Acetic Acid in Water B= 0.1% Acetic Acid in Acetonitrile 5-90% B in 2.6 min uv Black = apple juice Blue = peach juice Red = apricot juice Patulin elutes at 1.3 min min 5

6 Extraction Protocol: QuECHERS 5 ml juice/5ml ACN + QuEChERS salts (MgSO 4 and NaCl) Quick QuEChERS Draw sample into syringe Push sample through cartridge into vial 6

7 Extraction Protocol: SPE SPE using IRIS PLUS cartridges Condition 3 ml methanol 3 ml DI water Load sample 5 ml of juice Wash 2 ml 1% sodium bicarbonate or 2 ml DI water 2 ml 1% acetic acid Dry 10 min under full vacuum Elute 3 ml n-hexane/acetone (7:3) v/v or 3 ml methanol 7

8 Patulin and HMF Separation: SPP (Superficially Porous Particle) compared to TPP (totally porous particle) Peak height, µv TPP C18, 2.1 x 150 mm, 5 µm HMF Patulin Both of these are samples of apple juice that have been spiked at 50 ng/ml The HALO column shows a sharper peak for patulin Time, min HALO SPP Biphenyl, 2.1 x 100 mm, 2.7 µm 4.0E E E E E+00 Patulin -1.0E Time, min Adapted from Fig.3b in Food Anal. Methods (2016) 9:

9 Peak Area Patulin Calibration Curve y = x R² = Patulin Concentration (ng/ml) 9

10 Repeatability Across 3 different Spiked Samples uv Patulin min 10

11 Peak Area Recovery using Spiked Samples % recovery at 200 ng/ml % recovery at 50 ng/ml Patulin Concentration (ng/ml) 11