Development of isotope dilution–liquid chromatography/tandem mass spectrometry as a candidate reference method for the accurate determination of patulin in apple products

• Published in: Analytical and bioanalytical chemistry Vol. 414; no. 5; pp. 1867 - 1879
• Main Authors: Gab-Allah, Mohamed A., Choi, Kihwan, Kim, Byungjoo
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.02.2022; Springer; Springer Nature B.V
• Subjects: Acetic acid; Analysis; Analytical Chemistry; Apple; Apples; Biochemistry; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Chromatography; Chromatography, Liquid - methods; Dilution; Ethyl acetate; Extraction (Chemistry); Food Science; Health risks; Identification and classification; Imprinted polymers; Indicator Dilution Techniques; Laboratory Medicine; Liquid chromatography; Malus - chemistry; Mass spectrometry; Mass spectroscopy; Methods; Monitoring/Environmental Analysis; Patulin; Patulin - analysis; Patulin - standards; Performance enhancement; Polyketides; Polymers; Reference Standards; Research Paper; Sample preparation; Scientific imaging; Sharpness; Solid phases; Spectroscopy; Tandem Mass Spectrometry - methods; South Korea; Egypt; Isotope dilution–mass spectrometry; Higher-order reference method; Patulin; Mycotoxin; Apple-based products
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-021-03817-3

The incidence of patulin (PAT) in fruit products is a worldwide concern due to its acute and chronic toxic effects. Therefore, accurate and reliable PAT measurements are important for preventing consumer health risks. Our previous method, which was based on a common technique that uses ethyl acetate extraction and liquid chromatography–tandem mass spectrometry with isotope dilution (ID–LC–MS/MS), has shown great performance for the determination of PAT in apple products. However, prolonged extraction times and multistep clean-up processes were required to sufficiently eliminate the matrix interferences. Herein, a feasible alternative ID–LC–MS/MS method was successfully established, employing simplified and reliable sample preparation steps. The clean-up process was performed using molecularly imprinted polymer–solid-phase extraction (MIP–SPE) cartridges, which eliminated matrix interferences and facilitated the trace quantification. While the previous method used a multimode LC column for the retention of polar patulin, the current method used a UPLC HSS T3 column, which further improved the peak sharpness and reduced the run time. The method was validated by measuring fortified samples in the concentration range of 5‒100 µg/kg. The accuracy varied between 97.8 and 102.0%, with relative standard deviation for interday and intraday precision being below 3%. The measurement uncertainty was lower than 4% (at a 95% level of confidence). Therefore, this method demonstrated adequate metrological quality with greatly enhanced performance over various reported methods. Additional key benefits of this method are easy manipulation, short preparation time, and lower consumption of hazardous solvents. Finally, the method was successfully applied to commercially available apple-based products. Graphical abstract