Development and validation of a liquid chromatographic tandem mass spectrometric method for the analysis of patulin in apple and apple juice

Raviraj Shinde, Manisha Dhanshetty, Ankita Lakade, Christopher T. Elliott, Kaushik Banerjee*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


This study reports a robust and sensitive method for rapid testing of patulin in apple and apple juice. The method involved extraction of homogenised samples (10 g) with ethyl acetate (10 mL) and clean up by dispersive-solid phase extraction using primary secondary amine (25 mg/mL). Prior to the LC-MS/MS analysis, the cleaned extract was reconstituted in methanol/water (2:8). The optimised LC-MS condition provided a symmetric peak of patulin within a short LC-runtime of 5 min. The recoveries at the limit of quantification (0.005 mg/kg) and higher levels were satisfactory (> 80%), with the precision-RSDr (< 11%). In an inter-laboratory comparison study involving 13 accredited laboratories, the reproducibility-RSDR and HorRat values ranged between 4.80 and 6.08% and between 0.18 and 0.23 respectively, indicating a satisfactory method-precision. The z-scores of the participating laboratories were within ± 2. When the method was applied to incurred samples, the contamination range was 0.008–0.225 mg/kg and 0.018–0.034 mg/kg for apple and juice respectively, demonstrating a satisfactory performance in terms of precision. Based on the solvent standard, matrix-matched standard and standard-addition approaches, the calibration graphs provided similar quantitative performances. Because of its reliability, robustness and time-effectiveness, the method can be recommended for regulatory testing purposes.

Original languageEnglish
Pages (from-to)119-127
Number of pages9
JournalMycotoxin Research
Issue number2
Publication statusPublished - 22 Feb 2021

Bibliographical note

Funding Information:
We received partial funding from the Food Safety Standards Authority of India (FSSAI) under the National Referral Laboratory (NRL) scheme.

Publisher Copyright:
© 2021, Society for Mycotoxin (Research Gesellschaft für Mykotoxinforschung e.V.) and Springer-Verlag GmbH Germany, part of Springer Nature.

Copyright 2021 Elsevier B.V., All rights reserved.


  • Apple
  • Apple juice
  • Ethyl acetate extraction
  • LC–MS/MS
  • Method validation
  • Patulin analysis

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology
  • Toxicology


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