Development and Method Validation of Ochratoxin A Analysis in Cereals using High Performance Liquid Chromatography - High Resolution Mass Spectrometry
Keywords:cereal; HPLC-HRMS; method validation; Ochratoxin A; wheat
The objective of this study was to develop an analytical method for ochratoxin A in cereals, modified from AOAC official method 2000.03 using high performance liquid chromatography - high resolution mass spectrometry (HPLC-HRMS). The ion polarity (negative and positive mode) and the different ratios between 0.1% formic acid in water (v/v) and acetonitrile in mobile phase at 20:80, 30:70 and 40:60 were studied. We found that optimal conditions for ochratoxin A analysis using HPLC-HRMS were negative mode for ion polarity and 0.1% formic acid in water – acetonitrile ratio of 30: 70 as HPLC mobile phase. The method was then validated to ensure that it would deliver reliable results. The validation results demonstrated that the working range, limit of detection and limit of quantitation were 0.50 – 20 μ g/kg, 0.20 μ g/kg and 0.50 μ g/kg, respectively. Moreover, the accuracy and precision of the method are within acceptance criteria. (%Recovery 72.54 - 116.60, %RSD 13.10 - 13.69). All results demonstrated that the developed method could detect the presence of ochratoxin A in cereals at the level below permitted levels of the importing countries’ regulation. The method is accredited ISO/IEC 17025 by Bureau of Laboratory Standards, Department of Medical Sciences. and is being used as a standard method for ochratoxin A analysis in cereal at laboratory of Plant Standard and Certification Division, Department of Agriculture.
Bertil Magnusson and Ulf Örnemark. 2014. EURACHEM Guide:The Fitness for Purpose of Analytical Methods, A Laboratory Method Validation and Related Topics, Second edition. Available from www.eurachem.org
Codex Alimentarius. 1995. Codex General Standard for Contaminants and Toxin in Food and Feed (CODEX STAN 193-1995). 41p.
Codex Alimentarius. 2018. GENERAL STANDARD FOR CONTAMINANTS AND TOXINS IN FOOD AND FEED CODEX STAN 193-1995. Amended in 2018
European Commision. 2006. COMMISSION REGULATION (EC) No 1881/2006 of 19 December 2006 setting maximum levels for certain contaminants in foodstuffs. Official Journal of the European Union. 364: 5-24.
European Commision. 2017. Guidance document on analytical quality control and method validation procedures for pesticide residues and analysis in food and feed SANTE/11813/2017. Implemented by 01/01/2018
George W. Latimer, Jr. 2016. Official Method of Analysis of AOAC International 20thedition. AOAC International. Chapter 49: 74 -76, Appendix F: 1-18
IARC (International Agency for Research on Cancer). 1993. IARC Monographs on the Evaluation of Carcinogenic Risks to Humans: Some Naturally Occurring Substances: Food Items and Constituents, Heterocyclic Aromatic Amines and Mycotoxins. 56: 489–599
JECFA (Joint FAO/WHO Expert Committee on Food Additives). 2008.Safety evaluation of certain food additives and contaminants. TRS 947-JECFA 68/59
Lund, F., Frisvad, J.C. 2003. Penicillium verrucosum in wheat and barley indicates presence of ochratoxin A. Journal of Applied Microbiology. 95: 1117-1123
WHO (World Health Organization). 2008. Ochratoxin A. Safety evaluations of certain 1 food additives and contaminants. WHO food additives series. 59: 357-429
How to Cite
อ้างอิงแหล่งที่มา ห้ามนำไปใช้เพื่อการค้า และห้ามดัดแปลง