Development of a corn flour certified reference material for the accurate determination of zearalenone.

A certified reference material (CRM, KRISS 108-01-002) for zearalenone in corn flour was developed to assure reliable and accurate measurements in testing laboratories. Commercially available corn flour underwent freeze-drying, pulverization, sieving, and homogenization. The final product was packed in amber bottles, approximately 14 g per unit, and preserved at -70 °C. 13C18-Zearalenone was used as an internal standard (IS) for the certification of zearalenone by isotope-dilution liquid chromatography-tandem mass spectrometry (ID-LC‒MS/MS) and for the analysis of α-zearalenol, ß-zearalenol, and zearalanone by LC‒MS/MS. The prepared CRM was sufficiently homogeneous, as the among-unit relative standard deviation for each mycotoxin ranged from 2.2 to 5.7 %. Additionally, the stability of the mycotoxins in the CRM was evaluated under different temperature conditions and scheduled test periods, including storage at -70°C, -20°C, and 4°C and room temperature for up to 12 months, 6 months, and 1 month, respectively. The content of each target mycotoxin in the CRM remained stable throughout the monitoring period at each temperature. Zearalenone content (153.6 ± 8.0 µg/kg) was assigned as the certified value. Meanwhile, the contents of α-zearalenol (1.30 ± 0.17 µg/kg), ß-zearalenol (4.75 ± 0.33 µg/kg), and zearalanone (2.09 ± 0.16 µg/kg) were provided as informative values.

Occurrence of zearalenone and its major metabolites in cereal flour from Korea.

The occurrence of zearalenone (ZEN) and its metabolites (α-zearalenol (α-ZEL), ß-zearalenol (ß-ZEL), α-zearalanol (α-ZAL), ß-zearalanol (ß-ZAL), and zearalanone (ZAN)) was investigated in 78 cereal flour from Korea using UHPLC-MS/MS. Among these mycotoxins, ZEN was the most abundant in the analyzed samples at an incidence rate of 41% and concentration range of 0.5-536 µg/kg. The highest contamination and incidence rate of ZEN were found in corn flour samples, while oat flour samples showed the lowest contamination and incidence rate of this mycotoxin. α-ZEL, ß-ZEL, and ZAN were detected only in corn flour samples but at lower frequencies of 23%, 17%, and 15%, respectively, while α-ZAL and ß-ZAL were not detected in any sample. To the best of our knowledge, this is the first investigation of the simultaneous occurrence of ZEN and its major metabolites in commercially available cereal flour from Korea. Among the tested samples, only four were contaminated with ZEN at levels exceeding the maximum regulatory level established in Korea. The co-occurrence of ZEN, α-ZEL, ß-ZEL, and ZAN was observed in 14% of all samples. Although ZEN metabolites were detected at relatively lower levels than ZEN, the relatively high co-occurrence rate of those mycotoxins is of significant concern from a food safety perspective, since they can synergistically contribute to the overall toxicity and estrogenic effects.

Accurate determination of four tetracycline residues in chicken meat by isotope dilution-liquid chromatography/tandem mass spectrometry.

An analytical method based on isotope dilution-liquid chromatography-tandem mass spectrometry (ID-LC‒MS/MS) was developed to accurately determine four representative tetracyclines (tetracycline, chlortetracycline, doxycycline, and oxytetracycline) in chicken meat. Tetracyclines are known to have a great tendency for epimerization and keto-enol tautomerism, which often provoke major challenges in their determination. Since this isomerization was found to be unavoidable during the whole chain of the current analysis, the total content (µg kg‒1) of individual tetracycline was quantified as a sum of each parent compound and its respective isomeric forms. Using this approach in combination with IDMS analysis, more consistent, accurate, and reproducible measurement results for the four tetracyclines in chicken meat were acquired. LC-MS/MS conditions and sample preparation processes were comprehensively optimized to minimize the chelating effect of tetracyclines and possible co-extracted interferences. Details of the sample preparation scheme, LC‒MS/MS detection, calculation equation, and method validation are described in this article. The method provided very good accuracy (97.7-102.6%) for all analytes across the concentration range of 10-200 µg kg‒1, with relative standard deviations for intra-day and inter-day precision of less than 4%. The limits of quantification were below 0.2 µg kg‒1, demonstrating the high sensitivity of the method. Furthermore, the measurement uncertainty was generally below 5.5%. Hence, the established method exhibits high-order metrological quality with superior performance over various existing methodologies. Moreover, this method can provide references for general food testing laboratories close to and far below the established maximum residue limits (100 µg kg‒1) for animal muscle tissues.

Development of isotope dilution-liquid chromatography/tandem mass spectrometry for the accurate determination of zearalenone and its metabolites in corn.

A method using isotope dilution-liquid chromatography-tandem mass spectrometry (ID-LC/MS/MS) was developed for the accurate determination of zearalenone (ZEN) and its five major metabolites in corn. 13C- or 2H-labeled analogues of the target mycotoxins were used as internal standards. As the immunoaffinity columns used demonstrated selectivity to a specific chiral isomer of a racemic mixture of zearalanone-d6, a clean-up cartridge without stereoselectivity (Mycosep 226 column) was selected for the same recovery of the analyte and its internal standard with adequate elimination of matrix interferences. The method demonstrated sufficient selectivity, sensitivity, accuracy and precision over a concentration range of 20-400 µg/kg. The limit of detections and limit of quantifications were 0.14-0.33 µg/kg and 0.45-1.11 µg/kg, respectively. The accuracy values were 96.7%-103.6%, with intra and inter-day precisions of less than 3% and 4%, respectively. The expanded measurement uncertainty was less than 7% (with a 95% confidence level).