Modelling uncertainty estimation for the determination of aflatoxin M₁in milk by visual and densitometric thin-layer chromatography with immunoaffinity column clean-up

Abstract

The uncertainty of aflatoxin M(1) concentration in milk, determined by thin-layer chromatography (TLC) with visual and densitometric quantification of the fluorescence intensities of the spots, was estimated using the cause-and-effect approach proposed by ISO GUM (Guide to the expression of uncertainty in measurement) following its main four steps. The sources of uncertainties due to volume measurements, visual and densitometric TLC calibration curve, allowed range for recovery variation and intermediary precision to be taken into account in the uncertainty budget. For volume measurements the sources of uncertainties due to calibration, resolution, laboratory temperature variation and repeatability were considered. For the quantification by visual readings of the intensity of the aflatoxin M(1) in the TLC the uncertainty arising from resolution calibration curves was modelled based on the intervals of concentrations between pairs of the calibration standard solutions. The uncertainty of the densitometric TLC quantification arising from the calibration curve was obtained by weighted least square (WLS) regression. Finally, the repeatability uncertainty of the densitometric peak areas or of the visual readings for the test sample solutions was considered. For the test samples with aflatoxin M(1) concentration between 0.02 and 0.5 µg l(-1), the relative expanded uncertainties, with approximately 95% of coverage probability, obtained for visual TLC readings were between 60% and 130% of the values predicted by the Horwitz model. For the densitometric TLC determination they were about 20% lower. The main sources of uncertainties in both visual and densitometric TLC quantification were the intermediary precision, calibration curve and recovery. The main source of uncertainty in the calibration curve in the visual TLC analysis was due to the resolution of the visual readings, whereas in the densitometric analysis it was due to the peak areas of test sample solutions followed by the intercept and slope uncertainties of the calibration line.

Regulation of fungal community and the quality formation and safety control of Pu‐erh tea

“…Pu-erh tea grown in a regular processing environment contains less aflatoxin, but it is easy to produce aflatoxin if it is piled or stored in an environment with a high humidity (Haas et al, 2013). The main methods for detecting mycotoxins in Pu-erh tea are listed in Table 3, including liquid chromatographytandem mass spectrometry (LC-MS/MS) (Ge et al, 2011;Li, Tan, et al, 2015;Su et al, 2016), liquid chromatography (HPLC) (Campone et al, 2011;Li, Xie, et al, 2015), enzymelinked immunosorbent assay (ELISA) (Rosi et al, 2007;Tan et al, 2014), ultra-high performance liquid chromatography (UPLC) (Fu et al, 2009), thin-layer chromatography (TIC) (Carvalho et al, 2012;Rizzo et al, 2004), gas chromatography (GC) (Ibanez-Vea et al, 2011;Mao et al, 2015) and nanoprobe (Li et al, 2013;Wu, Duan, et al, 2013). Liu et al (2017) established the high-throughput detection method of aflatoxin B1(AFB1), aflatoxin B2(AFB2), deoxynivalenin Alcohol (DON), 3-Acetyl Deoxynivalenol (3-AcDON), Zearalenone (ZEN), T-2 Toxin (T-2), HT-2 Toxin (HT-2), ochratoxin A (OTA), and other mycotoxins in Pu-erh tea using UPLC-MS/MS.…”

Aflatoxin Occurrence, Detection, and Novel Strategies to Reduce Toxicity in Poultry Species

“…The AFs have been detected in food and feed samples according to the method of Official Analytical chemists (AOAC) [134]. The most commonly used methods are based on emission and absorption characteristics such as liquid chromatography mass spectroscopy (LC-MS) [135,136], thin layer chromatography [137], highperformance liquid chromatography (HPLC) [138], gas chromatography (GC) [139], and enzyme-linked immunosorbent assay (ELISA) [140]. However, the drawbacks of these commonly used methods are that these methods are tiresome, time-consuming and require skilled technical persons for operation.…”

Characteristics, Occurrence, Detection and Detoxification of Aflatoxins in Foods and Feeds

“…The detection of AFs is performed by several conventional methods based on the emission and absorption characteristics, such as liquid chromatography mass spectroscopy (LC-MS) [126], thin layer chromatography (TLC) [127], gas chromatography (GC) [128], high-performance liquid chromatography (HPLC) [129], immunoaffinity column assay (ICA) [130], and enzyme-linked immunosorbent assay (ELISA) [131].…”