Abstract
Culture methods supplemented with high-performance liquid chromatography (HPLC) technique provide a rapid and simple tool for detecting levels of aflatoxins (AFs) produced by fungi. This study presents a robust method for simultaneous quantification of aflatoxin (AF) B1, B2, G1, and G2 levels in several fungal cultivation states: submerged shake culture, liquid slant culture, and solid-state culture. The recovery of the method was evaluated by spiking a mixture of AFs at several concentrations to the test medium. The applicability of the method was evaluated by using aflatoxigenic and non-aflatoxigenic Aspergilli. A HPLC coupled with the diode array (DAD) and fluorescence (FLD) detectors was used to determine the presence and amounts of AFs. Both detectors showed high sensitivity in detecting spiked AFs or AFs produced in situ by toxigenic fungi. Our methods showed 76%–88% recovery from medium spiked with 2.5, 10, 50, 100, and 500 ng/mL AFs. The limit of quantification (LOQ) for AFs were 2.5 to 5.0 ng/mL with DAD and 0.025 to 2.5 ng/mL with FLD. In this work, we described in detail a protocol, which can be considered the foremost and only verified method, to extract, detect, and quantify AFs employing both aflatoxigenic and non-toxigenic Aspergilli.
Highlights
Aflatoxins (AFs) are a group of mycotoxins that are toxic, carcinogenic, and mutagenic
We described in detail a protocol, which can be considered the foremost and only verified method, to extract, detect, and quantify AFs employing both aflatoxigenic and non-toxigenic Aspergilli
In most cases, aflatoxigenic fungi are being identified by culture methods coupled with chromatography (TLC) or high-performance liquid chromatography (HPLC)
Summary
Aflatoxins (AFs) are a group of mycotoxins that are toxic, carcinogenic, and mutagenic. This ability to fluoresce has paved the way for most analytical methods for the detection and known as coumarinic nucleus, AFs have natural fluorescence properties (Figure 1A,B) This ability to quantification of these toxins [12]. In most cases, aflatoxigenic fungi are being identified by culture methods coupled with chromatography (TLC) or high-performance liquid chromatography (HPLC). Gellvalidated and Carbone have used quantifications of aflatoxin cocktail (AFB1,ofAFB2, AFG2)mycelium in the fungal cultures. 3.9 available ng/mL, respectively a number of detection advantages overand others, it being inexpensive, rapid, in most labs, Culture method has technical a numberskills. Due we report new of verifications of these methods, they are generally regarded being less precise than the other method for a rapid, sensitive, and simultaneous detection of AFB1,as. We have optimized and validated the method through a series of experiments to meet the research laboratory needs for a robust, fast, easy to use, cheap, and environmentally friendly protocol with minimum organic solvents waste
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