A Rapid, Sensitive Thin Layer Chromatography Procedure for the Detection of Fumonisin B₁ and B₂

Abstract

Abstract.A thin layer chromatography (TLC) method was developed for the detection of fumonisin B 1 and B 2 in corn and corn-based feedstuffs. Finely ground samples were extracted with acetonitrile : water (1: 1), filtered, and applied to C 18 cleanup columns. The columns were washed with 1% aqueous KCl followed by acetonitrile : 1% aqueous KC1 (1:9), and the fumonisins were eluted with acetonitrile : water (7:3). The eluants were concentrated and spotted on reverse-phase C 18 TLC plates along with fumonisin B 1 and B 2 standards, and the plates were developed in methanol : 4% aqueous KCl (3:2). The fumonisins were visualized by spraying the TLC plates successively with 0.1 M sodium borate buffer, fluorescamine, and 0.01 M boric acid. The plates were then dried and examined under longwave ultraviolet light. Fumonisin B 1 and B 2 appeared as bright yellowish-green fluorescent bands at R f s of 0.5 and 0.1, respectively. The detection limit for the fumonisins on the TLC plate was 0.1 ppm in corn. Recoveries from spiked samples averaged >80%. The identification of the fumonisins was confirmed by hydrolyzing the parent compounds of B 1 and B 2 to their respective C 22 amino-alcohols and reexamining by TLC with the same visualizing reagents. This procedure was used to survey 193 corn samples collected from University of Missouri test plots in 1990 for fumonisin B,. Fumonisin B 1 was detected in 15% of the corn samples.The fumonisins, a new class of mycotoxins, have been isolated and characterized from toxic cultures of Fusarium moniloforme 1,2 and Fusarium proliferatum. 9 One of the first reports of the natural occurrence of the fumonisins described the identification of the fumonisins in corn from Transkei, Republic of South Africa. 12 Recent studies have shown fumonisin B 1 to be responsible for equine leukoencephalomalacia (ELEM) 5,6 and swine pulmonary edema. 3 In the United States, fumonisin-contaminated corn screenings have been associated with cases of ELEM 7-9,13 and swine pulmonary edema, 3,9 but only limited information is available on natural occurrence and levels of incidence of the fumonisins. A number of analytical procedures for fumonisins in corn and corn products, including thin layer chromatography (TLC), high performance liquid chromatography (HPLC), and gas chromatography/ mass spectroscopy (GC/MS), 8,10,12,13 are available, but all are of limited value or not readily available for extensive monitoring of fumonisin contamination in corn. In this report, a rapid and sensitive TLC procedure for detecting fumonisin B 1 and B 2 in corn and corn-based feeds is described. This procedure was used to document the natural occurrence of fumonisin B 1 and B 2 in Missouri's 1990 corn harvest. Received for publication October 7, 1991. Materials and methodsSample collection. Corn samples (approximately 450 g) were collected, as part of an ongoing mycotoxin monitoring program, from the University of Missouri experimental test plots. Samples were labeled by location and hybrid. Dry land test plots were located in ...

Urchin-like gold nanoparticle-based immunochromatographic strip test for rapid detection of fumonisin B1 in grains

“…S4. FB 1 positivity was obtained for samples 2,9,14, and 24 of wheat and samples 4, 12, 15, and 19 of maize at levels of 1.0 to 3.53 mg/kg by ELISA and HPLC; these samples also gave a positive result with one red line on the ICS test (Fig. 3).…”

“…The former includes thin-layer chromatography, gas chromatography (GC), HPLC, capillary electrophoresis (CE), and liquid chromatography-mass spectrometry (LC-MS) [9][10][11][12][13][14], and the latter includes enzyme-linked immunosorbent assay (ELISA) and membrane matrix immunoassays [15][16][17]. To effectively measure the occurrence of FB 1 in food at low contamination levels, rapid, reliable, sensitive, low-cost, and simple analytical technique is required.…”

Coordinated and Distinct Functions of Velvet Proteins in Fusarium verticillioides

“…Full-scan spectra were acquired over a scan range of m/z 300 to 800. Fumonisins were identified by comparison to an FB1 standard, as well as the previously described molecular weight and fragmentation spectrum (35,36).…”

“…Extracts from cultures of the wild type, Fvve1, FvvelB, and FvvelC deletion mutants, and FvvelB [FvVELB] complemented strains grown on cracked corn medium in constant darkness for 11 days were analyzed by high-performance liquid chromatography-mass spectrometry (HPLC-MS). Fumonisins were first extracted according to the acetonitrile method described by Rottinghaus et al (35). HPLC-MS analysis of fumonisins was performed as described previously by Silva et al (36) with some modifications.…”

Immunochemical Methods for Fumonisins in Corn

“…Methods for fumonisin determination include thin-layer, liquid, and gas chromatography, as well as mass spectrometry (14)(15)(16). All these methods entail sample preparation, extraction, solid phase purification, chromatographic separation, and derivitization prior to quantitation.…”