Development of a multi‐mycotoxin liquid chromatography/tandem mass spectrometry method for sweet pepper analysis

Abstract

A multi-mycotoxin method was developed for the simultaneous determination of trichothecenes (nivalenol, deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, neosolaniol, fusarenon-X, diacetoxyscirpenol, HT-2 toxin, T-2 toxin), aflatoxins (aflatoxin-B(1), aflatoxin-B(2), aflatoxin-G(1) and aflatoxin-G(2)), Alternaria toxins (alternariol, alternariol methyl ether and altenuene), fumonisins (fumonisin-B(1), fumonisin-B(2) and fumonisin-B(3)), ochratoxin A, zearalenone, beauvericin and sterigmatocystin in sweet pepper. Sweet pepper was extracted with ethyl acetate/formic acid (99:1, v/v). After splitting up the extract, two-thirds of the extract was cleaned up using an aminopropyl column followed by an octadecyl column. The remaining part was cleaned up using a strong anion-exchange column. After recombination of both cleaned parts of the sample extract, the combined solvents were evaporated and the residue was dissolved in mobile phase; 20 microL was injected into the chromatographic system, so only one run was used to separate and detect the mycotoxins in positive electrospray ionization using selected reaction monitoring. The samples were analyzed with a Micromass Quattro Micro triple quadrupole mass spectrometer (Waters, Milford, MA, USA). The mobile phase consisted of variable mixtures of water and methanol, 1% acetic acid and 5 mM ammonium acetate. The limits of detection of the multi-mycotoxin method varied from 0.32 microg kg(-1) to 42.48 microg kg(-1). The multi-mycotoxin liquid chromatography/tandem mass spectrometry (LC/MS/MS) method fulfilled the method performance criteria required by the Commission Regulation (EC) No 401/2006. Sweet peppers inoculated by Fusarium species were analyzed using the developed method. Beauvericin (9-484 microg kg(-1)) and fumonisins (fumonisin-B(1) up to 4330 microg kg(-1), fumonisin-B(2) up to 4900 microg kg(-1), and fumonisin-B(3) up to 299 microg kg(-1)) were detected.

A direct assessment of mycotoxin biomarkers in human urine samples by liquid chromatography tandem mass spectrometry

“…For this reason there was the need to perform SPE clean-up using SAX SPE cartridges with the aim of obtaining a satisfactory recovery for this analyte. Data from the literature revealed that prior to SAX SPE clean-up, complete ionization of FB 1 was achieved when the pH of the sample extract (in this case, sweet pepper extract) was adjusted to between 5.8 to 6 [44]. However, in this study, complete ionization of FB 1 was achieved at pH 6.5-7, which highlights once more the influence of sample matrix on the ionization of FB 1 [45].…”

Scientific Opinion on the risks for animal and public health related to the presence ofAlternariatoxins in feed and food

“…The combination of LLE with SPE is applied for carrots (Solfrizzo et al, 2004), food supplements (Diana di Mavungu et al, 2009), sweet peppers (Monbaliu et al, 2009), cereal, fruit and vegetable products (Asam et al, 2010) and irradiated soya beans ). Nawaz et al (1997 combined LLE with gel permeation chromatography (GPC) for the detection of Alternaria toxins in oilseed rape meal and sunflower seed meal.…”

LC-MS/MS multi-analyte method for mycotoxin determination in food supplements

“…LC-MS/MS appears to be most promising as a highly specific, broadly applicable detection method that provides both qualitative and quantitative data. Considering the possible contamination of foodstuffs by several mycotoxin producing fungal species and the production of different types of mycotoxins by one mould, a trend is to develop methods suitable for the determination of several mycotoxins in a single run (Sewram et al 1999;Monti et al 2000;Rundberget and Wilkins 2002;Royer et al 2004;Berthiller et al 2005;Cavalière et al 2005;Kokkonen et al 2005;Sorensen and Elbaek 2005;Abbas et al 2006;Delmulle et al 2006;Monbaliu et al;Sulyok et al 2006;Spanjer et al 2008). Currently published protocols, however, have been developed and optimized for different foods and feeds.…”