Development and Validation of a UPLC-MS/MS and UPLC-HR-MS Method for the Determination of Fumonisin B1 and Its Hydrolysed Metabolites and Fumonisin B2 in Broiler Chicken Plasma.

Siegrid De Baere,Gerlinde Bichl,Barbara Novak,Siska Croubels,Gunther Antonissen

doi:10.3390/toxins10020062

Abstract

A sensitive and specific method for the quantitative determination of Fumonisin B1 (FB1), its partially hydrolysed metabolites pHFB1a+b and hydrolysed metabolite HFB1, and Fumonisin B2 (FB2) in broiler chicken plasma using ultra-performance liquid chromatography combined with tandem mass spectrometry (UPLC-MS/MS) was developed. The sample preparation was rapid, straightforward and consisted of a deproteinization and phospholipid removal step using an Oasis® OstroTM 96-well plate. Chromatography was performed on an Acquity HSS-T3 column, using 0.3% formic acid and 10 mM ammonium formate in water, and acetonitrile as mobile phases. The MS/MS instrument was operated in the positive electrospray ionization mode and the two multiple reaction monitoring transitions were monitored for each component for quantification and identification, respectively. The method was validated in-house: matrix-matched calibration graphs were prepared and good linearity (r ≥ 0.99) was achieved over the concentration ranges tested (1–500 ng/mL for FB1 and FB2; 0.86–860 ng/mL for pHFB1a; 0.72–1430 ng/mL for pHFB1b and 2.5–2500 ng/mL for HFB1). Limits of quantification (LOQ) and detection (LOD) in plasma ranged between 0.72 to 2.5 ng/mL and 0.03 to 0.17 ng/mL, respectively. The results for the within-day and between-day precision and accuracy fell within the specified ranges. Moreover, the method was transferred to an UPLC high-resolution mass spectrometry (HR-MS) instrument in order to determine potential metabolites of HFB1, such as N-acyl-HFB1s and phase II metabolites. The method has been successfully applied to investigate the toxicokinetics and biotransformation of HFB1 in broiler chickens.

Highlights

Fumonisins (FBs) are secondary metabolites of fungi, such as Fusarium verticillioides, Fusarium proliferatum and Aspergillus niger, which frequently contaminate maize and maize-based products [1,2,3,4]
To be able to analyse all these samples within 24 h, the UPLC-MS/MS analysis had to be accomplished within a short run-time (≤10 min)
This study describes the development and in‐house validation of a sensitive and specific

Summary

Introduction

Fumonisins (FBs) are secondary metabolites of fungi, such as Fusarium verticillioides, Fusarium proliferatum and Aspergillus niger, which frequently contaminate maize and maize-based products [1,2,3,4]. Fumonisin B1 (FB1) is the most prevalent component of the family and is hepato- and nephrotoxic in a variety of animal species, it is poorly absorbed and rapidly excreted [1,4,5,6,7]. Toxins 2018, 10, x FOR PEER REVIEW [1,2,3,4]. Fumonisin B1 (FB1) is the most prevalent component of the family and is hepato‐ and Toxins. 2018, 10, 62 in a variety of animal species, it is poorly absorbed and rapidly excreted [1,4– 2 of 20 nephrotoxic. FB1 is associated with a number of mycotoxicoses, such as equine leukoencephalomalacia [5], porcine pulmonary oedema [5,6] and hepatocarcinoma in rats [8,9]

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