Assessment of Citrinin in Spices and Infant Cereals Using Immunoaffinity Column Clean-Up with HPLC-Fluorescence Detection.

Christopher Mair,Dave Leeman,Claire Milligan,Phyllis Brown,Naomi Mackay,Elaine Marley,Michael Norris,Carol Donnelly

doi:10.3390/toxins13100715

Abstract

Historically, the analysis of citrinin has mainly been performed on cereals such as red yeast rice; however, in recent years, more complex and abnormal commodities such as spices and infant foods are becoming more widely assessed. The aim of this study was to develop and validate clean-up methods for spices and cereal-based infant foods using a citrinin immunoaffinity column before HPLC analysis with fluorescence detection. Each method developed was validated with a representative matrix, spiked at various citrinin concentrations, based around European Union (EU) regulations set for ochratoxin A (OTA), with recoveries >80% and % RSD < 9% in all cases. The limit of detection (LOD) and the limit of quantification (LOQ) were established at 1 and 3 µg/kg for spices and 0.1 and 0.25 µg/kg for infant cereals, respectively. These methods were then tested across a variety of spices and infant food products to establish efficacy with high recoveries >75% and % RSD < 5% across all matrices assessed. Therefore, these methods proved suitable for providing effective clean-up of spices and infant cereals, enabling reliable quantification of citrinin detected. Samples such as nutmeg and infant multigrain porridge had higher levels of citrinin contamination than anticipated, indicating that citrinin could be a concern for public health. This highlighted the need for close monitoring of citrinin contamination in these commodities, which may become regulated in the future.

Highlights

Citrinin (CIT) is a secondary fungal metabolite produced by several species of the genera Aspergillus, Penicillium, and Monascus [1]
The spice method was validated with “blank” and spiked chilli powder samples, spiked at 0.5×, 1× and 2× the legislative level (LL) that is set for ochratoxin A (OTA) in spices by European Union (EU) Regulation (EC) 1881/2006 (7.5, 15 and 30 μg/kg, respectively) as well as at a limit of quantification (LOQ) of 3 μg/kg (1/5th LL) and a limit of detection (LOD) of 1 μg/kg (1/3rd LOQ)
The spice method was validated with “blank” and spiked chilli powder samples, spiked at 0.5×, 1× and 2× the legislative level (LL) that is set for OTA in spices by EU 4Roefg1-1 ulation (EC) 1881/2006 (7.5, 15 and 30 μg/kg, respectively) as well as at a LOQ of 3 μg/kg (1/5th LL) and a LOD of 1 μg/kg (1/3rd LOQ)

Summary

Introduction

Citrinin (CIT) is a secondary fungal metabolite produced by several species of the genera Aspergillus, Penicillium, and Monascus [1]. It is known to be rapidly absorbed by the liver and kidney, with a recent CIT toxicokinetic study in humans showing that only 40% was excreted in urine, indicating that its absorption was greater than 40% [1,4]. CIT is often found to co-occur with ochratoxin A (OTA), showing similar structural and toxicological properties, which is a cause for concern for both human and animal health [5]. With the combined toxic effects of OTA, it is important that CIT exposure in humas is assessed across a variety of commodities in which OTA is likely to occur. Limited occurrence data for CIT in food matrices are currently available to establish reliable dietary exposure assessments, with only a few preliminary studies having taken place such as in Belgium in 2020 (Meerpoel, Vidal) [5,7]

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