Validation of a method for determination of mycotoxins subjected to the EU regulations in spices: The UHPLC–HESI–MS/MS analysis of the crude extracts

Abstract

Validation study of the method, previously developed for multimycotoxin analysis of cereals crude extracts by ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC–MS/MS), was performed to establish a high-throughput multi-matrix procedure for simultaneous determination of mycotoxins included in the European Union's (EU) regulations for spices. Two kinds of the spice matrices were included: red pepper (Capsicum species) and black pepper (Piper nigrum). They were fortified with aflatoxins (AFB1, AFB2, AFG1 and AFG2) and ochratoxin A (OTA) at levels that are equal or below the maximum allowable contents for spices according to the Commission Regulations (EU) No. 165/2010 and No. 105/2010. The obtained validation data suggested that the analysis of red pepper crude extracts is feasible and at the same time sensitive enough for determining AFs and OTA levels set in the EU Regulations. The analysis of 17 spice samples by the validated method did not reveal the occurrence of AFs and OTA in the products from the local markets in Novi Sad, capitol of the Vojvodina Province. To confirm the results obtained by UHPLC–MS/MS extracts were also screened by a method based on accurate mass measurements using high-resolution MS (HRMS) system with Orbitrap technology. However, validation parameters for analysis of some mycotoxins in black pepper crude extracts did not comply with the EU regulation, particularly with respect to the recoveries and LODs for AFG1, AFG2 and OTA, due to significant matrix interferences.