Discriminant analysis of pyrrolizidine alkaloid contamination in bee pollen based on near-infrared data from lab-stationary and portable spectrometers

Bee pollen may be contaminated with pyrrolizidine alkaloids (PAs) and their N -oxides (PANOs), which are mainly detected by liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS), even though the use of fast near-infrared (NIR) spectroscopy is an ongoing alternative. Therefore, the main challenge of this study was to assess the feasibility of both a lab-stationary (Foss) and a portable (Polispec) NIR spectrometer in 60 dehydrated bee pollen samples. After an ANOVA-feature selection of the most informative NIR spectral data, canonical discriminant analysis (CDA) was performed to distinguish three quantitative PA/PANO classes (µg/kg): < LOQ (0.4), low; 0.4–400, moderate; > 400, high. According to the LC–MS/MS analysis, 77% of the samples were contaminated with PAs/PANOs and the sum content of the 17 target analytes was higher than 400 µg/kg in 28% of the samples. CDA was carried out on a pool of 18 (Foss) and 22 (Polispec) selected spectral variables and allowed accurate classification of samples from the low class as confirmed by the high values of Matthews correlation coefficient (≥ 0.91) for both NIR spectrometers. Leave-one-out cross-validation highlighted precise recognition of samples characterised by a high PA/PANO content with a low misclassification rate (0.02) as false negatives. The most informative wavelengths were within the < 1000, 1000–1660 and > 2400 nm regions for Foss and > 1500 nm for Polispec that could be associated with cyclic amines, and epoxide chemical structures of PAs/PANOs. In sum, both lab-stationary and portable NIR systems are reliable and fast techniques for detecting PA/PANO contamination in bee pollen.