Bee pollen collected by honeybees (Apis mellifera) is one of the bee products, and it is as valuable as honey, propolis,
royal jelly, or beebread. Its quality varies according to its geographic
location or plant sources. This study aimed to apply rapid, simple,
and accurate analytical methods such as attenuated total reflectance
Fourier transform infrared spectroscopy (ATR–FTIR) and high-performance
liquid chromatography (HPLC) along with chemometrics analysis to construct
a model aimed at discriminating between different pollen samples.
In total, 33 samples were collected and analyzed using principal component
analysis (PCA), hierarchical clustering analysis (HCA), and partial
least squares regression (PLS) to assess the differences and similarities
between them. The PCA score plot based on both HPLC and ATR–FTIR
revealed the same discriminatory pattern, and the samples were divided
into four major classes depending on their total content of polyphenols.
The results revealed that spectral data obtained from ATR–FTIR
acquired in the region (4000–500 cm–1) were
further subjected to a standard normal variable (SNV) method that
removes scattering effects from spectra. However, PCA, HCA, and PLS
showed that the best PLS model was obtained with a regression coefficient
(R
2) of 0.9001, root-mean-square estimation
error (RMSEE) of 0.0304, and root-mean-squared error cross-validation
(RMSEcv) of 0.036. Discrimination between the three species has also
been possible by combining the pre-processed ATR–FTIR spectra
with PCA and PLS. Additionally, the HPLC chromatograms after pre-treatment
(SNV) were subjected to unsupervised analysis (PCA–HCA) and
supervised analysis (PLS). The PLS model confers good results by factors
(R
2 = 0.98, RMSEE = 8.22, and RMSEcv =
27.86). Prospects for devising bee pollen quality assessment methods
include utilizing ATR–FTIR and HPLC in combination with multivariate
methods for rapid authentication of the geographic location or plant
sources of bee pollen.
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