CamSep 3 S

DETERMINATION OF BISPHENOL A AND RELATED EDCs COMPOUNDS
IN MILK USING micro-TLC AND TEMPERATURE-DEPENDENT
INCLUSION CHROMATOGRAPHY
Paweł K. ZARZYCKI a , Elżbieta WŁODARCZYK a , Deborah HODGSON b ,
Vicki L. CLIFTON c
/Stosowana w pracy technika rozdzielania: micro-TLC, HPLC/
a Section of Toxicology and Bioanalytics, Department of Civil and Environmental Engineering, Koszalin
University of Technology, Śniadeckich 2,75-453 Koszalin, Poland,
b School of Psychology, Faculty Science and IT, The University of Newcastle, Callaghan,
NSW 2308, Australia,
c The Robinson Institute, School of Paediatrics and Reproductive Health, Lyell MCEwin Hospital, University of
Adelaide, Haydown Rd, Elizabethvale 5112 SA, Australia
This research communication summarizes the results of our preliminary work concerning
optimization of simple analytical protocol for fast screening of bisphenol A and related endocrine
disrupting compounds (EDCs), which are present in milk samples. Particularly, we adapted a solidphase
extraction (SPE) protocol that has previously been used by our group for quantification of
wide range of low-molecular mass substances, mainly steroids, from highly organic compounds
loaded materials like cord blood or untreated/treated sewage waters [1-3]. In present work we tested
if proposed SPE protocol can be effective for purification of fresh human and/or UHT milk matrices
as well as quantification of bisphenol A. Resulted extracts were separated using planar and column
isocratic systems working with UV-Vis, fluorescence and PMA detection. Particularly, we tested
micro-TLC platform [4] involving HPTLC RP18W plates and binary water/organic and
organic/organic mobile phases (Figure 1). It should be noted that contrary to column
chromatography and due to planar chromatography method principles, all sample components
including substances that are strongly retarded or chemisorbed by stationary phase on the start line,
can be visualized after plate development. This approach allows fast fractionation and detection of
main milk components for the future chemometric investigations, together with data generated by
the column technique (HPLC). Temperature-dependent inclusion chromatography involving C-18
HPLC analytical column was optimized for quantification of bisphenol A and for fingerprinting of
wide range of chemicals with polarity ranging from estetrol to progesterone. It has been found that
such isocratic separation using -cyclodextrin modified mobile phase and UV-DAD detection can
be simple methodology allowing sensitive determination of bisphenol A from different milk
matrices. Due to the low consumption of the mobile phase components for micro-TLC and
application of rich in water HPLC eluent, both described chromatographic techniques can be
considered as environmentally friendly and green chemistry analytical tools.
Figure 1. Fractionation of milk SPE extracts using different micro-TLC separation and detection conditions.
References:
[1] P.K. Zarzycki, K.M. Kulhanek, R. Smith, V.L. Clifton, Determination of steroids in human plasma using
temperature-dependent inclusion chromatography for metabolomic investigations, J. Chromatogr. A, 1104 (2006) 203-
208. [2] V. L. Clifton, A. Bisits, P.K. Zarzycki. Characterization of human fetal cord blood steroid profiles in relation to
fetal sex and mode of delivery using temperature-dependent inclusion chromatography and principal component
analysis (PCA), J. Chromatogr. B, 855(2) (2007) 249-254. [3] P.K. Zarzycki, E. Włodarczyk, M.J. Baran,
Determination of endocrine disrupting compounds using temperature-dependent inclusion chromatography I.
Optimization of separation protocol, J. Chromatogr. A, 1216 (2009) 7602-7611. [4] P.K. Zarzycki, Simple horizontal
chamber for thermostated micro-thin-layer chromatography, J. Chromatogr. A, 1187 (2008) 250-259.
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