Analysis of oxysterols and vitamin D metabolites in mouse brain and cell line samples by ultra-high-performance liquid chromatography-atmospheric pressure photoionization-mass spectrometry

Linda Ahonen¹, Florian B R Maire², Mari Savolainen³, Jaakko Kopra⁴, Rob J Vreeken⁵, Thomas Hankemeier⁶, Timo Myöhänen⁷, Petri Kylli⁸, Risto Kostiainen⁹

Affiliations

¹ Division of Pharmaceutical Chemistry and Technology, Faculty of Pharmacy, University of Helsinki, P.O. Box 56, FI-00014, Finland. Electronic address: linda.ahonen@helsinki.fi.
² Division of Analytical Biosciences, Leiden Academic Centre for Drug Research, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands. Electronic address: f.b.r.maire@lacdr.leidenuniv.nl.
³ Division of Pharmacology and Pharmacotherapy, Faculty of Pharmacy, University of Helsinki, P.O. Box 56, FI-00014, Finland. Electronic address: mari.savolainen@helsinki.fi.
⁴ Division of Pharmacology and Pharmacotherapy, Faculty of Pharmacy, University of Helsinki, P.O. Box 56, FI-00014, Finland. Electronic address: jaakko.kopra@helsinki.fi.
⁵ Division of Analytical Biosciences, Leiden Academic Centre for Drug Research, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands; Netherlands Metabolomics Centre, Leiden University, P.O. Box 9502, 22300 RA Leiden, The Netherlands. Electronic address: r.vreeken@lacdr.leidenuniv.nl.
⁶ Division of Analytical Biosciences, Leiden Academic Centre for Drug Research, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands; Netherlands Metabolomics Centre, Leiden University, P.O. Box 9502, 22300 RA Leiden, The Netherlands. Electronic address: hankemeier@lacdr.leidenuniv.nl.
⁷ Division of Pharmacology and Pharmacotherapy, Faculty of Pharmacy, University of Helsinki, P.O. Box 56, FI-00014, Finland. Electronic address: timo.myohanen@helsinki.fi.
⁸ Division of Pharmaceutical Chemistry and Technology, Faculty of Pharmacy, University of Helsinki, P.O. Box 56, FI-00014, Finland. Electronic address: petri.kylli@helsinki.fi.
⁹ Division of Pharmaceutical Chemistry and Technology, Faculty of Pharmacy, University of Helsinki, P.O. Box 56, FI-00014, Finland. Electronic address: risto.kostiainen@helsinki.fi.

PMID: 25204266
DOI: 10.1016/j.chroma.2014.08.088

Analysis of oxysterols and vitamin D metabolites in mouse brain and cell line samples by ultra-high-performance liquid chromatography-atmospheric pressure photoionization-mass spectrometry

Linda Ahonen et al. J Chromatogr A. 2014.

. 2014 Oct 17:1364:214-22.

doi: 10.1016/j.chroma.2014.08.088. Epub 2014 Sep 2.

Authors

Linda Ahonen¹, Florian B R Maire², Mari Savolainen³, Jaakko Kopra⁴, Rob J Vreeken⁵, Thomas Hankemeier⁶, Timo Myöhänen⁷, Petri Kylli⁸, Risto Kostiainen⁹

Affiliations

¹ Division of Pharmaceutical Chemistry and Technology, Faculty of Pharmacy, University of Helsinki, P.O. Box 56, FI-00014, Finland. Electronic address: linda.ahonen@helsinki.fi.
² Division of Analytical Biosciences, Leiden Academic Centre for Drug Research, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands. Electronic address: f.b.r.maire@lacdr.leidenuniv.nl.
³ Division of Pharmacology and Pharmacotherapy, Faculty of Pharmacy, University of Helsinki, P.O. Box 56, FI-00014, Finland. Electronic address: mari.savolainen@helsinki.fi.
⁴ Division of Pharmacology and Pharmacotherapy, Faculty of Pharmacy, University of Helsinki, P.O. Box 56, FI-00014, Finland. Electronic address: jaakko.kopra@helsinki.fi.
⁵ Division of Analytical Biosciences, Leiden Academic Centre for Drug Research, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands; Netherlands Metabolomics Centre, Leiden University, P.O. Box 9502, 22300 RA Leiden, The Netherlands. Electronic address: r.vreeken@lacdr.leidenuniv.nl.
⁶ Division of Analytical Biosciences, Leiden Academic Centre for Drug Research, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands; Netherlands Metabolomics Centre, Leiden University, P.O. Box 9502, 22300 RA Leiden, The Netherlands. Electronic address: hankemeier@lacdr.leidenuniv.nl.
⁷ Division of Pharmacology and Pharmacotherapy, Faculty of Pharmacy, University of Helsinki, P.O. Box 56, FI-00014, Finland. Electronic address: timo.myohanen@helsinki.fi.
⁸ Division of Pharmaceutical Chemistry and Technology, Faculty of Pharmacy, University of Helsinki, P.O. Box 56, FI-00014, Finland. Electronic address: petri.kylli@helsinki.fi.
⁹ Division of Pharmaceutical Chemistry and Technology, Faculty of Pharmacy, University of Helsinki, P.O. Box 56, FI-00014, Finland. Electronic address: risto.kostiainen@helsinki.fi.

PMID: 25204266
DOI: 10.1016/j.chroma.2014.08.088

Abstract

We have developed an ultra-high-performance liquid chromatography-atmospheric pressure photoionization-tandem mass spectrometric (UHPLC-APPI-MS/MS) method for the simultaneous quantitative analyses of several oxysterols and vitamin D metabolites in mouse brain and cell line samples. An UHPLC-APPI-high resolution mass spectrometric (UHPLC-APPI-HRMS) method that uses a quadrupole-time of flight mass spectrometer was also developed for confirmatory analysis and for the identification of non-targeted oxysterols. Both methods showed good quantitative performance. Furthermore, APPI provides high ionization efficiency for determining oxysterols and vitamin D related compounds without the time consuming derivatization step needed in the conventionally used electrospray ionization method to achieve acceptable sensitivity. Several oxysterols were quantified in mouse brain and cell line samples. Additionally, 25-hydroxyvitamin D3 was detected in mouse brain samples for the first time.

Keywords: Atmospheric pressure photoionization; Cell line samples; Liquid chromatography–mass spectrometry; Mouse brain samples; Oxysterols; Vitamin D.

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Analysis of oxysterols and vitamin D metabolites in mouse brain and cell line samples by ultra-high-performance liquid chromatography-atmospheric pressure photoionization-mass spectrometry

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Analysis of oxysterols and vitamin D metabolites in mouse brain and cell line samples by ultra-high-performance liquid chromatography-atmospheric pressure photoionization-mass spectrometry

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