Skip to Content
Merck
  • 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.

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.

Journal of chromatography. A (2014-09-11)
Linda Ahonen, Florian B R Maire, Mari Savolainen, Jaakko Kopra, Rob J Vreeken, Thomas Hankemeier, Timo Myöhänen, Petri Kylli, Risto Kostiainen
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Acetonitrile, ≥99.9% (GC)
Sigma-Aldrich
2-Propanol, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.8% (GC)
Sigma-Aldrich
Acetonitrile, suitable for DNA synthesis, ≥99.9% (GC)
Sigma-Aldrich
Acetonitrile, electronic grade, 99.999% trace metals basis
Sigma-Aldrich
Acetonitrile, HPLC Plus, ≥99.9%
Sigma-Aldrich
Methanol solution, 4% in methanol-d4 (99.8 atom % D), NMR tube size 3 mm × 8 in.
Sigma-Aldrich
Dichloromethane, suitable for HPLC, ≥99.8%, contains amylene as stabilizer
Supelco
Dichloromethane, Selectophore, ≥99.5%
Sigma-Aldrich
Vitamin D3(6,19,19-d3) solution, 100 μg/mL in ethanol, 97 atom % D, 97% (CP)
Sigma-Aldrich
2-Propanol, electronic grade, 99.999% trace metals basis
Sigma-Aldrich
Vitamin D3(6,19,19-d3) solution, 1 mg/mL in ethanol, 97 atom % D, 97% (CP)
Supelco
2-Propanol, Pharmaceutical Secondary Standard; Certified Reference Material
USP
Methyl alcohol, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Vitamin D3 solution, 100 μg/mL in ethanol, 97% (CP)
Ergocalciferol, European Pharmacopoeia (EP) Reference Standard
Supelco
Methylene Chloride, Pharmaceutical Secondary Standard; Certified Reference Material
USP
Ergocalciferol, United States Pharmacopeia (USP) Reference Standard
Supelco
Ergocalciferol (Vitamin D2), Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Cholecalciferol (Vitamin D3), Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Toluene, ACS reagent, ≥99.5%
Sigma-Aldrich
2-Propanol, Laboratory Reagent, ≥99.5%
Sigma-Aldrich
Acetonitrile, ReagentPlus®, 99%
Sigma-Aldrich
Dichloromethane, ACS reagent, ≥99.5%, contains 40-150 ppm amylene as stabilizer
Sigma-Aldrich
Acetonitrile, suitable for HPLC-GC, ≥99.8% (GC)
Supelco
Residual Solvent - Toluene, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Toluene, HPLC Plus, for HPLC, GC, and residue analysis, ≥99.9%
Supelco
Toluene, Pharmaceutical Secondary Standard; Certified Reference Material
USP
Residual Solvent Class 2 - Acetonitrile, United States Pharmacopeia (USP) Reference Standard
Supelco
Toluene, analytical standard
Sigma-Aldrich
Acetonitrile, suitable for HPLC, gradient grade, ≥99.9%