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

PMID25204266

ABSTRAKT

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.

MATERIAŁY

Numer produktu

Marka

Opis produktu

731285

Sigma-Aldrich

Vitamin D₃(6,19,19-d₃) solution, 1 mg/mL in ethanol, 97 atom % D, 97% (CP)

PHR1072

Supelco

2-Propanol, Pharmaceutical Secondary Standard; Certified Reference Material

606723

Sigma-Aldrich

Methanol-¹²C, 99.95 atom % ¹²C

721956

Sigma-Aldrich

Methanol solution, 4% in methanol-d₄ (99.8 atom % D), NMR tube size 3 mm × 8 in.

740284

Sigma-Aldrich

Vitamin D₃(6,19,19-d₃) solution, 100 μg/mL in ethanol, 97 atom % D, 97% (CP)

740292

Sigma-Aldrich

Vitamin D₃ solution, 100 μg/mL in ethanol, 97% (CP)

H4014

Sigma-Aldrich

25-Hydroxycholecalciferol, ≥98% (HPLC)

I9516

Sigma-Aldrich

2-Propanol, BioReagent, ≥99.5%

E5750

Sigma-Aldrich

Ergocalciferol, 40,000,000 USP units/g

439223

Sigma-Aldrich

Dichloromethane, suitable for HPLC, ≥99.9%, contains 40-150 ppm amylene as stabilizer

95220

Sigma-Aldrich

Ergocalciferol, ≥98.0% (sum of enantiomers, HPLC)

02575

Supelco

Dichloromethane, analytical standard

30800

Sigma-Aldrich

7-Dehydrocholesterol, ≥95.0% (HPLC)

89680

Supelco

Toluene, analytical standard

59304

Sigma-Aldrich

2-Propanol, BioUltra, ≥99.5% (GC)

91237

Supelco

2-Propanol, analytical standard

45983

Supelco

Acetonitrile, analytical standard

271004

Sigma-Aldrich

Acetonitrile, anhydrous, 99.8%

733466

Sigma-Aldrich

Acetonitrile, electronic grade, 99.999% trace metals basis

733458

Sigma-Aldrich

2-Propanol, electronic grade, 99.999% trace metals basis

E0900000

Ergocalciferol, European Pharmacopoeia (EP) Reference Standard

H-083

Supelco

25-Hydroxyvitamin D3 solution, 100 μg/mL in ethanol, ampule of 1 mL, certified reference material, Cerilliant^®

V-025

Supelco

Vitamin D3 solution, 1 mg/mL in ethanol, ampule of 1 mL, certified reference material, Cerilliant^®

V-024

Supelco

Vitamin D2 solution, 1 mg/mL in ethanol, ampule of 1 mL, certified reference material, Cerilliant^®

650188

Sigma-Aldrich

Methanol solution, (Methanol:Dimethyl sulfoxide 1:1 (v/v))

66742

Supelco

Dichloromethane, Selectophore^™, ≥99.5%

34442

Sigma-Aldrich

Acetonitrile, suitable for DNA synthesis, ≥99.9% (GC)

34856

Sigma-Aldrich

Dichloromethane, suitable for HPLC, ≥99.8%, contains amylene as stabilizer

PHR1311

Supelco

Toluene, Pharmaceutical Secondary Standard; Certified Reference Material

PHR1317

Supelco

Residual Solvent - Toluene, Pharmaceutical Secondary Standard; Certified Reference Material