Skip to Content
Merck
  • The use of hydrazine-based derivatization reagents for improved sensitivity and detection of carbonyl containing compounds using MALDI-MSI.

The use of hydrazine-based derivatization reagents for improved sensitivity and detection of carbonyl containing compounds using MALDI-MSI.

Analytical and bioanalytical chemistry (2014-11-05)
Bryn Flinders, Josie Morrell, Peter S Marshall, Lisa E Ranshaw, Malcolm R Clench
ABSTRACT

Hydrazine-based derivatization reagents have been used to detect the presence of the carbonyl containing glucocorticoid fluticasone proprionate in rat lung tissue by MALDI-MSI. Such reagents also act as a matrix for analysis by MALDI-MS and have been termed "reactive matrices". Cryosections of rat lung tissue (12 μm), spotted with a range of concentrations of fluticasone proprionate, were derivatized in situ with 2,4-dinitrophenylhydrazine (DNPH) and 4-dimethylamino-6-(4-methoxy-1-naphthyl)-1,3,5-triazine-2-hydrazine (DMNTH) by the use of an acoustic reagent spotter. It has been demonstrated that DMNTH gave superior results compared to DNPH and that analysis of samples immediately after application of DMNTH resulted in the detection of the protonated hydrazone derivative ([MD + H](+)) of fluticasone propionate at a concentration of 500 ng/μL. It has been further shown that a prolonged reaction time (~48 h) improves the detection limit of the protonated hydrazone derivative to 50 ng/μL and that improvements in sensitivity and limits of detection are obtained when a conventional MALDI matrix CHCA is employed in conjunction with the DNPH/DMNTH reactive matrix.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Acetonitrile, HPLC Plus, ≥99.9%, poly-coated bottles
Sigma-Aldrich
Acetonitrile, ≥99.5%, ACS reagent
Sigma-Aldrich
Ethyl alcohol, Pure, 140 proof, Excise Tax-free, Permit for use required
Dexamethasone, European Pharmacopoeia (EP) Reference Standard
Fluticasone propionate, European Pharmacopoeia (EP) Reference Standard
Beclomethasone dipropionate, anhydrous, European Pharmacopoeia (EP) Reference Standard
Dexamethasone for system suitability, European Pharmacopoeia (EP) Reference Standard
Beclometasone dipropionate for peak identification, European Pharmacopoeia (EP) Reference Standard
Supelco
Dexamethasone, VETRANAL®, analytical standard
Supelco
Beclomethasone dipropionate, analytical standard, for drug analysis
Sigma-Aldrich
Dexamethasone, meets USP testing specifications
Sigma-Aldrich
Trifluoroacetic acid, suitable for HPLC, ≥99.0%
Sigma-Aldrich
Trifluoroacetic acid, ReagentPlus®, 99%
Sigma-Aldrich
Trifluoroacetic acid, ≥99%, for protein sequencing
Supelco
Trifluoroacetic acid, analytical standard
USP
Beclomethasone dipropionate, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Acetonitrile, biotech. grade, ≥99.93%
Sigma-Aldrich
Acetonitrile, anhydrous, 99.8%
Supelco
Acetonitrile, analytical standard
Sigma-Aldrich
Acetonitrile, suitable for HPLC, gradient grade, ≥99.9%
Sigma-Aldrich
Acetonitrile, ReagentPlus®, 99%
Sigma-Aldrich
Acetonitrile, HPLC Plus, ≥99.9%
Sigma-Aldrich
Acetonitrile, suitable for HPLC, gradient grade, ≥99.9%
Sigma-Aldrich
Acetonitrile, ACS reagent, ≥99.5%
Sigma-Aldrich
Acetonitrile, electronic grade, 99.999% trace metals basis
Sigma-Aldrich
Fluticasone propionate, ≥98% (HPLC), powder
Sigma-Aldrich
Ethanol, BioUltra, ≥99.8%, (absolute alcohol, without additive, A15 o1)
Sigma-Aldrich
Ethanol, purum, fine spirit, denaturated with 2% 2-butanone, F25 MEK1, ~96% (based on denaturant-free substance)
Supelco
Dexamethasone, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Ethyl alcohol, Pure, 200 proof, anhydrous, ≥99.5%