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Anal Bioanal Chem. 2017 May;409(12):3145-3151. doi: 10.1007/s00216-017-0255-4. Epub 2017 Feb 21.

Signal Enhancement in the HPLC-ESI-MS/MS analysis of spironolactone and its metabolites using HFIP and NH4F as eluent additives.

Author information

1
Institute of Chemistry, University of Tartu, 14a Ravila Street, Tartu, 50411, Estonia.
2
Children's Clinic, Tartu University Hospital, N. Lunini 6, Tartu, 51014, Estonia.
3
Institute of Biomedicine and Translational Medicine, Department of Pharmacology, University of Tartu, Ravila 19, Tartu, 50411, Estonia.
4
Pharmacy Department, Tartu University Hospital, L. Puusepa 8, Tartu, 51014, Estonia.
5
Institute of Biomedicine and Translational Medicine, Department of Microbiology, University of Tartu, Ravila 19, Tartu, 50411, Estonia.
6
Institute of Chemistry, University of Tartu, 14a Ravila Street, Tartu, 50411, Estonia. karin.kipper@gmail.com.
7
Paediatric Infectious Diseases Research Group, Institute for Infection and Immunity, St. George's, University of London, Cranmer Terrace, London, SW17 0RE, UK. karin.kipper@gmail.com.

Abstract

This paper describes an LC-MS/MS method to determine the concentration of spironolactone and its metabolites 7-alpha-methylthiospironolactone and canrenone in blood plasma samples. The resulting assay is simple (using protein precipitation for sample preparation) and sensitive (the lower limit of quantification is close to 0.5 ng/ml) while requiring only 50 μl of plasma, making it especially suitable for analyzing samples obtained from pediatric and neonatal patients where sample sizes are limited. The sensitivity is achieved by using ammonium fluoride as an eluent additive, which in our case amplifies the signal from our analytes in the plasma solution on average about 70 times. The method is fully validated according to the European Medicines Agency's guideline and used for the measurement of pediatric patients' samples in clinical trials for evaluating oral spironolactone's and its metabolites' pharmacokinetics in children up to 2 years of age.

KEYWORDS:

Ammonium fluoride; Hexafluoroisopropanol; Ionization efficiency; Liquid chromatography-mass spectrometry; Signal enhancement; Spironolactone

PMID:
28224249
PMCID:
PMC5395588
DOI:
10.1007/s00216-017-0255-4
[Indexed for MEDLINE]
Free PMC Article

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