Pushing quantitation limits in micro UHPLC–MS/MS analysis of steroid hormones by sample dilution using high volume injection

Zoltán Márta, Balázs Bobály, J. Fekete, Balázs Magda, T. Imre, Katalin Viola Mészáros, P. Szabó

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Ultratrace analysis of sample components requires excellent analytical performance in terms of limits of quantitation (LoQ). Micro UHPLC coupling with sensitive tandem mass spectrometry provides state of the art solutions for such analytical problems. Decreased column volume in micro LC limits the injectable sample volume. However, if analyte concentration is extremely low, it might be necessary to inject high sample volumes. This is particularly critical for strong sample solvents and weakly retained analytes, which are often the case when preparing biological samples (protein precipitation, sample extraction, etc.). In that case, high injection volumes may cause band broadening, peak distortion or even elution in dead volume. In this study, we evaluated possibilities of high volume injection onto microbore RP-LC columns, when sample solvent is diluted. The presented micro RP-LC–MS/MS method was optimized for the analysis of steroid hormones from human plasma after protein precipitation with organic solvents. A proper sample dilution procedure helps to increase the injection volume without compromising peak shapes. Finally, due to increased injection volume, the limit of quantitation can be decreased by a factor of 2–5, depending on the analytes and the experimental conditions.

Original languageEnglish
Pages (from-to)135-141
Number of pages7
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume129
DOIs
Publication statusPublished - Sep 10 2016

Fingerprint

Steroid hormones
Dilution
Steroids
Hormones
Plasma (human)
Injections
Organic solvents
Mass spectrometry
Proteins
Tandem Mass Spectrometry
Blood Proteins

Keywords

  • High volume injection
  • LC–MS/MS
  • Microflow liquid chromatography
  • Steroid hormone

ASJC Scopus subject areas

  • Analytical Chemistry
  • Drug Discovery
  • Pharmaceutical Science
  • Spectroscopy
  • Clinical Biochemistry

Cite this

Pushing quantitation limits in micro UHPLC–MS/MS analysis of steroid hormones by sample dilution using high volume injection. / Márta, Zoltán; Bobály, Balázs; Fekete, J.; Magda, Balázs; Imre, T.; Mészáros, Katalin Viola; Szabó, P.

In: Journal of Pharmaceutical and Biomedical Analysis, Vol. 129, 10.09.2016, p. 135-141.

Research output: Contribution to journalArticle

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