title: Liquid chromatography quadrupole linear ion trap mass spectrometry for quantitative steroid hormone analysis in plasma, urine, saliva and hair.
abstract: Steroid analysis is being conquered by liquid chromatography tandem mass spectrometry (LC-MS/MS) benefiting from higher standardization, selectivity and diversity. Regarding high throughput in routine diagnostics rapid chromatography is mandatory. Introducing MS(3) (MS/MS/MS), specificity of mass spectrometric detection can be enhanced without sacrificing analysis time. 100mL of human plasma/serum, saliva, urine and 10-20mg of hair are used for the simultaneous quantification of 17α-hydroxyprogesterone, aldosterone, androstenedione, cortisol, cortisone, dehydroepiandrosterone sulfate (DHEAS), estradiol, progesterone, and testosterone using online solid phase extraction (SPE) LC-MS/MS or LC-MS(3). Steroids can be analyzed in 4min after a single manual dilution and protein precipitation step. In complex sample matrices like hair MS(3) detection was found to be appropriate for quantitation. Lower limits of quantitation ranged from 37pmol/L (estradiol) up to 3.1nmol/L (DHEAS). General accuracy was 89-107% with between-run imprecision ≤10%. Comparison to immunoassays revealed significant differences in quantitation for urinary cortisol (-71% mean), aldosterone (-40% mean) and plasma aldosterone (-45% mean). The comparison of MS(2) and MS(3) quantitation of hair cortisol also revealed significant differences. In general, quantitation via MS(3) was not applicable for a long time. But with the current generation of mass spectrometers quantitation via MS(3) can be superior to MS(2) regarding specificity and accuracy when dealing with matrix issues. However, drawbacks regarding flexibility and precision have to be taken into account. Concludingly, simple protein precipitation combined with rapid online SPE LC-MS/MS/MS allows us to quantify over broad, essential concentration ranges in human serum, saliva, urine and hair.
authors.affiliation: Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, Leipzig University, Liebigstraße 27, 04103 Leipzig, Germany. Electronic address: Alexander.Gaudl@medizin.uni-leipzig.de.
authors.forename: Alexander
authors.initials: A
authors.lastname: Gaudl
authors.affiliation: Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, Leipzig University, Liebigstraße 27, 04103 Leipzig, Germany. Electronic address: Juergen.Kratzsch@medizin.uni-leipzig.de.
authors.forename: Juergen
authors.initials: J
authors.lastname: Kratzsch
authors.affiliation: Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, Leipzig University, Liebigstraße 27, 04103 Leipzig, Germany. Electronic address: YoonJu.Bae@medizin.uni-leipzig.de.
authors.forename: Yoon Ju
authors.initials: YJ
authors.lastname: Bae
authors.affiliation: Hospital for Children and Adolescents, University Hospital, Leipzig University, Liebigstraße 20a, 04103 Leipzig, Germany; LIFE - Leipzig Research Center for Civilization Diseases, Leipzig University, Philipp-Rosenthal-Straße 27, 04103 Leipzig, Germany. Electronic address: Wieland.Kiess@medizin.uni-leipzig.de.
authors.forename: Wieland
authors.initials: W
authors.lastname: Kiess
authors.affiliation: Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, Leipzig University, Liebigstraße 27, 04103 Leipzig, Germany; LIFE - Leipzig Research Center for Civilization Diseases, Leipzig University, Philipp-Rosenthal-Straße 27, 04103 Leipzig, Germany. Electronic address: Joachim.Thiery@medizin.uni-leipzig.de.
authors.forename: Joachim
authors.initials: J
authors.lastname: Thiery
authors.affiliation: Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, Leipzig University, Liebigstraße 27, 04103 Leipzig, Germany; LIFE - Leipzig Research Center for Civilization Diseases, Leipzig University, Philipp-Rosenthal-Straße 27, 04103 Leipzig, Germany. Electronic address: Uta.Ceglarek@medizin.uni-leipzig.de.
authors.forename: Uta
authors.initials: U
authors.lastname: Ceglarek
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D010949:Plasma / Q000737:chemistry; D012463:Saliva / Q000737:chemistry*; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction / Q000379:methods; D013256:Steroids / Q000097:blood* / Q000302:isolation & purification / Q000652:urine*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014556:Urine / Q000737:chemistry
chemical_list: D013256:Steroids
keywords: Linear ion trap; Liquid chromatography tandem mass spectrometry; Quantitation; Steroid analysis