title: Highly selective and automated online SPE LC-MS3 method for determination of cortisol and cortisone in human hair as biomarker for stress related diseases.
abstract: Hair analysis has been increasingly used to establish long-term biomarkers of exposure to both endogenous and exogenous substances, with a special emphasis on steroidal hormones. Hair cortisol and cortisone have been associated to physiological and psychological strains, anxiety and depression. Hair is a very complex matrix, which might jeopardize analyte detection at low concentrations. A new, highly selective and sensitive method based on fragments of second order, MS(3) (MS/MS/MS), was developed and validated for the analysis of hair cortisol and cortisone. An online solid phase extraction was performed on a C8 restricted access material (RAM) phase following by separation on a reversed-phase C18 column using methanol and 0.02% ammonium hydroxide as mobile phase. The developed method required minimal sample preparation and the injection of only 50 µL of sample leading to a LOQ of 2 pg mg(-1). Good linear responses were observed in the range 2-200 pg mg(-1) (R(2)>0.99) and extraction recoveries ranged between 77-125% and 70-123% for cortisol and cortisone, respectively. Intra- and inter-assay coefficients of variation were between 1.4 and 14%. In order to evaluate the applicability of the method, preliminary tests (N=33) were conducted in 3 cm hair samples (close to scalp) of healthy volunteers with an age range of 4-63. Average concentrations in hair were 12.7±14 pg mg(-1) and 41.6±42 pg mg(-1) for cortisol and cortisone, respectively. Further investigations on cortisol and cortisone as biomarkers for chronic psychological strain will be assessed as a next step.
authors.affiliation: Institute for Occupational and Social Medicine, Medical Faculty, RWTH Aachen University, Pauwelsstrasse 30, D-52074 Aachen, Germany. Electronic address: nsoaresquine@ukaachen.de.
authors.forename: Natalia
authors.initials: N
authors.lastname: Quinete
authors.affiliation: Institute for Occupational and Social Medicine, Medical Faculty, RWTH Aachen University, Pauwelsstrasse 30, D-52074 Aachen, Germany.
authors.forename: Jens
authors.initials: J
authors.lastname: Bertram
authors.affiliation: Institute for Occupational and Social Medicine, Medical Faculty, RWTH Aachen University, Pauwelsstrasse 30, D-52074 Aachen, Germany.
authors.forename: Marcus
authors.initials: M
authors.lastname: Reska
authors.affiliation: Institute for Occupational and Social Medicine, Medical Faculty, RWTH Aachen University, Pauwelsstrasse 30, D-52074 Aachen, Germany.
authors.forename: Jessica
authors.initials: J
authors.lastname: Lang
authors.affiliation: Institute for Occupational and Social Medicine, Medical Faculty, RWTH Aachen University, Pauwelsstrasse 30, D-52074 Aachen, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Kraus
mesh_terms: D000293:Adolescent; D000328:Adult; D000367:Age Factors; D015415:Biomarkers / Q000032:analysis; D002648:Child; D002675:Child, Preschool; D002853:Chromatography, Liquid; D003348:Cortisone / Q000032:analysis*; D005260:Female; D006197:Hair / Q000737:chemistry*; D006801:Humans; D006854:Hydrocortisone / Q000032:analysis*; D008297:Male; D008875:Middle Aged; D015203:Reproducibility of Results; D052616:Solid Phase Extraction; D013315:Stress, Psychological; D053719:Tandem Mass Spectrometry; D055815:Young Adult
chemical_list: D015415:Biomarkers; D003348:Cortisone; D006854:Hydrocortisone
keywords: Cortisol; Cortisone; Hair analysis; LC–MS(3); Stress biomarker