title: Simultaneous quantification of 17-hydroxyprogesterone, androstenedione, testosterone and cortisol in human serum by LC-MS/MS using TurboFlow online sample extraction.
abstract: BACKGROUND
  The simultaneous quantification of a steroid hormones panel provides more clinical information than a single steroid assay. Traditionally, steroids have been quantified with immunoassays which are characterized by high rate of positive results. Aim of this work, was to develop a TurboFlow-LC-MS/MS method for the simultaneous quantification of 17-hydroxyprogesterone, androstenedione, cortisol and testosterone in serum.
  
  
  METHODS
  To 100μL of serum, 100μL of internal standard solution in methanol were added; after centrifugation the supernatant was injected in the TurboFlow for further purification. Steroids were determined using a TSQ Vantage operating with an atmospheric pressure chemical ionization source. Method was fully validated and results compared with immunoassay methods.
  
  
  RESULTS
  Limit of quantification ranged from 0.02ng/mL to 1ng/mL. The precision was lower than 11% and accuracy ranged from 93.5 to 121.6%. The correlation was acceptable for all analytes except for low levels of testosterone. However, the Bland-Altman plots display a positive bias for androstenedione and 17-hydroxyprogesterone, and a negative bias for cortisol and testosterone.
  
  
  CONCLUSIONS
  TurboFlow analysis provides a simple and effective clean-up procedure minimizing the interference of the matrix. The presented method is selective, precise, and sensitive being suitable in a clinical laboratory.
authors.affiliation: Fondazione Policlinico Universitario A. Gemelli, Rome, Italy. Electronic address: jacopo.gervasoni@policlinicogemelli.it.
authors.forename: Jacopo
authors.initials: J
authors.lastname: Gervasoni
authors.affiliation: Fondazione Policlinico Universitario A. Gemelli, Rome, Italy.
authors.forename: Arcangelo
authors.initials: A
authors.lastname: Schiattarella
authors.affiliation: Fondazione Policlinico Universitario A. Gemelli, Rome, Italy.
authors.forename: Aniello
authors.initials: A
authors.lastname: Primiano
authors.affiliation: Fondazione Policlinico Universitario A. Gemelli, Rome, Italy.
authors.forename: Ilaria
authors.initials: I
authors.lastname: D'Addurno
authors.affiliation: Fondazione Policlinico Universitario A. Gemelli, Rome, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Cocci
authors.affiliation: Fondazione Policlinico Universitario A. Gemelli, Rome, Italy.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Zuppi
authors.affiliation: Fondazione Policlinico Universitario A. Gemelli, Rome, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Persichilli
mesh_terms: D019326:17-alpha-Hydroxyprogesterone / Q000097:blood*; D000735:Androstenedione / Q000097:blood*; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D006854:Hydrocortisone / Q000097:blood*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013739:Testosterone / Q000097:blood*
chemical_list: D013739:Testosterone; D000735:Androstenedione; D019326:17-alpha-Hydroxyprogesterone; D006854:Hydrocortisone
keywords: LC-MS/MS; Method validation; Steroids; TurboFlow