| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 1968440 | Clinical Biochemistry | 2016 | 6 Pages |
•Mass spectrometry is considered the gold standard for steroid hormone analysis (because of its accuracy and sensitivity)•An LC-MS/MS method for analysis of 17-hydroxyprogesterone, androstenedione, testosterone and cortisol is proposed•Turboflow chromatography guarantee an efficient purification from the matrix component•the comparison with immunoassay indicates an acceptable correlation except for low level of testosterone•The presented method is suitable for routine clinical laboratory
BackgroundThe 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.MethodsTo 100 μL of serum, 100 μL 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.ResultsLimit of quantification ranged from 0.02 ng/mL to 1 ng/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.ConclusionsTurboFlow 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.
