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Simultaneous Quantification of 17-hydroxyprogesterone, Androstenedione, Testosterone and Cortisol in Human Serum by LC-MS/MS Using TurboFlow Online Sample Extraction

Simultaneous Quantification of 17-hydroxyprogesterone, Androstenedione, Testosterone and Cortisol in Human Serum by LC-MS/MS Using TurboFlow Online Sample Extraction

Jacopo Gervasoni, Arcangelo Schiattarella, Aniello Primiano, Ilaria D'Addurno, Andrea Cocci, Cecilia Zuppi, Silvia Persichilli

Clin Biochem. 2016 Sep;49(13-14):998-1003.

PMID: 27208555

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP68962-B	17α-Hydroxyprogesterone solution		68-96-2	Price

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