Development and performance evaluation of a tandem mass spectrometry assay for 4 adrenal steroids

Author(s): Kushnir MM, Rockwood AL, Roberts WL, Pattison EG, Owen WE, et al.


Background:Congenital adrenal hyperplasia is a group of autosomal recessive disorders caused by a deficiency of 1 of 4 enzymes required for the synthesis of glucocorticoids, mineralocorticoids, and sex hormones. Analysis of 11-deoxycortisol (11DC), 17-hydroxyprogesterone (17OHP), 17-hydroxypregnenolone (17OHPr), and pregnenolone (Pr) in blood allows detection of these enzyme defects.

Methods:The steroids were extracted from 200 microL of serum or plasma by solid-phase extraction, derivatized to form oximes, and extracted again with methyl t-butyl ether. Instrumental analysis was performed on an API 4000 tandem mass spectrometer with electrospray ionization in positive mode and multiple reaction-monitoring acquisition.

Results:The limits of detection were 0.025 microg/L for 11DC, 17OHP, and Pr and 0.10 microg/L for 17OHPr. The method was linear to 100 microg/L for 11DC, 17OHP, and Pr, respectively, and to 40 microg/L for 17OHPr. Within- and between-run (total) imprecision (CVs) were <7.1% and 11%, respectively. Reference intervals for children in Tanner stages 1 through 5 and adult males and females for 17OHP, 11DC, Pr, and 17OHPr were established. Prepared samples were stable for >72 h.

Conclusions:The detection limit and selectivity of this method and its small sample volume requirement allow analysis of endogenous concentrations of adrenal steroids in serum or plasma from children and adults. The method thus has an important potential role in the evaluation of the status of 4 of the enzymes involved in adrenal steroid biosynthesis.

Similar Articles

DHEA: a comprehensive review

Author(s): Thijssen JHH, Nieuwenhuyse H

24-Hour secretory pattern of dehydroisoandrosterone and dehydroisoandrosterone sulfate

Author(s): Rosenfeld RS, Rosenberg BJ, Fukushima DK, Hellman L

Decreased cerebrospinal fluid allopregnanolone levels in women with posttraumatic stress disorder

Author(s): Rasmusson AM, Pinna G, Paliwal P, Weisman D, Gottschalk C, et al.

Neurosteroids in depression: a review

Author(s): van Broekhoven F, Verkes RJ

Liquid chromatography tandem mass spectrometry for analysis of steroids in clinical laboratories

Author(s): Kushnir MM, Rockwood AL, Roberts WL, Yue B, Bergquist J, et al.

Beck Depression Inventory

Author(s): Beck A, Brown G

Beck Depression Inventory

Author(s): Beck A, Brown G

Diagnostic and statistical manual of mental disorders

Author(s): American Psychiatric Association

High-sensitivity tandem mass spectrometry assay for serum estrone and estradiol

Author(s): Kushnir MM, Rockwood AL, Bergquist J, Varshavsky M, Roberts WL, et al.

Assessing analytical specificity in quantitative analysis using tandem mass spectrometry

Author(s): Kushnir MM, Rockwood AL, Nelson GJ, Yue B, Urry FM

Pharmacokinetic and behavioral effects of allopregnanolone in healthy women

Author(s): Timby E, Balgard M, Nyberg S, Spigset O, Andersson A, et al.

Radioimmunoassay of 3 alpha-hydroxy-5 alpha-pregnan-20-one in rat and human plasma

Author(s): Purdy RH, Moore PH, Rao PN, Hagino N, Yamaguchi T, et al.

Serum profiles of free and conjugated neuroactive pregnanolone isomers in nonpregnant women of fertile age

Author(s): Havlikova H, Hill M, Kancheva L, Vrbikova J, Pouzar V, et al.