Ann Lab Med.  2015 Nov;35(6):578-585. 10.3343/alm.2015.35.6.578.

Dried Blood Spot Testing for Seven Steroids Using Liquid Chromatography-Tandem Mass Spectrometry With Reference Interval Determination in the Korean Population

• Kim B
• Lee MN
• Park HD
• Kim JW
• Chang YS
• Park WS
• Lee SY

• KMID: 2363260
• DOI: http://doi.org/10.3343/alm.2015.35.6.578

Abstract

BACKGROUND
Conventional screening for congenital adrenal hyperplasia (CAH) using immunoassays generates a large number of false-positive results. A more specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been introduced to minimize unnecessary follow-ups. However, because of limited data on its use in the Korean population, LC-MS/MS has not yet been incorporated into newborn screening programs in this region. The present study aims to develop and validate an LC-MS/MS method for the simultaneous determination of seven steroids in dried blood spots (DBS) for CAH screening, and to define age-specific reference intervals in the Korean population.
METHODS
We developed and validated an LC-MS/MS method to determine the reference intervals of cortisol, 17-hydroxyprogesterone, 11-deoxycortisol, 21-deoxycortisol, androstenedione, corticosterone, and 11-deoxycorticosterone simultaneously in 453 DBS samples. The samples were from Korean subjects stratified by age group (78 full-term neonates, 76 premature neonates, 89 children, and 100 adults).
RESULTS
The accuracy, precision, matrix effects, and extraction recovery were satisfactory for all the steroids at three concentrations; values of intra- and inter-day precision coefficients of variance, bias, and recovery were 0.7-7.7%, -1.5-9.8%, and 49.3-97.5%, respectively. The linearity range was 1-100 ng/mL for cortisol and 0.5-50 ng/mL for other steroids (R2>0.99). The reference intervals were in agreement with the previous reports.
CONCLUSIONS
This LC-MS/MS method and the reference intervals validated in the Korean population can be successfully applied to analyze seven steroids in DBS for the diagnosis of CAH.

Keyword

Mass spectrometry; Method validation; Congenital adrenal hyperplasia; Steroid; Korea; Reference interval

MeSH Terms

Adult
Asian Continental Ancestry Group
Child
*Chromatography, High Pressure Liquid/standards
Dried Blood Spot Testing
Female
Humans
Infant, Newborn
Infant, Premature
Male
Reference Values
Republic of Korea
Steroids/*blood/standards
*Tandem Mass Spectrometry/standards
Steroids

Figure

• 

  Fig. 1 Multiple reaction monitoring (MRM) transition of the respective steroids; intensity vs. time (min). The black peaks represent the target steroids. White peaks are steroids that have the same molecular weight with the target steroids.

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