Dried Blood Spot Multiplexed Steroid Profiling Using Liquid Chromatography Tandem Mass Spectrometry in Korean Neonates

Overview

Journal Ann Lab Med

Specialty Pathology

Date 2019 Jan 10

PMID 30623618

Citations 12

Authors

Rihwa Choi

Hyung Doo Park

Hyeon Ju Oh

Kyounghoon Lee

Junghan Song

Soo Youn Lee

Affiliations

Soon will be listed here.

Abstract

Background: Screening for congenital adrenal hyperplasia (CAH) using immunoassays for 17α-hydroxyprogesterone generates many false-positive results. We developed and validated a liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for simultaneous quantification of nine steroid hormones in dried blood spot (DBS) samples, and established reference intervals for these hormones.

Methods: We examined our method for linearity, precision, accuracy, extraction recovery, and matrix effects and determined the reference intervals of cortisol, 17α-hydroxyproges-terone, 11-deoxycortisol, 21-deoxycortisol, androstenedione, corticosterone, 11-deoxycorticosterone, testosterone, and progesterone in 1,146 DBS samples (from 272 preterm and 874 full-term neonates). Immunoassay and LC-MS/MS methods were compared for 17α-hydroxyprogesterone. Fourteen additional samples were tested to validate the clinical applicability of the LC-MS/MS method.

Results: The linearity range was 2.8-828.0 nmol/L for cortisol and 0.9-40.0 nmol/L for the other steroids (R²>0.99). Intra-day and inter-day precision CVs were 2.52-12.26% and 3.53-17.12%, respectively. Accuracy was 80.81-99.94%, and extraction recovery and matrix effects were 88.0-125.4% and 61.7-74.2%, respectively. There was a negative bias, with higher values measured by immunoassay compared with LC-MS/MS (=0.8104, <0.0001). The LC-MS/MS method was successfully applied to the analysis of nine steroids in DBS for screening and diagnosis of CAH using the 14 additional samples.

Conclusions: Our method enables highly sensitive and specific assessment of nine steroids from DBS and is a promising tool for clinical analysis of CAH.

Citing Articles

Development and evaluation of a candidate reference measurement procedure for detecting 17α-hydroxyprogesterone in dried blood spots using isotope dilution liquid chromatography tandem mass spectrometry.

He Z, Dai H, Shen J, Huang Y, Liu J, Yan R Anal Bioanal Chem. 2024; 416(20):4635-4645.

PMID: 38949681 PMC: 11294408. DOI: 10.1007/s00216-024-05411-9.

Best Practice for Identification of Classical 21-Hydroxylase Deficiency Should Include 21 Deoxycortisol Analysis with Appropriate Isomeric Steroid Separation.

Greaves R, Kumar M, Mawad N, Francescon A, Le C, OConnell M Int J Neonatal Screen. 2023; 9(4).

PMID: 37873849 PMC: 10594498. DOI: 10.3390/ijns9040058.

Comprehensive Evaluation of the NeoBase 2 Non-derivatized MSMS Assay and Exploration of Analytes With Significantly Different Concentrations Between Term and Preterm Neonates.

Lee B, Heo W, Kim J, Lee H, Hwang N, Park H Ann Lab Med. 2022; 43(2):153-166.

PMID: 36281509 PMC: 9618896. DOI: 10.3343/alm.2023.43.2.153.

Clinical Usefulness of Ultraperformance Liquid Chromatography-Tandem Mass Spectrometry Method for Low Serum Testosterone Measurement.

Cho S, Han J, Park J, Park E, Kim G, Lee J Ann Lab Med. 2022; 43(1):19-28.

PMID: 36045053 PMC: 9467846. DOI: 10.3343/alm.2023.43.1.19.

Back to the Basics of Liquid Chromatography-Mass Spectrometry.

Kim Y, Lee S, Hur M Ann Lab Med. 2021; 42(2):119-120.

PMID: 34635605 PMC: 8548245. DOI: 10.3343/alm.2022.42.2.119.

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