Abstract

Defective synthesis of the steroid hormones by the adrenal cortex has profound effects on human development andhomeostasis. Due to the time-consuming chromatography procedure combined with mass spectrometry, the matrix-assisted laserdesorption ionization method coupled to the linear ion-trap tandem mass spectrometry (MALDI-LTQ-MS/MS) was developedfor quantitative analysis of 10 adrenal steroids in human serum. Although MALDI-MS can be introduced for its applicability asa high-throughput screening method, it has a limitation on reproducibility within and between samples, which renders poorreproducibility for quantification. For quantitative MALDI-MS/MS analysis, the stable-isotope labeled internal standards wereused and the conditions of crystallization were tested. The precision and accuracy were 3.1~35.5% and 83.8~138.5%, respectively,when a mixture of 10 mg/mL α-cyano-4-hydroxycinnamic acid in 0.2% TFA of 70% acetonitrile was used as the MALDImatrix. The limit of quantification ranged from 5 to 340 ng/mL, and the linearity as a correlation coefficient was higher than0.988 for all analytes in the calibration range. Clinical applications include quantitative analyses of patients with congenital adrenalhyperplasia. The devised MALDI-MS/MS technique could be successfully applied to diagnosis of clinical samples.