Metabolite profiling of sex developmental steroid conjugates reveals an association between decreased levels of steroid sulfates and adiposity in obese girls. Issue 162 (September 2016)

• Record Type: Journal Article
• Authors: Lee, Su Hyeon; Kim, Shin Hye; Lee, Won-Yong; Chung, Bong Chul; Park, Mi Jung; Choi, Man Ho
• Is Part Of: Journal of steroid biochemistry and molecular biology. Issue 162(2016)
• Page Start: 100
• Page End: 109
• Publication Date: 2016-09
• Subjects: Steroids -- Glucuronide -- Sulfate -- LC–MS -- Obesity -- Adiposity; Steroid hormones -- Periodicals; Biochemistry -- Periodicals; Hormones -- Periodicals; Molecular Biology -- Periodicals; Hormones stéroïdes -- Périodiques; Steroid hormones; Periodicals; 572.579
• DOI: 10.1016/j.jsbmb.2016.04.020
• Languages: English
• ISSNs: 0960-0760

Abstract

Highlights: LC–MS-based analysis of 16 steroid sulfates and glucuronides was developed. LC-SIM-MS method with ion-trap analyzer was fully validated. Serum steroid sulfates were significantly altered in obese girls. Changed serum levels of steroid sulfates were correlated with adiposity. Abstract: Free and conjugated steroids coexist in a dynamic equilibrium due to complex biosynthetic and metabolic processes. This may have clinical significance related to various physiological conditions, including sex development involving the reproductive system. Therefore, we performed quantitative profiling of 16 serum steroids conjugated with glucuronic and sulfuric acids using liquid chromatography-mass spectrometry (LC–MS). All steroid conjugates were purified by solid-phase extraction and then separated through a 3-μm particle size C18 column (150 mm × 2.1 mm) at a flow rate of 0.3 mL/min in the negative ionization mode. The LC–MS-based analysis was found to be linear ( r 2 > 0.99), and all steroid conjugates had a limit-of-quantification (LOQ) of 10 ng/mL, except for cholesterol sulfate and 17β-estradiol-3, 17-disulfate (20 ng/mL). The extraction recoveries of all steroid conjugates ranged from 97.9% to 110.7%, while the overall precision (% CV) and accuracy (% bias) ranged from 4.8% to 10.9% and from 94.4% to 112.9% at four different concentrations, respectively. Profiling of steroid conjugates corrected by adiposity revealed decreased levels of steroid sulfates ( P < 0.01) in overweight and obese girls compared to normal girls. The suggested technique can be used for evaluating metabolic changes in steroid conjugates and for understanding the pathophysiology and relative contributions of adiposity in childhood obesity.