122nd General Meeting of the KCS

Type Poster Presentation
Area Analytical Chemistry
Room No. Grand Ballroom
Time 10월 19일 (금요일) 11:00~12:30
Code ANAL.P-342
Subject Simultaneous Determination of 39 Steroid Hormones in Human Urine Sample Using Liquid Chromatography-electrospray Tandem Mass Spectrometry
Authors Hyuck Ho Son, Wan Soo Yun1, Sung-Hee Cho2,*
Center for chemical analysis, Korea Research Institute of Chemical Technology, Korea
1Department of Chemistry, Sungkyunkwan University, Korea
2Center for Chemical Analysis, Korea Research Institute of Chemical Technology, Korea
Abstract Abnormal production or metabolism of steroid hormones are responsible for the development and prevention of endocrine diseases. Accurate quantification of steroid hormones is necessary for both research into clinical conditions as well as for diagnostic monitoring. In this research, an improved analytical method was developed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the analysis of 39 steroid hormones including corticosteroids, progestins, estrogens, and androgens in urine sample. As a pre-treatment procedure prior to LC-MS/MS analysis, hydrolyzation of the human urine samples with β-glucuronidase and solid-phase extraction for purifying the samples was performed. Steroids were separated using Waters ACQUITY @ BEH C18 column (2.1 x 100 mm, 1.7 µm) and a mobile phase consisting of eluent A (0.01% formic acid and 1 mM ammonium formate in water) and eluent B (0.01% formic acid and 1 mM ammonium formate in methanol) with a gradient program at a flow rate of 0.4 mL/min. The [M+H]+, [M+NH4]+ and [M+H-H2O]+ ions were used as precursor ions to detect endogenous steroid hormones in MS. All steroids were monitored in multiple reaction monitoring (MRM) mode within 30.0 min by tandem mass spectrometry (MS/MS) with electrospray ionization in positive mode. The linearity of this method was over 0.991. The limits of detection at signal to noise (S/N) ratio of 3 were 0.1 ~ 300 ng/L. The coefficients of variation were in the range of 2 -25% for within-day variation and 2 -25% for day-to-day variation, respectively. The present method could be applied to clinical research such as diagnosis, monitoring, and biomedical investigations.
E-mail sonhh@krict.re.kr