Open AccessMicroscale serum extraction method for the simultaneous analysis of corticosterone and lipids
Author(s) -
Alana L. Rister,
Katie L Bidne,
Jennifer R. Wood,
Eric D. Dodds
Publication year - 2019
Publication title -
analytical methods
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.615
H-Index - 67
eISSN - 1759-9679
pISSN - 1759-9660
DOI - 10.1039/c9ay01757g
Subject(s) - corticosterone , analyte , chromatography , chemistry , microscale chemistry , radioimmunoassay , detection limit , extraction (chemistry) , steroid , biochemistry , hormone , mathematics education , mathematics
Corticosterone is an important steroid for the regulation of metabolism and stress response. Existing methods for the measurement of corticosterone include radioimmunoassay (RIA), enzyme linked immunosorbent assay (ELISA), and liquid chromatography-mass spectrometry (LC-MS). While each of these approaches have their advantages, RIAs use radioactive isotopes that necessitate specially regulated usage and disposal. Furthermore, both ELISAs and RIAs require expensive kits and can only measure a single analyte. In this study, we establish a new sample preparation technique based on a modified Folch extraction that allows for the simultaneous isolation of corticosterone and lipids from serum. The extract is then analyzed by LC-MS. Using only 5 µL of serum, quantification of corticosterone was achieved with coefficients of variation at 3% or less and a detection limit of 0.12 µM. Overall, the results of this study should be beneficial to the measurement of circulating corticosterone and lipids for a variety of studies using small volumes of samples.