We have developed a workflow to extract, separate, and semi-quantify bioactive oxysterols
from mouse colon tissues and fecal matters using solid- and liquid-phase extractions, enzymatic and
chemical modifications, and stable-isotope dilution LC/MS/MS. The method was applied to a dextran
sodium sulphate (DSS)-induced mouse colitis model, which revealed that one particular dihydroxycholesterol
(diOHC), 7α,25-diOHC, was significantly elevated in both colon tissue and fecal matters
of mice with colitis compared to that in naïve mice. The extent of 7α,25-diOHC elevation was positively
correlated with colitis severity.