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The Carbohydrate-responsive element binding protein(ChREBP)is a glucose-regulated transcription factor that acts as a key regulator of enzymes involved in fatty acid synthesis.In this article,the influence of ChREBP on the level of free fatty acids in mouse liver was investigated.Briefly,liver samples were collected from wild-type and ChREBP knockout mice at 9 months which were treated with N-nitrosodiethylamine at 2 weeks old,fed with high fat diet since 4 weeks old,and homogenized in methanol,acidified with hydrochloric acid,and extracted with isooctane.The isooctane layer was evaporated to dryness under nitrogen flow.Free fatty acids were derivatized with N,O-bis(trimethylsilyl)trifluoroacetamide(BSTFA)/1%trimethylchlorosilane(TMCS)/pyridine into their trimethylsilyl derivatives and analyzed by gas chromatography-mass spectrometry in selective ion monitoring(SIM)with electron impact ionization(EI).C14:0-d27 was used as internal standard(IS).The levels of free fatty acids in ChREBP knockout mouse liver were significantly higher than those in wild-type mouse liver,such as C6:0(p<0.01),C8:0(p<0.01),C10:0(p<0.05),C12:0(p<0.05),C14:0(p<0.01),C17:0(p<0.01),C18:2(p<0.05),C18:3(p<0.05),C20:0(p<0.01),C22:0(p<0.01),C24:0(p<0.05),except for C7:0(p=0.47),C16:0(p=0.07),C18:0(p=0.75),C18:1(p=0.31),C20:4(p=0.39).In conclusion,ChREBP deficiency increases the levels of most free fatty acids in mouse liver.