A B S T R A C TThis study aimed to investigate the effect of microwave (MW) and microwave-chemical (MWC) pretreatment on barley straw and to identify the acidic, basic, or oxidative chemicals that provide the highest sugar conversion for subsequent enzymatic hydrolysis. The MW and MWC processes were applied as a pretreatment step before fermentation. MW radiation at 200 and 300 W and MW radiation plus a chemical (H 2 SO 4 or NaOH or H 2 O 2 ) as catalyst were applied, and total sugar, total phenol, and Klason and acid-soluble lignin were measured. Although the MWC pretreatment produced a higher total sugar concentration than the MW pretreatment, the addition of an NaOH solution produced the best results in terms of all parameters. Fourier transform infrared analysis was also performed to observe the deterioration of molecular structures after the application of MW and MWC.