Ethylene glycol in the presence of sodium hydroxide was utilised as pretreatment for effective delignification and reduced the recalcitrance of lignocellulosic biomass which ramified the exposure of cellulose. Two-staged acid hydrolysis was also investigated which demonstrated its synergistic efficiency by minimising the deficiency of single stage acid hydrolysis. The operating parameters including acid concentration, temperature, residence time and cellulose loading for two-staged acid hydrolysis were studied by using ethylene glycol delignified degraded oil palm empty fruit bunch (DEFB) to recover the sugar based substrates for potential biofuels and other bio-chemicals production. In this study, stage I 45 wt% acid at 65 °C for 30 min coupled with high cellulose loading 21.25 w/v% and 12 wt% acid at 100 °C for 120 min was able to release a total of 89.8% optimum sugar yield with minimal formation of degradation products including 0.058 g/L furfural, 0.0251 g/L hydroxymethylfurfural and 0.200 g/L phenolic compounds.