This paper presents a high-throughout method for simultaneous determination of acetic acid and furfural in hydrothermal hydrolysate of lignocellulose using a full evaporation headspace gas chromatographic technique. In this method, the sample was acidified to pH 1.0-3.5 to a complete acetic acid release from the medium, which also avoids the risk for an acidized furfural decomposition at an elevated temperature. With a maximum sampling volume of 5 mu L, full evaporation for both acetic acid and furfural at the headspace equilibration temperature (100 degrees C) can be achieved in 3 min. The results showed that the present method has a good precision [relative standard deviation (RSD) < 4.67%] and accuracy (apparent recoveries of 95.0-104.8% for acetic acid and 95.4-104.3% for furfural. The limits of quantitation in the acetic acid and furfural detection were 0.339 and 0.208 mu g, respectively. The present method is simple, rapid, and accurate, which is suitable for rapid analysis of acetic acid and furfural in lignocellulosic hydrolysate for research of biotechnology screening and biofuel production.