Objective To establish a method for determining methanol in urine, blood and wine during emergency scenarios using headspace gas chromatography (HS-GC) and gas chromatography-mass spectrometry (GC-MS). This method was applied to samples obtained from a poisoning incident to identify the source of intoxication; thereby providing a validated analytical approach for laboratory detection to support the prevention of similar incidents. Methods The appropriate amount of samples was added into a 20 mL headspace vial, equilibrated at 70 °C for 15 min, separated by capillary column DB-FFAP，quantitatively detected by gas chromatography（hydrogen flame ionization detector; and separated by capillary column DB-5MS, qualitatively detected by gas chromatography-mass spectrometry(SCAN).Results The methanol showed good linearity in the range of 1.58~506.88 μg/mL, the correlation coefficient r was 0.999 6, the detection limit was calculated with S/N as 3, and the quantitative limit was calculated with S/N as 10. The detection limit and quantitative limit of methanol in urine and wine were 0.53 μg/mL and 1.58 μg/mL, the detection limit and quantitative limit of methanol in the blood were 2.64 μg/mL and 7.92 μg/mL, and the relative standard deviation (RSD) of methanol in urine and wine were 6.6~2.3 % (n=7) and 3.7~6.7 % (n=7), the RSD of methanol in the blood is 2.8~3.0 % (n=7).The recoveries of three levels of spiking in urine, alcohol and blood were 98.4~ 111.0 %, 83.1~ 86.0 % and 91.9~ 98.9 %(n=7).The concentration of methanol in the blood was 788.0 μg/mL, the concentration of methanol in the urine was 66.4 μg/mL, the concentration of methanol in the liquid of unknown residence was 542000.0 μg/mL, and the concentration of methanol in the remaining self-made wine was 71225.0 μg/mL.The blood methanol content of the patient was higher than the urine. The concentration of methanol in bulk wine in surrounding shops was lower than the national standard limit. Conclusion The methanol detection method established in our laboratory has the characteristics of rapid and high sensitivity, and can be used for the qualitatively and quantitatively determination of methanol in various samples of methanol poisoning event, this poisoning event was caused by methanol.