A GC method for the separation and determination of three diasteromers in neuroprotectant 3β，5α，6β-androstriol(YC-6) bulk drug was developed.The separation conditions such as column type，injection port temperature，column temperature and sample concentrate were optimized，and method validation was developed.The results showed that the method had a good specificity（resolution（R） ≥1.5） and a good precision（RSD≤2.0%，n=6）.The calibration curve showed a good linearity over the concentration range of 6.0-30.0 μg/mL for YC-6.The response factors of three diasteromers were 1.59，1.32 and 1.41，respectively，and their contents were detected by principal component self-comparison with calibration factor and internal standard method.The method was simple，sensitive and valid，and was successfully applied in the analysis of diasteromers in neuroprotectant 3β，5α，6β-androstriol bulk drug.