The identification of bloodstain is one of the most important approaches in obtaining evidence in criminalistics. A threshold method based on spectral co-efficient and interclass variance is proposed in this paper, it is a non-contact, non-destructive method for quickly identifying bloodstains. The spectra of bloodstains and other suspected substances were all extracted from their hyper-spectral image. Then calculate the correlation coefficients of these spectral and interclass variances, analyze the differences between substances. The best blood recognition threshold was determined as 0.9. After preprocessing for eliminating systematic errors, experiments with the threshold 0.9 are carried out to identify bloodstains on the calico and red T-shirt. The method can remarkably identify the bloodstain from other non-blood substances both quickly and efficiently. The blood extraction rate can reach to 93.35% and 89.19%, respectively. It is an important step toward the implementation of bloodstain non-contact and non-destructive identification in forensic casework.