Blood transfusion is a critical medical treatment, which is performed to save patients’ lives. Chylous blood had high fats. The transfusion of chylous blood into a patient can cause the blockage of micro-vessels. Most blood collection stations are not equipped with the equipment for the detection of chylous blood, and the detection is usually performed with direct observation through the human naked eye, which is prone to certain human errors. Only a few large blood collection stations use the equipment for the detection of chylous blood. In this study, plasma hyperspectral data were collected to detect and identify chylous plasma. The data were preprocessed using the multiple scattering correction (MSC) method and then classified using four classification algorithms, including random forest (RF), K-nearest neighbor KNN), Perceptron, and stochastic gradient descent (SGD) algorithms. First, the healthy and chylous plasma samples were classified into simple dichotomies. The best algorithm was identified by comparing the results of classification algorithms. The results showed that the random forest algorithm-based classification model had the best effect.Then, the chylous plasma was subdivided into different degrees of chylous plasma, which were less separable.A random forest algorithm-based plasma chylous degree detection model was established. Finally, 10 important spectral bands, including 1192.45 nm, 1182.9 nm, 946.98 nm, 1202.01 nm, 1080.93 nm, 1278.41 nm, 1237.03 nm, 991.65 nm, 1020.35 nm, and 1697.8 nm, were selected by band selection. After adjusting the parameters to optimize the model, the prediction accuracy of the whole band was 0.89. This study suggested that hyperspectral technology could identify chylous plasma and could be used to improve its detection efficiency in biomedicine, blood donation centers, human function tests, and other aspects. Filling the gap between machine learning and hyperspectral technology.To provide a new method for the diagnosis of chylous plasma.