In this paper,the rapid and nondestructive method for quantitative detection of cholecalciferol chol esterol content is proposed and realized by using the full Raman spectrum data analysis and partial least square s method.Raman spectra of 39groups of samples with different cholecalciferol ch olesterol contents were collected by self-made portable Raman spectrometer,26g roups were used as training set to establish partial least squares prediction model,and 13groups were used as testing set to validate the model.The linear correlation coefficient of the predicted content and the actual content of the test set is 0.9994.The results of the study demonstrate t he feasibility of total spectral analysis of Raman spectrum combined with partial least squares method in quantitative detect ion of cholecalciferol cholesterol content.