The identification of the type and the writing time of black signature pen handwriting has always been a hot topic in the field of forensic science at home and abroad. Based on the advantages of Raman spectroscopy, such as high resolution, good stability, high efficiency and nondestructive testing, 16 brands of black signature pen handwriting samples were tested. The samples of the same brand at the same time with different storage conditions (darkroom and lighting) and different paper background (copy paper and notebook) handwriting samples were completed. The handwriting samples were clipped and fixed on the glass slide with the double-sided adhesive, and stored in the storage box to protect them from light, and then the samples were tested at intervals. The test lasted for one year to form a Raman spectrum database. The optimum conditions were selected based on the investigation of the influence of experimental conditions, such as measuring point, laser power and confocal type. Based on the optimum conditions, the handwriting samples of black signature pen were determined under different writing time, storage environment and paper background. The experimental results indicated that: (1) Under different conditions of storage conditions (darkroom and lighting) and paper background (copy paper and notebook), the same kind of black signature pen handwriting had the same Raman shift, and the repeatability was good, indicating the less disturbance by the background of water and paper. (2) According to the Raman shift, 16 types of black signature pen could be classified into five categories. (3) Taking 2# sample as example, a characteristic peak of Raman spectrum at 1 140 cm-1 was assigned to the C—O stretching vibration ν(CO) of the ester compound, the content of which decreasing over time. Therefore, the relative intensity of the characteristic peak was increased with the writing time (8→1). The relative area value I was calculated by the fitting method, and it indicated that the I value was increased with the writing time of the sample. Our strategy can be applied in quick and nondestructive determination of the types and relative writing times of the black signature pen handwriting.