The invention provides a method for determining related substances in a
vitamin B6
drug, which comprises the following steps: dissolving a
vitamin B6
drug sample in a prefabricated solution in a dark environment, filtering, and diluting to a
target concentration interval to obtain a sample solution; placing the blank
solvent, the contrast solution and the sample solution in a
chromatographic column, introducing the mobile phase A and the mobile phase B, and sequentially carrying out
gradient elution to obtain a chromatogram; and identifying and separating
impurity chromatographic peaks in the chromatogram, performing quantitative or semi-quantitative analysis on each
impurity chromatographic peak, and calculating the content proportion of each
impurity chromatographic peak relative to the main chromatographic peak of the
vitamin B6. Through
collaborative design of the sample pretreatment state, the
gradient elution path and the peak interpretation mode, known impurities and unknown impurities in the
vitamin B6
medicine are in a separable state before entering a chromatographic
system, and are fully expanded in the
chromatographic separation process, so that accurate identification and effective separation of various impurities are realized.