A method for performing linear or stepped gradient elution on a glatiramer acetate with a high-performance liquid chromatography and determining whether the content of each component of the glatiramer acetate is qualified, comprising the following steps: step: preparing a solution of the glatiramer acetate to be tested; step: performing gradient elution on a sample to be tested with a reversed-phase liquid chromatography; step: determining a peak area corresponding to each component of the glatiramer acetate, comparing the peak area with to a peak area of a reference substance to determine whether the content of each component of the sample to be tested is in a qualified range, wherein in step, chromatographic conditions for the reversed-phase liquid chromatography are as follows: C18-bonded, C12-bonded, C8-bonded, or C4-bonded silica particle with a particle diameter of 1.7-10 μm is used as a packing material for a chromatographic column; acetonitrile is used as mobile phase A; a solution containing 30-80 mM ammonium sulfate is used as mobile phase B, wherein pH of the mobile phase B is adjusted to 2-3 with a phosphoric acid solution; or 0.1% trifluoroacetic solution is used as the mobile phase B; injection volume is 5-50 uL, the concentration of the sample is 1-20 mg/mL; a flow rate is 0.5-1.5 mL/min, the time for the gradient elution is 50-250 min; gradient of the gradient elution is as follows: a total time of the gradient elution is divided into N steps uniformly for gradient elution sequentially, wherein 2 ≤ N ≤ 20, in the first N-1 steps, the proportion of the mobile phase A is gradually increased from 5% to 40%, and the proportion of the mobile phase B is gradually reduced from 95% to 60%, in the N-th step, the proportion of the mobile phase A is 5%, and the proportion of the mobile phase B is 95%; column temperature is 25-50°C. step: preparing a solution of the glatiramer acetate to be tested; step: performing gradient elution on a sample to be tested with a reversed-phase liquid chromatography; step: determining a peak area corresponding to each component of the glatiramer acetate, comparing the peak area with to a peak area of a reference substance to determine whether the content of each component of the sample to be tested is in a qualified range, C18-bonded, C12-bonded, C8-bonded, or C4-bonded silica particle with a particle diameter of 1.7-10 μm is used as a packing material for a chromatographic column; acetonitrile is used as mobile phase A; a solution containing 30-80 mM ammonium sulfate is used as mobile phase B, wherein pH of the mobile phase B is adjusted to 2-3 with a phosphoric acid solution; or 0.1% trifluoroacetic solution is used as the mobile phase B; injection volume is 5-50 uL, the concentration of the sample is 1-20 mg/mL; a flow rate is 0.5-1.5 mL/min, the time for the gradient elution is 50-250 min; gradient of the gradient elution is as follows: a total time of the gradient elution is divided into N steps uniformly for gradient elution sequentially, wherein 2 ≤ N ≤ 20, in the first N-1 steps, the proportion of the mobile phase A is gradually increased from 5% to 40%, and the proportion of the mobile phase B is gradually reduced from 95% to 60%, in the N-th step, the proportion of the mobile phase A is 5%, and the proportion of the mobile phase B is 95%; column temperature is 25-50°C. a total time of the gradient elution is divided into N steps uniformly for gradient elution sequentially, wherein 2 ≤ N ≤ 20, in the first N-1 steps, the proportion of the mobile phase A is gradually increased from 5% to 40%, and the proportion of the mobile phase B is gradually reduced from 95% to 60%, in the N-th step, the proportion of the mobile phase A is 5%, and the proportion of the mobile phase B is 95%; column temperature is 25-50°C.