D-p-hydroxyphenylglycine (D-HPG) is an important intermediate for the synthesis of sweeteners and drugs. An innovative continuous chromatographic process was developed to recover D-HPG from crystallization mother liquor, with the aim of reducing the process cost and improving the quality of D-HPG. The thickening properties of different nanofiltration membranes on mother liquor were investigated by pre-treatment test, and the Applexion NF200 nanofiltration membrane was selected to enrich D-HPG. The separation properties of different resins were analyzed by single column experiment, and the separation degree of Applexion XA945 resin to D-HPG and ammonium sulfate could reach 16. Based on the results, the way of sequential simulated moving bed separation was further studied. The results showed that after separation by the sequential simulated moving bed, the average purity of D-HPG in feeding was increased from 42.2% to 96.3%, and the average yield was 91%. The continuous chromatographic system is a promising method for the recovery of D-HPG from crystallization mother liquor. Compared with the traditional process, the new process not only ensures the total yield of D-HPG, but also greatly reduces the process cost and improves the product quality.