[目的]建立柱前手性衍生化-反相高效液相色谱法拆分D,L-草铵膦。[方法]以邻苯二甲醛/N-乙酰基-L-半胱氨酸(OPA/NAC)为手性衍生化试剂,反应生成具有荧光吸收的一对非对映异构体衍生物,采用戴安C18色谱柱,流速1.0 mL/min,50 mmol/L醋酸铵缓冲溶液(pH值5.7)-甲醇(体积比90∶10)流动相,柱温35℃,激发波长350 nm,发射波长450 nm。[结果]L-草铵膦与其光学异构体分离度大于4,最低检测质量浓度为0.1μg/L(S/N≥3),线性范围为1~100 mg/L(r2为0.9995),方法重复性好。[结论]采用邻苯二甲醛/N-乙酰基-L-半胱氨酸(OPA/NAC)经柱前手性衍生化-RP-HPLC法可用于L-草铵膦的光学纯度控制。 [Objective] To establish a pre-column chiral derivatization-RP-HPLC method for the resolution of D, L-glufosinate. [Method] OPA / NAC was used as a chiral derivatization reagent to generate a pair of diastereoisomer derivatives with fluorescence absorption by using Diao C18 column with a flow rate of 1.0 mL / min and a mobile phase of 50 mmol / L ammonium acetate buffer solution (pH 5.7) -methanol (90:10 by volume) with a column temperature of 35 ℃, excitation wavelength of 350 nm and emission wavelength of 450 nm. [Result] The results showed that the separation of L-glufosinate and its optical isomers was more than 4 with the lowest detection concentration of 0.1μg / L (S / N≥3) and the linear range was 1-100 mg / L (r2 was 0.9995) Method repeatability is good. [Conclusion] The optical purity of L-glufosinate could be controlled by OP-N-acetyl-L-cysteine ​​(OPA / NAC)