高效液相色谱测定D,L-对羟基苯甘氨酸

Determination of D,L-p-hydroxyphenylglycine by HPLC

建立了乙醛酸为原料合成D,L-对羟基苯甘氨酸反应体系的分析方法,采用高效液相色谱法(HPLC)测定,以ZORBA Eclipse Plus C_(18)色谱柱(4.6×250 mm,5μm)为色谱柱,水/甲醇为流动相梯度洗脱,探讨了各种色谱条件对分离度的影响,在优化后的色谱条件下(流速为0.8 mL·min^(-1),柱温为25℃,检测波长为254 nm),D,L-对羟基苯甘氨酸与杂质A分离度大于1.5,D,L-对羟基苯甘氨酸的线性方程为y=7 602.4x+254.55,线性相关系数0.999 6,检出限(S/N=3)为0.05 mg·kg^(-1),测定下限(S/N=10)为0.18 mg·kg^(-1)。加标回收率为93.4%~101.0%,相对标准偏差(n=6)为0.7%~2.0%。该方法与薄层色谱(TLC)测定结果基本一致,但更为简单,且灵敏度高。

A HPLC determination method for analysing D,L-p-hydroxyphenylglycine,reacted from glyoxylic acid was established.The effects of various chromatographic conditions on the chromatographic separation were discussed,using ZORBA Eclipse Plus C_(18) column(4.6×250 mm,5 μm) as chromatographic column,water/methanol as mobile phase.Under the optimized chromatographic conditions(gradient elution with flow rate of 0.8 mL·min^(-1),25 ℃ as column temperature,254 nm as detection wavelength),the resolution of D,L-p-hydroxyphenylglycine and impurity A was more than 1.5.The linear equation of D,L-p-hydroxyphenylglycine was y=7 602.4x+254.55,the linear correlation coefficient was 0.999 6,the detection limit(S/N=3) was 0.05 mg·kg^(-1),and the detection limit(S/N=3) was 0.18 mg·kg^(-1).The standard recovery rate was 93.4% to 101.0%,and relative standard deviations(n=6) were from 0.7% to 2.0%.The method was basically consistent with that of thin layer chromatography(TLC),but it was more simple and sensitive.