HPLC测定D-对羟基苯甘氨酸甲酯中的手性杂质

Determination of Chiral Impurities in D-hydroxyphenylglycine Methyl Ester by HPLC

关键词:  D-对羟基苯甘氨酸甲酯  手性杂质  高效液相色谱法

Key words:  D-hydroxyphenylglycine methyl ester  chiral impurity  HPLC

摘要:

目的 建立HPLC测定D-对羟基苯甘氨酸甲酯中潜在的手性杂质[杂质1：L-对羟基苯甘氨酸，杂质2：L-对羟基苯甘氨酸甲酯，杂质3：D-对羟基苯甘氨酸]的含量和限度。方法 采用Crownpak CR（-）手性色谱柱（150 mm×4.0 mm，5 μm）；流动相：高氯酸溶液（pH 1.7）-甲醇（90：10）；检测波长为226 nm；流速：0.7 mL·min^-1；柱温：25℃。结果 杂质1、杂质2和杂质3均在其定量限浓度~2.400 μg·mL^-1内线性良好（r分别为1.000 0，0.999 9，0.999 9），平均回收率分别为99.97%，100.30%和103.18%，RSD分别为0.30%，0.64%和0.62%（n=9）。结论 该方法专属性强，准确、方便，可以作为D-对羟基苯甘氨酸甲酯中手性杂质1、杂质2和杂质3的液相分析方法。

Abstract:

OBJECTIVE To establish a method of HPLC to determine the limit and content of potential chiral impurities:L-hydroxy phenyl glycine (impurity 1), L-hydroxyphenylglycine methyl ester (impurity 2) and D-hydroxyphenylglycine (impurity 3) in active pharmaceutical substance D-hydroxyphenylglycine methyl ester qualitatively and quantitatively. METHODS A Crownpak CR(-) chiral column (150 mm×4.0 mm, 5 μm) was used, with perchlorate solution (pH 1.7) -methanol (90:10) as the mobile phase with a flow rate at 0.7 mL·min^-1. The detection wavelength was 226 nm, and the temperature of column was 25℃. RESULTS The calibration curve for impurity 1, impurity 2 and impurity 3 were well linear in the range of LOQ concentration-2.400 μg·mL^-1 (r were 1.000 0, 0.999 9 and 0.999 9, respectively). Average recoveries for impurity 1, impurity 2 and impurity 3 were 99.97%, 100.30% and 103.18%, and RSDs were 0.30%, 0.64% and 0.62% (n=9), separately. CONCLUSION The method is highly specific, accurate and convenient, it can be used for the quality control of the chiral impurities 1, impurities 2 and impurities 3 of D-hydroxyphenylglycine methyl ester.