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目的:用反相高效液相色谱法测定N-甘氨酰-L-谷氨酰胺一水合物含量。方法:以2,4-二硝基氟苯作柱前衍生化试剂,在碱性条件下制得2,4-二硝基苯基-N-甘氨酰-L-谷氨酰胺,上ODS色谱柱,置日本岛津SCL-10AVP高效液相色谱仪上进行分析。流动相为0.05mol·L~(-1)醋酸钠水溶液(用冰醋酸调pH6.4)-乙腈(80:20),检测波长为360nm,进样量20μL,流速1mL·min~(-1)。结果:N-甘氨酰-L-谷氨酰胺-水合物在16～80μg·mL~(-1)范围内线性良好,r=0.994 3,回归方程Y=1.2436×10~(-4)X,重现性RSD=0.31％(n=5)。结论:本法可用于测定N-甘氨酰-L-谷氨酸胺-水合物的含量。
Objective: To determine the content of N-glycyl-L-glutamine monohydrate by reversed-phase high performance liquid chromatography. Methods: 2,4-dinitrofluorobenzene was used as pre-column derivatization reagent to prepare 2,4-dinitrophenyl-N-glycyl-L-glutamine under basic conditions. Column, set Japan Shimadzu SCL-10AVP high-performance liquid chromatography for analysis. The mobile phase consisted of 0.05 mol·L -1 sodium acetate solution (adjusted to pH6.4 with glacial acetic acid) -acetonitrile (80:20), the detection wavelength was 360 nm, the injection volume was 20 μL, the flow rate was 1 mL · min -1 ). Results: The linearity of N-glycyl-L-glutamine-hydrate was good in the range of 16-80 μg · mL -1, r = 0.994 3. The regression equation was Y = 1.2436 × 10 -4 , Reproducibility RSD = 0.31% (n = 5). Conclusion: This method can be used to determine the content of N-glycyl-L-glutamine amine-hydrate.