HPLC测定L-肌肽中L-组氨酸及其中间体的含量

Determination of L-histidine and Intermediate in L-carnosine by HPLC

目的建立高效液相色谱法(HPLC)测定L-肌肽中L-组氨酸及其中间体含量的方法。方法采用Atlantis HILIC Silica(150 mm×4.6 mm,5μm)色谱柱,柱温35℃,流动相为0.01 mol/L醋酸铵溶液(加冰醋酸调p H4.0)-乙腈(28:72),流速1.0 ml/min,检测波长220 nm。结果 L-组氨酸在0.5093~10.1856μg/ml范围内与峰面积有良好线性关系(r=0.9995),平均回收率为99.30%(RSD=0.99%);中间体在0.4810~9.6206μg/ml范围内与峰面积有良好线性关系(r=0.9991),平均回收率为98.99%(RSD=1.85%)。结论方法专属性强、重现性好、灵敏度高,可用于L-肌肽中L-组氨酸及中间体含量的测定。

Objective To establish a method for determination of L-histidine and intermediate in L-carnosine by HPLC.Methods An Atlantis HILIC Silica column(150 mm×4.6 mm,5μm)with DAD detecter was adopted,using 0.01 mol/L ammonium acetate(adjust pH to 4.0 with glacial acetic acid)-acetonitrile(28:72,v/v)as mobile phase.The detection wavelength was 220 nm,the flow rate was 1.0 ml/min and the column temperature was 35℃.Results A good linear relationship was obtained in the concentration rang of 0.5093-10.1856μg/ml for histidine,with the correlation coefficient of 0.9995.The average recovery was 99.30%(n=6),with RSD of 0.99%.There was a good linear relationship whithin the rang of 0.4810-9.6206μg/ml for intermediate,with the correlation coefficient of 0.9995.The average recovery was 98.99%(n=6),with RSD of 1.85%.Conclusion The method has good specificity,repeatability and high sensitivity.It can be used for determination of the content of L-histidine and intermediate in L-carnosine.