Determination of glycine in human thrombin by HPLC / 药学实践杂志

Objective To establish a HPLC method for glycine assay in the human thrombin. Methods The sample was derivatized with 2,4-dinitrofluorobenzene (DNFB). HPLC was performed on an ODS-C18 column with 50% acetonitrile –0.05mol/L sodium acetate buffer (35:65) as the mobile phase. The flow rate was 1.0 ml/min and detection wavelength at 360 nm. Results The linear range of glycine was 0.006-0.030 mg/ml (r=0.999 6). The average recovery was 100.4%, with RSD 0.44% (n=9). Conclusion This method is simple, accurate and specific. It is suitable for glycine assay in human thrombin.