HPLC-SEC法测定中药注射剂中残留大分子蛋白含量的研究OA

Objective:To establish a HPLC-SEC method for determination of macromolecular protein residues in traditional Chinese medicine injections(TCMIs).Methods:The analysis was performed on the TSK GEL G2000SWxl(7.8 mm ×300 mm,5 μm)column at the temperature of 35 ℃.The mobile phase was composed of acetonitrile and 0.08％trifluoroacetic acid(5∶95,v/v).The flow rate was 0.8 mL·min-1 and the detection wavelength was set at 214 nm.Results:The protein reference substances were used to validate the method,including serum albumin(bovine),lysozyme,insulin human and thymic peptide α1.The molecular weight range of the proteins was 66 400-3 108 Da.The linear relationship was good in the range of 0.381 2-3.812 μg(r＞0.999)as lysozyme was used as the reference substance to calculate the relative content.The precision,repeatability,and stability of the method were good(RSD≤2.0％).The relative content of macromolecular protein was detected in 2 out of 5 batches of TCMIs and the content was 25.7 μg·mL-1 and 32.6 μg·mL-1.Conclusion:The HPLC-SEC method was simple,sensitive and reproducible,and can be used for the determination of macromolecular protein residues in TCMIs.It provided an effective method for governing the quality of TCMIs.