Determination of phenol content in Neurotropin injection by HPLC

WANG Shuo; ZHANG Hua-jie; XUE Yan-hua; MA Xiao

doi:10.20251/j.cnki.1003-3734.2025.11.012

Abstract：Objective:To establish a high performance liquid chromatography method for the determination of phenol content in Neurotropin injection.Methods:The chromatography was performed on a ZORBAX 300 Extend-C18 column(150mm×4.6 mm,5 μm)using mobile phase of 0.01 mol·L-1 potassium dihydrogen phosphate and 20%methanol at a flow rate of 1 mL·min-1.The column temperature was set at 35℃.The detection wavelength was 270 nm and the injection volume of sample was 10 μL.The specificity,precision,repeatability,recovery rate and robustness of the method were verified,and the linear range of the method was determined.The phenol contents in 3 batches of Neurotropin injection were determined by the developed HPLC method,and the results are compared with those obtained by the bromine titration method.Results:In the linear range of 100～500 μg·mL-1,the concentrations of phenol showed a good linear relationship with the peak area(R2=1).The RSD of peak area and retention time of phenol were 0.19%and 0.19%,respectively,when the control solution was injected for 6 consecutive times.The RSD of peak area and retention time of phenol were 0.26%and 0.19%,respectively,when the samples was injected for 6 consecutive times.The average recovery of phenol was 100.73%,with a RSD of 0.14%.Small variations in detection conditions such as column temperature and flow rate do not affect the detection results,and the results of the two methods for the determination of phenol contents in Neurotropin were basically consistent.Conclusion:The method established in this study can accurately measure phenol contents.The method is simple and reproducible,and can avoid the interference of amino acids,nucleotides,polypeptides and other substances in the product.It is suitable for the determination of phenol content in the Neurotropin injection,and can also be used for the determination of phenol content in the product containing a variety of amino acids and other complex components.