Determination of related substances in tranexamic acid injection by reversed-phase and weak cation-exchange mixed-mode column
Objective:To establish a new HPLC method for the determination of the related substances in tranex-amic acid injection by screening new type of HPLC columns,which can avoid the use of ion pair reagents in cur-rent methods and reduce salt concentration.Methods:The method was carried out using a Thermo Mixed-mode WCX column(150 mm × 4.6 mm,5 μm),the mobile phase consisted of 10 mmol·L-1 sodium dihydrogen phosphate solution(adjust the pH value to 5.2±0.05 with sodium hydroxide solution)-water-acetonitrile(50∶5∶45),the flow rate was 1.0 mL·min-1,the column temperature was 25 ℃,the detection wavelength was 210 nm and the injection volume was 20 μL.Results:The chromatographic peaks of tranexamic acid and its four impurities B,C,D and E were all separated.Good linear relationship was shown between the concentration of all the five compounds and their corresponding peak areas(r ≥ 0.999).The LODs were 0.34,0.50,0.005 6,0.002 1,0.12 μg·mL-1.The average recoveries(n=9)were 97.4％,100.5％,98.4％and 96.6％with RSDs of 3.9％,0.24％,0.52％and 1.4％,respectively.The test solution and standard solution were all stable within 22 h.The detection results of 5 batches of tranexamic acid injection showed that the number of unspecified impurities and the total impurities content by using the new method were better than the current ChP method.Conclusion:The established method is specified and sensitive.Good separation can be achieved with low concen-tration of phosphate.Its applicable to the determination of related substances in tranexamic acid injection.
tranexamic acidrelated substancesreversed-phase and weak cation-exchange mixed-mode col-umnquality controlhigh performance liquid chromatography methodoptimization of chromatographic methods
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