Determination of residual acetic acid in the raw materials of β-lactam antibiotics using high performance liquid chromatography
Objective To establish high performance liquid chromatography(HPLC)for the determination of residual acetic acid in β-lactam raw materials.Methods A multifactorial within-group design method was used to determine the composition,ratio,and pH of the chromatographic mobile phase.The established method was validated in terms of specificity,limit of detection,limit of quantification,linearity,accuracy,precision,and robustness with reference to ICH Q2.This developed method identified 19 β-lactam antibiotic raw materials.Results The separation was carried out on a C18 column with the mobile phase of methanol-0.02 mol/L potassium dihydrogen phosphate solution in gradient elution;the column temperature was 25 □,and the detection wavelength was 220 nm.There was no obvious interference with the determination of acetic acid by the test samples,solvent,or mobile phase.The lowest detection limit of acetic acid was about 0.029 μg and the lowest limit of quantification was about 0.12 μg.The linearity of the method was good in the range of concentration from the limit of quantification to 2 times the limit concentration of acetic acid.And the fitted linear equation was y=388866.7186x-177.5720,r2=0.9999.The average recoveries of the spiked samples at low,medium,and high concentrations were 93％,96％and 99％respectively;the repeatability and intermediate precision of the method were 2.37％and 2.11％respectively.The robustness of the method was good on the three different kinds of C18 columns.Residual acetic acid was tested in three out of the 19 β-lactam antibiotics raw materials.Conclusion The proposed method could achieve accurate quantification of residual acetic acid in β-lactam raw materials,which provided a reference for process control and the content assignment of standard substances for such raw materials.
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