Determination of the content and related substances in isepamicin sulfate injection by post-column derivatization HPLC
Pages: 363-368
Year: 2017 Issue: 5
Journal: Chinese Journal of Antibiotics
Abstract: Objective To compare the methods of HPLC with post-column derivatization and HPLC-ELSD for the determination of the content and related substances in isepamicin sulfate injection,and to establish a reasonable method for its quality control.Methods A YMC-pack ODS AM column (4.6mm×250mm,3μm) was used,the mobile phase was sodium sulfate solution (containing hexane sulfonate sodium salt and glacial acetic acid)-acetonitrile(99∶1),the column temperature was 35 ℃,and the flow rate was 0.8mL/min.The OPA solution was used for the post-column derivatization,at a flow rate of 0.8mL/min.The temperature of the post-column reaction was 45℃.The fluorescence was detected by an excitation and emission wavelength of 360nm and 440nm,respectively.Results The resolution between D-isepamicin,1N,3N-isepamicin,isepamicin and 3N-isepamicin is satisfied.The calibration curve was linear in the range of 124.8～291.2μg/mL (r=1.0000).The average recovery was 100.9％,and the RSD value was 0.9％ (n=9).The detection limit of isepamicin was 0.57ng,which was much lower than that of the method of HPLC-ELSD.Conclusion The method can make up the deficiencies of coelution of D-isepamicin with isepamicin,and low sensitivity recorded in Chinese Pharmacopoeia.It can also resolve the problems of coelution of 1N,3N-isepamicin with isepamicin,and bad resolution between isepamicin and 3N-isepamicin recorded in Japanese Pharmacopoeia.The method is rapid,inexpensive,versatile,and non-laborious.In addtion it ensures excellent results in terms of simplicity,sensitivity,specificity,reproducibility,and enables the quality control of isepamicin sulfate injection and its related substances.
Year: 2017 Issue: 5
Journal: Chinese Journal of Antibiotics
Abstract: Objective To compare the methods of HPLC with post-column derivatization and HPLC-ELSD for the determination of the content and related substances in isepamicin sulfate injection,and to establish a reasonable method for its quality control.Methods A YMC-pack ODS AM column (4.6mm×250mm,3μm) was used,the mobile phase was sodium sulfate solution (containing hexane sulfonate sodium salt and glacial acetic acid)-acetonitrile(99∶1),the column temperature was 35 ℃,and the flow rate was 0.8mL/min.The OPA solution was used for the post-column derivatization,at a flow rate of 0.8mL/min.The temperature of the post-column reaction was 45℃.The fluorescence was detected by an excitation and emission wavelength of 360nm and 440nm,respectively.Results The resolution between D-isepamicin,1N,3N-isepamicin,isepamicin and 3N-isepamicin is satisfied.The calibration curve was linear in the range of 124.8～291.2μg/mL (r=1.0000).The average recovery was 100.9％,and the RSD value was 0.9％ (n=9).The detection limit of isepamicin was 0.57ng,which was much lower than that of the method of HPLC-ELSD.Conclusion The method can make up the deficiencies of coelution of D-isepamicin with isepamicin,and low sensitivity recorded in Chinese Pharmacopoeia.It can also resolve the problems of coelution of 1N,3N-isepamicin with isepamicin,and bad resolution between isepamicin and 3N-isepamicin recorded in Japanese Pharmacopoeia.The method is rapid,inexpensive,versatile,and non-laborious.In addtion it ensures excellent results in terms of simplicity,sensitivity,specificity,reproducibility,and enables the quality control of isepamicin sulfate injection and its related substances.
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