Analysis Method Research Of Cefonicid Sodium For Injection

Objectives:To create a test method of HPLC for the assay of Cefonicid Sodium for injection, and to create a test method of HPLC for the related substance of Cefonicid Sodium for injection, and to create a test method of HPSEC for the Polymer in Cefonicid Sodium for injection.Method:Applying HPLC method, with Thermo BDS column (4.6mm×250mm,5μm) as solid phase, detection wave length272nm, flow speed1.0mL· min-1, Column temperature35℃, and Acetonitrile-0.01mol·L-1of Potassium Dihydrogen Phosphate (Regulating the PH to3.5with Phosphoric Acid)(13:87) as mobile phase, to test the assay of Cefonicid Sodium for injection; applying gradient elution partition chromatography with Acetonitrile-0.01mol· L-1of Potassium Dihydrogen Phosphate (Regulating the PH to3.5with Phosphoric Acid)(13:87) as mobile phase, to test the related substance of Cefonicid Sodium for injection. Applying HPLC method, with Sephadex G-10(40cm) as solid phase, detection wave length254nm, flow speed1.2mL· min-1, and0.1mol· L-1Disodium Hydrogen Phosphate-0.1mol· L-1Sodium Dihydrogen Phosphate (69:31) as mobile phase A, and water as mobile phase B, to test the Polymer in Cefonicid Sodium for injection.Results:With the foresaid HPLC method, the exclusion for assay and related substance test is acceptable, and the resolution of unknown impurity and7-ACA,3-TCA, and active substance Cefonicid Sodium is acceptable. The linearity of Cefonicid Sodium, within the range of1.25-12.5μg· mL-1(r=1)、0.05-0.8mg·mL-1(r=1) are acceptable. The accuracy of assay is RSD=0.76%(n=6), the repeatability is RSD=0.70%(n=6), the precision is100.5%(n=9), the stability of the solution within12hrs is acceptable(RSD=0.34%); the repeatability of related substance test RSD=1.12%(n=6), lowest limit of test is10ng, it is indicated that the known impurity3-TSA would be remarkably increasing whilst keeping up the sample solution of Cefonicid Sodium, so it have to be prepared just before testing. With the HPLC method of the test of Polymer, the linearity of Cefonicid Sodium is acceptable within0.010-0.20mg· mL-1(r=0.9999), the rate between the concentration of control solution and peak area, i.e. F, the relative standard variance within daytime is2.89%(n=6), the relative standard variance between days is2.40%(n=6), minimum test limit is0.15mg, the correlation between the concentration of test solution and test results of Polymer is acceptable(r=0.9995).Conclusion:The test method of Assay and Polymer are concise, quick, precise, sensitive, the method of related substance is concise, and able to identify impurities in Cefonicid Sodium for Injection. The entire method can effectively control the quality of product.