RP-HPLC法测定替加环素的有关物质

目的建立高效液相色谱法测定替加环素的有关物质.方法方法 1:色谱柱:采用安捷伦Agilent ZORBAXSB-C8柱(250 mm×4.6 mm,5μm);流动相为0.2 mol爛L-1醋酸铵溶液(含0.01 mol爛L-1乙二胺四乙酸二钠)-N,N-二甲基甲酰胺-四氢呋喃(67∶29∶4)(用三乙胺调pH值至7.9);检测波长为254 nm.方法 2:色谱柱:采用岛津Shim-pack VP-ODS C18柱(250 mm×4.6 mm,5μm);流动相为0.01 mol爛L-1磷酸二氢钠缓冲液(含0.02 mol爛L-1辛烷磺酸钠及0.5%三乙胺,以磷酸调节pH值至3.0)-乙腈(75∶25);检测波长为246 nm.流速均为1 mL爛min-1;进样体积均为20μL.结果使用方法 1,杂质与替加环素能达到基线分离,替加环素与相邻峰的分离度为3.4,理论塔板数为3 446.线性范围为0.49～19.42μg爛mL-1,r=0.999 9.结论采用反相高效液相色谱法,专属性强,能准确、快速检测替加环素的有关物质,可用于替加环素的质量控制.

Determination of related substances in Tigecycline by RP -HPLC

Objective To establish an HPLC method for the determination of related substances in Tigecycline.Methods Method 1:Agilent ZORBAX SB-C8 column(250 mm×4.6 mm,5 μm) was adopted with 0.2 mol·L-1 solution of ammonium acetate(containing 0.01 mol·L-1 EDTA)-DMF-THF(67∶ 29∶ 4)(adjusted pH7.9 with TEA) as mobile phase.The detection wavelength was 254 nm.Method 2:Shim-pack VP-ODS C18 column(250 mm×4.6 mm,5 μm) was adopted with 0.01 mol·L-1 sodium phosphate monobasic buffer fluid(containing 0.02 mol·L-1 sodium octanesulfonate and 0.5% TEA,adjusted pH3.0 with phosphoric acid)-acetonitrile(75∶ 25) as mobile phase.The detection wavelength was 246 nm.The flow rate was 1.0 mL·min-1 and the injection volume was 20 μL for both methods.Results Tigecycline and impurities can be separated with baseline though method 1,with the resolution 3.4,the number of theoretical plate 3 446 and the linear range 0.49~19.42 μg·mL-1(r=0.999 9).Conclusion This RP-HPLC method was specific,accurate and rapid in determination of related substances in Tigecycline,and it can be used for the quality control of Tigecycline.