HPLC法测定克林霉素磷酸酯及其有关物质的方法学研究

Research on Determination of the Content and the Related Substances of Clindamycin Phosphate by HPLC

目的:建立克林霉素磷酸酯及其有关物质含量的测定方法并进行方法学研究。方法:采用HPLC法,色谱柱为Li-chrospher C_(18)柱(250mm×46mm,5μm);柱温:25℃;流动相为0.1mol·L^(-1)的磷酸二氢钾缓冲液(用40％氢氧化钾溶液调节pH=5.6)-乙腈(76:24);流速:1.2mL·min^(-1);检测波长:210nm。结果:与BP1998和EP1997相比,表克林霉素-2-磷酸酯及其另一未知杂质和克林霉素磷酸酯得到很好分离;线性范围;克林霉素-2-磷酸酯在0.1～5.0mg·mL^(-1)呈良好线性,r=0.9996;克林霉素B-2-磷酸酯在0.0013～0.13mg·mL^(-1)呈良好线性,r=0.9981;表克林霉素-2-磷酸酯在0.0014～0.14mg·mL^(-1)呈良好线性,r=0.9972;克林霉素在0.0011～0.11mg·mL^(-1)呈良好线性,r=0.9983;最低检测限为0.1μg;最低定量限为0.33μg。结论:本法比英国药典和欧洲药典更能准确地测定克林霉素磷酸酯及其有关物质含量。

Objective: To establish a HPLC method for determination of clinadamycin phosphate and its related substances. Methods: The column: Lichrospher C18 (250 mm×4. 6 mm, 5 μm) ; The mobile phase: 0. 1 mol · L-1 KH2PO4 buffer (adjust pH to 5. 6 using 40% KOH solution) - acetonitrile (76: 24) ; The flow rate: 1.2 mL · min-1 ; The detective wavelength; 210 nm. Results: Epiclindamycin phosphate and a unknown impu-rity could be separated from clindamycin phosphate, but these two impurities were overlapped with clindamycin phosphate according to BP and EP method. The linear range of clindamycin - 2 - phosphate was 0. 1 - 5. 0 mg · mL-1 , r = 0. 999 6; The linear range of clindamycin B - 2 - phosphate was 0. 001 3 - 0. 13 mg · mL-1 , r = 0. 998 1 ; The linear range of epiclindamycin -2 -phosphate was 0. 001 4 - 0. 14 mg ·mL-1 , r = 0. 997 2; The linear range of clindamycin was 0. 001 l - 0. 11 mg · mL-1, r = 0. 998 3; The LOD was 0. 1 μg; The LOQ was 0. 33 μg. Conclusions: The method is more accurate to determination than BP and EP method.