HPLC法测定盐酸多西环素及其制剂含量和有关物质

目的: 克服现有色谱流动相易使分析系统堵塞等问题,建立新的盐酸多西环素及其制剂含量和有关物质测定的HPLC方法。 方法: 用十八烷基硅烷键合硅胶为填充剂,以醋酸盐缓冲液 10∶ 1),用冰醋酸或氨水调节pH至8.8]-乙腈(85∶ 15)为流动相;柱温35 ℃;检测波长280 nm,流速1.0 mL·min^-1;进样体积20 μL。 结果: 新建色谱条件克服了堵塞问题,可使多西环素与相邻杂质基线分离,在0.07~122.556 μg·mL^-1范围内线性良好(r=1.0),检测限和定量限分别为0.016和0.063 μg·mL^-1,方法精密度良好,片剂和胶囊的回收率分别为99.2%和99.4%,盐酸多西环素在0.01 mol·L^-1的盐酸溶液中24 h内稳定。该色谱条件微调后,可用于土霉素、四环素、金霉素、米诺环素和美他环素的含量和有关物质的测定。 结论: 新建色谱条件优于中国药典2005年版、美国药典32版、英国药典2008年版、欧洲药典6.0版和日本药局方XV版,被中国药典2010年版采用。

Objective: To solve the blocking problem of the current official pharmacopeia and establish a new HPLC method for the determination of doxycycline hyclate and its preparations. Methods: The meithod was carried using a column packed with octadecysilane bonded silica gel and a mixture of a buffer solution containing 100 volumes of 0.25 mol·L^-1 ammonium acetate solution,10 volumes of 0.1 mol·L^-1 ethyldiaminetetraacetic acid disodium salt solution and 1 volume of triethyamine adjusted pH to 8.8 with glacial acetic acid-acetonitrile (85∶ 15) as the mobile phase,and with a column temperature of 35 ℃,a detection wavelength of 280 nm,a flow rate of 1 mL·min^-1,and the injection volume was 20 μL. Results: The blocking problem of the current official pharmacopeia was overcame and a good resolution with baseline separation was obtained by the established chromatographic system.The linear range was 0.07-122.56 μg·mL^-1 (r=1.0),the limit of detection was 0.016 μg·mL^-1,and the limit of quantity was 0.063 μg·mL^-1.The average recovery data of tablet and capsule was 99.2%(n=9,RSD=0.2%) and 99.4% (n=9,RSD=0.2%),respectively.This chromatographic system can be applied to the determination of oxytetracycline,tetracycline,chlorotetracycline,minocycline and metacycline by adjusting the mobile phase ratio of acetate buffer and acetonitrile. Conclusions: The established method is better than the methods of ChP 2005,USP 31,BP 2008,EP 6.0 and JPⅩⅤ and was adopted by ChP 2010.