HPLC法测定注射用丹参酮Ⅱ_A磺酸钠含量及其有关物质

Determination of tanshinone Ⅱ_A sulfonate and related substances in sulfotanshinone sodium injection by HPLC

目的:建立注射用丹参酮ⅡA磺酸钠中丹参酮ⅡA磺酸钠的含量及其有关物质丹参酮Ⅰ磺酸钠及丹参酮ⅡA的HPLC测定方法。方法:色谱柱为RestekC18(4.6mm×200mm,5μm);流动相为A相(0.02mol/L磷酸氢二钠缓冲溶液,用磷酸调pH为7.0)-B相(甲醇)进行梯度洗脱(B相起始比例为50%,10min内上升到70%,保持5min,30minB相占95%);流速为1mL/min;检测波长为271nm。结果:被测定峰与其他峰可达到基线分离,丹参酮ⅡA磺酸钠、丹参酮Ⅰ磺酸钠及丹参酮ⅡA的线性范围分别为19.98～119.88μg/mL(r=0.9999),0.202～1.212μg/mL(r=0.9999)及0.202～1.212μg/mL(r=0.9998);平均回收率分别为99.8%,99.6%,99.7%;RSD分别为2.2%,2.0%,2.3%(n=9)。结论:该法简便、准确、分离效果好,适用于该制剂的质量控制及有关物质研究。

Aim ： To establish the HPLC method for determination of tanshinone Ⅱ A sulfonate and its related substances （tanshinone Ⅰ sulfonate and tanshinone Ⅱ A）in sulfotanshinone sodium injection. Methods： Restek C18 column（4.6 mm × 200 mm, 5 μm）was used with a mobile phase of A（0.02 mol/L dibasic sodium phosphate, adjusted to pH 7.0）- B（methanol） ,with a linear gradient from 50% to 70% of B phase mixed with A phase in 10 min, retained for 5 min, and the proportion of B phase reached 95 % in 30 min. The flowing rate was 1 mL/min. The detection wavelength was set at 271 nm. Results： A good resolution with baseline separation was obtained. The linear ranges of tanshinone 11 A sulfonate, tanshinone Ⅰ sulfonate and tanshinone ⅡA were 19.98 - 119.88 μg/mL（ r = 0.999 9）,0.202 - 1.212 μg/ mL（r = 0.999 9）and 0.202- 1.212 μg/mL（ r = 0.999 8）, respectively. The average recoveries were 99.8%, 99.6 % and 99.7 % with RSDs of 2.2 %, 2.0 % and 2.3 % （ n = 9）, respectively. Conclusion： The method is simple, accurate, selective, and reliable for quality control and determination of related substances of sulfotanshinone sodium injection.