HPLC测定穿琥宁中的聚合物

Research of polymer in potassium dehydroandrogapholide succinate

目的采用HPLC法测定穿琥宁聚合物的含量,并研究聚合物的性质及成因。方法采用依利特Sephadex G-10色谱柱(10 mm×300 mm,40～120μm),流动相A为pH7 0.05 mol.L-1磷酸盐缓冲液[0.05 mol.L-1磷酸氢二钠溶液-0.05 mol.L-1磷酸二氢钠溶液(61∶39)],流动相B为水,流速1.0 mL.min-1,检测波长251 nm。用HPLC法分析温度、pH、湿度、灭菌时间对聚合物产生的影响。结果对照品、供试品溶液的线性范围分别为0.011～22.20μg(r=0.9999)、0.0163～2.98 mg(r=0.9988),最低检测限为2.2 ng(S/N=3),最低定量限为11 ng(S/N=10),进样RSD=1.13%,日内RSD=0.56%,对照品溶液在10 h内较稳定,供试品溶液不稳定。穿琥宁聚合后极性增加;高温、高湿、pH、长灭菌时间使聚合物的含量增加。结论所用方法简便、灵敏、准确、重复性好,可用于穿琥宁聚合物的检查和含量测定;生产中应严格控制温度、湿度、pH及灭菌时间。

OBJECTIVE To establish an HPLC method for determination of polymer in potassium dehydroandrographohde succinate （PDS） ,and study the nature and the causes of polymer. METHODS An optimal HPLC method was set up. The samples were separa- ted by Elite Sephadex G - 10 column（10 mm×150 mm, 40 - 120 μm） with 0.05 mol·L^-1,phosphate buffer [0.05 mol·L^-1 disodium hydrogen phosphate - 0.05 mol·L^-1 sodium dihydrogen phosphate（61：39） ] at pH 7.0 as mobile phase A,water as mobile phase B, and detected with UV 251 nm with a flow rate of 1.0 mL·min^-1. The effect of temperature, pH, humidity, and sterilization time on poly-mer was analyzed by HPLC. RESULTS The calibration curve of the control solution was linear in the range of 0.011 - 22.20 μg（ r = 0.9999） and the calibration curve of the test solution was linear in the range of 0.0163 - 2.98 mg（ r = 0.9988 ）. LOD was 2.2 ng （S/N =3） and LOQ was 11 ng（ S/N = 10）. The RSD of injection precision was 1.13% and the RSD of intra - day precision was 0.56%. The control solution was stable within 10 h, but the test solution was not stable. After polymerization, the polarity of PDS was increased. The content of polymer was inereased with high temperature, high humidity,high pH and long sterilization time. CONCLU- SION The method is good at the aspect of simplicity, sensitivity, reproducibility and it is applicable for determination of the content of polmyer in PDS. Seriously control of temperature,humidity, !oH and sterilization time through nharmaeeutical production is important.