痰热清注射液中微量蛋白检测方法的改进

Improvement in detection of trace protein in Tanreqing Injection

目的对痰热清注射液(黄芩、金银花、连翘、山羊角、熊胆粉)及其工艺中间体中微量蛋白的检测方法进行系统研究并选择注射剂中灵敏度高和专属强的蛋白质的检测方法。方法以凝胶电泳为检测手段考察了丙酮法、沉淀试剂法、超滤法、凝胶小柱法、沉淀法和固相萃取法对痰热清注射液及其中间体中微量蛋白的提取效果。结果以固相萃取除杂结合沉淀法富集的纯化方法为优,凝胶电泳法在痰热清注射液成品中未检出蛋白条带,但在中间体中检出蛋白条带,其检出限为2.03μg/mL。结论本法有效解决了药典方法中假阳性率高的问题,且灵敏度提高了25倍,更适宜于中药注射液中蛋白质的检测.

AIM To systematically study the detection of trace protein in Tanreqing Injection and its intermediates,and to select an analytic method with high sensitivity and specificity for protein in injection. METHODS Gel electrophoresis was used to compare the extraction efficiency of trace protein in injection and its intermediates among acetone,precipitation,ultrafiltration,gel column and solid phase extraction combined with precipitation.RESULTS The experiment showed solid phase extraction combined with precipitation ranked first in all the protein extractions. There was no protein detected in Tanreqing Injection,but the intermediates had proteins with the detection limit of 2. 03 μg /mL by gel electrophoresis. CONCLUSION The developed method can rule out the existence of false positive reaction and raise the sensitivity of up to 25 times,and be applied to detecting the trace protein in injection containing herbs.