注射用炎琥宁稳定性和相关产物的化学分析及毒性

The degraded products and toxicity of potassium sodium pehydroandrographolide succinate for injection

目的研究注射用炎琥宁(脱水穿心莲内酯双琥珀酸半酯钠钾盐)的稳定性,分析其降解产物的化学组成,以在体实验考察极端条件获得降解物的毒性作用,为临床应用出现的不良反应分析提供借鉴。方法建立HPLC分析条件,分析炎琥宁分别在酸、碱条件下、常规生理盐水、葡萄注射液配制条件下的化学稳定性和成分变化情况,以LC-TOF-MS/MS法分析产物化学组成;采用HE染色法观察炎琥宁酸处理产物对实验动物对血管刺激性,对心、肝、肾等脏器的损伤程度,检测血清中的尿素氮(BUN)、肌酐(CRE)、谷丙转氨酶(GPT)、和谷草转氨酶(GOT)的含量来评价转化炎琥宁对实验动物的肝、肾损伤程度。结果炎琥宁/生理盐水注射液在日间24hr内稳定性良好;在pH5-8的溶液环境下相对稳定,在pH<5和pH>8的条件下降解程度加大。毒性实验结果表明,与正常小鼠相比,炎琥宁酸处理产物组血清中的BUN显著降低(P<0.01),CRE、GOT、和GPT显著升高(P<0.01),而常规注射用炎琥宁组血清中的CRE也显著升高(P<0.01)。常规炎琥宁组对肾脏均有不同程度的损伤,对血管刺激所导致的损伤能够随着时间恢复正常。结论本结果为临床应用注射用炎琥宁提供实验依据。

Objective The stability of potassium sodium dehydroandroandrographolide succinate for injection(PSPSI), dehydrated andrographolide diacid succinic sodium potassium salt) was studied and the chemical composition of the degradation products was analyzed. The toxicity of the degrading substances was obtained in the extreme conditions of the experiment. Methods The HPLC analysis conditions were established to analyze the chemical stability and composition changes of PSPSI succinate under the conditions of acid, alkali, normal saline and grape injection. The chemical composition of the products was analyzed by LC-TOF-MS/MS method. HE staining was used to observe the damage on the vascular irritation of the experimental animals. The contents of urea nitrogen(BUN), creatinine(CRE), glutamic-pyruvic transaminase(GPT), and glutamic-oxaloacetic transaminase(GOT) were determined to evaluate the degree of injury to organs. Results The stability of PSPSI in daytime 24 hr was good, and the content of PSPSI with 5% glucose injection was changed with time, the content of products decreased. PSPSI was relatively stable under pH 5-8 solution environment, and the degradation increased in pH<5 and pH>8 conditions. The degradation products include the open ring product of the succinyllactone, the single succinyl product, dehydration product, decarboxylation product in extreme conditions of pH 3 and pH 12, with a single succinyl product of m/z 432, and the product of internal lipid ring opening. Compared with normal mice, the serum BUN level was significantly decreased(P<0.01), the values of CRE, GOT, GPT and CRE content in serum were significantly increased in PSPSI group were significantly increased(P<0.01).The kidney had different degrees of damage to the kidney, and the damage caused by vascular stimulation could be restored to normal with time, and the group of acid treatment products was far greater than the normal group, and the injury of the heart was obvious. Conclusion The present study would provide experimental basis for the use of PSPSI in clinic.