目的 建立RP-HPLC法測定穿心蓮內(nèi)酯中有關(guān)物質(zhì)，并對穿心蓮內(nèi)酯原料進行不同影響因素的穩(wěn)定性考察，了解影響其穩(wěn)定性的具體因素。方法 采用反相高效液相色譜法，菲羅門Gemini C18色譜柱（250 mm×4.6 mm，5 μm），檢測波長為224 nm，柱溫為30 ℃，體積流量為1.0 mL/min，流動相為乙腈–甲醇–水梯度洗脫。考察穿心蓮內(nèi)酯原料藥在不同pH值、氧化、高溫、高濕、強光照條件下的穩(wěn)定性。結(jié)果 穿心蓮內(nèi)酯及其3個有關(guān)物質(zhì)得到了很好地分離，所建立的有關(guān)物質(zhì)方法經(jīng)驗證，表明專屬性、重復(fù)性及穩(wěn)定性均良好。穿心蓮內(nèi)酯固體狀態(tài)較為穩(wěn)定；在溶液狀態(tài)對氧化和pH 3～5條件較為穩(wěn)定，在堿性介質(zhì)、高溫、光照條件下均不穩(wěn)定。結(jié)論 該方法可以很好地用于穿心蓮內(nèi)酯的質(zhì)量控制，穿心蓮內(nèi)酯在溶液狀態(tài)下總體穩(wěn)定性較差，該結(jié)果為穿心蓮內(nèi)酯及其相關(guān)制劑研究提供一定參考。

Objective To establish a selective RP-HPLC method for the determination of related substances in andrographolide, and to investigate the stability of them affected by different factors. Methods RP-HPLC method was used on Phenomenex Gemini C18 column (250 mm × 4.6 mm, 5 μm) at the temperature of 30 ℃. The mobile phases consisted of acetonitrile, methanol, and water. The flow rate was set at 1.0 mL/min and the UV detection was performed at 224 nm. This method was applied to evaluate the stability of andrographolide in solid and liquid forms by the following conditions: different pH values, oxidation, high temperature, high humidity, and strong light. Results The andrographolide and three related substances were separated. The developed method was further validated and proved to be sensitive, precise, and stable. The andrographolide in solid state was stable relatively, and its liquid state was also stable relatively in oxidation and pH 3－5 conditions; however in the conditions of alkaline, high temperature and light, andrographolide was proved not to be stable. Conclusion The method is successfully applied to the quality control of andrographolide. The andrographolide in liquid state has poor stability, and the results could give some references for the preparation study.