高效液相色谱法同时检测4种金线莲黄酮苷元及其在提取工艺评价中的应用

Simultaneous Determination of Flavonoid Glycosides from Anoectochilus roxburghii by High Perfomance Liquid Chromatogrophy and Its Application for the Assessment of Extraction Technology

基于高效液相色谱(HPLC)和提取离子流色谱-质谱(EIC-MS),建立了金线莲提取成分定性定量分析及提取效率评价新方法。方法采用20RBAX Ecipse XDB-C18色谱柱,以乙腈-0.03%乙酸水溶液为流动相,采用梯度洗脱,同时分离测定金线莲提取液中芦丁、山奈酚、异鼠李素和槲皮素4种黄酮苷元的含量,经EIC-MS验证,结果可靠。以黄酮提取率为指标,评价酶解-醇提、回流-醇提、搅拌-醇提、超声-醇提、超声-微波-醇提和煎煮6种提取工艺,结果显示,最佳提取工艺为回流-醇提法,提取时间短,醇用量少,总提取率最高,可达14.96mg/g。

Based on high perfomance liquid chromatogrophy（HPLC） and extracted ion chromatogram- mass spectrometry（EIC-MS）, a method for the determiantion of flavonoid glycosides, including rutin, quercetin, kaempferol and isorhamnetin was proposed and applied for the assessment of extraction technology. Acetonitrile and acetic acid solution（0.03% ,v/v） were used as mobile phase. Four kinds of flavonoid glycosides from the extract of Anoectochilus ro：cburghii could be identified respectively by EIC- MS and determined by using gradient progrem. Six kinds of extraction technology, including enzymic- alcohol extraction, reflux-ethanol extraction, rotatory-ethanol extraction, ultrasonic extraction and ultrasonic-microwave extraction were used. The efficiency of extraction technology was assessed by the total content of flavonoid glycosides. The optimum extraction technology was reflux-ethanol extraction for its high-speed,the least usage of ethanol and the maximum extraction efficiency（i, e. , 14. 9 mg/g）.