液-质联用分析金线莲中4种黄酮类化合物

Analysis of the Extraction of the Four Kinds of Flavonoids from Anoectochilus roxburghi by HPLC-ESI/MS

目的：测定和鉴别福建和广西产地金线莲中4种黄酮类化合物（异鼠李素‐3‐O‐新橙皮糖苷、异槲皮苷、槲皮素和异鼠李素）。方法应用高效液相色谱‐电喷雾电离／离子阱质谱法（HPLC‐ESI／MS）测定和鉴别4种黄酮类化合物。色谱条件：Agilent TC‐C18柱（4．6 mm ×250 mm ，5μm）；流动相：A为0．8％乙酸＋0．2％氨水溶液，B为甲醇；流速：1．0 mL／min；梯度洗脱：0～10 min ，50％～58％ B；10．01～30 min ，65％ B。检测波长368 nm ，柱温30℃，进样量20μL。质谱条件：电喷雾离子源（ESI源），正离子监测。结果不同产地金线莲中异鼠李素‐3‐O‐新橙皮糖苷、异槲皮苷、槲皮素和异鼠李素含量分别是：福建产地为 0．094，0．053，0．039和0．070 mg／g ；广西产地为0．089，0．067，0．042和0．061 mg／g。结论 HPLC‐ESI／MS法分析金线莲中4种黄酮类物质具有快速、简便的特点，且不同产地金线莲中这4种具有重要药理活性黄酮类化合物含量有差异，首次发现金线莲中含有异鼠李素‐3‐O‐新橙皮糖苷。

Objective To determine and identify the four kinds of flavonoids （isorhamnetin‐3‐O‐neohesperidoside ,isoquercitrin ,quercetin and isorhamnetin） in A noectochilus roxburghii （A .roxburghii） by high performance liquid chromatography‐electrospray ionization/ion traps mass spectrometry （HPLC‐ESI/MS） . Methods The four kinds of flavonoids were extracted from A .roxburghii by ultrasonic‐assis‐ted extraction ,and determined by HPLC . Chromatographic condition：Agilent TC‐C18 column （4 .6 mm × 250 mm ,5 μm） . The mobile phase was composed of solvent A （0 .8% acetic acid aqueous solution and 0 .2% ammonia）and solvent B （methanol） . Linear gradient program was performed as follows：0~10 min , linear gradient 50% ~58% B ,10 .01~30 min ,isocratic elution 65% B . The flow rate was 1 .0 mL/min . The injection volume was 20 μL . The column temperature and wavelength were set at 30 ℃ and 368 nm ,respectively . Results The content of the four kinds of flavonoids （isorhamnetin‐3‐O‐neohesper‐idoside ,isoquercitrin ,quercetin and isorhamnetin） in different origin of A .roxburghii of Fujian origin and Guangxi origin were 0 .094 and 0 .089 mg/g ,0 .053 and 0 .067 mg/g ,0 .039 and 0 .042 mg/g ,0 .070 and 0 .061 mg/g ,respectively . Conclusion The method used for analysis of the four kinds of flavonoids from A .roxburghii has fast and convenient characteristics . There is a difference in the content of four kinds of flavonoids in A .roxburghii from different origins .This is the first report that isorhamnetin‐3‐O‐neohes‐peridosid coexist in A .roxburghii .