目的 建立同時(shí)測(cè)定黃芪中5種黃酮類(lèi)成分含量的高效液相色譜（HPLC）分析方法并進(jìn)行黃芪的指紋圖譜研究。方法 采用HPLC-DAD，Waters SymmetryShield^TM RP₁₈色譜柱（250 mm×4.6 mm，5 μm），乙腈（A）-水（0.05%磷酸）（B）為流動(dòng)相梯度洗脫，體積流量1.0 mL/min，檢測(cè)波長(zhǎng)246、206 nm，柱溫25℃，進(jìn)樣量20 μL，對(duì)黃芪中毛蕊異黃酮葡萄糖苷、芒柄花苷、（6aR，11aR）-9，10-二甲氧基紫檀烷-3-O-β-D-葡萄糖苷、毛蕊異黃酮、7-羥基-4'-甲氧基異黃酮進(jìn)行含量測(cè)定。結(jié)果 方法學(xué)考察結(jié)果顯示，黃芪中5種黃酮類(lèi)成分線(xiàn)性范圍，相關(guān)系數(shù)良好（r=0.999 8~0.999 9，n=9），各色譜峰間分離度均達(dá)到定量要求。精密度（RSD<1.86%），穩(wěn)定性（RSD<2.01%），重復(fù)性（RSD<1.97%），加樣回收率（RSD<2.90%，n=9）等參數(shù)均達(dá)到定量分析要求。建立了黃芪的HPLC指紋圖譜，標(biāo)定了27個(gè)共有峰，樣品相似度在0.808~0.971，存在一定差異。結(jié)論 本實(shí)驗(yàn)建立的方法準(zhǔn)確、簡(jiǎn)便、重復(fù)性好，可為黃芪藥材的質(zhì)量控制提供參考。

Objective To establish a method to determine the components of five flavonoids in Radix Astragali and built fingerprint of Radix Astragali by HPLC. Methods HPLC-DAD method was used to determine five flavonoids simultaneously, including calycosin-7-glucoside, ononin, (6aR, 11aR)-9, 10-dimethoxypterocarpan-3-O-β-D-glucoside, calycosin and 7-hydroxyl-4'-methoxyisoflavone. The separation was carried on a Waters Symmetry Shield^TM RP₁₈ column (250 mm×4.6 mm, 5 μm) with the mobile phase consisting of acetonitrile (A)-water (0.05% phosphoric acid) (B) by using gradient elution. The flow rate was 1.0 mL. min^-1 and the detection wavelengths were at 246 and 206 nm. The column temperature was set at 25℃ and the injection volume was 20 μL.Results Methodology examination showed that the five flavonoids showed a good linear relationship (r=0.999 8-0.999 9, n=9), precision (RSD < 1.86%), stability (RSD < 2.01%) and repeatability (RSD < 1.97%) conformed to requirements. The fingerprint of Radix Astragali was built. 27 mutual peaks were labeled and the similarity was more than 0.808. Conclusion The established method is accurate, simple and reproducible, which can provide reference for quality control of Radix Astragali.

國(guó)家自然科學(xué)基金資助項(xiàng)目（21502142）