HPLC同时测定雪松不同部位中4个黄酮类化合物的含量

Simultaneous Determination of Four Flavonoids in Different Parts of Cedrus Deodara by HPLC

目的 建立同时测定雪松不同部位4个黄酮类化合物（杨梅素、槲皮素、山柰酚和异鼠李素）含量的高效液相色谱法。方法 雪松样品用60%甲醇回流提取,提取液加10%盐酸水解。采用Agilent TC-C18（250 mm×4.6 mm,5μm）色谱柱,流动相为乙腈-0.1%磷酸水溶液,梯度洗脱,流速1.0 m L·min^-1,检测波长360 nm,柱温30℃。结果 雪松不同部位中杨梅素、槲皮素和异鼠李素浓度都在0.5~10μg·m L^-1、山柰酚浓度在1~24μg·m L-1内呈良好线性关系（r≥0.999 6）,加样回收率（n=6）分别为98.9%,100.1%,99.5%和101.7%。雪松各部位中4个黄酮类化合物的含量存在明显差异,其中松针中槲皮素和山柰酚的含量最高分别达0.285 mg·g^-1和1.493 mg·g^-1,小枝中杨梅素和异鼠李素的含量最高分别达0.365 mg·g^-1和0.476 mg·g^-1;雪松不同部位中4个黄酮类化合物含量之和依次为松针〉小枝〉花序〉木质〉树皮。结论 该方法简单、快速、高效,可用于雪松不同部位中4个黄酮类化合物的含量测定。

OBJECTIVE To develop an HPLC method for simultaneous determination of four flavonoids（myricetin, quercetin, kaempferol, isorhamnetin） in different parts of Cedrus deodara. METHODS The sample were pretreated, including reflux extraction with 60% methanol for 1 h, and then extracting solution hydrolysis with 10% hydrochloric acid for another 1 h. Agilent TC-C18（250 mm×4.6 mm, 5 μm） column was adopted. The mobile phase was acetonitrile-0.1% phosphoric acid solution with the gradient elution at the flow rate of 1.0 m L·min^-1. The column temperature was 30 ℃ and the detection wavelength was 360 nm. RESULTS Good linear correlation were found in the ranges of 0.5-10.0 μg·m L^-1 for myricetin, quercetin and isorhamnetin and 1-24.0 μg·m L^-1 for kaempferol（r≥0.999 6）. The average recoveries（n=6） of the four flavonoids were 98.9%, 100.1%, 99.5% and 101.7%, respectively. The results showed that the concents of four flavonoids in different parts of C. deodara were significant different, the highest contents of quercetin and kaempferol of pine needles were 0.285 mg·g^-1 and 1.493 mg·g^-1, the highest contents of myricetin and isorhamnetin of tender branchlets were 0.365 mg·g^-1 and 0.476 mg·g^-1 respectively. The order of total contents of four flavonoids from highest to lowest were pine needles, tender branchlets, inflorescences, woods, barks. CONCLUSION The method is simple, rapid and efficient, and applicable for determination of four flavonoids in different parts of C. deodara.