HPLC法同时测定紫花地丁提取物中绿原酸、秦皮乙素、芦丁、金合欢素-7-O-β-D-葡萄糖苷、槲皮素和木犀草素含量

Simultaneous Determination of Chlorogenic Acid,Aesculetin,Rutin,Acacetin-7-O-β-D-glucoside,Quercetin and Luteolin in the Extract of Viola Yedoensis Makino by HPLC

目的:建立同时测定紫花地丁提取物中绿原酸、秦皮乙素、芦丁、金合欢素-7-O-β-D-葡萄糖苷、槲皮素和木犀草素的HPLC法。方法:采用HPLC法,以Hypersil ODS C18柱(250 mm×4.6 mm,5μm)为色谱柱,以0.1%磷酸水(A)-甲醇(B)为流动相进行梯度洗脱,检测波长为345 nm,流速为1.0 ml·min^(-1),柱温为35℃。结果:绿原酸、秦皮乙素、芦丁、金合欢素-7-O-β-D-葡萄糖苷、槲皮素和木犀草素分别在4.317 5~172.700 0 mg·L^(-1)、2.7350~109.400 0 mg·L^(-1)、6.980 0~279.200 0 mg·L^(-1)、3.720 0~148.800 0 mg·L^(-1)、4.135 0~165.400 0 mg·L^(-1)和3.395 0~135.800 0 mg·L^(-1)范围内线性关系良好,平均回收率分别为99.06%,98.84%,98.77%,99.40%,98.53%和98.71%,测得紫花地丁提取物中绿原酸、秦皮乙素、芦丁、金合欢素-7-O-β-D-葡萄糖苷、槲皮素和木犀草素的含量分别为1.629 0,1.191 0,4.585 0,2.281 0,3.179 0,1.971 0 mg·g^(-1)。结论:该方法准确、可靠,重复性好,可用于紫花地丁提取物的质量控制。

Objective： To establish an HPLC method for the simultaneous determination of chlorogenic acid, aesculetin, rutin, acacetin-7-O-β-D-glucoside, quercetin and lute01in in the extract of Viola yedoensis Makino. Methods： The HPLC analysis was carried out on a Hypersil ODS C18 column （250 mm ×4.6mm,5μm） with 0.1% phosphoric acid （A）- methanol （B） as the mobile phase with gradient elution at the flow rate of 1.0 ml min-I The detection wavelength was 345 nm and the column temperature was 35 ℃. Results： Good linear relationship was found within the range of 4. 317 5-172. 700 0 mg L-1 for chlorogenic acid, 2.735 0-109. 400 0 mg. L-1 for aeseuletin, 6.980 0-279.200 0 mg L-1 for rutin, 3.720 0-148.800 0 mg L-1 for acacetin-7-O-β-D-glucoside, 4. 135 0-165.400 0 mg L-1 for quercetin, and 3. 395 0-135. 800 0 mg L-I for luteolin. The average recovery was 99.06%, 98.84%, 98.77%, 99.40%, 98.53% and 98.71%, respectively. The content of chlorogenic acid, aesculetin, rutin, acacetin-7-O-β-D-glucoside, quercetin and luteolin in the extract of Viola yedoensis Makino was 1. 629 0, 1. 191 0, 4.585 0, 2.281 0, 3. 179 0 and 1. 971 0 mg g-1, respectively. Conclusion： The method is accurate and reliable with good reproducibility, which can be used for the quality control of the extract of Viola yedoensis Makino.