HPLC法同时测定白花蛇舌草中7种活性成分的含量  

Simultaneous Determination of 7 Active Components in Hedyotis Diffuse by HPLC

作　　者：翟学佳[1] 吕永宁[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院药剂科,武汉430022

出　　处：《中国药师》2016年第1期70-72,81,共4页China Pharmacist

摘　　要：目的:建立同时测定白花蛇舌草中去乙酰车草酸酐、车叶草苷、秦皮乙素、4-香豆酸、阿魏酸、槲皮素和山奈酚7种有效成分含量的高效液相色谱方法。方法:采用Thermo ODS-2 HYPERSIL C_(18)色谱柱(250 mm×4.6 mm,5μm);流动相为0.05%醋酸水溶液(A)-0.05%醋酸乙腈(B),线性梯度洗脱[0~50 min,5%B#80%B],流速为1.0 ml·min^(-1);柱温为35℃;检测波长分别为238 nm(乙酰车草酸酐和车叶草苷)、347 nm(秦皮乙素)、306 nm(4-香豆酸)、322 nm(阿魏酸)、369 nm(槲皮素和山奈酚)。结果:去乙酰车草酸酐、车叶草苷、秦皮乙素、4-香豆酸、阿魏酸、槲皮素和山奈酚的浓度分别在2.940~58.700μg·ml^(-1),2.650~52.900μg·ml^(-1),0.762~15.200μg·ml^(-1),0.448~8.960μg·ml^(-1),0.794~15.900μg·ml^(-1),0.897~17.900μg·ml^(-1),0.489~9.780μg·ml^(-1)范围内均呈良好线性关系(r≥0.999 7);日内、日间精密度良好,RSD均小于1.95%(n=6);各组分低、中、高浓度的平均加样回收率均在98.16%~101.88%之间,RSD均小于1.88%(n=6);白花蛇舌草中去乙酰车草酸酐、车叶草苷、秦皮乙素、4-香豆酸、阿魏酸、槲皮素和山奈酚含量分别为(42.48±1.43)μg·g^(-1)、(63.76±1.01)μg·g^(-1)、(1 765±0.69)μg·g^(-1)、(881.9±0.74)μg·g^(-1)、(86.99±1.65)μg·g^(-1)、(1 395±0.731)μg·g^(-1)和(902.2±0.82)μg·g^(-1)。结论:本文方法简便可靠,重复性好,可全面反映白花蛇舌草中各种极性较高的成分的含量,适用于白花蛇舌草质量控制。Objective： To develop an HPLC method for the determination of deacetylation plantain anhydride, asperuloside, aes- culetin, 4-coumaric, ferulie acid, quercetin and kaempferol acid in Hedyotis Diffuse. Methods： A Thermo ODS-2 HYPERSIL C18 column （250 mm× 4.6 mm, 5 μm） was used. The mobile phase consisted of 0.05% aqueous acetic acid （A） and 0.05% acetic acid - acetonitrile （B） with linear gradient elution （0 - 50 min, 5% - 80% B） , and the flow rate was 1.0 ml ·min-1. DAD detection was used and the detection wavelength was set at 238 nm for deacetylation plantain anhydride and asperuloside, 347 nm for aesculetin, 306 nm for 4-coumaric, 322 nm for ferulie acid, and 369 nm for quereetin and kaempferol acid, respectively. The temperature of column was 35℃. Results： Deacetylation plantain anhydride, asperuloside, aesculetin, 4-coumaric, ferulic acid, quereetin and kaempferol acid showed good linearity （r≥0.999 7） respectively within the range of 2. 940-58. 700 μg · ml-1,2. 650-52. 900 μg· ml-1 ,0. 762- 15.200 μg· ml-1 ,0.448-8.960 μg· ml-1 ,0.794-15.900 μg · ml-1 ,0.897-17.900 μg· ml-1 and 0.489-9.780 μg · ml-1. The average recovery of the seven investigated compounds at low, medium and high concentration levels was within the range of 98.16%- 101.88% with RSD below 1.88% （n = 6）. The intra- and inter-day RSD was below 1.95% （ n = 6）. The contents of deacetylation plantain anhydride, asperuloside, aesculetin, 4-coumaric, ferulie acid, quercetin and kaempferol acid in Hedyotis Diffuse was （42.48 ±1.43）μg·g-1,（63.76 ±1.01）μg·g-1,（1 765±0.69）μg·g-1,（881.9±0.74）μg· g-1,（86.99±1.65）μg· g-1（1 395 ± 0.731 ） μg · g -1 and （902.2 ± 0.82 ） μg · g - 1 , respectively. Conclusion： The developed method is simple, accurate anti reproducible, and can be used to control the quality of Hedyotis Diffuse.

关 键 词：高效液相色谱法 白花蛇舌草 去乙酰车草酸酐 车叶草苷 秦皮乙素 4.香豆酸 阿魏酸 槲皮素 山奈酚 

分 类 号：R927.2[医药卫生—药学]