Bhandari Pamita, Kumar Neeraj, Gupta Ajai P, Singh Bikram, Kaul Vijay K
Natural Plant Products Division, Institute of Himalayan Bioresource Technology, Palampur, HP, India.
J Sep Sci. 2007 Aug;30(13):2092-6. doi: 10.1002/jssc.200700066.
A simple, sensitive, selective, precise, and robust high-performance TLC (HPTLC) method was developed and validated for determination of flavonoids in herbal extracts Bauhinia variegata, Bacopa monnieri, Centella asiatica, Ginkgo biloba, Lonicera japonica, Rosa bourboniana, Rosa brunonii, and Rosa damascena. The HPTLC of flavonoids was performed on RP-18 F(254) TLC plates with dual run, water (5% formic acid)/methanol (70:30) and water (5% formic acid)/methanol (50:50) as mobile phases. Densitometric determination of flavonoids was performed at lambda = 280 nm in reflectance/absorbance mode. The linear regression analysis data for the calibration plots showed a good linear relationship with r(2 )= 0.998 +/- 0.0003 in the concentration range of 150-800 ng/spot for apigenin and rutin and 200-1000 ng/spot for quercetin, luteolin, and quercitrin with respect to peak area. The average recovery for apigenin, quercetin, rutin, luteolin, and quercitrin was 97-99.8% indicating the excellent reproducibility. Statistical analysis of the data showed that the method is reproducible and selective for determination of flavonoids.
开发并验证了一种简单、灵敏、选择性好、精确且稳健的高效薄层色谱(HPTLC)方法,用于测定草药提取物羊蹄甲、积雪草、亚洲积雪草、银杏、忍冬、波旁蔷薇、布氏蔷薇和大马士革蔷薇中的黄酮类化合物。黄酮类化合物的HPTLC在RP - 18 F(254) TLC板上进行,采用双次展开,以水(5%甲酸)/甲醇(70:30)和水(5%甲酸)/甲醇(50:50)作为流动相。黄酮类化合物的密度测定在反射率/吸光度模式下于λ = 280 nm处进行。校准曲线的线性回归分析数据表明,芹菜素、芦丁在150 - 800 ng/斑点的浓度范围内,槲皮素、木犀草素和槲皮苷在200 - 1000 ng/斑点的浓度范围内,相对于峰面积具有良好的线性关系,r(2 ) = 0.998 ± 0.0003。芹菜素、槲皮素、芦丁、木犀草素和槲皮苷的平均回收率为97 - 99.8%,表明具有出色的重现性。数据的统计分析表明,该方法对于黄酮类化合物的测定具有可重复性和选择性。