HPLC法同时测定和评价“黔中金荞麦”中4种成分含量

Simultaneous Determination and Evaluation of the Contents of 4 Components in "Qianzhong Golden Buckwheat" by HPLC

关键词:  黔中金荞麦  芦丁  槲皮素  表儿茶素  原花青素B2  高效液相色谱

Key words:  Qianzhong golden buckwheat  rutinum  quercetin  L-epicatechin  procyanidin B2  high performance liquid chromatography

摘要:

“黔中金荞麦”是贵州省农业科学院畜牧兽医研究所选育的优质牧草（品种登记号：2019581）。为建立同时测定该药材中表儿茶素、原花青素B2、芦丁和槲皮素的检测方法，考察其含量多样性。采用高效液相色谱法建立检测方法，测定并分析其含量与野生金荞麦的差异。结果得出检测方法为：C18柱（250×4.6 mm，5 μm），乙腈-0.4%磷酸水梯度洗脱，进样量10 μL，流速0.7 mL/min，检测波长254 nm，温度30 ℃；原花青素B2、表儿茶素、芦丁和槲皮素分别在5～85 μg/mL（r2=0.9998）、5～80 μg/mL（r2=0.9998）、0.5～8 μg/mL（r2=0.9996）、0.125~2.0 μg/mL（r2=0.9999）范围内呈现良好线性关系，加样回收率高；黔西和麦坪的“黔中金荞麦”4种成分总含量高达1189.70μg/g、1108.01μg/g，其中原花青素B2、芦丁和槲皮素含量最高，丹寨野生金荞麦表儿茶素含量最高。试验结果说明本法准确可靠，适合“黔中金荞麦”的内控质量监测，“黔中金荞麦”中活性物质含量高且质量稳定，优于野生金荞麦。因此本试验为该药材的开发应用提供了理论基础。

Abstract:

"Qianzhong Golden Buckwheat" is a high-quality forage selected by the Animal Husbandry and Veterinary Research Institute of the Guizhou Academy of Agricultural Sciences (variety registration number: 2019581). In order to establish a detection method for the simultaneous determination of epicatechin, procyanidin B2, rutin and quercetin in this medicinal material, the content diversity was investigated. High performance liquid chromatography was used to establish a detection method, and the difference between its content and that of wild golden buckwheat was determined and analyzed. The results showed that the detection method was as follows: C18 column (250×4.6 mm, 5 μm), acetonitrile-0.4% phosphoric acid water gradient elution, injection volume 10 μL, flow rate 0.7 mL/min, detection wavelength 254 nm, temperature 30℃; Procyanidin B2, epicatechin, rutin and quercetin were at 5-85μg/mL (r2=0.9998), 5-80μg/mL (r2=0.9998), 0.5-8μg/mL (r2=0.9996), 0.125~2.0μg/mL (r2=0.9999) showing a good linear relationship respectively. And the recovery rate of sample addition is high; The total content of the four components of "Qianzhong Golden Buckwheat" collected from Qianxi and Maiping was as high as 1189.70μg/g and 1108.01μg/g. The content of procyanidin B2, rutin and quercetin was the highest, and the content of epicatechin in Danzhai wild golden buckwheat was the highest. The test results show that this method is accurate and reliable, and is suitable for the internal quality monitoring of "Qianzhong Golden Buckwheat". "Qianzhong Golden Buckwheat" has high content of active substances and stable quality, which is better than wild Golden Buckwheat. Therefore. This experiment provides a theoretical basis for the development and application of this medicinal material.