| 于凤琳,袁丹 陈宇,李美芽,等.高效液相色谱法同时测定牡蒿叶中4个绿原酸类成分含量[J].浙江中医药大学学报,2021,45(6):675-684. |
| 高效液相色谱法同时测定牡蒿叶中4个绿原酸类成分含量 |
| Simultaneous Determination of Four Chlorogenic Acids in Artemisia Japonica Thunb. Leaves by HPLC |
| DOI：10.16466/j.issn1005-5509.2021.06.021 |
| 中文关键词: 牡蒿 绿原酸 异绿原酸 高效液相色谱法 响应面法 超声提取 提取工艺 含量测定 |
| 英文关键词: Artemisia japonica Thunb. chlorogenic acid isochlorogenic acid high performance liquid chromatography response surface method ultrasonic extraction extraction process content determination |
| 基金项目:2020年浙江中医大学学生发展性资助项目(2020-64、2020-66) |
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| 中文摘要: |
| [目的]通过响应面法优化提取工艺，并结合高效液相色谱(high performance liquid chromatography，HPLC)法测定牡蒿叶中4个绿原酸类成分的含量，探究其开发利用价值。[方法]采用Thermo Scientific Syncronis Amino色谱柱(4.6mm×250mm，5μm)，色谱条件为流动相乙腈.0.1%甲酸水(40.60，V/V)，流速1mL·min-1，柱温25℃，检测波长325nm；以绿原酸、异绿原酸A、异绿原酸B和异绿原酸C的总含量为响应值，乙醇浓度、液料比、超声功率为主要因素，采用Box-Behnken响应面法对牡蒿叶中4个绿原酸类成分进行优化提取，并测定其含量。[结果]以乙腈.0.1%甲酸水(40.60，V/V)等度洗脱，流速1mL·min-1，柱温25℃，检测波长325nm，牡蒿叶中绿原酸、异绿原酸A、异绿原酸B和异绿原酸C分离度良好，在6.25～200.00μg·mL-1浓度范围内线性关系良好，加样回收、精密度、稳定性和重复性均符合要求。优化得到的最佳提取工艺为:乙醇浓度60%，液料比40.1(单位:mL·g-1)，超声功率250W，频率53kHz，室温超声20min。通过HPLC法测得绿原酸、异绿原酸A、异绿原酸B和异绿原酸C含量分别为14.317 0mg·g-1、27.945 2mg·g-1、1.230 4mg·g-1和5.066 5mg·g-1，合计总量达48.559 0mg·g-1。[结论] HPLC法测得牡蒿叶中绿原酸类成分含量较为丰富。采用响应面法优化提取工艺，能够显著提高牡蒿叶中4个绿原酸类成分提取率，具有较好潜在开发应用价值。 |
| 英文摘要: |
| [Objective] To determine the contents of four chlorogenic acids in Artemisia japonica Thunb. leaves and analyze its utilization value by optimization of the extraction process with Box-Behnken response surface method and high performance liquid chromatography(HPLC) method. [Methods] Thermo Scientific Syncronis Amino column(4.6mm×250mm， 5μm) was carried out， acetonitrile. 0.1% formic acid water(40.60， V/V) was taken as the mobile phase， the flow rate was 1mL·min-1， the column temperature was 25℃， the detection wavelength was 325nm. And taking the total contents of chlorogenic acid， isochlorogenic acid A， isochlorogenic acid B and isochlorogenic acid C as response values， the extraction process of four components from Artemisia japonica Thunb. leaves was optimized by Box-Behnken response surface method with ethanol concentration， liquid-solid ratio and ultrasonic power as the main factors. Then the contents of the four compounds were determined by HPLC method. [Results] Chlorogenic acid， isochlorogenic acid A， isochlorogenic acid B and isochlorogenic acid C in Artemisia japonica Thunb. leaves could be well separated by isocratic elution with acetonitrile and 0.1% formic acid water at the flow rate of 1mL·min-1 and detection at 32 nm under 25℃. And the four components showed good linear relationship with their corresponding peak areas in the concentration range of 6.25～200.00μg·mL-1. Moreover， the recovery， precision， stability and repeatability of the method were in line with the requirements. The optimal extraction conditions were as follows. Ethanol concentration 60%， liquid-solid ratio 40.1(mL·g-1)， ultrasonic power 250W， ultrasonic frequency 53kHz and ultrasonic 20min at room temperature. Under these conditions， the content of chlorogenic acid， isochlorogenic acid A， isochlorogenic acid B and isochlorogenic acid C in Artemisia japonica Thunb. leaves were 14.317 0mg·g-1， 27.945 2mg·g-1， 1.230 4mg·g-1 and 5.066 5mg·g-1， respectively， with a total amount of 48.559 0mg·g-1. [Conclusion]The content of chlorogenic acids in the leaves of Artemisia japonica is abundant detected by HPLC， which has good potential development and application value. Response surface method optimized extraction process significantly improves the extraction rate of the four chlorogenic acids in Artemisia japonica Thunb. leaves. |
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