采收期、生长年限和加工方法对黄花乌头不同部位中关附甲素含量的影响

Effects of Harvesting Time,Growing Years and Processing Methods on the Content of Guanfu Base A in Different Parts of Aconitum coreanum

目的:为规范黄花乌头的人工栽培,通过优化高效液相法,对不同采收时期、生长年限黄花乌头不同部位中关附甲素的含量进行测定,比较不同加工方法对关附甲素含量的影响,为制定黄花乌头合理的采收时间和加工方式提供依据。方法:采用反相离子对高效液相色谱法,测定黄花乌头中不同部位中关附甲素的含量;色谱柱为安捷伦Zorbax SB-C18柱(250 mm×4.6 mm,5μm),流动相A为2 mg/mL庚烷磺酸钠水溶液(含0.2%三乙胺并以H_(3)PO_(4)调pH值至3.0),流动相B为乙腈,采用梯度洗脱,流速为1.0 mL/min,检测波长为205 nm,柱温20℃。结果:关附甲素的质量浓度在0.0251~0.2513 mg/mL^(2)范围内线性关系良好(R^(2)=1),精密度、重复性、稳定性试验结果的RSD均≤1.0%;平均加样回收率为97.83%,RSD为3.5%。不同采收期黄花乌头不同部位中9月份采集的样品子根含量最高,说明9月份为最佳采收期。不同生产年限样品中,三年生和三年以上生黄花乌头母根含量相差不大,但均大于两年生母根,地上部分中所含关附甲素都明显小于根部。不同加工方法所得关白附中关附甲素的含量大小依次为:阴干>晒干>炕干,说明不同条件对含量的影响较大。结论:建立的含量测定方法专属性强、精密度、重复性和稳定性较好,结果可靠,黄花乌头的最佳采收期为9月份,生长期限为3年,本研究为黄花乌头科学的人工栽培采收提供了实验依据。

Objective:To establish an efficient HPLC method to determine the content of Guanfu base A(GFA)in different parts of Aconitum coreanum(Lèvl.)Rapaics(ACR)at different harvesting periods and growth stages.And the effects of different processing methods on the content of GFA were compared.This would provide reference for formulating the reasonable harvesting time and processing method of ACR.Methods:Using reversed-phaseion-paired high-performance liquid chromatography(HPLC),the content of GFA in ACR was determined.The chromatographic column used was an Agilent Zorbax SB-C18 column(250 mm×4.6 mm,5μm).The mobile phase A consisted of 2 mg/mL aqueous solution of sodium heptane sulfonate(containing 0.2%triethylamine and adjusted to pH 3.0 with phosphoric acid),while the mobile phase B was acetonitrile.Gradient elution was employed at a flow rate of 1.0 mL/min,with detection wavelength set at 205 nm and column temperature maintained at 20℃.Results:The quality concentration of GFA ranged from 0.0251 to 0.2513 mg/mL,showing a good linear relationship 2(R^(2)=1).The precision,repeatability,and stability test results were good with RSD≤1.0%.The average recovery rate was 97.83%,with an RSD of 3.5%.Among different harvesting periods,the highest content of GFA in ACR was found in the sub-roots collected in September,indicating that September is the optimal harvest period.There was no significant difference in GFA content between wild rhizomes and cultivated rhizomes.The aerial parts contained significantly less GFA than the roots.The content of GFA obtained from different processing methods ranked as follows:shade-drying>sun-drying>kiln-drying.This indicates that different mothods had a significant impact on the GFA content,making shade-drying the best method.Conclusion:The established HPLC method for GFA content determination exhibited strong specificity,precision,repeatability and stability.The best harvest time for Aconitum coreanum was September,and the growth period was recommended to be three years.This study provided experimental basis for the scientific artificial cultivation and harvesting of Aconitum coreanum.