高效液相色谱法测定甘草植物体内脱落酸含量的研究

目的：建立甘草植物抗旱信号传导物质脱落酸的含量测定方法,为揭示甘草抗旱性与有效成分积累相关机制奠定理论依据。方法：甘草须根中的脱落酸用80%的甲醇抽提,抽提液经过盐酸酸化、乙酸乙酯萃取最终得到供试品溶液,应用Waters-C18色谱柱（4.6mm×250mm,5μm）,以乙腈-0.1%磷酸水为流动相,进行二元梯度洗脱;流速：0.6mL.min-1,紫外检测波长为260nm,柱温：25℃。结果：脱落酸在0.0336～0.672μg线性关系良好（r=0.9999）,甘草样品平均回收率90.35%,RSD为1.70%,甘草须根中的ABA含量为0.077～0.188mg.g-1。结论：该法灵敏度高、专一性强、重复性好,为进一步开展甘草抗旱性与有效成分积累相关机制的研究提供依据。

Determination of Abscisic acid in Fibrous root of Glycyrrhiza uralensis Fisch. by HPLC

Objective：To establish an HPLC method for determination the content of abscisic acid （ABA） in fibrous root of Glycyrrhiza uralensis Fisch.Methods：The phytohormone in glycyrrhizae fibrous root was extracted with methanol—water（80∶20）,and the extract was acidized by hydrochloric acid,then ABA was partitioned between acetoacetate and water,finally ABA was separated and quantified by the analytical column.The analytical column was waters C18（4.6 mm×250 mm,5 μm）.The mobile phase was acetonirile-0.1% orthophosphoric acid in a gradient elution.The flow rate was 0.6 mL·min-1,the UV detector wavelength was set at 260 nm,and the column temperature was 25 ℃. Results：The calibration curve was linear in the range of 0.0336~0.672μg （r = 0. 9999）.The average recovery rate of ABA in fibrous root of G. uralensis Fisch was 90.35% with RSD 1.70%,the content of ABA in fibrous root of G. uralensis Fisch was 0.077～ 0.188 mg·g-1. Conclusion：This method is high sensitivity,specificity,and reproducibility,and can be used to assay the content of ABA in in fibrous root of G. uralensis Fisch.It provide a basis mechanism for further study of drought resistance associated with the accumulation of active ingredients in G. uralensis Fisch.