固相萃取-HPLC测定外生菌根真菌产生植物激素IAA和GA_3

Determination of Plant Hormone IAA and GA_3 by Ectomycorrhizal Fungi in Pure Culture by Solid Phase Extraction and HPLC

离体发酵培养条件下,外生菌根真菌产生的植物激素释放到培养液中,由于菌液成分复杂,高效液相直接测定培养液中的植物激素有一定困难,应用C18萃取小柱提取纯化培养液中菌根真菌产生的2种植物激素,选择出合适的液相色谱测定条件。结果表明,经过纯化富集,采用Waters SymmetryShieldTMRP18(4.6 mm×250 mm,5μm)色谱柱,以体积分数为35%的乙腈/水溶液(磷酸调pH至3.0)为流动相,流速1.0 mL/min,柱温35℃,波长210 nm下检测,植物激素IAA和GA3分离效果理想,回收率分别大于96.8%和95.4%,最低检出限(S/N=3)分别为0.053和0.072 mg/L。

Plant hormone was produced by ectomycorrhizal fungi under in vitro culture condition.It is difficult to determine plant hormone directly by high performance liquid chromatography（HPLC） because of complicated components of liquid culture.In this paper,C18 solid phase column was used to extract and purify two plant hormone produced by ectomycorrhizal fungi and an analytical method for reversed-phase high performance liquid chromatography（RP-HPLC） was developed.The results indicated that,good isolation of indole-acetic acid（IAA） and gibberellin acid（GA3） was obtained on a Waters SymmetryShieldTM RP18（4.6 mm×250 mm,5 μm） column.35% acetonitrile（pH 3.0） was used as mobile phases at a flow rate of 1.0 mL/min.The column temperature was 35 ℃ and UV wavelength was 210 nm.The recoveries of IAA and GA3 were more than 96.8% and 95.4% respectively.The detection limits of two plant hormones were 0.053 mg/L and 0.072 mg/L based on the signal-to-noise ratio of 3.