固相萃取-高效液相色谱法测定植物根际促生菌发酵产物中3种植物激素的含量

发布时间：2018-08-19 11:25

【摘要】：建立了固相萃取-高效液相色谱法同时测定植物根际促生菌发酵产物中吲哚-3-乙酸、赤霉素和玉米素3种植物激素含量的方法。固相萃取采用Agilent Bond Elut C18固相萃取柱,6 mL甲醇+6 mL 10%甲醇活化;以5 mL 10%甲醇淋洗2次;以5mL 80%甲醇洗脱2次。采用Agilent ZORBAX Eclipse Plus C18色谱柱(250×4.6mm,5μm)分离,流动相采用体积比为2∶3的甲醇-水(含0.2%冰乙酸)二元混合溶剂,流速为1mL·min~(-1),柱温为30℃,进样量为20μL,检测波长为210nm。结果表明:10株供试菌株发酵产物中均能检测到3种植物激素,除菌株003NXZ4外,其余菌株均呈现出玉米素含量最高,赤霉素次之,吲哚-3-乙酸含量最低的趋势。3种植物激素在12min内分离完全,样品的加标回收率达90.3%~104.3%,相对标准偏差(n=6)在1.04%~2.98%之间。该方法简便、准确,可同时检测植物根际促生菌发酵产物中吲哚-3-乙酸、赤霉素和玉米素3种植物激素含量。
[Abstract]:A method for simultaneous determination of indole-3-acetic acid, gibberellin and zeatin in the fermentation products of rhizosphere growth-promoting bacteria by solid-phase extraction (SPE) and high performance liquid chromatography (HPLC) was established. The solid phase extraction was performed on Agilent Bond Elut C18 solid phase extraction column with 6 mL methanol 6 mL 10% methanol, 2 times elution with 5 mL 10% methanol and 2 times elution with 5mL 80% methanol. A Agilent ZORBAX Eclipse Plus C18 column (250 脳 4.6 mm ~ 5 渭 m) was used. The mobile phase consisted of a mixture of methanol and water (containing 0.2% glacial acetic acid) at 2:3, the flow rate was 1mL min ~ (-1), the column temperature was 30 鈩，

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