反相高效液相色谱法测定植物组织培养基中的外源激素

建立了高效液相色谱-光电二极管阵列检测器测定植物组织培养基中6种外源激素的分析方法。采用Waters公司μBondaPak C18柱(3.9 mm×300 mm, 10 μm),用140 mmol/L乙酸钠-三乙胺缓冲液(pH 4.95)和乙腈作流动相,以75:25的比例等度洗脱,柱温37 ℃,流速1.0 mL/min,在波长285 nm下检测。在9 min内6种激素均达到基线分离,其进样量在4~200 ng之间具有良好的相关性(r2>0.9995)。培养基中的激素在减压烘干后用甲醇提取,各种激素的回收率均在85%以上。该方法可以用于植物组织培养基中外源激素的分析检测,或对培养基中未知激素的比例和种类进行分析测定。

Determination of exogenous hormones in plant tissue culture media by reversed-phase high performance liquid chromatography

A method of high performance liquid chromatography coupled with photodiode array detector (PAD) was established for the determination of the content ratio and type of 6 exogenous hormones in plant tissue culture media. The column was a microBondaPak C18 (3.9 mm x 300 mm, 10 microm, Waters), the mobile phase was 140 mmol/L sodium acetate in triethyl amine buffer (pH 4.95)-acetonitrile (75:25, v/v), and the flow rate was 1.0 mL/min. The column temperature was 37 degrees C, and the detection wavelength was 285 nm. The six hormones reached the baseline separation in 9 minutes. The linear relationship was very good in the range of 4-200 ng (r2 > 0.9995). The exogenous hormones in the medium were extracted by methanol after vacuum dried. The average recoveries of the exogenous hormones were more than 85%. The method can be used for the analysis of exogenous hormones of plant tissue culture media, or of unknown hormone ratio and the type of media.