高效液相色谱法测定水稻穗部的GA_3含量

Determination of gibberellin A_3 in rice panicle by high-performance liquid chromatography

以水稻穗部为材料,探索内源赤霉酸(Gibberellin A3,GA3)的提取和纯化方法,利用高效液相色谱法建立分析测定方法。针对水稻样品富含色素和多酚类等杂质的特点,用氯仿去除色素,然后采用三步萃取法纯化GA3。在以往用高效液相色谱法分析植物内源激素的研究基础上,试验对色谱条件进行了改进。研究发现,最优色谱条件包括紫外检测器在254 nm波长下进行检测,以乙腈—水—乙酸(体积比为20∶79∶1)溶液为流动相,流速为1.0 mL.min-1等条件下峰形良好且保留时间稳定。结果表明,按照本试验流程,样品的添标回收率在80%以上,且相对标准偏差小于3%,较高的回收率和精密度表明本试验流程可以准确地分析测定水稻穗部GA3。

This paper aims to find out a suitable method for extracting,purifying and analyzing endogenous gibberellin A3 of rice panicle by high-performance liquid chromatography method.To deal with the existence of large amount of pigments,polyphenols and other impurities,pigment was eliminated by chloroform and then gibberellin A3 was purified by liquid-liquid extraction of three steps.On the basis of former reports,the high-performance liquid chromatographic conditions for analyzing endogenous hormone of plant were improved.It was found that the optimal chromatographic conditions included that detection wave length was made by the UV detector at 254 nm and the mobile phase was composed of aceto-nitrile-water-acetic acid liquid（volume ratio 20∶79∶1） at the velocity of 1.0 mL·min-1,et al.The peak shape was good and retention time was stable under these chromatographic conditions.The results showed that the add mark recovery rate of gibberellin A3 was above 80% and the relative standard deviation was less than 3% according to this test procedure,which demonstrated that the test procedure could detect endogenous gibberellin A3 of rice panicle excisely.