以烟草叶片为材料,研究利用气相色谱-质谱联用仪(GC-MS)测定植物激素茉莉酸(JA)含量的方法。利用甲醇和乙酸乙酯提取JA,提取物进一步用C18小柱纯化,然后用过量重氮甲烷甲酯化,最后用GC-MS对其进行定量分析。结果表明,JA和内标的标准品均可用该方法进行分析。通过对烟草叶片中不同浓度C18小柱洗脱物进行分析表明,40%和60%甲醇洗脱液中含有JA。GC-MS定量分析结果表明,烟草响应机械伤害后,内源JA含量呈现先升高后降低的规律。说明应用GC-MS可以比较准确地测定烟草内源JA的含量,重复性较好。
In this study, an effective method for quantitative analysis of jasmonic acid (JA)using gas chromatography mass spectrometry (GC-MS)was reported. The tobacco leaves were used as materials in this study. Samples were extracted with methanol and ethyl acetate. Then the extracts were further purified with C18 cartridges. The methyl esters of each fraction were performed with diazomethane and then measured by GC-MS. The results showed that the authentic MeJA and internal standard could be measured using GC-MS after extraction and purification were performed. Sequential elution from the Cls cartridges were analyzed with GC-MS and the results showed that JA was eluted in 40% and 60% methanol. In response to mechanical wounding, JA accumulation in tobacco leaves exhibited a rapid and transient increase, and subsequently declined to the levels of unwounded leaves. The results indicated that JA content was measured accurately using this method, and the repeatability was good.