水稻生物硝化抑制剂1,9-癸二醇的定量方法优化

Optimized Determination of Biological Nitrification Inhibitor 1,9-decanediol in Rice Root Exudates

选取水稻生物硝化抑制剂1,9-癸二醇作为研究对象,对比旋转蒸发法和固相萃取法对该物质的回收率(80.17%和82.97%)及效率,确定固相萃取法为水稻根系分泌物收集液的前处理方法,且该方法具有高效省时的特点。在气相色谱(GC)分析方法的基础上,对衍生化试剂和衍生化条件进行了优化,结果表明,使用N,O-双(三甲基硅基)三氟乙酰胺(BSTFA)作为1,9-癸二醇的衍生化试剂时,衍生化产物具有较高的响应值;衍生化过程中加入200μl BSTFA,在60℃条件下反应30 min时,1,9-癸二醇经过GC方法得到的分析效果最好;且本方法仪器的日内精密度为2.18%,日间精密度为3.01%,线性方程为y=34.77x–0.90,r=0.9993,最小检出限为0.05μg/ml,此方法可为水稻根系分泌物中生物硝化抑制剂1,9-癸二醇的定量研究提供参考。

The optimized pre-treatment and derivatization methods were developed in this study for the determination of 1,9-decanediol by gas chromatograph in the rice root exudates.Results showed that the C18 SPE column could retain 1,9-decanediol efficiently and its relative recovery rate(82.97%)had no significant difference with that of the rotary evaporation(80.17%).This method is time-saving and fit for concentrating the identified hydrophobic BNI-compound,1,9-decanediol,in large amount of root exudates solutions.In addition,the use of N,O-bis(trimethylsily)trifluoroacetamide(BSTFA,200μl)as derivatization reagent,together with 30 min heating in the oven at 60℃,was found to be the most efficient method for stable derivatives and high sensitivity.The linear equation was y=34.77x-0.90(r=0.9993);the 1,9-decanediol detection limit of the method was 0.05μg/ml;and the method exhibited good accuracy(recoveries of 94.42%)and precision(within-day precision of 2.18%and day-to-day precision of 3.01%)which could provide reference for the subsequent analysis of 1,9-decanediol in rice root exudates.