GPC-HPLC-MS/MS测定植物组织中IAA与ABA的方法

Abstract

植物激素检测的有效性和高效性是开展植物研究的前提。本试验采用净化-定量-浓缩多联机系统(GPC)纯化样品、利用HPLC-MS/MS检测以及内标法定量测定植物组织内源IAA和ABA含量。通过正交试验设计优化适于激素提取和纯化的条件，结果表明：取样量0.5 g、80%甲醇提取液、浓缩温度35℃和C18 SPE柱为最优组合。IAA和ABA的含量分别为20.34 ng/g和789.93 ng/g；最低检出限分别为2.36 ng/g和31.95 ng/g；回收率为70.43%和80.17%；变异系数分别为1.87%和2.26%。 The effectiveness and efficiency of the plant hormone detection is a prerequisite for plant research. The sample was purified with the GPC system, and the method employed HPLC-MS/MS for multiple reaction monitoring of concentrations of IAA, ABA, and deuterated IAA and ABA analogs. The condition of extraction and purification of hormones were optimized by orthogonal design. The results show that the composite of 0.5 g samples, 80% methanol extraction solvent, concentration temperature 35˚C and C18 cartridge column for solid-phase extraction (SPE), were optimal extraction procedure for each type of plant tissue. The contents of IAA and ABA as high as 20.34 ng/g and 789.3 ng/g were achieved by this method, respectively; the detectable limits were 2.36 ng/g and 31.95 ng/g; the recoveries were 70.43% and 80.17%; the RSD were 1.87% and 2.26%.