【摘要】 为了探讨和开发凝胶渗透色谱(GPC)--高压液相色谱(HPLC)法定量测定植物组织中的细胞分裂素,以群众杨叶片和烟草组培苗为试验材料,根据细胞分裂素标准品回收率的测定,得出最佳试验方法:80%甲醇研磨后4℃浸提24h,抽滤,滤液离心,上清液加2滴氨水,40℃浓缩至干,用含3%甲醇的CH2Cl2溶解样品,经0.22μm滤膜过滤后过GPC纯化,最后HPLC外标曲线法定量。用液相--质谱联用仪(LC-MS)对所检测的细胞分裂素定性。测试表明:群众杨叶片中玉米素鲜质量含量439.5018ng/g,烟草叶片中玉米素鲜质量含量为617.9957ng/g。在植物样品中检测到的激动素(鲜质量含量49.4739～124.7124ng/g)应为外源引入。研究结果表明,GPC纯化植物细胞分裂素后用HPLC定量是可行的。更多还原

【Abstract】 Using leaves originated from Populus popularis cuttings and Nicotiana tabacum plantlets, the research aimed to develop a GPC-HPLC method to determine the concentration of cytokinin in plant tissues. According to the reclaim efficiency of standard cytokinin, the recommended procedures were as follows: plant samples were grinded with 80% methanol and extracted at 4℃ overnight. Crude extract was filtrated and then centrifugated. The liquid was added with two drops of ammonia, and then vacuum dried at 40℃. The samples were resolved by 3% methanol-CH2Cl2 and filtrated through 0.22 μm filter. The filtrate was purified through GPC (Gel permeation chromatography). The GPC-purified samples were quantified with HPLC by means of external standard curves. The cytokinins detected in plant tissues were confirmed with LC-MS. The results showed that the contents of zeatin in poplar and tobacco leaves were 439.501 8 and 617.995 7 ng/g FW, respectively. Kinetin (49.473 9-124.712 4 ng/g FW) was found in plant tissues, presumably resulting from the uptake from tissue culture medium of tabacco and insects feeding (aphid) in poplar.更多还原