Sapp Melanie, Ertunç Tanya, Bringmann Inken, Schäffer Andreas, Schmidt Burkhard
Department of Biology V, RWTH Aachen University, D-52056 Aachen, Germany.
Pest Manag Sci. 2004 Jan;60(1):65-74. doi: 10.1002/ps.787.
The non-extractable residues of the fungicide cyprodinil formed in heterotrophic cell suspension cultures of wheat were studied by application of [2-pyrimidyl-14C] or [2-pyrimidyl-13C]cyprodinil. The main objective was to examine whether solid-state and liquid 13C NMR spectroscopy can be used to examine plant bound residues of pesticides. For 14C experiments, wheat suspensions grown on glucose as carbon source were treated with 10 mg litre(-1) of 14C-cyprodinil. After incubation for 12 days, 20% of applied 14C was detected as non-extractable residues. The cell debris were treated with 0.1 M HCl (reflux), 1.0 M HCl (reflux), buffer, or 2 M NaOH (50 degrees C); Björkman lignin and acidolysis lignin fractions were also prepared from the debris. Radioactivity liberated and solubilized by these procedures was examined by thin-layer chromatography and high-performance liquid chromatography. The results showed that cyprodinil and primary metabolites contributed to the fungicide's bound residues. Most of the residues (12% of applied 14C) remained associated with polar or polymeric/oligomeric endogenous cell materials in a stable manner. For the study with 13C-cyprodinil, wheat suspensions were cultivated on 13C-depleted glucose for four growth cycles, resulting in maximum 13C depletion of the natural cell components to about 0.10%. During the fourth cycle, 13C-labelled cyprodinil was applied, and cells were incubated (12 days). Cell debris was prepared and examined by solid-state 13C NMR spectroscopy. Debris was then treated as described above in the 14C experiment. Solubilized fractions were analyzed by liquid 13C NMR spectroscopy. However, none of the 13C NMR spectra recorded gave utilizable or unambiguous results, and all exhibited large inconsistencies, especially concerning the data from the conventional 14C experiment.
通过应用[2 - 嘧啶基 - ¹⁴C]或[2 - 嘧啶基 - ¹³C]嘧菌环胺，研究了在小麦异养细胞悬浮培养物中形成的杀菌剂嘧菌环胺的不可提取残留物。主要目的是检验固态和液体¹³C核磁共振光谱是否可用于检测植物中结合的农药残留。对于¹⁴C实验，以葡萄糖为碳源培养的小麦悬浮液用10毫克/升的¹⁴C - 嘧菌环胺处理。培养12天后，检测到20%的施用¹⁴C为不可提取残留物。细胞碎片用0.1 M盐酸（回流）、1.0 M盐酸（回流）、缓冲液或2 M氢氧化钠（50℃）处理；还从碎片中制备了比约克曼木质素和酸解木质素级分。通过薄层色谱和高效液相色谱检查这些程序释放和溶解的放射性。结果表明，嘧菌环胺及其主要代谢产物构成了杀菌剂的结合残留物。大多数残留物（施用¹⁴C的12%）以稳定的方式与极性或聚合/寡聚内源性细胞物质结合。对于¹³C - 嘧菌环胺的研究，小麦悬浮液在¹³C贫化的葡萄糖上培养四个生长周期，导致天然细胞成分的最大¹³C贫化至约0.10%。在第四个周期中，施用¹³C标记的嘧菌环胺，并将细胞培养（12天）。制备细胞碎片并通过固态¹³C核磁共振光谱检查。然后将碎片按上述¹⁴C实验中的方法处理。对溶解级分进行液体¹³C核磁共振光谱分析。然而，记录的所有¹³C核磁共振光谱都没有给出可用或明确的结果，并且都表现出很大的不一致性，特别是关于传统¹⁴C实验的数据。
