Characterization of the bound residues of the fungicide cyprodinil formed in plant cell suspension cultures of wheat.
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.
Sapp
Melanie
M
Department of Biology V, RWTH Aachen University, D-52056 Aachen, Germany.
Ertunç
Tanya
T
Bringmann
Inken
I
Schäffer
Andreas
A
Schmidt
Burkhard
B
D001665:Binding Sites; D002247:Carbon Isotopes / Q000378:metabolism; D002250:Carbon Radioisotopes / Q000378:metabolism; D002478:Cells, Cultured; D005659:Fungicides, Industrial / Q000378:metabolism*; D008031:Lignin / Q000032:analysis; D009682:Magnetic Resonance Spectroscopy; D010573:Pesticide Residues / Q000378:metabolism*; D011743:Pyrimidines / Q000378:metabolism*; D014908:Triticum / Q000378:metabolism*
D002247:Carbon Isotopes; D002250:Carbon Radioisotopes; D005659:Fungicides, Industrial; D010573:Pesticide Residues; D011743:Pyrimidines; C108338:cyprodinil; D008031:Lignin