Development of an enzyme-linked immunosorbent assay (ELISA) for residue analysis of the fungicide azoxystrobin in agricultural products.
A direct competitive enzyme-linked immunosorbent assay (dc-ELISA) was developed for residue analysis of azoxystrobin in garden crops, for which the maximum residue limits (MRLs) are 0.5-50 mg/kg in Japan. For hapten synthesis, an ethyl carboxyl group was introduced to the 4-position of the 2-cyanophenoxy group in azoxystrobin, and its cyano group was changed to a methyl group. An anti-azoxystrobin monoclonal antibody was prepared from mice immunized with hapten-keyhole limpet hemocyanin conjugate. The dc-ELISA using prepared antibody showed 50-250-fold higher sensitivity compared to the MRLs. The working range of the dc-ELISA was 10-200 ng/mL. The dc-ELISA showed high specificity to azoxystrobin. When methanol extracts from nine kinds of garden crops spiked with azoxystrobin ranging near the MRLs were analyzed, the determined results by the dc-ELISA agreed well with the results of their controls. In addition, azoxystrobin spiked in garden crops homogenates was satisfactorily extracted by methanol solution and easily analyzed. The recovery rate of dc-ELISA was 96-109% and correlated well with the results obtained by HPLC analysis.
Research and Development Division, HORIBA, Ltd., Minami-ku, Kyoto, Japan.
Mika
M
Kondo
Kazuyuki
K
Tsuzuki
Hiroshi
H
Hamada
Yukie
Y
Yamaguchi Murakami
Mikiko
M
Uchigashima
Machiko
M
Saka
Eiki
E
Watanabe
Seiji
S
Iwasa
Hiroshi
H
Narita
Shiro
S
Miyake
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D018556:Crops, Agricultural / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005659:Fungicides, Industrial / Q000032:analysis*; D007564:Japan; D008689:Methacrylates / Q000032:analysis*; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D010573:Pesticide Residues / Q000032:analysis*; D010936:Plant Extracts / Q000737:chemistry; D011743:Pyrimidines / Q000032:analysis*; D000073739:Strobilurins
D000911:Antibodies, Monoclonal; D005659:Fungicides, Industrial; D008689:Methacrylates; D010573:Pesticide Residues; D010936:Plant Extracts; D011743:Pyrimidines; D000073739:Strobilurins; C087670:azoxystrobin