|
|||||
|
|
Modified use of a commercial ELISA kit for deoxynivalenol determination in rice and corn silage |
|
| Authors: | Hisaaki Hiraoka Katsumi Yamamoto Yukiko Mori Naoki Asao Rie Fukunaka Kenzaburo Deguchi Kenzi Iida Shigeru Miyazaki Tetsuhisa Goto |
| Institution: | 1. Kobe Regional Center, Food and Agricultural Materials Inspection Center, 1-3-7 Minatojima-minami-machi, Chuo-ku, Kobe, 650-0047, Japan 2. Headquarters, Food and Agricultural Materials Inspection Center, 2-1 Shintoshin, Chuo-ku, Saitama, 330-9731, Japan 3. Sapporo Regional Center, Food and Agricultural Materials Inspection Center, Kita 10-jo Nishi 4-1-13, Kita-ku, Sapporo, 001-0010, Japan 4. Hokkaido Animal Research Center, 5-39 Shintokunishi, Shintoku-cho, Kamikawa, Hokkaido, 081-0038, Japan 5. National Institute of Animal Health Hokkaido Research Station, National Agriculture and Food Research Organization, 4 Hitsujigaoka, Toyohira, Sapporo, 062-0045, Japan 6. Faculty of Agriculture, Shinshu University, 8304 Minami-minowa-Mura, Kamiina, Nagano, 399-4598, Japan |
| Abstract: | The analysis of deoxynivalenol (DON) in silage samples using enzyme-linked immunosorbent assay (ELISA) often leads to an overestimation. To better analyze DON in rice and corn silages using a commercially available ELISA kit, a cleanup method using a MultiSep #226 column was developed. As a result, overestimation of DON by the influence of specific cross-reaction with acetyldeoxynivalenol (AcDON) was confirmed. In samples where AcDON was not detected by liquid chromatography with mass spectrometry (LC-MS), no samples showed a significant difference (P?<?0.05) in DON amounts between ELISA with cleanup and LC-MS analysis. For the recovery study, blank silage was spiked with 0.5 or 1.0 mg/kg DON. The mean recoveries of DON determined by ELISA with cleanup and LC-MS analysis were 112 and 96 %, respectively, and the relative standard deviation for the repeatability (RSDr) were 8.2 and 9.8 %, respectively. No samples showed a significant difference (P?<?0.05) in DON concentration determined by either ELISA or LC-MS analysis. A collaborative study to validate this rapid method was carried out using four samples, two rice and two corn silage, by 10 participating laboratories. Each sample was analyzed using blind duplicates. The mean values of DON detected were 1.5–2.3 mg/kg, RSDr and the relative standard deviation for the reproducibility (RSDR) were 4.1–12.7 and 7.6–23.4 %, respectively, and the HorRat values were 0.5–1.6. Therefore, the overestimation of DON by the influence of nonspecific cross-reaction with sample matrix was reduced by the cleanup method using a MultiSep #226 column, and analysis of DON in silage was improved. This use of this method for estimation of DON contamination in silage allows rapid detection at the place of use that is likely to result in improved animal health. |
| Keywords: | |
| 本文献已被 SpringerLink 等数据库收录！ | |