A Chemiluminescence Enzyme-Linked Immuno-Sorbent Assay (CLELISA) for determination and quantification of the fungicide thiram in honeybees was developed in an indirect competitive format. The assay was optimized by determining: the optimal coating conjugate concentration and anti-thiram antiserum dilution, the effect of the incubation time on the competitive step, the tolerance to organic solvents. The IC50 and the limit of detection (LOD) values were 60 ng mL21 and 9 ng mL21, respectively, similar to those of colorimetric ELISA with a calibration range of 9–15,000 ng mL21. Cross reactivity of some related compounds such as some dithiocarbamates, a thiocarbamate, the ethylenethiourea and the tetramethylthiourea were tested. The assay was then applied to honeybees sample extracts obtained by using the liquid-liquid extraction or the graphitized carbon-based solid phase extraction. The calibration curves in honeybee extracts from liquid-liquid procedure gave an IC50 of 141 ng mL21 and a LOD of 17 ng mL21. In case of extracts obtained by SPE these values were 139 ng mL21 and 15 ng mL21, respectively. The average recovery value from honeybee extracts spiked with 75 ng mL21 of thiram was 72%
Quantification of Thiram in Honeybees: Development of a Chemiluminescent ELISA / S.Girotti; E.Maiolini; S.Ghini; E.N.Ferri; F.Fini; P.Nodet; S.Eremin.. - In: ANALYTICAL LETTERS. - ISSN 0003-2719. - STAMPA. - 41(1):(2008), pp. 46-55. [10.1080/00032710701748291]
Quantification of Thiram in Honeybees: Development of a Chemiluminescent ELISA
GIROTTI, STEFANO;MAIOLINI, ELISABETTA;GHINI, SEVERINO;FERRI, ELIDA NORA;FINI, FABIANA;
2008
Abstract
A Chemiluminescence Enzyme-Linked Immuno-Sorbent Assay (CLELISA) for determination and quantification of the fungicide thiram in honeybees was developed in an indirect competitive format. The assay was optimized by determining: the optimal coating conjugate concentration and anti-thiram antiserum dilution, the effect of the incubation time on the competitive step, the tolerance to organic solvents. The IC50 and the limit of detection (LOD) values were 60 ng mL21 and 9 ng mL21, respectively, similar to those of colorimetric ELISA with a calibration range of 9–15,000 ng mL21. Cross reactivity of some related compounds such as some dithiocarbamates, a thiocarbamate, the ethylenethiourea and the tetramethylthiourea were tested. The assay was then applied to honeybees sample extracts obtained by using the liquid-liquid extraction or the graphitized carbon-based solid phase extraction. The calibration curves in honeybee extracts from liquid-liquid procedure gave an IC50 of 141 ng mL21 and a LOD of 17 ng mL21. In case of extracts obtained by SPE these values were 139 ng mL21 and 15 ng mL21, respectively. The average recovery value from honeybee extracts spiked with 75 ng mL21 of thiram was 72%I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
