Abstract
Pyrethroids are widely used in agriculture as insecticides. In this study, we describe a simple one-step homogeneous fluoroimmunoassay for the glycine conjugate of phenoxybenzoic acid (PBAG), a putative pyrethroid metabolite that may be used as a biomarker of exposure to pyrethroids. Quenching fluoroimmunoassay (QFIA) is based on the competition of labeled and non-labeled pesticide for binding with antibodies and the resulting calibration curve is based on the relationship between analyte concentration and fluorescence quenching of labeled pesticide by specific antibodies. We developed a QFIA for PBAG in aqueous solution using fluorescein-labeled PBAG and polyclonal antibodies. The estimated IC50 (analyte concentration giving 50% inhibition of quenching) for PBAG was 4.5 nM. The detection limit (DL) was 0.9 nM. The dynamic range of the calibration curve was 2-50 nM. The average analytical recovery obtained by applying the method to urine samples (400- or 1000-fold urine dilution) was 85-111%. This demonstrates the QFIA to be a very simple and rapid detection method for PBAG; no washing steps and no enzyme conjugates were required.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 103-117 |
| Number of pages | 15 |
| Journal | Analytica Chimica Acta |
| Volume | 444 |
| Issue number | 1 |
| DOIs | |
| State | Published - Oct 12 2001 |
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Keywords
- Fluorescence quenching
- Homogeneous fluoroimmunoassay
- Polyclonal antibodies
- Pyrethroid metabolite
- Urine
ASJC Scopus subject areas
- Biochemistry
- Analytical Chemistry
- Spectroscopy
- Environmental Chemistry
Cite this
Homogeneous fluoroimmunoassay of a pyrethroid metabolite in urine. / Matveeva, Evgenia G.; Shan, Guomin; Kennedy, Ian M.; Gee, Shirley J.; Stoutamire, Donald W.; Hammock, Bruce D.
In: Analytica Chimica Acta, Vol. 444, No. 1, 12.10.2001, p. 103-117.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Homogeneous fluoroimmunoassay of a pyrethroid metabolite in urine
AU - Matveeva, Evgenia G.
AU - Shan, Guomin
AU - Kennedy, Ian M.
AU - Gee, Shirley J.
AU - Stoutamire, Donald W.
AU - Hammock, Bruce D.
PY - 2001/10/12
Y1 - 2001/10/12
N2 - Pyrethroids are widely used in agriculture as insecticides. In this study, we describe a simple one-step homogeneous fluoroimmunoassay for the glycine conjugate of phenoxybenzoic acid (PBAG), a putative pyrethroid metabolite that may be used as a biomarker of exposure to pyrethroids. Quenching fluoroimmunoassay (QFIA) is based on the competition of labeled and non-labeled pesticide for binding with antibodies and the resulting calibration curve is based on the relationship between analyte concentration and fluorescence quenching of labeled pesticide by specific antibodies. We developed a QFIA for PBAG in aqueous solution using fluorescein-labeled PBAG and polyclonal antibodies. The estimated IC50 (analyte concentration giving 50% inhibition of quenching) for PBAG was 4.5 nM. The detection limit (DL) was 0.9 nM. The dynamic range of the calibration curve was 2-50 nM. The average analytical recovery obtained by applying the method to urine samples (400- or 1000-fold urine dilution) was 85-111%. This demonstrates the QFIA to be a very simple and rapid detection method for PBAG; no washing steps and no enzyme conjugates were required.
AB - Pyrethroids are widely used in agriculture as insecticides. In this study, we describe a simple one-step homogeneous fluoroimmunoassay for the glycine conjugate of phenoxybenzoic acid (PBAG), a putative pyrethroid metabolite that may be used as a biomarker of exposure to pyrethroids. Quenching fluoroimmunoassay (QFIA) is based on the competition of labeled and non-labeled pesticide for binding with antibodies and the resulting calibration curve is based on the relationship between analyte concentration and fluorescence quenching of labeled pesticide by specific antibodies. We developed a QFIA for PBAG in aqueous solution using fluorescein-labeled PBAG and polyclonal antibodies. The estimated IC50 (analyte concentration giving 50% inhibition of quenching) for PBAG was 4.5 nM. The detection limit (DL) was 0.9 nM. The dynamic range of the calibration curve was 2-50 nM. The average analytical recovery obtained by applying the method to urine samples (400- or 1000-fold urine dilution) was 85-111%. This demonstrates the QFIA to be a very simple and rapid detection method for PBAG; no washing steps and no enzyme conjugates were required.
KW - Fluorescence quenching
KW - Homogeneous fluoroimmunoassay
KW - Polyclonal antibodies
KW - Pyrethroid metabolite
KW - Urine
UR - http://www.scopus.com/inward/record.url?scp=0035850971&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0035850971&partnerID=8YFLogxK
U2 - 10.1016/S0003-2670(01)01161-8
DO - 10.1016/S0003-2670(01)01161-8
M3 - Article
AN - SCOPUS:0035850971
VL - 444
SP - 103
EP - 117
JO - Analytica Chimica Acta
JF - Analytica Chimica Acta
SN - 0003-2670
IS - 1
ER -