PMID- 12530822
OWN - NLM
STAT- MEDLINE
DCOM- 20030429
LR  - 20190717
IS  - 0003-2700 (Print)
IS  - 0003-2700 (Linking)
VI  - 75
IP  - 1
DP  - 2003 Jan 1
TI  - Competitive quenching fluorescence immunoassay for chlorophenols based on 
      laser-induced fluorescence detection in microdroplets.
PG  - 83-90
AB  - An improved biomonitoring system for the analysis of 2,4,6-trichlorophenol (TCP) 
      in urine samples has been developed. The principle of the biosensor device is the 
      detection of laser-induced fluorescence (LIF) in single microdroplets by a 
      homogeneous quenching fluorescence immunoassay (QFIA). The competitive 
      immunoassay occurs in microdroplets (d = 58,4 microm) produced by a piezoelectric 
      generator system with 10-microm-diameter orifice. A continuous Ar ion laser (488 
      nm) excites the fluorescent tracer; its fluorescence is detected by a 
      spectrometer attached to a 512 x 512 cooled, charge-coupled device camera. 
      Fluorescence is quenched by specific binding of TCP polyclonal antibodies to the 
      fluorescent tracer (hapten A-fluorescein); the quenching effect is diminished by 
      the presence of the analyte. Thus, an increase in the signal is produced in a 
      positive dose-dependent manner when TCP is present in the sample. In 10 mM PBS 
      buffer, the IC50 of the LIF-microdroplet QFIA is 0.45 microg L(-1) reaching a LOD 
      of 0.04 microg L(-1). The QFIA with the same reagents performed in microtiter 
      plate format achieved a LOD of 0.36 microg L(-1) in buffer solution. Performance 
      in human urine was similar to that observed in the buffer. A LOD of 1.6 ,g L(-1), 
      with a dynamic range between 4 and 149.5 microg L(-1) in urine, was obtained 
      without any sample treatment other than dilution with the assay buffer. The 
      detectability achieved is sufficient for occupational exposure risk assessment.
FAU - Nichkova, Mikaela
AU  - Nichkova M
AD  - Department of Biological Organic Chemistry. IIQAB-CSIC, Jorge Girona, 18-26, 
      08034-Barcelona, Spain.
FAU - Feng, Jun
AU  - Feng J
FAU - Sanchez-Baeza, Francisco
AU  - Sanchez-Baeza F
FAU - Marco, M Pilar
AU  - Marco MP
FAU - Hammock, Bruce D
AU  - Hammock BD
FAU - Kennedy, Ian M
AU  - Kennedy IM
LA  - eng
GR  - 5P42ES04699/ES/NIEHS NIH HHS/United States
GR  - P30 ES05707/ES/NIEHS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Anal Chem
JT  - Analytical chemistry
JID - 0370536
RN  - 0 (Chlorophenols)
RN  - MHS8C5BAUZ (2,4,6-trichlorophenol)
SB  - IM
MH  - Binding, Competitive
MH  - Calibration
MH  - Chlorophenols/*urine
MH  - Fluorescence
MH  - Fluoroimmunoassay/*methods
MH  - Humans
MH  - Kinetics
MH  - *Lasers
MH  - Molecular Structure
MH  - Occupational Exposure
MH  - Sensitivity and Specificity
EDAT- 2003/01/18 04:00
MHDA- 2003/04/30 05:00
CRDT- 2003/01/18 04:00
PHST- 2003/01/18 04:00 [pubmed]
PHST- 2003/04/30 05:00 [medline]
PHST- 2003/01/18 04:00 [entrez]
AID - 10.1021/ac025933n [doi]
PST - ppublish
SO  - Anal Chem. 2003 Jan 1;75(1):83-90. doi: 10.1021/ac025933n.