ELISA on a microchip with a photodiode for detection of amphetamine in plasma and urine.
A rapid and sensitive assay was developed for the detection of amphetamine in plasma and urine. The method relies on the principle of competitive ELISA (enzyme-linked immunosorbent assay). A flow microchip with a total volume of 7 microL was used for the development of a chemiluminescent ELISA technique. Solutions, samples, and the chemiluminescence substrate were injected by a flow system, and a photodiode detector was used to measure the light intensity. The incubation time of the competitor (competition phase) was reduced to 10 min. Calibration curves corresponding to analyte concentrations ranging from 40 to 1,000 microg/L in urine samples and from 6 to 96 microg/L in plasma samples were obtained. The detection limits were in the region of 20 and 6 microg/L in urine and plasma, respectively. The main focus of the work was on speed, reliability, reproducibility, and operational stability of the assay. This method was proven readily adaptable to automation and provided reproducible results.
Mobini Far
Hamid Reza
HR
Department of Pure and Applied Biochemistry, Center for Chemistry and Chemical Engineering, Lund University, P.O. Box 124, Sweden. hamid.mobini-far@surgsci.uu.se
Torabi
Fereidon
F
Danielsson
Bengt
B
Khayyami
Masoud
M
D000661:Amphetamine / Q000032:analysis* / Q000097:blood / Q000652:urine; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D008163:Luminescent Measurements; D046208:Microchip Analytical Procedures / Q000379:methods*; D015203:Reproducibility of Results
D000661:Amphetamine