[Use of ultrasensitive time-resolved fluoroimmunoassay for rapid detection of clenbuterol hydrochloride in urine].
OBJECTIVE
In order to provide a rapid and selectivity method for the determination of clenbuterol(CBL), an indirect competitive time-resolved fluoroimmunoassay (TRFIA) was developed.


METHODS
Anti-CBL antibody, was raised by immunization against CBL-BSA in rabbits. CBL-OVA was coated by physical adsorption onto the microtitre plate, CBL or sample with CBL as a competitor. Both them were incubated with limited anti- CBL antibody. and a goat antirabbit IgG-Eu3+ conjugate was used as a tracer.


RESULTS
The sensitivity of CBL-TRFIA was 0.01microg/L, and the recovery rate was 99.7%. RSD of CBL-TRFIA was 3.9% . The sensitivity of CBL-TRFIA provided a linear response from 0.01 - 25microg/L, with ED50 of (1.47+/-0.11) microg/L or ED80 of (0.07+/-0.01)microg/L and ED, of (23.6+/- 0.56) microg/L. The cross reactivity of the CBL-TRFIA with salbutamol, epinephrine hydrochloride and epinephrine bitartrate was negligible, while that with isoprenaline hydrochloride was 0.01% . Both CBL-TRFIA and CBL-ELISA test were applied for the quantitative measurement of CBL in the same urine, and the coefficient of correlation was 0.932.


CONCLUSIONS
The CBL-TRFIA could be applied to detect the CBL in urine and it is useful to screening easily for CBL contamination in meat or foods.
Shi
Jin
J
School of Biotechnology, Southern Yangtze University, Wuxi 214036, China.
Huang
Biao
B
Zhang
Lian-fen
LF
Mi
Xiao-li
XL
Xu
Gan-rong
GR
Jin
Jian
J
D000818:Animals; D002976:Clenbuterol / Q000652:urine*; D015200:Fluoroimmunoassay / Q000379:methods*; D008297:Male; D011817:Rabbits; D012680:Sensitivity and Specificity
D002976:Clenbuterol