Skip to Content
MilliporeSigma
  • Simultaneous determination of malachite green, brilliant green and crystal violet in grass carp tissues by a broad-specificity indirect competitive enzyme-linked immunosorbent assay.

Simultaneous determination of malachite green, brilliant green and crystal violet in grass carp tissues by a broad-specificity indirect competitive enzyme-linked immunosorbent assay.

Analytica chimica acta (2011-10-27)
Yu-Dong Shen, Xing-Fei Deng, Zhen-Lin Xu, Yu Wang, Hong-Tao Lei, Hong Wang, Jin-Yi Yang, Zhi-Li Xiao, Yuan-Ming Sun
ABSTRACT

An immunizing hapten (4-(carboxymethoxy)phenyl)bis(4-(diethylamino)phenyl)methylium for brilliant green (BG), a triphenylmethane dye with a potential illegal use in fish feeding, was synthesized and used to produce polyclonal antibody (PcAb) against BG. Unexpectedly, the obtained PcAb showed high cross-reactivity (CR) to malachite green (MG) and crystal violet (CV) in an indirect competitive enzyme-linked immunosorbent assay (icELISA). After screening against three heterologous coating antigens, the icELISA exhibited good sensitivity and uniform response to BG (IC(50) of 1.98 ng mL(-1) and CR of 100%), MG (IC(50) of 1.61 ng mL(-1) and CR of 105%) and CV (IC(50) of 1.34 ng mL(-1) and CR of 142%) when using (4-(carboxymethoxy)phenyl)bis(4-(dimethylamino)phenyl)methylium as the coating hapten. Therefore, a broad-specificity icELISA for simultaneous determination of BG, MG and CV was developed. The recoveries of single analyte and mixture of three analytes from spiked grass carp tissues were estimated ranging from 74.94% to 110.39%. A statistically significant correlation of results was obtained between the developed icELISA and previously established HPLC approaches with the food-relevant three triphenylmethane dyes concentration range 1.83-200 ng mL(-1) (R(2)=0.9224), indicating good accuracy of the icELISA and suitability for the broad-specific detection of the three triphenylmethane dyes in grass carp tissues.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Brilliant Green, certified by the Biological Stain Commission
Sigma-Aldrich
Brilliant Green, Dye content ≥85 %
Supelco
Brilliant Green, analytical standard