Zhuokun Li

Simultaneous and sensitive determination of multiplex chemical residues based on multicolor quantum dot probes.

Biotinylated denatured bovine serum albumin (Bt-dBSA)-coated cadmium telluride (CdTe) quantum dot (QD) conjugates were prepared and used to develop the multiplexed fluoroimmunoassay for the simultaneous determination of five chemical residues. An immune complex was formed using avidin as the bridge to link the Bt-dBSA-QDs with the antibodies. Primarily, individual quantitative determinations of five representative chemical residues were carried out based on the different emission properties of the QDs. Five antibodies were then conjugated with the corresponding QDs to establish the indirect competition fluorescent-linked immunosorbent assay (ic-FLISA) for the simultaneous detection of five chemicals in one well of a microplate. The linear range for dexamethason (DEX) was from 0.33 microg/kg to 10 microg/kg, 0.28 microg/kg to 10 microg/kg for gentamicin (GM), 0.16 microg/kg to 25 microg/kg for clonazepam (CZP), 0.17 microg/kg to 10 microg/kg for medroxyprogesterone acetate (MPA) and 0.32 microg/kg to 25 microg/kg for ceftiofur (CEF), respectively. The limit of detection (LOD) for the simultaneous determination of DEX, GM, CZP, MPA and CEF were as low as 0.13 microg/kg, 0.16 microg/kg, 0.07 microg/kg, 0.06 microg/kg and 0.14 microg/kg, respectively. This detection method was used to analyze samples of pork muscle and the recoveries ranged from 61.3% to 80.3% for DEX and from 74.0% to 87.2% for MPA. Further more, good correlation between the novel ic-FLISA and traditional ELISA was demonstrated during the determination of DEX and MPA residues in real samples. The QD-based protocol described here is less time consuming than the classical method and it may be sufficiently flexible to be used in other systems for the simultaneous multicolor detection of the drugs.

Rapid and sensitive detection of microcystin by immunosensor based on nuclear magnetic resonance.

A stable and sensitive toxin residues immunosensor based on the relaxation of magnetic nanoparticles was developed. The method was performed in one reaction and offered sensitive, fast detection of target toxin residues in water. The target analyte, microcystin-LR (MC-LR) in Tai lake water, competed with the antigens on the surface of the magnetic nanoparticles and then influenced the formation of aggregates of the magnetic nanoparticles. Accordingly, the magnetic relaxation time of the magnetic nanoparticles was changed under the effect of the target analyte. The calibration curve was deduced at different concentrations of the target analyte. The limit of detection (LOD) of MC-LR was 0.6 ng g(-1) and the detection range was 1-18 ng g(-1). Another important feature of the developed method was the easy operation: only two steps were needed (1) to mix the magnetic nanoparticle solution with the sample solution and (2) read the results through the instrument. Therefore, the developed method may be a useful tool for toxin residues sensing and may find widespread applications.

Development of an Indirect Enzyme-Linked Immunosorbent Assay for the Organophosphorus Pesticide Paraoxon-Methyl

In the present study, the synthesis of hapten for the organophosphorus (OP) pesticide paraoxon-methyl was developed, with a spacer arm (aminocarboxylic acid) attached at the aromatic ring. It was conjugated to bovine serum albumin (BSA) for use as an immunogen and to ovalbumin (OVA) for coating antigen for ELISA testing. Rabbits were immunized with the immunogen and two polyclonal antisera were produced and screened against the coating antigen using competitive indirect enzyme-linked immunosorbent assay (ELISA). For application to textile samples, the influence of several factors such as organic solvent, ionic strength, and pH on the ELISA results were studied. Under optimized conditions, the quantitative working range was 0.012–1.158 μg/mL with a limit of detection (LOD) of 0.005 μg/mL and the IC50 was 0.115 μg/mL.There was negligible cross reactivity (CR) with other OP pesticides. The recoveries obtained by standard paraoxon-methyl addition to the different textile samples such as cotton, wool and muslin delaine were all from 86.0% to 108.0%. Therefore, the optimized ELISA may become a new convenient and economical analytical tool for monitoring paraoxon-methyl residues in textile samples.

Development of an enzyme-linked immunosorbent assay for octylphenol

An indirect enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative analysis of octylphenol. A linear carboxylated analogue of 4-nonylphenol was synthesised and characterised as hapten. The hapten was coupled to the carrier protein to produce a specific antibody. An indirect competitive ELISA method was established based on the polyclonal antibody, which exhibited an IC50 value of 51 ng/mL for octylphenol. A monoclonal antibody was produced with an IC50 value of 76 ng/mL for octylphenol. The immunoassay used with the monoclonal antibodies was applied to analyse water samples from Tai Lake in China.

Development of an ultrasensitive immunoassay for detecting tartrazine.

We have developed an ultrasensitive indirect competitive enzyme-linked immunosorbent assay for the determination of tartrazine. Two carboxylated analogues of tartrazine with different spacer lengths, and one derivative from commercial tartrazine after a little chemical modification, were synthesized as haptens in order to produce antibodies specific to tartrazine. The effect of sulfonic acid groups on the hapten structure of tartrazine was also studied carefully for the first time. A most specific monoclonal antibody against tartrazine was created and exhibited an IC50 value of 0.105 ng/mL and a limit of detection of 0.014 ng/mL, with no cross-reactivity to other structurally-related pigments. The established immunoassay was applied to the determination of tartrazine in fortified samples of orange juice and in real positive samples of carbonated beverages.