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A flow-injection ultrafiltration sampling chemiluminescence system for on-line determination of drug–protein interaction

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Abstract

A flow-injection ultrafiltration sampling chemiluminescence system for on-line determination of cimetidine–bovine serum albumin (BSA) interaction is proposed in this paper. Cimetidine can be oxidized by N-bromosuccinimide (NBS) and sensitized by fluorescein to produce high chemiluminescence emission in basic media. The concentration of cimetidine is linear with the CL intensity in the range 3×10−7–1×10−4 mol L−1 with a detection limit of 1×10−7 mol L−1 (3σ). The drug and protein were mixed in different molar ratios in 0.067 mol L−1 phosphate buffer, pH 7.4, and incubated at 37 °C in a water bath. The ultrafiltration probe was utilized to sample the mixed solution at a flow rate of 5 µL min−1. The data obtained by the proposed ultrafiltration flow-injection chemiluminescence method was analyzed with Scrathard analysis and a Klotz plot. The estimated association constant (K) and the number of the binding site (n) on one molecule of BSA by Scrathard analysis and Klotz plot were 3.15×104 L mol−1 and 0.95, 3.25×104 L mol−1 and 0.92, respectively. The proposed system proved that flow-injection chemiluminescence analysis coupled with on-line ultrafiltration sampling is a simple and reliable technique for the study of drug–protein interaction.

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Acknowledgements

This study was support by the National Natural Science Foundation of China (No. 20175039).

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Authors and Affiliations

  1. Department of Chemistry, Institute of Analytical Science, Southwest China Normal University, 400715, Chongqing, P.R. China

    Zhouping Wang, Zhujun Zhang, Zhifeng Fu, Yan Xiong & Xiao Zhang

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  1. Zhouping Wang
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  2. Zhujun Zhang
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  3. Zhifeng Fu
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  4. Yan Xiong
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  5. Xiao Zhang
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Correspondence to Zhujun Zhang.

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Wang, Z., Zhang, Z., Fu, Z. et al. A flow-injection ultrafiltration sampling chemiluminescence system for on-line determination of drug–protein interaction. Anal Bioanal Chem 377, 660–665 (2003). https://doi.org/10.1007/s00216-003-2125-5

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  • DOI: https://doi.org/10.1007/s00216-003-2125-5

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