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Improvement of reproducibility and sensitivity of CE analysis by using the capillary coated dynamically with carboxymethyl chitosan

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Abstract

Analysis reproducibility and detection sensitivity of capillary electrophoresis (CE) are often questioned by applied scientists, which has hindered its application as a routine method. To address these issues, a simple, precise, and reproducible dynamic coating method was developed by applying carboxymethyl chitosan (CMC) dynamic coating on fused silica capillary. The proposed coating was accomplished by simply rinsing the capillary with CMC solution for 1 min in between runs, with no regeneration procedure or buffer additives needed. Electroosmotic flow could be well controlled by adjusting the pH of background electrolyte, and the adsorption of analytes onto the capillary inner wall was effectively eliminated. The main parameters of the coating condition were optimized, and extensive applications of these CMC-dynamically coated capillaries in CE separations were then firmly confirmed. By using proteins, aristolochic acids, and inorganic anions as model analytes, the coating showed a good stability, high reproducibility, as well as improved sensitivity. Baseline separations could be obtained with high efficiency. The reduced adsorption was impressively effective for basic proteins, with an average plate number of 90,000/m for each protein, apart from the good resolution on the chromatogram. A high sensitive detection of α-lactalbumin was achieved with a limit of detection (S/N = 3) of 3.5 nM, and the number of theoretical plates was as high as 1,200,000/m. In addition, the combination of the CMC coating with nonaqueous CE and CE-mass spectrometry proved to be practical. All results showed that the CMC-dynamically coated capillary has special properties and obvious superiority over the uncoated ones for CE analysis.

 

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Acknowledgements

This project was financially supported by NSFC (grant No. 90717002, 20775090, and 20975005) and the Fundamental Research Funds for the Central Universities. Agilent Technologies is thanked for providing the Agilent 3DCE system.

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Authors and Affiliations

  1. Beijing National Laboratory for Molecular Sciences, Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, College of Chemistry and Molecular Engineering, Peking University, Beijing, 100871, China

    Yi Liu, Xiaofang Fu, Yu Bai, Yiping Liao & Huwei Liu

  2. Institute of Applied Chemistry, College of Chemistry and Molecular Engineering, Peking University, Beijing, 100871, China

    Maolin Zhai

  3. Medical Experiment & Analysis Center, General Hospital of Chinese PLA, Beijing, 100853, China

    Jie Liao

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  1. Yi Liu
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  2. Xiaofang Fu
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  3. Yu Bai
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  5. Yiping Liao
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  6. Jie Liao
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  7. Huwei Liu
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Correspondence to Huwei Liu.

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Liu, Y., Fu, X., Bai, Y. et al. Improvement of reproducibility and sensitivity of CE analysis by using the capillary coated dynamically with carboxymethyl chitosan. Anal Bioanal Chem 399, 2821–2829 (2011). https://doi.org/10.1007/s00216-011-4659-2

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  • DOI: https://doi.org/10.1007/s00216-011-4659-2

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