Chem. J. Chinese Universities ›› 2009, Vol. 30 ›› Issue (10): 1945.

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Adsorption and Separation of Cytochrome c on Silk Fibroin

CHEN Ming-Li, YAN Xiao-Rui, WANG Jian-Hua*   

  1. Research Center for Analytical Sciences, Northeastern University, Shenyang 110004, China
  • Received:2009-06-22 Online:2009-10-10 Published:2009-10-10
  • Contact: WANG Jian-Hua. E-mail: jianhuajrz@mail.neu.edu.cn
  • Supported by:

    国家自然科学基金面上项目(批准号: 20805004), 国家自然科学基金重点项目(批准号: 20635010)以及国家杰出青年科学基金(批准号: 20725517)资助.

Abstract:

Selective adsorption of cytochrome c using silk fibroin as adsorbent was observed under certain experimental conditions. A novel separation and preconcentration approach for cytochrome c was thus developed via a silk fibroin packed micro-column in a sequential injection system. The entire process of separation and elution was monitored by spectrophotometry at 410 nm. At pH 5.6, cytochrome c in 2 mL of aqueous solution at 5 g/mL were quantitatively adsorbed by the silk fibroin micro-column, and a NaCl solution of 1.0 mol/L suffices the complete elution of the adsorbed cytochrome c from the micro-column. An enrichment factor of 10 was achieved by fixing a sample volume of 2 mL and an eluent volume of 200 μL. A linear calibration graph within 1.0—10.0 μg/mL cytochrome c was obtained with a detection limit of 0.33 μg/mL and a precision(RSD) of 2.5%(5 μg/mL, n=9). The practical applicability of this system was demonstrated by processing human whole blood for preconcentration of cytochrome c, and the results were verified by assay with SDS-PAGE.

Key words: Silk fibroin; Solid phase extraction; Cytochrome c

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