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Related Experiment Video

Updated: Dec 2, 2025

Organic Solvent-Based Protein Precipitation for Robust Proteome Purification Ahead of Mass Spectrometry
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A deep eutectic solvent modified magnetic β-cyclodextrin particle for solid-phase extraction of trypsin.

Fangting Xu1, Yuzhi Wang1, Ziwei Liu1

  • 1State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082, PR China.

Analytica Chimica Acta
|November 6, 2020
PubMed
Summary
This summary is machine-generated.

A novel magnetic bead-based deep eutectic solvent (DES) modified with cyclodextrin (CD) offers efficient and eco-friendly solid-phase extraction of trypsin. This new material demonstrates high capacity and reusability for trypsin purification from complex samples.

Keywords:
Deep eutectic solventMagnetic solid phase extractionTrypsinβ-cyclodextrin

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Area of Science:

  • Materials Science
  • Analytical Chemistry
  • Biochemistry

Background:

  • Solid-phase extraction (SPE) is crucial for protein purification.
  • Developing efficient and environmentally friendly SPE materials is an ongoing challenge.
  • Trypsin is a key enzyme with significant applications in biotechnology and medicine.

Purpose of the Study:

  • To synthesize and characterize a novel magnetic bead-based deep eutectic solvent (DES) modified with cyclodextrin (CD) for trypsin extraction.
  • To evaluate the extraction efficiency and capacity of the synthesized material for trypsin.
  • To compare the performance of the new material with existing trypsin extractants.

Main Methods:

  • Synthesis of magnetic beads grafted with β-cyclodextrin and modified with a DES (trifluoroacetamide and benzyltrimethylammonium chloride).
  • Solid-phase extraction (SPE) of trypsin using the synthesized MB-NH2@CD@DES.
  • Adsorption isotherm studies (Langmuir and Freundlich) to understand the adsorption mechanism.
  • Comparison of extraction capacity with other proteins and assessment of reusability.
  • Validation using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).

Main Results:

  • The MB-NH2@CD@DES material exhibited a high extraction capacity for trypsin, reaching up to 549.87 mg·g⁻¹.
  • Langmuir adsorption model provided a better fit for the equilibrium data.
  • The material showed superior selectivity for trypsin over other common proteins.
  • The extraction process was efficient, with a high elution rate (74.32%) and good reusability over ten cycles.
  • Low detection and quantitation limits (LOD: 0.072 mg·mL⁻¹, LOQ: 0.240 mg·mL⁻¹) were achieved.

Conclusions:

  • The fabricated MB-NH2@CD@DES is a highly efficient, selective, and reusable material for trypsin solid-phase extraction.
  • This novel material offers a safer and more environmentally friendly alternative to conventional extractants.
  • The developed method shows great potential for purifying trypsin from complex biological samples.