Capillary Electrophoresis (CE)

Overview

Source: Laboratory of Dr. B. Jill Venton - University of Virginia

Capillary electrophoresis (CE) is a separation technique that separates molecules in an electric field according to size and charge. CE is performed in a small glass tube called a capillary that is filled with an electrolyte solution. Analytes are separated due to differences in electrophoretic mobility, which varies with charge, solvent viscosity, and size. Traditional electrophoresis in gels is limited in the amount of voltage that can be applied because Joule heating effects will ruin the gel and the separation. Capillaries have a large surface area-to-volume ratio and thus dissipate heat better. Therefore, the voltages applied for a capillary electrophoresis experiment are quite large, often 10,000–20,000 V.

Capillary electrophoresis is useful for high-performance separations. Compared to liquid chromatography, CE separations are often faster and more efficient. However, capillary electrophoresis works best to separate charged molecules, which is not a limitation of liquid chromatography. CE has a greater peak capacity than high-performance liquid chromatography (HPLC), meaning the separations are more efficient and more peaks can be detected. The instrumentation can be very simple. However, HPLC is more versatile and many stationary and mobile phases have been developed for different types of molecules.

Procedure

1. CE Instrumentation Setup

  1. Turn on the CE instrument and computer. Using the computer software, turn on the light source for UV analysis to allow it to warm up. Some software has an indicator when the lamp is ready for use (lamp icon turns color).
  2. Make a methods file. Set the important parameters for running the CE. In this analysis the temperatures of the cartridge and sample storage are 35 °C. The wavelength for UV detection is 214 nm.
  3. Write a time program. The program general

Log in or to access full content. Learn more about your institution’s access to JoVE content here

Results

Electropherograms collected for diet Pepsi and Pepsi samples are shown in Figures 1 and 2, respectively. The three peaks for caffeine, aspartame, and benzoic acid are observed in diet Pepsi and have similar migration times as the standards. For regular Pepsi, the caffeine peak is present but not the aspartame and benzoic acid peaks. The CE analysis is fast as the migration times are only 3–4 min.

The calibration curve for caffeine is shown in Fig

Log in or to access full content. Learn more about your institution’s access to JoVE content here

Application and Summary

Capillary electrophoresis is used for many specialty separations. For example, it is used in the pharmaceutical industry for quality testing, to make sure there are no side products or interferents. CE is particularly useful for separating drugs with a basic amino group, as the walls of the capillary can be made neutral with an acidic pH and thus the drug will not stick to the capillary.

A mode of CE was also used to sequence the human genome and separate DNA. This mode of CE is capillary gel

Log in or to access full content. Learn more about your institution’s access to JoVE content here

Tags
Capillary ElectrophoresisCEChemical AnalysisSeparate MoleculesElectric FieldSizeChargeSub millimeter Diameter TubeCapillaryFlowing Electrolyte SolutionVelocityChargeSizeSolvent s ViscosityResolutionHigh performance Liquid ChromatographyEfficiencySensitivity

건너뛰기...

0:00

Overview

1:06

Principles of Capillary Electrophoresis

3:19

CE Instrumentation Setup

4:26

Preparation of the Standards and Soda Samples

5:23

Running the Samples

6:50

Applications

8:26

Summary

이 컬렉션의 비디오:

article

Now Playing

Capillary Electrophoresis (CE)

Analytical Chemistry

93.0K Views

article

분석 특성화를 위한 샘플 준비

Analytical Chemistry

83.7K Views

article

내부 표준

Analytical Chemistry

203.5K Views

article

표준 추가 방법

Analytical Chemistry

318.9K Views

article

교정 곡선

Analytical Chemistry

789.1K Views

article

자외선 눈에 보이는 (UV-Vis) 분광법

Analytical Chemistry

616.6K Views

article

화학 분석을 위한 라만 분광법

Analytical Chemistry

50.8K Views

article

엑스레이 형광 (XRF)

Analytical Chemistry

25.3K Views

article

화염 이온화 감지를 갖춘 가스 크로마토그래피(GC)

Analytical Chemistry

279.9K Views

article

고성능 액체 크로마토그래피 (HPLC)

Analytical Chemistry

381.9K Views

article

이온 교환 크로마토그래피

Analytical Chemistry

263.0K Views

article

질량 분광법 소개

Analytical Chemistry

111.4K Views

article

스캐닝 전자 현미경 검사법 (SEM)

Analytical Chemistry

86.6K Views

article

전위요스타트/갈바노스타트를 사용하여 지원되는 촉매의 전기화학적 측정

Analytical Chemistry

51.2K Views

article

순환 볼탐량 (CV)

Analytical Chemistry

123.4K Views

JoVE Logo

개인 정보 보호

이용 약관

정책

연구

교육

JoVE 소개

Copyright © 2025 MyJoVE Corporation. 판권 소유