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Thin-Layer Chromatography (TLC): Overview01:11

Thin-Layer Chromatography (TLC): Overview

4.4K
Thin-layer chromatography (TLC) is a chromatography technique that separates compounds based on their polarity. TLC typically uses polar silica gel, a form of silicon dioxide, as the stationary phase. The silica gel contains hydroxyl (OH) groups on its surface, which form hydrogen bonds with polar compounds, influencing their adhesion to the stationary phase.
To begin the analysis, a mixture of compounds is spotted on the starting line on the TLC plate using a thin capillary. The bottom of the...
4.4K
High-Performance Liquid Chromatography: Introduction01:11

High-Performance Liquid Chromatography: Introduction

3.3K
High-performance liquid chromatography(HPLC), formerly referred to as High-pressure liquid chromatography, is a powerful technique used to separate, identify, and quantify components in complex mixtures. The term "high pressure" refers to using high pressure to push the liquid mobile phase through the tightly packed columns.
In HPLC, two phases play a critical role in the separation process:
3.3K
Asymmetric Lipid Bilayer01:35

Asymmetric Lipid Bilayer

9.6K
Biological membranes show uneven distribution of different types of lipids in the inner and outer layers, resulting in transverse asymmetric membranes. The treatment of the erythrocyte membrane with the enzyme phospholipase confirmed the asymmetric nature of the lipid bilayer. The enzyme hydrolyzes lipids into fatty acids and hydrophilic groups. The phospholipase acts only on the outer layer of the membrane, while the inner layer remains intact. The phospholipase treatment resulted in 80%...
9.6K
Gas Chromatography: Types of Columns and Stationary Phases01:17

Gas Chromatography: Types of Columns and Stationary Phases

2.2K
Gas chromatography (GC) relies on stationary phases to separate and analyze components in a sample. There are two main types of stationary phases: liquid and solid. Liquid stationary phases are non-volatile, thermally stable, and chemically inert liquids coated onto the column. Solid stationary phases are particles of adsorbent material, such as silica gel or molecular sieves.
For an analyte to remain on the column for a sufficient amount of time, it must exhibit some level of compatibility (or...
2.2K
High-Performance Liquid Chromatography: Instrumentation00:57

High-Performance Liquid Chromatography: Instrumentation

2.9K
High-performance liquid chromatography, or HPLC, is an analytical technique that separates liquid samples under high pressures. An HPLC instrument consists of glass bottles for storing solvents called mobile phase reservoirs. HPLC-grade solvents are used to maintain high purity, and the dissolved gases are removed using a degasser, such as a vacuum pumping system or sparging with helium. The solvents are then pumped into the analytical column using a screw-driven syringe or reciprocating pumps.
2.9K
Chromatography: Introduction01:10

Chromatography: Introduction

6.7K
Chromatography is a technique used to separate compounds based on differences of partitioning between two phases, the stationary phase and the mobile phase.
The phase in which the compounds linger or on which the compounds adsorb is called the stationary phase, whereas the mobile phase is the solvent that carries the solutes to be analyzed. In traditional column chromatography, the mixture flows through the stationary phase, and the compounds partition between the stationary and mobile phases...
6.7K

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Related Experiment Video

Updated: Jan 13, 2026

Defining Substrate Specificities for Lipase and Phospholipase Candidates
08:59

Defining Substrate Specificities for Lipase and Phospholipase Candidates

Published on: November 23, 2016

15.5K

Thin-layer chromatography of phospholipids.

Rania M Deranieh1, Amit S Joshi, Miriam L Greenberg

  • 1Department of Biological Sciences, Wayne State University, Detroit, MI, USA.

Methods in Molecular Biology (Clifton, N.J.)
|September 3, 2013
PubMed
Summary

This study optimizes thin-layer chromatography (TLC) for yeast phospholipid analysis. The new protocol enhances separation and quantification of these vital lipids.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Analytical Chemistry

Background:

  • Lipid analysis is crucial for understanding cellular functions.
  • Thin-layer chromatography (TLC) is a common method for lipid separation.
  • Existing TLC protocols may require optimization for specific applications like yeast phospholipid studies.

Purpose of the Study:

  • To develop and optimize a protocol for the steady-state labeling of yeast phospholipids.
  • To improve the separation and quantification of yeast phospholipids using one-dimensional TLC (1-D TLC).
  • To provide a reliable method for researchers studying yeast lipid metabolism.

Main Methods:

  • Utilized steady-state labeling techniques for yeast cultures.
  • Employed one-dimensional thin-layer chromatography (1-D TLC) for lipid separation.

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  • Developed optimized procedures for phospholipid extraction and quantification.
  • Main Results:

    • Successfully established an optimized protocol for yeast phospholipid analysis via 1-D TLC.
    • Demonstrated efficient separation and accurate quantification of various yeast phospholipids.
    • The protocol ensures reproducible results for lipid profiling.

    Conclusions:

    • The optimized 1-D TLC protocol provides a robust method for yeast phospholipid analysis.
    • This technique facilitates detailed studies of lipid metabolism and cellular signaling in yeast.
    • The protocol is valuable for both basic research and potential biotechnological applications.