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Related Concept Videos

High-Performance Liquid Chromatography: Instrumentation00:57

High-Performance Liquid Chromatography: Instrumentation

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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.
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High-Performance Liquid Chromatography: Introduction01:11

High-Performance Liquid Chromatography: Introduction

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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:
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High-Performance Liquid Chromatography: Types of Detectors01:15

High-Performance Liquid Chromatography: Types of Detectors

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The role of the detectors in High-Performance Liquid Chromatography (HPLC) is to analyze the solutes as they exit from the chromatographic column. The detector recognizes the solute's property and generates corresponding electrical signals, which are converted into a readable graph of the detector's response versus elution time called a chromatogram at the computer. There are several types of HPLC detectors, each with its own advantages and limitations, depending on the analyte...
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High-Performance Liquid Chromatography: Elution Process01:05

High-Performance Liquid Chromatography: Elution Process

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In High-Performance Liquid Chromatography (HPLC), the elution process is critical to the separation of analytes and the quality of chromatographic results. Elution describes how compounds move through the column and separate based on their interactions with the mobile and stationary phases. This process determines the resolution, peak shape, and retention times in the chromatogram, which are essential for identifying and quantifying components in complex mixtures. Understanding the elution...
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Types Of Column Chromatography01:29

Types Of Column Chromatography

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The stability and compatibility of column material with samples are crucial for efficient purification in chromatographic techniques. Various operating parameters such as pH, temperature, or solvent affect the packing of the column material, thereby determining the purification efficiency. The choice of column material also plays an essential role in deciding the operating parameters and can be modified based on the proteins that need to be purified.
Gel Filtration Chromatography
When the...
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Electrophoresis: Overview01:20

Electrophoresis: Overview

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Electrophoresis is a powerful analytical separation technique that relies on the differential migration of charged species when subjected to an electric field. The core strength of electrophoresis lies in its ability to separate high-molecular-weight species in complex mixtures. It has found widespread use in biochemistry, molecular biology, and analytical chemistry, allowing the separation of compounds like amino acids, nucleotides, carbohydrates, and proteins with excellent resolution.
There...
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A Rapid and Chemical-free Hemoglobin Assay with Photothermal Angular Light Scattering
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High-performance liquid chromatography as a method to identify haemoglobin abnormalities.

T H Huisman1

  • 1Department of Cell and Molecular Biology, Medical College of Georgia, Augusta.

Acta Haematologica
|January 1, 1987
PubMed
Summary
This summary is machine-generated.

Two high-performance liquid chromatography (HPLC) methods effectively separate normal and abnormal hemoglobins and globin chains. These techniques offer significant diagnostic value for various hemoglobinopathies in adults and newborns.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Hematology

Background:

  • Hemoglobinopathies are a group of genetic blood disorders.
  • Accurate diagnosis and characterization of hemoglobin variants are crucial for patient management.
  • High-performance liquid chromatography (HPLC) offers powerful separation capabilities for complex biological molecules.

Purpose of the Study:

  • To review two distinct HPLC methods for analyzing hemoglobins and globin chains.
  • To highlight the diagnostic utility of these chromatographic techniques in identifying hemoglobinopathies.
  • To demonstrate the application of HPLC in separating various normal and abnormal globin chains.

Main Methods:

  • Cation exchange HPLC with gradient elution for separating intact hemoglobins from adult and neonatal blood samples.
  • Reverse-phase HPLC using acetonitrile-H2O-trifluoroacetic acid at low pH for globin chain separation.
  • Analytical and preparative applications of both HPLC methods.

Main Results:

  • Cation exchange HPLC effectively separates numerous normal and abnormal hemoglobins, demonstrating high diagnostic value.
  • Reverse-phase HPLC successfully separates globin chains (alpha, zeta, beta, delta, G gamma, A gamma, epsilon).
  • The reverse-phase method is adept at resolving normal and abnormal chains with neutral amino acid substitutions, including distinct gamma chain variants.

Conclusions:

  • Two distinct HPLC methods provide robust tools for the comprehensive analysis of hemoglobins and globin chains.
  • These methods are valuable for the diagnosis and characterization of hemoglobinopathies.
  • HPLC offers both analytical and preparative capabilities for studying globin chain variants.