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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.
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The high insolubility of some precipitates can result in an unfavorable relative supersaturation. This can lead to colloidal particles with a large surface-to-mass ratio, where adsorption is promoted. For instance, in the precipitation of silver chloride, silver ions are adsorbed on the surface of the colloidal particles, forming a primary layer. This layer attracts ions of opposite charge (such as nitrate ions), forming a diffuse secondary layer of adsorbed ions. This electric double layer...
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Metal ions can be separated from one another by complexation with organic ligands–the chelating agent– to form uncharged chelates. Here, the chelating agent must contain hydrophobic groups and behave as a weak acid, losing a proton to bind with the metal. Since most organic ligands used in this process are insoluble or undergo oxidation in the aqueous phase, the chelating agent is initially added to the organic phase and extracted into the aqueous phase. The metal-ligand complex is...
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The phase of a given substance depends on the pressure and temperature. Thus, plots of pressure versus temperature showing the phase in each region provide considerable insights into the thermal properties of substances. Such plots are known as phase diagrams. For instance, in the phase diagram for water (Figure 1), the solid curve boundaries between the phases indicate phase transitions (i.e., temperatures and pressures at which the phases coexist).
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Extraction: Effects of pH00:53

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Consider a neutral form of an amine, B, with a partition coefficient, K, in a liquid mixture containing organic and aqueous phases. The pH of the aqueous phase affects the charge on acidic and basic solutes, and the charged form is usually more soluble in the aqueous phase. Suppose the conjugate acid form of the amine is soluble only in the aqueous phase while the base form is soluble in both phases. Then the distribution coefficient, D, can be given as the ratio of amine concentration in the...
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Liquid Phase Separation Controlled by pH.

Omar Adame-Arana1, Christoph A Weber2, Vasily Zaburdaev3

  • 1Max-Planck-Institut für Physik komplexer Systeme, Dresden, Germany.

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|October 3, 2020
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This summary is machine-generated.

This study introduces a minimal model for macromolecule liquid phase separation, revealing pH-dependent phase diagrams and reentrant behavior. The model explains in vitro and in vivo protein phase separation, particularly near the isoelectric point.

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Area of Science:

  • Biophysics
  • Physical Chemistry
  • Soft Matter Physics

Background:

  • Liquid phase separation of macromolecules is crucial in biological systems.
  • pH significantly influences macromolecule behavior and interactions.
  • Understanding pH effects on phase separation is key for cellular function and biomaterial design.

Purpose of the Study:

  • To develop a minimal model investigating the impact of pH on macromolecule liquid phase separation.
  • To explore the relationship between pH, macromolecule charge states, and phase behavior.
  • To predict and analyze phase diagram topologies under varying pH conditions.

Main Methods:

  • A minimal model incorporating macromolecule protonation/deprotonation and water self-ionization.
  • Utilizing Flory-Huggins interaction parameters for short Debye screening lengths.
  • Thermodynamic analysis of effective free energy at fixed pH and chemical equilibrium.

Main Results:

  • Identified diverse phase diagram topologies, including critical and triple points.
  • Demonstrated pH-dependent phase diagrams, influenced by charge interactions.
  • Predicted reentrant phase separation behavior and broader separation regions at the isoelectric point.

Conclusions:

  • The model successfully accounts for pH-driven liquid phase separation in macromolecules.
  • Findings are relevant to in vitro protein phase separation and in vivo cellular processes.
  • The study provides insights into the role of pH in regulating macromolecular condensates.