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High-Performance Liquid Chromatography: Elution Process01:05

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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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High-Performance Liquid Chromatography: Introduction01:11

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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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High-Performance Liquid Chromatography: Instrumentation00:57

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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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Chromatography: Introduction01:10

Chromatography: Introduction

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Chromatography is a technique used to separate compounds based on differences of partitioning between two phases, the stationary phase and the mobile phase.
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The chromatography technique was first invented in 1901 by Michael S. Tswett, a Russian botanist, to separate plant pigments using organic solvents. Further, in 1941, Archer John Porter Martin and R. L. M. Synge modified the technique by packing silica gel into a column. A mixture of amino acids was then separated on the packed column using chloroform and water mixture as the mobile phase. This was the first report on column chromatography. At present, column chromatography is a widely used...
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Chromatographic techniques are classified in three ways: the classification is based on the physical state of the stationary and mobile phases, how the mobile phase and the stationary phase contact each other, or through the chemical or physical processes that isolate the components of the sample. Typically, the mobile phase is either a liquid or gas, while the stationary phase is either a solid or a liquid layer applied to a solid surface.
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Isolation and Preparation of Bacterial Cell Walls for Compositional Analysis by Ultra Performance Liquid Chromatography
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Functionality-type and chemical-composition separation of poly(lactide-co-glycolide) using gradient elution

Masashi Serizawa1, Jeroen Reekers2, Pieter van Delft3

  • 1Van't Hoff Institute for Molecular Sciences, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, the Netherlands; Centre for Analytical Sciences Amsterdam, Science Park 904, 1098 XH Amsterdam, the Netherlands; Material Characterization laboratory, Mitsubishi Chemical Corporation, 1000 Kamoshida-cho, Aoba-ku, Yokohama-shi, Kanagawa 227-8502, Japan.

Journal of Chromatography. A
|July 12, 2024
PubMed
Summary

A new liquid chromatography method effectively analyzes poly(Lactide-co-glycolide) (PLGA) end groups up to 183 kDa. This technique improves quality control and understanding of PLGA polymer properties for drug delivery applications.

Keywords:
Chemical-composition distribution (CCD)Functionality-type distribution (FTD)Gradient NPLCPoly(lactic acid) (PLA)Poly(lactic-co-glycolic acid) (PLGA)

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Area of Science:

  • Polymer Chemistry
  • Materials Science
  • Analytical Chemistry

Background:

  • End groups of poly(Lactide-co-glycolide) (PLGA) significantly influence polymer properties for drug delivery.
  • Existing methods for analyzing PLGA end groups have limitations in molecular weight range or throughput.
  • Understanding end-group functionality is crucial for predicting degradation and encapsulation efficiency.

Purpose of the Study:

  • To develop a novel, high-throughput method for analyzing PLGA end groups.
  • To characterize functionality type distribution (FTD) and chemical composition distribution (CCD) in PLGA polymers.
  • To enable simultaneous separation of Lactic acid (LA)/Glycolic acid (GA) ratios.

Main Methods:

  • Normal-phase liquid chromatography (LC) utilizing a cross-linked diol column.
  • A ternary gradient elution system with Hexane, Ethyl Acetate, and Tetrahydrofuran.
  • Separation of mono-ester and di-acid terminated PLGA, and ester-terminated PLGA with varying LA/GA ratios.

Main Results:

  • The LC method successfully resolved FTD and partially CCD in commercial PLGA.
  • Separation was achieved for PLGA polymers up to 183.0 kDa molecular weight.
  • Simultaneous separation of different Lactic acid (LA)/Glycolic acid (GA) ratios was demonstrated.

Conclusions:

  • The developed LC method offers a significant advancement for PLGA characterization.
  • This technique facilitates a deeper understanding of the correlation between PLGA's FTD, CCD, and physical properties.
  • The method is suitable for product development and quality control of PLGA in drug delivery systems.