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Related Concept Videos

High-Performance Liquid Chromatography: Introduction01:11

High-Performance Liquid Chromatography: Introduction

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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Mass Spectrometry: Complex Analysis01:21

Mass Spectrometry: Complex Analysis

Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
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High-Performance Liquid Chromatography: Instrumentation00:57

High-Performance Liquid Chromatography: Instrumentation

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.
Ion-Exchange Chromatography01:09

Ion-Exchange Chromatography

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2D NMR: Heteronuclear Single-Quantum Correlation Spectroscopy (HSQC)01:19

2D NMR: Heteronuclear Single-Quantum Correlation Spectroscopy (HSQC)

Heteronuclear single-quantum correlation spectroscopy (HSQC) is a 2D NMR technique that reveals one-bond correlations between hydrogen and a heteronucleus. The HSQC experiment is similar to the heteronuclear correlation experiment (HETCOR) but is more sensitive. In the HSQC spectrum, the proton chemical shift is plotted on the horizontal F2 axis, while the 13C chemical shift is plotted on the vertical F1 axis. The corresponding proton and 13C spectra are also shown. The HSQC contour plot does...

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2D-HPLC-MS Technology Combined with Molecular Network for the Identification of Components in Tibetan Medicine Aconitum pendulum
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Simultaneous, sequential quantitative achiral-chiral analysis by two-dimensional liquid chromatography.

Cadapakam J Venkatramani1, Larry Wigman, Kavita Mistry

  • 1Small Molecules Pharmaceutical Science, Genentech Inc., South San Francisco, CA 94080, USA. cadapakv@gene.com

Journal of Separation Science
|July 19, 2012
PubMed
Summary
This summary is machine-generated.

This study introduces heart-cutting (LC-LC) and comprehensive two-dimensional liquid chromatography (LC × LC) for analyzing chiral compounds. These methods accurately quantify minor enantiomers, achieving high sensitivity and accuracy for chiral purity assessment.

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Area of Science:

  • Analytical Chemistry
  • Chromatography
  • Chiral Separations

Background:

  • Chiral compound analysis typically requires separate achiral and chiral chromatographic methods.
  • Achiral methods assess assay and impurities, while chiral methods determine enantiomeric purity.
  • Reversed-phase chromatography is common for achiral analysis; various modes suit chiral separations.

Purpose of the Study:

  • To demonstrate the application of heart-cutting (LC-LC) and comprehensive two-dimensional liquid chromatography (LC × LC) for simultaneous achiral and chiral analysis.
  • To quantify minor, undesired enantiomers in the presence of major enantiomers using phenylalanine as a model compound.
  • To compare the performance of LC-LC and LC × LC for chiral purity determination.

Main Methods:

  • Utilized heart-cutting (LC-LC) and comprehensive two-dimensional liquid chromatography (LC × LC).
  • Employed a reversed-phase C18 achiral column for the primary dimension and a Chirobiotic Tag chiral column for the secondary dimension.
  • Developed a compatible mobile phase for both dimensions to enable sequential achiral and chiral separation.

Main Results:

  • Achieved simultaneous achiral and chiral analysis and quantitation of enantiomers.
  • LC-LC and LC × LC provided comparable results, with LC-LC showing enhanced sensitivity and accuracy.
  • Quantified undesired D-phenylalanine at ~0.3% in L-phenylalanine (99.7%) over three orders of magnitude, with limits of quantitation comparable to conventional HPLC.

Conclusions:

  • LC-LC and LC × LC are effective for simultaneous, sequential achiral and chiral analysis of enantiomers.
  • These two-dimensional chromatography techniques offer high sensitivity and accuracy for chiral purity assessment.
  • The methods are suitable for quantifying trace enantiomers, comparable to established high-performance liquid chromatography techniques.