Related Experiment Video
Updated: Jul 10, 2026
07:11
Automated HPLC Separation Using LC-Mate: An Integrated Repetitive Autosampler and Fraction Collector for Microscale Purification
Published on: February 27, 2026
Peak separation by adventitious or added water in normal-phase chiral HPLC
1Institut für Organische Chemie, RWTH Aachen, Landoltweg 1, D-52074 Aachen, Germany. lukas.hintermann@oc.rwth-aachen.de
The Journal of Organic Chemistry
|November 15, 2007
Summary
Water in normal-phase chiral HPLC eluents impacts analysis reproducibility. Deliberately adding water can improve peak resolution and reduce solvent use.
Area of Science:
- Analytical Chemistry
- Chromatography
Background:
- Normal-phase chiral High-Performance Liquid Chromatography (HPLC) is sensitive to eluent composition.
- Uncontrolled water content in eluents can lead to significant analytical challenges.
Purpose of the Study:
- To investigate the impact of trace water on normal-phase chiral HPLC.
- To evaluate the benefits of deliberate water addition to eluents.
Main Methods:
- Analysis of peak retention time, tailing, and resolution in normal-phase chiral HPLC.
- Comparison of results with adventitious versus deliberately added water in eluents.
Main Results:
- Trace water significantly affects retention time, peak tailing, and resolution.
- Adventitious water leads to irreproducible analytical results.
- Deliberate addition of water enhances peak resolution and reduces solvent consumption.
Conclusions:
- Controlling water content is crucial for reproducible normal-phase chiral HPLC.
- Strategic addition of water to eluents offers a method to improve chromatographic performance and efficiency.
Related Concept Videos
01:11High-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.
In HPLC, two phases play a critical role in the separation process:
In HPLC, two phases play a critical role in the separation process:
01:05High-Performance Liquid Chromatography: Elution Process
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...
00:57High-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.
01:09Ion-Exchange Chromatography
Ion-exchange chromatography, or IEC, is a technique for separating ions based on their affinity for the stationary phase. The stationary phase is a cross-linked polymer resin with covalently attached ionic functional groups. The functional groups can be either positively charged (cation exchangers) or negatively charged (anion exchangers). A cation exchanger consists of a polymeric anion and active cations, while an anion exchanger is a polymeric cation with active anions. The choice of...
01:08Size-Exclusion Chromatography
In size-exclusion chromatography (SEC), also known as molecular-exclusion or gel-permeation chromatography, molecules are separated based on their sizes. This technique is important for separating large molecules such as polymers and biomolecules. The two classes of micron-sized stationary phases encountered in SEC are silica particles and cross-linked polymer resin beads. Both materials are porous, but their pore sizes vary significantly.
Silica particles offer advantages such as rigidity,...
Silica particles offer advantages such as rigidity,...
01:10Chromatography: Introduction
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...
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...