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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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Updated: Jun 28, 2026

Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)

Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)

Published on: May 20, 2013

Dynamic analysis of on-line high-performance liquid chromatography for multivariate statistical process control.

Sila Kittiwachana¹, Diana L S Ferreira, Louise A Fido

¹Centre for Chemometrics, School of Chemistry, University of Bristol, Cantocks Close, Bristol BS8 1TS, UK.

Journal of Chromatography. A

|November 11, 2008

Summary

This study introduces a real-time multivariate statistical process control method for analyzing continuous chromatographic data. An "unlocked" peak table effectively identifies process deviations and contaminants, outperforming a "locked" table.

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Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)

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Published on: May 20, 2013

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Large Scale Non-targeted Metabolomic Profiling of Serum by Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS)

Large Scale Non-targeted Metabolomic Profiling of Serum by Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS)

Published on: March 14, 2013

Area of Science:

Analytical Chemistry
Process Engineering

Background:

Continuous manufacturing requires robust real-time monitoring.
High-performance liquid chromatography (HPLC) generates large datasets.
Statistical process control (SPC) is crucial for quality assurance.

Purpose of the Study:

To develop and evaluate a real-time SPC method for continuous HPLC processes.
To compare "unlocked" and "locked" peak table approaches for process monitoring.
To identify normal operating conditions and process contaminants.

Main Methods:

On-line HPLC data acquisition over 83.32 hours (252 chromatograms).
Multivariate statistical process control applied to peak tables.
Evolving principal components analysis (EPCA) to define the normal operating condition (NOC) region.
Comparison of "unlocked" (dynamic) and "locked" (static) peak table strategies.

Main Results:

The NOC region was identified between 5.77 and 8.13 hours.
19 out of 37 detected peaks were within the NOC; others indicated contaminants.
The "unlocked" peak table demonstrated superior detection of out-of-control samples.
The "unlocked" approach provided better diagnostic insights into process issues.

Conclusions:

Real-time SPC using an "unlocked" peak table is effective for continuous HPLC processes.
This method enhances the ability to detect process deviations and identify contaminants.
Dynamic updating of the peak table improves process monitoring and control.