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

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Single-injection calibration approach for high-performance liquid chromatography.

Eduardo Paredes¹, Salvador E Maestre, Ma Soledad Prats

¹Department of Analytical Chemistry, Nutrition and Food Sciences, University of Alicante, 03080 Alicante, Spain.

Journal of Chromatography. A

|February 27, 2008

Summary

A new single-injection calibration approach (SICA) for high-performance liquid chromatography significantly reduces analysis time. This validated method offers a faster, efficient way to calibrate for various organic compounds across multiple detection techniques.

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Area of Science:

Analytical Chemistry
Chromatography

Background:

Traditional calibration methods in chromatography can be time-consuming.
Accurate calibration is crucial for quantitative analysis of complex samples.

Purpose of the Study:

To validate a novel single-injection calibration approach (SICA) for high-performance liquid chromatography (HPLC).
To assess the efficiency and applicability of SICA across different detection methods and sample types.

Main Methods:

Developed and validated the single-injection calibration approach (SICA).
Applied SICA to analyze non-volatile and semi-volatile organic compounds including carboxylic acids, polyalcohols, carbohydrates, and vitamins.
Tested SICA with refractive index (RI), diode array detection (DAD), and inductively coupled plasma atomic emission detection (ICP-AED).
Validated the method using a reference non-fat milk powder and various real food samples.

Main Results:

SICA achieved a 1-7 fold reduction in analysis time compared to conventional methods.
Excellent linearity (r(2)>0.999) was observed for all three detection methods.
Inductively coupled plasma atomic emission detection (ICP-AED) demonstrated superior universality, with signal intensity dependent only on carbon mass content.
Signal correction was necessary for RI and DAD detection methods.

Conclusions:

The single-injection calibration approach (SICA) is a validated, time-saving method for HPLC.
ICP-AED offers the most universal detection for SICA due to its independence from compound-specific chemical properties.
SICA is applicable to diverse real-world samples, including food matrices.