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High-Performance Liquid Chromatography: Types of Detectors01:15

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Validated HPLC-Diode Array Detector Method for Simultaneous Evaluation of Six Quality Markers in Coffee.

Anastasia Gant1, Vanessa E Leyva, Ana E Gonzalez

  • 1Pontificia Universidad Catolica del Peru (PUCP), Departamento de Ciencias-Quimica, Av. Universitaria 1801, Lima 32, Peru.

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|April 11, 2015
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Summary

A new HPLC-diode array detector method enables simultaneous quantitation of key coffee compounds, including nicotinic acid and trigonelline. This rapid analysis aids in evaluating coffee quality and thermal decomposition during roasting.

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Area of Science:

  • Analytical Chemistry
  • Food Science
  • Biochemistry

Background:

  • Nicotinic acid, N-methylpyridinium ion, and trigonelline are key nutritional biomarkers in coffee.
  • These compounds indicate thermal decomposition during coffee roasting.
  • No existing method allows for the simultaneous determination of these and other coffee compounds.

Purpose of the Study:

  • To develop and validate a rapid High-Performance Liquid Chromatography (HPLC) method with a diode array detector (DAD).
  • To enable simultaneous quantitation of multiple coffee compounds: caffeine, trigonelline, nicotinic acid, N-methylpyridinium ion, 5-caffeoylquinic acid, and 5-hydroxymethyl furfural.
  • To apply the method to diverse coffee matrices: green, roasted, and instant coffee.

Main Methods:

  • Utilized a phenyl-hexyl reversed-phase (RP) column (250×4.6 mm, 5 μm).
  • Employed a mobile phase of 0.3% aqueous formic buffer (pH 2.4) and methanol.
  • Optimized flow rate at 1 mL/min and column temperature at 30°C for 30-minute separation.

Main Results:

  • Achieved baseline separation of all target compounds within 30 minutes.
  • Demonstrated excellent linearity (r² > 0.9985) for all analytes.
  • Confirmed high sensitivity (LOD=0.023-0.237 μg/mL; LOQ=0.069-0.711 μg/mL) and recovery (84-102%).

Conclusions:

  • The developed HPLC-DAD method is rapid, validated, and effective for simultaneous quantitation.
  • The method is applicable across various coffee types (green, roasted, instant).
  • This simplified approach facilitates comprehensive routine coffee analysis in the industry.