Determination of Biogenic Amines in Alcoholic Beverages by Ion Chromatography with Suppressed Conductivity and Integrated Pulsed Amperometric Detections

Importance of the Topic

Biogenic amines are nitrogenous compounds formed by microbial decarboxylation of amino acids and occur widely in plants, animals, and fermented foods. In alcoholic beverages, elevated levels of amines such as histamine and tyramine can trigger adverse reactions—headaches, hypertension, and allergic responses—and serve as indicators of spoilage or poor processing control. Reliable analysis of these compounds supports food safety, quality assurance, and regulatory compliance in the beverage industry.

Objectives and Study Overview

This work evaluated a robust ion chromatography (IC) method employing an IonPac CS18 cation-exchange column combined with suppressed conductivity detection, integrated pulsed amperometric detection (IPAD), and UV absorbance to determine underivatized biogenic amines in beer and wine. The study aimed to optimize separation, assess detector performance (linearity, limits of detection, precision, recovery), and monitor amine concentration changes during refrigerated storage.

Methodology and Instrumentation

Samples (beers, red, white, and rosé wines) were diluted or centrifuged and injected onto a Dionex ICS-3000 IC system. A gradient of online-generated methanesulfonic acid (3–45 mM MSA) at 0.30 mL/min, 40 °C, achieved separation of ten amines. Detection modes included:

• Suppressed conductivity with a CSRS® ULTRA II suppressor in external water mode
• IPAD with postcolumn addition of 100 mM NaOH and a conventional Au electrode
• UV absorbance at 276 nm for confirmation of aromatic amines (e.g., tyramine)

Main Results and Discussion

All target amines (dopamine, tyramine, putrescine, cadaverine, histamine, serotonin, agmatine, phenylethylamine, spermidine, spermine) were baseline-resolved, except certain coelutions requiring dual detection. Suppressed conductivity delivered the lowest detection limits (3–18 µg/L for most amines) and low noise (0.2–0.3 nS). IPAD showed linearity (r² > 0.997) over broad ranges (0.1–20 mg/L) with detection limits of 20–400 µg/L. UV detection selectively confirmed tyramine at 0.2–10 mg/L (LOD 110 µg/L). Method precision (n=10) yielded retention-time RSDs <0.1% and peak-area RSDs <1.3% (suppressed conductivity). Applications to commercial beers and wines revealed:

• Beers: putrescine (2–6 mg/L), histamine (0.2–0.7 mg/L), agmatine (6–15 mg/L)
• Wines: higher amine levels in red (putrescine 5–16 mg/L; histamine up to 5 mg/L) versus white/rosé (putrescine ~1 mg/L; histamine <0.7 mg/L)

Storage studies at 4 °C over 1–3 weeks showed increases in putrescine (+0–65%), histamine (+67–184%), cadaverine, and late-appearing agmatine, spermidine, and spermine, demonstrating dynamic amine profiles.

Benefits and Practical Applications

This IC approach avoids complex derivatization steps, reduces analysis time, and enables simultaneous multi-detector confirmation to enhance specificity. It supports quality control, microbial spoilage assessment (biogenic amine index), and compliance with safety guidelines for beverages.

Future Trends and Applications

Advances may include expanded analyte panels (other amines, peptides), miniaturized or field-deployable IC systems, hyphenation with mass spectrometry for structural confirmation, and real-time process monitoring during fermentation and storage.

Conclusion

The combination of an IonPac CS18 column with suppressed conductivity, IPAD, and UV absorbance provides a sensitive, precise, and versatile platform for biogenic amine analysis in complex alcoholic matrices. The method offers streamlined workflows, low detection limits, and robust performance for beverage safety and quality assurance.

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