Analysis of Biogenic Amines in Wine Using the Agilent 1290 Infinity II LC and Ultivo Triple Quadrupole LC/MS

Significance of the topic

Biogenic amines are key indicators of food quality and safety, arising from microbial activity and amino acid decarboxylation during fermentation. Elevated levels can signal poor hygiene and pose health risks such as headaches, hypertension, and allergic reactions. Reliable, rapid analytical methods are essential for regulatory compliance and consumer protection.

Objectives and study overview

This work aimed to establish a streamlined, high-throughput approach for direct quantification of underivatized biogenic amines in wine. The method integrates an Agilent 1290 Infinity II liquid chromatograph with an Ultivo triple quadrupole mass spectrometer. Performance was evaluated using Tempranillo red wine as a representative matrix.

Methodology and instrumentation

Ion-pair reversed-phase chromatography was performed on a Poroshell 120 EC-C18 column (2.1×50 mm, 2.7 μm) using 0.1 percent heptafluorobutyric acid in water (A) and acetonitrile (B). A gradient from 2 percent to 30 percent B over 7 minutes achieved baseline separation of 15 biogenic amines, including isomeric pairs. The Ultivo LC/TQ operated in positive electrospray mode with dynamic multiple reaction monitoring. Automated optimization of collision energy, fragmentor voltage, gas temperatures, and flows was performed via MassHunter software. Chromatographic parameters yielded moderate backpressure, enabling use on lower-end systems.

Main results and discussion

Calibration curves in 0.1 M HCl exhibited linearity with correlation coefficients above 0.98 across relevant ranges. Limits of detection ranged between 0.5 and 1.0 ng/mL. Precision studies (n=10) showed relative standard deviations below 1 percent for areas and retention times. Matrix effects in Tempranillo wine were minimal for most analytes (<10 percent), with agmatine and norepinephrine showing ~50 percent suppression. Analysis of diluted wine detected nine amines, notably putrescine at 11.8 mg/L, histamine at 2.5 mg/L, and tyramine at 2.1 mg/L, all below toxicological thresholds.

Benefits and practical applications of the method

This workflow eliminates the need for derivatization, reducing assay complexity, cost, and time. The compact LC/MS stack minimizes laboratory footprint, while moderate system pressure permits operation on benchtop LC instruments. Short runtimes (<8 minutes) and automated parameter tuning support high sample throughput in food safety laboratories.

Future trends and opportunities

Future developments may involve isotopically labeled internal standards to correct residual matrix effects, further miniaturization of LC/MS platforms, and integration with high-resolution mass spectrometers for comprehensive profiling. Advances in online sample preparation and automation could broaden applicability across diverse food matrices and enable near real-time monitoring.

Conclusion

The described method provides a rapid, sensitive, and reproducible solution for direct analysis of biogenic amines in wine. It offers a practical alternative to derivatization-based assays, supporting robust quality control and regulatory compliance in food testing laboratories.

References

• Scientific Opinion on Risk Based Control of Biogenic Amine Formation in Fermented Foods EFSA Journal 2011 9(10) 2393
• Sentellas S Núñez Ó Saurina J Recent Advances in Determination of Biogenic Amines in Food Samples by (U)HPLC J Agric Food Chem 2016 64(41) 7667-7678
• Kaufmann A Maden K Easy and Fast Method for the Determination of Biogenic Amines in Fish and Fish Products with Liquid Chromatography Coupled to Orbitrap Tandem Mass Spectrometry J AOAC Int 2018 101(2) 336-341