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HPLC/MS/MS Analysis of Bitter Acids in Hops and Beer

been done by simultaneously extracting the same beer sample with hexane as one sample and diluting the beer with distilled water as a second sample. A same dilution step of 1:20 has been used for both samples, where the sample evaporated from hexane has been diluted in mobile phase. Comparison of the results shows that while the intensity of alpha acids is the same, the levels of iso-alpha acids are higher for the water diluted sample. This reflects the higher solubility of the iso-alpha acids in water. Both preparation methods provide samples suitable for mass spectrometer analysis.

Based on the results from the water dilution method a quantitation limit estimate is approximately beer diluted 40,000 times. While the water dilution provides simplification of the sample preparation and handling (lower CO2 content after dilution allows consistent sample uptake with an autosampler syringe) it places greater emphasis on robustness of the LC/MS interface. The API 3000 interface has been tested with 12 hours of continuous injections (60) with results showing no loss in sensitivity.

Conclusion

LC/MS/MS method for detection of bitter acids in beer and hops has been presented. It offers a possible application in on-line quantitative analysis for quality control as well as in brewing process development. The qualitative work presented could be extended to impurity identification and structural elucidation of degradation product present in hop products.

Authors

Peter Kovarik, Yves Mouget, Takeo Sakuma, MDS SCIEX


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