Simultaneous and Direct Analysis of Biogenic Amines in Food by LC–MS-MS Using Hydrophilic Chromatography

Biogenic amines are present in protein-rich foods such as fish, meat, and milk. Their concentration increases during food decomposition. The reasons to monitor biogenic amines in food products are to determine that the food is suitable for consumption and to establish appropriate storage conditions. Analysis methods for biogenic amines include HPLC with fluorescence detection — using derivatization with dansyl chloride or o-phthalaldehyde. The drawbacks to these methods are a 40-min analysis time, sample pretreatment that requires fluorescence derivatization, and liquid phase extraction. In this study, we investigated a simple, highly sensitive, and direct analytical method that does not require derivatization and uses a TSKgel Amide-80 column under HILIC conditions followed by MS-MS detection (Figure 1).

Results and Discussion

Figure 2 and Table I detail the results of the analysis of six biogenic amines. Excellent separation of the analytes and linearity of the calibration curves were obtained. When compared to a fluorescence method using a reversed phase column (data not shown), this LC–MS-MS method can be completed in half the analysis time (20 min versus 40 min). The limits of quantitation for the analytes were 0.001–0.02 mg in a 100 g sample, an improvement of 5–1300 times compared with the fluorescence method.

Figure 1: Sample pretreatment procedure of LC–MS-MS method.

A study was conducted to compare the LC–MS-MS method to the fluorescence method in the evaluation of tuna samples under differing storage conditions. Table II lists the quantitative values of biogenic amines in tuna preserved for two days at freezing and room temperatures. Good correlation was obtained between the two methods.

Figure 2: Chromatograms of biogenic amines.

Conclusions

A new LC–MS-MS method for the analysis of biogenic amines was investigated by Tosoh scientists. Six biogenic amines were analyzed in 15 min under HILIC conditions using a TSKgel Amide-80 column without the need for a complex and time-consuming derivatization procedure. Limits of quantitation of the analytes were improved 5–1300 times using this new method compared with the conventional fluorescence method. Since good correlation between the two methods was obtained in a comparative study of quantitative values, the LC–MS-MS method is a proven alternative method that is both simple and sensitive for the analysis of biogenic amines in food products.

Table I: Comparison of limit of detection and quantitation values (LC–MS-MS versus fluorescence method)

Table II: Comparison of quantitative values of biogenic amines in tuna sample (mg/100 g)

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