Analysis of biogenic amines food and physiological samples


Courtesy of Pickering Laboratories, Inc.

Histamine and other polyamines such as Putrescine, Cadaverine, Spermidine and Spermine are products of decomposition. The level of histamine, in particular, has been used as a regulatory guideline for degree of decomposition in fish. Also, Histamine and Tyramine levels are of interest to producers of wine and cheese.

The polyamines, Spermidine, Spermine and Putrescine are elevated in neoplastic and damaged, e.g., burned, tissue. In fact, the level of circulating and urinary polyamines is an indicator of total body damage. The levels can be monitored for the effect of diet and other modalities on recovery.

This method employs ion-exchange chromatography so, after extraction, centrifugation/filtration are the only sample preparations necessary.

Extraction Procedure

Weigh 10 g of sample into a small glass blender cup (any fortifications should be added at this time). Add 50 mL of extraction solution, 80% HPLC Methanol and 20% 0.1N HCl, and homogenize for two minutes. Centrifuge for five minutes. Mix equal portions of the supernatant with the mobile phase (K600). Allow to coagulate at -4 °C for 15 minutes, then centrifuge for 5 minutes. The clear supernatant is filtered (0.45 μm Nylon) and placed in an autosampler vial.

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