4 °C - the National Sea Grant Library

DIPSTICK TESTS FOR BIOGENIC AMINES
Robert C. Bateman, Jr’ and Margot Hall2
1. Department of Chemistry and Biochemistry
University of Southern Mississippi
Hattiesburg, MS 39406-5043
and
2. Department of Medical Technology
University of Southern Mississippi
Hattiesburg, MS 39406-5 134
ABSTRACT
A rapid calorimetric dipstick test for histamine in tuna was developed based on the
coupling of diamine oxidase to a peroxidase/dye system. The assay was linear to 1.0 mM
histamine and the minimum detectable concentration was 0.07 mM corresponding to 2.3
mg% in tuna extracts. Histidine and cadaverine did not interfere with the assay whereas
putrescine reacted slightly with the dipstick. Histamine determinations in fresh and spoiled
tuna gave good agreement between the dipstick method and the AOAC fluorometric
method. The dipstick assay technique should be readily adaptable to putrescine or
cadaverine.
INTRODUCTION
A variety of assay methods exist for the determination of histamine in seafood
including spectrophotometric (l), chromatographic (2,3), and chemical chelation (4).
However, they are generally slow and require considerable training on the part of the user.
Enzymatic calorimetric assays for histamine using diamine oxidase (histaminase) have also
been reported (5,6). In the presence of oxygen, diamine oxidase (DAO) deaminates
histamine to form hydrogen peroxide, ammonia, and imidazolacetaldehyde.
DA0
Histamine + O2 -----------> H,O, + NH3 + irnidazolacetaldehyde
The liberated hydrogen peroxide, coupled with horseradish peroxidase (HRP), converts a
reduced dye to its oxidized form with accompanying color development.
HRP
H,O, + reduced dye ----------> H,O + oxidized dye
Dry chemistry tests exist for glucose (7) and cholesterol (8) using the appropriate
oxidase, HRP, a compatible dye, buffers and stabilizers embedded in a solid matrix such as
filter paper or silica. In combination with reflectance photometry, they give simple, rapid,
quantitative results. Alternatively, color development may be read visually and compared