ISSN 1063-0740, Russian Journal of Marine Biology, 2016, Vol. 42, No. 3, pp. 258–265. © Pleiades Publishing, Ltd., 2016.
Original Russian Text © P.A. Kameneva, T.Yu. Orlova, A.B. Imbs, 2016, published in Biologiya Morya.
The Seasonal Dynamics of Diarrhetic Shellfish Poisoning Toxins
in the Digestive Gland of Crenomytilus grayanus (Dunker, 1853)
Analyzed by High-Performance Liquid Chromatography
with Fluorometric Detection
P. A . K a m e n e v a
, T. Yu. Orlova
, and A. B. Imbs
Zhirmunsky Institute of Marine Biology, Far East Branch, Russian Academy of Sciences,
ul. Pal’chevskogo 17, Vladivostok, 690041 Russia
Far Eastern Federal University, ul. Sukhanova 8, Vladivostok, 690950 Russia
Received November 26, 2015
Abstract—The seasonal dynamics of diarrhetic shellfish poisoning (DSP) toxins in the digestive gland of the
mussel Crenomytilus grayanus (Dunker, 1853) from Peter the Great Bay in the Sea of Japan have been studied.
Toxins in the form of 4-bromomethyl-7-methoxycoumarin derivatives have been analyzed using high-perfor-
mance liquid chromatography (HPLC) with fluorometric detection. This method allows the determination
of the quantitative and qualitative composition of individual toxins. All the C. grayanus samples contained
DSP toxins, but their concentration varied substantially during the year. The highest concentration was
recorded from the mussels collected in May; the lowest concentration was recorded in February. The toxins
were represented predominantly by dinophysistoxin-2; in July-September and December, okadaic acid was
detected. An assumption has been made about the relationship between the potential producers of DSP toxins
in the waters of the Peter the Great Bay and the dynamics of these toxins in tissues of C. grayanus. The HPLC
method, as an alternative to the enzyme-linked immunosorbent assay, was used to assess the quantitative and
qualitative composition of DSP toxins in seafood for the first time in the Russian Federation. This method
can be recommended for the routine monitoring of toxins conducted by the controlling organizations of the
Russian Federal Service for Veterinary and Phytosanitary Surveillance.
Keywords: diarrhetic shellfish poisoning, okadaic acid, dinophysistoxin-2, Crenomytilus grayanus, HPLC,
Diarrhetic shellfish poisoning (DSP) toxins,
including okadaic acid (OA) and its derivatives, dino-
physistoxins (DTX), are a class of phytotoxins. These
substances cause symptoms of gastrointestinal disor-
ders [3, 22]. OA and DTX are known as inhibitors of
serin/threonine phosphatases in mammalian cells.
Due to blockage of these enzymes in the human gas-
trointestinal tract, epithelial cells become damaged,
which results in the diarrhetic syndrome . As well,
OA is a cancer promoter in mammalian cells that can
cause damage to the DNA structure . DSP toxins
are accumulated in the tissues of marine filter-feeding
bivalves and can be transferred to other aquatic organ-
isms up the food chains.
The substances that are DSP toxins differ in their
chemical structures; this fact greatly complicates their
study. In spite of the small difference in physical and
chemical properties between the two OA derivatives,
dinophysistoxin-1 (DTX-1) and dinophysistoxin-2
(DTX-2) (Fig. 1), their toxic effects differ substantially.
DTX-1 exhibits a stronger poisoning effect on mamma-
lian cells, as compared to OA and DTX-2 .
In tissues of bivalves, OA and DTX-1/2 acquire the
form of fatty acid esters (7-O-acyl derivatives). The
group of 7-O-acyl derivatives of OA and DTX-1/2 is
referred to as dinophysistoxin-3, or DTX-3 (Fig. 1)
. It is known that OA derivatives in the form of
esters lose their ability to bind with serin/threonine
phosphatases and thus do not have a toxic effect on
bivalves, i.e., they turn into an inactive form .
When DTX-3-containing bivalves are consumed as
food, in the human digestive tract the ester bonds are
hydrolyzed with the release of the active forms of tox-
ins, OA and/or DTX-1/2, which can have a poisoning
effect . In bivalves, OA, DTX-1/2, and DTX-3 are
accumulated mainly in the digestive gland . How-
ever, as has been shown in a number of works, DTX-3