TY - JOUR
T1 - Single-cell analysis of paralytic shellfish toxins in Alexandrium tamarense by HPLC with post-column fluorescent derivatization
AU - Cho, Yuko
AU - Ozeki, Ryoko
AU - Yotsu-Yamashita, Mari
AU - Oshima, Yasukatsu
N1 - Funding Information:
This work was supported by KAKENHI (Grant-in-Aid for Exploratory Research, No. 21658075 and 24580295) to Y. Cho and Funding Program for the Next Generation World-Leading Researchers (LS012) to M. Y. Yamashita from the Japan Society of Promotion and Science. We thank Dr. T. Omura and Dr. T. Ishimaru of Tokyo University of Marine Science and Technology , who kindly donated the A. tamarense clonal culture strains, Axat-2 and UAT-014-009.[SS]
PY - 2013/5
Y1 - 2013/5
N2 - We developed a methodology for analyzing the C-toxin (C2) content in single Alexandrium tamarense cells; this method was based on high performance liquid chromatography (HPLC). C2 is the main paralytic shellfish toxin (PST) detected in a clonal culture of A. tamarense, which is a common causative organism in cases of paralytic shellfish poisoning in Japan. This HPLC method employs post-column fluorescent derivatization (FL). Mobile phase, column size, flow rate, reagent concentrations, and lamp type for the fluorescent detector were all optimized for the detection of C2. With this improved methodology, we could measure 1 fmol of C2 with a signal to noise ratio (S/N)=2. Clonal heterogeneity within the toxic strain, which was maintained for 13 years after re-isolation from the original clonal culture, ranged from <1fmol to 700fmolcell-1. This report is the first to demonstrate definitively that PST content varies on a cell-by-cell basis in a clonal culture of a dinoflagellate that causes paralytic shellfish poisoning.
AB - We developed a methodology for analyzing the C-toxin (C2) content in single Alexandrium tamarense cells; this method was based on high performance liquid chromatography (HPLC). C2 is the main paralytic shellfish toxin (PST) detected in a clonal culture of A. tamarense, which is a common causative organism in cases of paralytic shellfish poisoning in Japan. This HPLC method employs post-column fluorescent derivatization (FL). Mobile phase, column size, flow rate, reagent concentrations, and lamp type for the fluorescent detector were all optimized for the detection of C2. With this improved methodology, we could measure 1 fmol of C2 with a signal to noise ratio (S/N)=2. Clonal heterogeneity within the toxic strain, which was maintained for 13 years after re-isolation from the original clonal culture, ranged from <1fmol to 700fmolcell-1. This report is the first to demonstrate definitively that PST content varies on a cell-by-cell basis in a clonal culture of a dinoflagellate that causes paralytic shellfish poisoning.
KW - Alexandrium tamarense
KW - Paralytic shellfish toxins
KW - Single-cell analysis
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U2 - 10.1016/j.hal.2013.02.005
DO - 10.1016/j.hal.2013.02.005
M3 - Article
AN - SCOPUS:84876314374
SN - 1568-9883
VL - 25
SP - 47
EP - 53
JO - Harmful Algae
JF - Harmful Algae
ER -