Detection of Selenocyanate in Biological Samples by HPLC with Fluorescence Detection Using König Reaction

Ryu Mochizuki,a Kyohei Higashi,b Yusuke Okamoto,a Hiroko Abe,c Hirotaro Iwase,c and To s h i h i ko Toi d a

a Graduate School of Pharmaceutical Sciences, Chiba University; 1–8–1 Inohana, Chuo-ku, Chiba 260–8675, Japan

b Faculty of Pharmaceutical Sciences, Tokyo University of Science; 2641 Yamazaki, Noda, Chiba 278–8510, Japan

c Graduate School of Medicine, Chiba University; 1–8–1 Inohana, Chuo-ku, Chiba 260–8670, Japan.


Chem. Pharm. Bull.67, 884–887 (2019). Received March 28, 2019; accepted May 22, 2019


Abstract: We developed a simple and sensitive HPLC method for the determination of selenocyanate (SeCN­). The König reaction, which is generally used for the determination of cyanide and thiocyanate, was applied for the post-column detection, and using barbituric acid as a fluorogenic reagent made it possible to detect SeCN­ with high sensitivity. The limits of detection (LOD) and quantification (LOQ) were 73.5 fmol and 245.1 fmol, respectively. Subsequently, the amounts of SeCN­ in human blood and in cultured cell samples were analyzed, and no SeCN­ was detected in human whole blood. Interestingly, we have found that some of the spiked SeCN­ decomposed to cyanide in human whole blood. Ascorbic acid suppressed the decomposition of SeCN­ to cyanide by reducing the ferric ion, which is typically involved in SeCN­ decomposition. Then, SeCN­ was detected in cultured HEK293 cells exposed to selenite. The established HPLC method with fluo-rescence detection of SeCN­ is useful for investigating small amounts of SeCN­ in biological samples.


Scherzo SS-C18