Determination of Thiocyanate Anion by High-Performance Liquid Chromatography with Fluorimetric Detection

Abstract

A simple and sensitive high-performance liquid chromatographic (HPLC) method was established for the trace determination of thiocyanate anion as a fluorogenic derivative. The method is based on the chemical derivatization of thiocyanate anion with 3-bromomethyl-7-methoxy-1,4-benzoxazin-2-one. The resulting derivative was separated by a Nova-Pak C18 reversed-phase column. Optimization conditions for the derivatization of thiocyanate anion were investigated by HPLC with fluorimetric detection. The linear range for the quantitation of thiocyanate anion was 1-0.05 nmol in 0.1 mL of sample; the detection limit (with a signal-to-noise ratio of 5) of a 20-microL injected aliquot was approximately 3.3 +/- 1.2 fmol. Application of the method to the analysis of thiocyanate anion in saliva and plasma proved to be feasible.

The Analysis of Cyanide and its Breakdown Products in Biological Samples

“…Although non-chromatographic fluorescence methods for the determination of thiocyanate, ATCA, or cyanide-protein adducts have not been published, the possibility exists to use such methods for ATCA and thiocyanate. For instance, 3-bromomethyl-7methoxy-1,4-benzoxazin-2-one ( Figure 4) and coumarin dyes have been used effectively for the HPLC-fluorometric determination of thiocyanate (217) and ATCA (92), respectively.…”

“…Thiocyanate is weakly spectrophotometrically active, and therefore is often derivatized with a strong absorber or fluorophore prior to analysis. For example, 3-bromomethyl-7-methoxy-1,4-benzoxazin-2-one has been effectively used (Figure 4) for the fluorometric determination of thiocyanate by HPLC (217). Others have also used variations of the König reaction to produce strong spectrophotometric absorption (157,162).…”

Thiocyanate Ion Selective Solid Contact Electrode Based on Mn Complex of N,N'-BIs-(4-Phenylazosalicylidene)-O-Phenylene Diamine Ionophore

“…Therefore, an accurate, simple, and rapid method for the determination of thiocyanate is significant in medicine and in the life sciences [4]. Several methods for the determination of thiocyanate ion such as spectrophotometry [5][6][7][8][9], chromatography [10][11][12][13][14][15], polarography [16], capillary zone electrophoresis [17], amperometry [18], potentiometric methods [19][20][21][22][23][24][25][26][27][28][29][30][31][32][33][34][35][36][37][38] etc., have been reported previously. Among the various methods, the thiocyanate ion selective electrodes (ISEs) are useful since it provides high sensitivity and a wide dynamic range.…”

Ion chromatographic quantification of cyanate in urea solutions: estimation of the efficiency of cyanate scavengers for use in recombinant protein manufacturing

“…HPLC methods involving cyanate modification to enhance detectability include use of thionitrobenzoic acid derivatization of cyanate with UV detection [25]. Several HPLC methods exist for analyzing thiocyanates including fluorescent derivatization or complexation analysis of cyanate-like compounds in body fluids clarified by precipitation and ion chromatography (IC) with amperometric detection [26][27][28][29]. A method combining capillary electrophoresis with indirect fluorescence detection for cyanate estimation in wastewater has also been reported, using fluorescein as the fluorophore [30].…”