Determination of azide in biological fluids by use of electron paramagnetic resonance

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Abstract

A simple and sensitive method has been developed for the determination of azide ion (N3 - ) in biological fluids and beverages. The procedure was based on the formation of a ternary complex Cu(N3)2(4-methylpyridine)x in benzene, followed by its detection by electron paramagnetic resonance. The complex in benzene showed a characteristic four-peak hyperfine structure with a g-value of 2.115 at room temperature. Cu2+ reacted with N3 - most strongly among common metals found in biological fluids. Several anions and metal ions in biological fluids did not interfere with the determination of N3 - in the presence of large amounts of Cu2+ and oxidants. In the present method, N3 - at the concentration from 5 μM to 2 mM in 100 μl solution could be determined with the detection limit of 20 ng. The recoveries were more than 95 % for N3 - added to 100 μl of blood, urine, milk and beverages at 200 μM. Our method is recommendable because it takes less than 10 min to determine N3 - and the produced complex is quite stable.

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