Analytical method development for the determination of nitrate and nitrite contents in Anatolian honey

All living systems require a certain amount of nitrogen for the production of nutrients like proteins, nucleic acids, vitamins, and enzymes etc. Due to the contamination in environment and use of certain artificial fertilizers the amounts of nitrates and nitrites have so far increased in eatables. Besides artifically incorporated nitrogen compounds, naturally occuring nitrates in foods can be converted into other chemicals in the gastrointestinal system of the living body. Some of these newly formed compounds might show some lethal, injurious, carcinogenic, and mutagenic effects. To be able to acquire appropriate knowledge of such constituents and their outcomes, it is imperative nowadays to develop valid, competent, reasonable, sensitive, and fast methods for analysis of such ingredients. In general, nitrates and nitrites are inorganic anions form naturally during the nitrification process of nitrogen cycle. Naturally they occur in vegetables and drinking waters and even added as an additive into certain meat products. The toxicity of nitrates initiates from when it starts converting into nitrites. Nitrites are known to be bringing carcinogenic nitrosamines to the skin and also causing deadly blue baby syndrome in infants. In the presented research, it was aimed to apply a capillary electrophoresis method for the concurrent quantification of nitrate and nitrite anions in the honey samples. These anions in high concentrations, become toxic after chemical reactions, even if occur as an additive or naturally. Also, an overview of consequences of thier ingestion and their safe ingestion estimations issued by World Health Organization (WHO) have been presented in this work. The naturally occuring amounts of these anions in honey are very low. As honey is consumed frequently by individuals belonging to every age group, the analysis of nitrate and nitrite ions amounts is a significant food control issue. At this time, capillary electrophoresis (CE) analytical techniques are the most widely and frequently used methods used to separate and detect a huge number of compounds. From small ions to large molecules such as DNA can be identified and quantified with applied high voltage in CE. With its emerging separation power and wide range of modes, it has become convenient to inspect samples of complex matrices such as food products. A capillary zone electrophoresis (CZE) technique has been employed here which have the easiest, quickest and lowest observable limits and least number of sample preparation steps. In this work, a capillary electrophoresis-sample stacking method was applied. This method has successfully provided us meaningful results for the qualitative and quantitative determination of nitrate and nitrite anions. The conditions for this separation system included using an acidic separation medium due to which the speed of EOF was reduced. Thus, these anions having very high electrophoretic mobilities were detected. The optimized method of analysis is based on, for example, dividing the sample and buffer into zones, concentrating the sample zone, making an electric field difference in the zones and making a high volume of injection. As a result, the sensitivity of the method was increased about 30 times for both anions compared to low-volume injections. The method was successfully applied in the determination of nitrite and nitrate anions in honey samples of rhododendron, heather, oak, chestnut, pine, and lavender honeys.