A Highly Sensitive and Selective Spectrofluorimetric Method for the Determination of Nitrite in Food Products
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A Highly Sensitive and Selective Spectrofluorimetric Method for the Determination of Nitrite in Food Products Qiuhua Wang 1,2 & Haiwei Huang 1 & Baoming Ning 1 & Minfeng Li 2 & Lan He 1
Received: 25 March 2015 / Accepted: 28 August 2015 # Springer Science+Business Media New York 2015
Abstract This paper describes a simple, sensitive, and selective spectrofluorimetric method to determine trace amount of nitrite in food products using 4-amino-5methylamino-2′,7′-difluorofluorescein diacetate (DAFFM DA) as a fluorescent probe. The reaction of DAFFM DA with nitrite in acidic medium resulted in triazolofluorescein (DAF-FM T), a highly fluorescent reagent in neutral medium. The fluorescence enhancement was proportional to nitrite concentration in the range of 5.0×10−8 to 1.5×10−6 mol L−1 with a detection limit of 3.3×10−8 mol L−1 (S/N=3). The proposed method has been applied to the determination of nitrite in real food samples, with relative standard deviation (RSD) (n = 6) less than 5.1 % and recoveries in the range of 86.4~102.9 %.
Keywords Nitrite . Spectrofluorimetric . Determination . 4-Amino-5-methylamino-2′,7′-difluorofluorescein diacetate (DAF-FM DA)
* Minfeng Li [email protected] * Lan He [email protected] Qiuhua Wang [email protected] 1
National Institutes for Food and Drug Control, Beijing 100050, China
2
College of Chemistry, Beijing Normal University, Beijing 100875, China
Introduction Nitrite, a chemically active substance, exists widely in the environment and food products. In order to suppress the propagation of Clostridium botulinum and to improve the flavor of meat, it has been widely used in meat preservation and processing (Li et al. 2003). Excessive amount of nitrite in food can be fatal, especially for pregnant women and infants. It can interfere with the body’s oxygen delivery system, decreasing the ability of hemoglobin to carry oxygen (Seike et al. 2004; Cotton and Wilkinson 1988). It can also react with the secondary amines and amides in the stomach to form carcinogenic N-nitrosamines (Burden 1961). These hazards of nitrite make its determination and quantification of great interest. A large number of analytical methods for the detection of nitrite have been developed, including spectrophotometric (Burakham et al. 2004; Nagaraja et al. 2010; Senra-Ferreiro et al. 2010; Tsikas 2007), electrochemical (Menart et al. 2015; Azad et al. 2014; Ramdane-Terbouche et al. 2014; Wang et al. 2013), capillary electrophoresis (Della Betta et al. 2014; Troška et al. 2013; Merusi et al. 2010; Wang et al. 2012b; Tanaka et al. 2004), chromatographic (Wang et al. 2012a; He et al. 2011; Akyüz and Ata 2009; Pagliano et al. 2014), and spectrofluorimetric methods (Lee and Field 1984; Guo et al. 2013; Liu et al. 2009; Huang et al. 2006; Wang et al. 2000). Each of them has its own merits, but each method also offers some drawbacks. Spectrophotometry is the most widely used method for the determination of nitrite. The most common approach to the spectrophotometric detection of nitrite is the Griess method which
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