Abstract
The present work reports a rapid reversed-phase high-performance liquid chromatography (RP-HPLC) method for the determination of seven volatile N-nitrosamines namely N-nitrosodimethylamine (NDMA), N-nitrosomethylethylamine (NMEA), N-nitrosodiethylamine (NDEA), N-nitrosodiproylamine (NDPA), N-nitrosodi butylamine (NDBA), N-nitrosopiperidine (NPIP) and N-nitrosopyrrolidine (NPYR) for monitoring food safety. A strategic experimental approach was implemented for the method development. The desired chromatographic separation was achieved on a Symmetry C18 (4.6 × 150 mm, 5 μm) column using gradient elution. The optimized mobile phase consisted of the 10 mM ammonium hydroxide pH = 8.9 and acetonitrile. The eluted compounds were monitored at 231 nm wavelength using spectrophotometric detector. The developed method separated seven compounds from each other within a run time of 10 min. The method is effective for the determination of presence of these carcinogenic compounds. The average extraction recovery of seven nitrosamines was found 84.5%; the precision of method was found less than 2.7% and accuracy was found between 95%-102.5%. The assay could be applied in food monitoring safety.
Highlights
Nitrosamines form a large group of genotoxic chemical carcinogens which occur in the human diet and otherHow to cite this paper: Al-Kaseem, M., et al (2014) A Rapid, Validated reversed-phase high-performance liquid chromatography (RP-HPLC) Method for the Determination of Seven Volatile N-Nitrosamines in Meat
In order to judge the suitability of method for determining the N-nitrosodimethylamine (NDMA), N-nitrosomethylethylamine (NMEA), N-nitrosodiethylamine (NDEA), N nitrosopyrrolidine (NPYR), N-nitrosodinpropylamine (NDPA), N-nitrosopiperidine (NPIP) and N-nitrosodi-n-butylamine (NDBA) traces in meat, the method was validated as per the International Conference on Harmonization (ICH) guideline for specificity, limit of detection, limit of quantification, linearity, accuracy and precision [33]
Meat samples were treated with 1 N NaOH for 10 min in autoclaved conditions and analyzed by HPLC-UV and the results of 25 samples of different meat products which examined for the evaluation of the analytical method shows the presence of some volatile nitrosamine in all different products, and the absence of NMEA NDPA, and nitrosodi butylamine (NDBA) in 5 sample tested which may related to the presence of these three nitrosamines in low amounts which may below under the quantitation limit of our procedure, or the real absence of these nitrosamine in the sample tested
Summary
Nitrosamines form a large group of genotoxic chemical carcinogens which occur in the human diet and otherHow to cite this paper: Al-Kaseem, M., et al (2014) A Rapid, Validated RP-HPLC Method for the Determination of Seven Volatile N-Nitrosamines in Meat. Nitrosamines are produced from nitrites and secondary amines, which often occur in the form of proteins [5] Their formation can occur only under certain conditions, including strongly acidic conditions such as that of the human stomach [6]. The literature reveals a very few methods which have been published for quantification of N-nitrosamines in meat products using HPLC with direct UV detection and all the reported methods were adopting pre-column or post-column photolysis units or additional derivatization reaction to enhance the light absorbance or to get fluorescent derivatives [3] [8] [20]. The N-nitroso bond is cleaved by UV photolysis with the formation of nitrite ion the nitrite is diazotized with sulfanilamide in an acid medium (according to the Griess reaction), and is coupled with N-(1-naphthyl) ethylenediamine dihydrochloride (NED) to form a purple-coloured azo dye that is quantitatively determined spectrophotometrically at a maximum wavelength, λmax, of 540 nm [1] [32]
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