Study On The Analytieal Methods Of 10 Kinds Of Pigments By HPLC And HPLC-MS/MS

The yellow color pigments was used in the foodstuff frequently, especially synthetic pigment sunset yellow and tartrazine. Much research has been done on the sunset yellow, and tartrazine, and basic orange, acid orange which is prohibited by national standards respectively, the synthetic pigments which is allowed was often have the excessive or beyond add; the prohibited pigments is often added secretly. At present, the detection of non-food coloring didn’t have a comprehensive national standards, therefore, the analysis on the pigments it have certain deficiencies in the detection. Development of a more simple and effective sample preparation methods, and establish a high performance liquid chromatography and mass spectrometry method can simultaneously detect a variety of synthetic pigments and non-food coloring is more urgently.In this issue selected ten kinds of yellow pigment contain one kind of natural pigments(curcumin), three kinds of allowed synthetic pigments(tartrazine, sunset yellow, quinoline yellow), and six kinds of common non-food yellow pigment(Metanil yellow, Auramine O, Basic Orange 2, Basic Orange 21, Basic Orange 22, Acid Orange Ⅱ), were established HPLC method for qualitative and quantitative detection, HPLC-MS/MS as a qualitative confirmation of 10 kinds of pigment mass spectrometry for qualitative analysis to ensure the accuracy of the results.The main contents are as follows:Ananalysis method for the determination of ten colorants in drinks, candy, cakes three products by high Performance liquid chromatography-diode array detector was developed. The samples was ectracted with methanol as extraction solvent, establishing optimum pretreatment conditions after processing optimization; select Macherey- Nagel C18 column(4.6 × 250 mm, 5 μm); injection volume was 20 μL; column temperature was 25 ℃; the flow rate was 1.0 m L/min; detection wavelength was 435 nm; mobile phase: methanol(A), 0.02 mol/L ammonium acetate solution(B), acetonitrile(C) as the mobile phase gradient. Under optimum chromatographic conditions optimized, 10 kinds of pigment’s the standard curve was linear in the range of 0.25-160.0 μg / m L, the correlation coefficient were more than 0.994. The limits of quantification varied from 0.125 to 0.25 μg/m L, the recoveries of drinks were 88.2 to 110.8%, the recoveries of candies were 85.8 to 109.9%, the recoveries of pastry were 85.1 to 109.2%. This method has reproducibility, sensitivity and accuracy.To establish a HPLC-MS / MS method for qualitative analysis the ten kinds of colors, the mobile phase selection was(0.1% formic acid + 0.005 mol/L ammonium formate) solution: acetonitrile = 70: 30(V/V), positive ion mode(ESI+), detection curcumin, Auramine O, Basic Orange 2, Basic Orange 21, Basic Orange 22; mobile phase selection was(0.01 mol/L ammonium formate) solution: acetonitrile = 70: 30(V/V), negative ion detection mode(ESI-), detection between acidic amine yellow, tartrazine, sunset yellow, quinoline yellow, Acid Orange Ⅱ.The results shows the method is suitable for the qualitative and quantitative analysis the ten kinds of pigments in drinks, candies and pastries, which has pre-processing step is simple, good extraction effect, high sensitivity, low detection limit and quantitative accurate. It can provide a rapid analysis method to detect whether the synthesis yellow pigments have the excessive or beyond add; at the same time provide the method whether the prohibited pigments was added.