Determination of Acid Orange II and Acidic Golden in Soy Products by High Performance Liquid Chromatography
The understanding is still superficial. This paper discusses the determination of acid orange II and acid golden by high performance liquid chromatography . This method has yet to be further improved.
Acid Orange Ⅱ soybean products crushed, [Abstract] This article discusses the measurement method of high performance liquid chromatography soy Acid Orange and Acid Orange Ⅱ content. The sample extract was extracted and filtered, and the sample extract was determined by high performance liquid chromatography. The mobile phase was 0.02 mol/L = 75+25 methanol ammonium chloride at a detection wavelength of 450 nm, and the flow rate was 1.0 ml/min. The column was Symmetry C18250 mm 4.6 mmid 5 μm.
[Key words] soy products; acid golden II; high performance liquid chromatography
Coloring agents for wooden furniture, wool, fabrics, etc., acid orange II and acid golden in soy products are used in tissue dyeing in medicine. However, illegal traders use their bright colors, stable color, and low price to illegally add to soy products, which seriously endangers the health of consumers. There is no corresponding test method in the country. This paper has initially established high performance liquid chromatography. The method for measuring acid orange II and acid golden color can quickly, accurately and simply detect acid orange II and acid golden, and can meet the needs of food inspection.
1 test local
1.1 Instruments and reagents
1.1.1 Instruments
Model 2487 UV detector, high performance liquid chromatography including Waters 1525 pump. Breez chromatography workstation and manual injector.
1.1.2 Standard Products
Acid Orange II (purity 86% and acid golden (industrial purity 90%)
1.1.3 Methanol
Chromatographically pure.
1.1.4 ultra pure water
High purity water supplied by the AKUPIII20 ultrapure water machine .
pH = 4.7 1.1.50.02 mol/L ammonium acetate solution.
1.1.6 alkaline extract
Anhydrous ethanol: ammonia: water = 70:20:10
1.2 chromatographic conditions
Detection wavelength 450nm injection volume 20μl liquid chromatography column is SymmetryC18250mm4.6mmid5μm mobile phase methanol: 0.02mol/L ammonium acetate solution=75+25 solution flow rate 1.0ml/min column temperature is normal temperature.
1.3 Preparation of standard products
The series was prepared to a concentration of 00.01 μg/ml 0.05 μg/ml 0.10 μg/ml 0.50 μg/ml 1.00 μg/ml. Draw a proper amount of 1.00 mg/ml of the mixed standard storage solution.
1.4 sample preparation
Add 50 ml of pH=6 water and mix well, and weigh the mixed pulverized sample from 5.00 g to 10.00 g in a 100 ml beaker. Filtration, adding 1.00g of polyamide powder to the filtrate, filtering by G3 funnel, rinsing 2 times to 3 times with pH=6 water, desorbing the alkaline extract 2 times~3 times, collecting desorption solution 5ml each time, draining in water bath, will The residue was dissolved in distilled water, made up to 5 ml and then passed through a 0.45 μm filter. The specification and sample filtrate were injected into the HPLC system and determined according to the chromatographic conditions described above. Characterize the storage time and quantify the peak area.
2 results
2.1 wavelength selection
An ultraviolet scan is performed and the standard solution is diluted with the mobile phase. It can be seen from the scanning spectrum that acid II and acid gold have a large absorption at 450 nm, so I chose 450 nm as the detection wavelength.
2.2 Working curve and detection limit
The results show that the chromatographic peak area Y is linearly regressed with the working fluid content X. The relationship between acid orange II and acid golden in the range of 0.11μg/ml~1.00μg/ml (Linearrang is good. Regression equation (Regressionequ linear range, correlation coefficient (Correlationcoeffici and detection limit (DetectionlimitS/N=3 see Table 1 Table 1 specification) Curve and detection limit items.
3 Discussion
SICHUAN UNIWELL BIOTECHNOLOGY CO.,LTD. , https://www.uniwellbio.com