Detection method of tyrosine in food
A detection method and tyrosine technology, applied in the field of amino acid detection in food, can solve the problems of inaccurate detection results, expensive derivatives, and low sensitivity.
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Embodiment 1
[0108] Weigh the tyrosine of the standard substance / reference substance (purity ≥ 99%), dissolve it with hydrochloric acid (0.1mol / L) to prepare a test solution with a concentration of 0.5, 1.0, 2.0, 5.0, 10.0 μg / mL, and use 0.45 After filtering through a μm organic filter membrane, the samples were analyzed by high performance liquid chromatography.
[0109] The solution to be tested is subjected to high-performance liquid chromatography analysis, and the chromatographic conditions are:
[0110] Analytical column: C-18 column;
[0111] Mobile phase: organic phase A: acetonitrile; aqueous phase B: sodium acetate buffer; the pH of the sodium acetate buffer is 4.2.
[0112] Flow rate: 1mL / min;
[0113] Elution method: Gradient elution, 0min, A:B=45:55, 10min, A:B=65:35, 12min, A:B=75:25, 18.5min, A:B=45:55;
[0114] Detection wavelength: 252nm;
[0115] Detection temperature: 40°C;
[0116] Injection volume: 20μL;
[0117] Detection time: 35min;
Embodiment 2
[0122] Take 3.3441g sample soybean paste, add 100mL, 0.1mol / L hydrochloric acid into the Erlenmeyer flask for 30min sonication, after cooling to room temperature, dilute to 50mL with 0.1mol / L hydrochloric acid, centrifuge at 3000rpm for 5min, then buffer with sodium carbonate Adjust the pH value of the solution to be neutral, then dilute with water to the concentration range of the standard curve and make the volume to 50mL. Under light-shielded conditions, derivatize 1.00 mL of the filtrate with 1.00 mL, 1.5 mg / mL dansyl chloride solution at room temperature for 2 h, then add 0.1 mL, 20 mg / L methylamine hydrochloride solution and vortex to terminate the reaction , The reaction product was separated after standing for 30 min, and the supernatant was filtered through a 0.45 μm organic filter membrane and then analyzed by high performance liquid chromatography.
[0123] Adopt the same high performance liquid chromatography detection condition as embodiment 1 to detect the liquid...
Embodiment 3
[0137] Using the same steps as in Example 2, the only difference is that the sample size is replaced by 3.4069g bean paste. The obtained HPLC chromatogram is as follows Figure 5 as shown, Figure 5 1 is tyrosine. Figure 5 The peak area of the middle peak 1 is 563.96954, and the concentration of tyrosine in the food is calculated by the external standard method with the standard equation.
[0138] Calculate the content of tyrosine in the sample soybean paste according to the standard equation provided by Example 1, and the calculation formula is as follows:
[0139] expressed in milligrams per hectogram
[0140]
[0141] In the formula:
[0142] X - the content of tyrosine in the sample, in milligrams per hectogram (mg / 100g);
[0143] m——weigh the sample amount, the unit is gram (g);
[0144] C——The measured concentration of tyrosine, in micrograms per milliliter (μg / mL);
[0145] C 0 ——The measured concentration of tyrosine in the blank sample, in micrograms per...
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