A method for detecting neu5gc in fermented red meat products by fluorescence high performance liquid chromatography
A technology of high performance liquid chromatography and meat products, applied in the field of detection of Neu5Gc in fermented red meat products by fluorescence high performance liquid chromatography, can solve the problems of ineffective separation, affecting detection efficiency, and long detection time, so as to reduce sample detection time, Improve detection efficiency and lower cost
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Embodiment 1
[0031] A method for detecting Neu5Gc in fermented red meat products by fluorescence high performance liquid chromatography, comprising the following steps:
[0032] (1) Preparation of Neu5Gc standard solution
[0033] Accurately weigh 0.0825g Neu5Gc standard substance, dissolve it in a 250ml volumetric flask with ultrapure water, and set the volume to the mark, which is 1mmol / L Neu5Gc standard substance solution.
[0034] (2) Neu5Gc standard curve drawing
[0035] Draw 1mmol / L Neu5Gc standard solution to make 10, 25, 50, 100, 200, 300, 400μmol / L Neu5Gc standard solution, take 900μL of different concentrations of Neu5Gc standard solution, add 100μL of DMB derivative reagent respectively, and place in a water bath at 50℃ After derivatization in the dark for 2 hours, put it into the ice-water mixture to quickly cool it through a 0.22 film, and then inject the sample for detection. The standard concentration is taken as the horizontal axis, and the chromatographic peak area is ta...
Embodiment 2
[0049] Detection of Neu5G content in traditionally fermented bream meat for 30 days by fluorescence high performance liquid chromatography:
[0050] (1) Sample solution preparation
[0051] Select the lean meat part of sour bream meat that has been traditionally fermented for 30 days, take 1 g of the middle meat sample below 1 cm, add 10 ml of 30% saturated ammonium sulfate solution to homogenize to precipitate protein, and freeze-dry to remove ammonium sulfate. In order to free Neu5Gc, add 10ml of 2mol / L glacial acetic acid to the freeze-dried sample and hydrolyze it in a water bath at 80°C for 3h, centrifuge at a speed of 12000r / min for 10min to get the supernatant, filter it through a 0.45μm membrane, and then freeze it again Dry to remove glacial acetic acid. The lyophilized powder was dissolved in 1ml of 0.1mol / L NaOH, bathed in water at 37°C for 30min, and filtered with a 0.45μm membrane to obtain a sample solution.
[0052] (2) Pre-column derivatives
[0053] Accurat...
Embodiment 3
[0063] Detection of Neu5G content in salami sausage by fluorescence high performance liquid chromatography:
[0064] (1) Sample solution preparation
[0065] Strip off the casing, accurately weigh 1g of minced meat and homogenize with 10ml of 30% saturated ammonium sulfate solution to precipitate protein, and freeze-dry to remove ammonium sulfate. In order to free Neu5Gc, add 10ml of 2mol / L glacial acetic acid to the freeze-dried sample and hydrolyze it in a water bath at 80°C for 3h, centrifuge at a speed of 12000r / min for 10min to get the supernatant, filter it through a 0.45μm membrane, and then freeze it again Dry to remove glacial acetic acid. The lyophilized powder was dissolved in 1ml of 0.1mol / L NaOH, bathed in water at 37°C for 30min, and filtered with a 0.45μm membrane to obtain a sample solution.
[0066] (2) Pre-column derivatives
[0067] Accurately draw 900 μL of sample solution, add 100 μL DMB derivatization reagent, derivatize in a water bath at 50°C in the ...
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