Method for detecting protein content in 2-keto-L-gulonic acid
A technology of protein content and gulon acid, which is applied in the field of chemical analysis, can solve the problems of inability to detect protein content, and achieve the effects of low detection equipment requirements, good reproducibility, and good time stability
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[0054] Example 1: Detection of protein content in sodium 2-keto-L-gulonate fermentation broth;
[0055] Centrifuge the fermentation broth of sodium 2-keto-L-gulonate at 2000r / min for 20min in production, transfer 2ml of the supernatant to a test tube, then add 2ml of 40% KOH solution, boil it in boiling water for 30 minutes, and wash it with deionized water Put into a 100ml volumetric flask, then add 4.5ml of 20% nitric acid solution and 10ml of pH 7 buffer solution, and make the volume constant with deionized water.
[0056] Pipette one part of 1mL deionized water and two parts of the test solution into three test tubes, add 5mL Coomassie Brilliant Blue reagent to the first two test tubes, add 5mL deionized water to the third test tube, and detect the deionized water at 595nm The absorbance of test tube 2 and test tube 2 are 0.037 and 0.058L / (g.cm), respectively. Adjust the absorbance of test tube 1 to 0. The absorbance of test tube 3 is 0.162L / (g.cm) within 30-60 minutes, which ...
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[0057] Example 2: Detection of protein content in 2-keto-L-gulonic sodium ultrafiltrate
[0058] Take 20ml of post ultrafiltrate to a 50ml volumetric flask, then add 10ml of pH 7 buffer solution, and make the volume constant with deionized water.
[0059] Pipette one part of 1mL deionized water and two parts of the test solution into three test tubes, add 5mL Coomassie Brilliant Blue reagent to the first two test tubes, add 5mL deionized water to the third test tube, and detect the deionized water at 595nm The absorbance of test tube 2 and test tube 2 are 0.037 and 0.046L / (g.cm), respectively. Adjust the absorbance of test tube 1 to 0. The absorbance of test tube 3 is 0.205L / (g.cm) within 30-60 minutes. The ultrafiltration can be obtained by calculation. The protein content in the liquid is 0.089mg / ml.
Example Embodiment
[0060] Example 3: Detection of protein content in 2-keto-L-gulonic acid dry product
[0061] Weigh 5g of 2-keto-L-gulonic acid dry product in production, dissolve in deionized water and transfer it to a 100ml volumetric flask, then add 6.5ml 20% KOH solution and 10ml pH 7 buffer solution, deionized water to a constant volume .
[0062] Pipette one part of 1mL deionized water and two parts of the test solution into three test tubes, add 5mL Coomassie Brilliant Blue reagent to the first two test tubes, add 5mL deionized water to the third test tube, and detect the deionized water at 595nm The absorbance of test tube 2 and test tube 2 are respectively 0.037 and 0.039L / (g.cm). Adjust the absorbance of test tube 1 to 0. The absorbance of test tube 3 is 0.115L / (g.cm) within 30 to 60 minutes, which can be calculated by formula 1. The protein content of 2-keto-L-gulonic acid dry product is 0.036%.
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