Determination method of free nucleic acid hydrolyzate in protein products and evaluation method of production process on nucleotide damage
A technology of free nucleic acid and determination method, which is applied in the field of quality control of single-cell protein production and determination of nucleic acid hydrolyzate, which can solve the problems of animal toxicity, yeast product quality decline, and function reduction.
Active Publication Date: 2017-01-11
江苏征泰饲料有限公司
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This will result in a decrease in the quality of the yeast product, and a reduction in the amount of nucleotides provided, which in turn can reduce the immune function of the animal. At the same time, the increased bases such as adenine will cause toxicity to animals.
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[0030] Following are the implementation steps of the present invention:
[0031] 1 Reagents and solutions
[0032] 1.1 Unless otherwise specified, the reagents used in this method are of analytical grade, and the water is deionized water, which complies with the provisions of GB / T 6682 for secondary water.
[0033] 1.2 Potassium dihydrogen phosphate: analytically pure.
[0034] 1.3 Potassium hydroxide: analytically pure.
[0035] 1.4 Methanol: chromatographically pure.
[0036] 1.5 Acetonitrile: chromatographically pure.
[0037] 1.6 20% w / v potassium hydroxide solution: weigh 100 grams of potassium hydroxide, dissolve it in 500 ml of water, and store it in a volumetric flask.
[0038] 1.7 Mobile phase A: 0.1M potassium dihydrogen phosphate solution, weigh 27.2 grams of potassium dihydrogen phosphate, dissolve it in 1000 ml of water, add 1.0 ml of 20% potassium hydroxide solution, add water to make up to 2 liters. 0.45um water membrane filter and then reserve.
[0039] 1...
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Abstract
The invention discloses a method for determining a free nucleic acid hydrolysate in protein products. The method comprise the following steps: (1) respectively preparing a nucleotide mixed standard solution and a bases mixed standard solution; (2) preparing a solution of a sample to be tested, and using the nucleotide mixed standard solution and the base mixed standard solution described in step 1 as the reference solution, and employing high performance liquid chromatography through washing peak area method comparison to obtain the content of free nucleotides and free base in the sample. According to the invention, high performance liquid chromatography is used for determining 5 kinds of bases in a single-cell protein raw material (cytosine, uracil, guanine, thymine and adenine), 5 kinds of nucleotides (cytidine monophosphate, uridylic acid, guanylic acid, inosinic acid and adenylic acid), and 5 kinds of nucleosides (cytidine, uridine, inosine, guanosine and adenosine); and then the destroy degree on nucleotide of the production process is determined according to t the relationships between the bases and nucleotides, and between the bases and nucleosides.
Description
technical field [0001] The invention belongs to the technical field of nucleic acid hydrolyzate measurement, in particular to the field of single-cell protein production quality control. Background technique [0002] Single cell protein (SCP for short), also known as microbial protein or bacterial protein, uses industrial wastewater, waste gas, natural gas, petroleum alkanes, agricultural and sideline processing products, and organic waste as culture media to cultivate yeast, non-pathogenic Bacteria, micro-bacteria, fungi and other single-celled organisms, which are then purified and dried, are important protein sources for the food industry and feed industry. [0003] Single-cell protein products are rich in nucleotides, and yeast products on the market are the main nucleotide providers for the feed industry. Due to the characteristics of the process, products such as autolyzed yeast, enzymatic yeast, and hydrolyzed yeast contain more free nucleic acid hydrolysates. Studie...
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IPC IPC(8): G01N30/02
Inventor 许建刚
Owner 江苏征泰饲料有限公司