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Method for simultaneously determining five nucleic acid bases in dried yeast

A nucleic acid base, yeast technology, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of poor peak shape and response value, unsuitable for batch testing, complicated testing process, etc., to achieve simple and fast operation and excellent repeatability and reproducible, easy-to-equip results

Pending Publication Date: 2022-04-29
珠海天祥粤澳质量技术服务有限公司
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Problems solved by technology

[0003] The papers of the Second National Fermentation Engineering Symposium published the extraction of yeast RNA from yeast powder, and the use of high-performance liquid chromatography for high-sensitivity analysis to simultaneously separate cytosine, uracil, guanine and Research on the four alkaloids of adenine, but the research is old, the test process is complicated, it is not suitable for batch testing, and the peak shape and response value are poor

Method used

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  • Method for simultaneously determining five nucleic acid bases in dried yeast
  • Method for simultaneously determining five nucleic acid bases in dried yeast
  • Method for simultaneously determining five nucleic acid bases in dried yeast

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preparation example Construction

[0075] S23, the preparation of standard working solution: take by weighing 0.01-0.02g of the standard substance of five kinds of nucleic acid bases respectively in the volumetric flask of 10mL, use methanol to constant volume, obtain standard stock solution; Dilutions were performed to obtain standard working solutions.

[0076] As described in the above step S23, the preparation of the standard working solution: respectively weigh 0.01-0.02 g of the standard products of the five nucleic acid bases in a 10 mL volumetric flask, and dilute to the volume with methanol to obtain a standard stock solution; The stock solution is diluted step by step with methanol to obtain a standard working solution, wherein the five nucleic acid bases include cytosine, uracil, guanine, adenine and hypoxanthine, and the five nucleic acid bases including cytosine , 0.01-0.02 g of standard substances of uracil, guanine, adenine and hypoxanthine in a 10mL volumetric flask, and use chromatographic grad...

Embodiment 1

[0115] Weigh 0.03g of sample into a 25mL stoppered glass graduated tube, add 2mL of pure perchloric acid, vortex, heat in a water bath at 90°C for 65min, and vortex once every 15min. Cool, then add 5mL of 23.7g / 100mL ammonium dihydrogen phosphate solution, add water to 14mL, shake well and filter, and measure with HPLC-DAD; weigh 0.01g of standard substances of five nucleic acid bases in 10mL capacity Dilute the standard stock solution with methanol step by step to obtain a standard working solution, which is detected by HPLC-DAD.

Embodiment 2

[0117] Weigh 0.05g of sample into a 25mL stoppered glass graduated tube, add 3mL of pure perchloric acid, vortex, place in a water bath at 98°C for 80min, shake once every 15min, cool in an ice-water bath after taking it out, add 6mL of 23.7g / 100mL Ammonium dihydrogen phosphate solution, add water to 14mL, shake well and filter, and measure with HPLC-DAD; weigh 0.01g of standard substances of five kinds of nucleic acid bases in 10mL volumetric flasks, dilute to volume with methanol to obtain standard Stock solution; the standard stock solution was diluted step by step with methanol respectively to obtain a standard working solution, which was detected by HPLC-DAD.

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Abstract

The invention provides a method for simultaneously determining five nucleic acid bases in dried yeast, the five nucleic acid bases comprise cytosine, uracil, guanine, adenine and hypoxanthine, and the method comprises the following steps: sampling; pre-treating and measuring the pre-treated sample solution; wherein the pretreatment step comprises the following steps: extraction: mixing a sample with pure perchloric acid, and heating in a water bath at a first specified temperature for a first specified time; purification: adding a second specified amount of ammonium dihydrogen phosphate solution, adding water to a third specified amount, and filtering to obtain a pretreated sample solution; wherein the step of measuring the pretreated sample solution comprises the following steps: detection: detecting the pretreated sample solution by using HPLC-DAD (High Performance Liquid Chromatography-Digital Amplification Detection). According to the present invention, cytosine, uracil, guanine, adenine and hypoxanthine in the dried yeast can be simultaneously determined, the method has advantages of excellent repeatability, excellent reproducibility, simple and rapid operation, efficient separation, stable base line, easily available equipment, and high generalizability.

Description

technical field [0001] The application relates to the technical field of nucleic acid base detection, in particular to a method for simultaneously determining five nucleic acid bases in dried yeast. Background technique [0002] Dried yeast is specially cultivated fresh yeast that still maintains strong fermenting ability after being squeezed, dried and dehydrated. Yeast nucleic acid is ribonucleic acid extracted from natural yeast. Through modern biological extraction technology, the nucleic acid in yeast is separated and purified, so that the purity of nucleic acid can reach more than 85%. Yeast nucleic acid is mainly used in medicine, health food and infant food. With the development of modern biological extraction technology, the application of yeast nucleic acid It tends to be widespread, especially in new anti-cancer drugs, health care products and infant food, and has made many breakthroughs. The development of yeast nucleic acid industry will become a bright spot in...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/027
Inventor 赖蕾李庚陈祎明黎冠
Owner 珠海天祥粤澳质量技术服务有限公司
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