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A method for one-time separation and detection of five compounds including adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, adenosine and deoxynucleotide
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A technology of adenosine triphosphate and adenosine diphosphate, which is applied in material separation, measuring devices, and analysis materials, can solve the problems of inability to separate and detect at one time, poor repeatability, and low sensitivity, and achieve reduced damage, good repeatability, and The effect of high sensitivity
Inactive Publication Date: 2021-03-12
浙江省农药检定管理所
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However, the existing technologies are all aimed at detecting and separating the three compounds of adenosine triphosphate, adenosine diphosphate and adenosine monophosphate, and cannot separate and detect adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, adenosine and deoxynucleotide at one time. Five compounds, and there are still problems of troublesome operation, low sensitivity and poor repeatability
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Embodiment 1
[0047] 1) Prepare the sample to be tested;
[0048] 2) The sample in step 1) is subjected to HPLC detection, and the detection conditions are:
[0065] Results The three adenosine phosphate peaks were relatively early and dense, and the five mixed standards were completely separated and detected within 40 minutes. The spectrum is shown in figure 2 .
Embodiment 3
[0067] 1) Prepare the sample to be tested;
[0068] 2) The sample in step 1) is subjected to HPLC detection, and the detection conditions are:
[0069] Equilibrate the column with 30mmol / l phosphate buffer solution with a pH value of 6.7, and the parameters are as follows:
[0075] The five mixed standards were successfully separated and detected within 40 minutes, see the spectrum image 3 .
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Abstract
A method for one-time separation and detection of five compounds including adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, adenosine and deoxynucleotides, comprising the following steps: 1) preparing a sample to be tested; The samples were detected by HPLC, the detection conditions were: the chromatographic column was an Inertsil ODS-3 column, the sample volume of the detection sample was 10 μl, the column temperature was 26±2°C, and the mobile phase was: gradient elution or 80% to 98% phosphate Mixture of buffer solution and 2%~20% methanol, the flow rate is 0.6~1.41mL / min; gradient elution: 95%~100% phosphate buffer and 0%~5% methanol in 0~10min solution; 85% to 95% phosphatebuffer solution or a mixture of water and 5% to 15% methanol during 11 to 40 minutes; the concentration of the phosphate buffer solution is 20 to 60mmol / l, and the pH value is 6.3 to 6.8 ; Compared with the prior art, the present invention has the characteristics of simple method, high sensitivity and good repeatability.
Description
technical field [0001] The invention belongs to the technical field of detection and analysis, and in particular relates to a method for one-time separation and detection of five compounds including adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, adenosine and deoxynucleotides. Background technique [0002] Adenosine triphosphate (ATP) is the direct source of energy required for all life activities of tissue cells in the living body. It is known as the "molecular currency" of intracellular energy. All diseases are highly targeted. ATP can be decomposed into adenosine diphosphate (ADP) and adenosine monophosphate (AMP), adenosine (Adenosine) is an important intermediate for the synthesis of ATP, 2-deoxynucleotide (2-AD) is the synthesis of DNA One of the important elements, the above five compounds coexist in some plants and fungi. In the analysis and detection of some crops, it is of great significance to separate and detect adenosine triphosphate, ...
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