CN111537461A - Method for detecting adenine and guanine in solution by using boron cluster nanogold - Google Patents
Method for detecting adenine and guanine in solution by using boron cluster nanogold Download PDFInfo
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Abstract
The invention is suitable for the field of nano-gold identification and detection, and provides a method for detecting adenine and guanine in a solution by using boron cluster nano-gold. Preparing boron cluster reduced nanogold according to a conventional process, and diluting the boron cluster reduced nanogold to obtain a diluted solution; then, adjusting the pH value of the diluted solution by using acid, and adding a sample aqueous solution to realize the specific recognition of adenine; if no adenine exists in the system, acid is continuously added to adjust the pH value of the solution, so that the specific recognition of guanine can be realized; finally, the obtained nano-gold sample solution is subjected to ultraviolet visible absorption spectrum scanning according to nano-gold A650/A525The signal of (2) is strong or weak, and the concentration of adenine or guanine can be measured. Compared with the traditional detection method, the method has the advantages of convenience, rapidness, low detection cost, less raw material loss, low instrument requirement and the like.
Description
Technical Field
The invention belongs to the field of nano-gold identification and detection, and particularly relates to a method for detecting adenine and guanine in a solution by using boron cluster nano-gold.
Background
Purines are important contributors to human health and play a vital role in supplying energy, composing coenzymes, regulating metabolism, and the like. Vitamin for adenineB4DNA and RNA, which promote the proliferation of leukocytes, are frequently used for the treatment of leukopenia and acute granulocytopenia, and are also used for the production of vitamin B4Drugs such as plant growth hormone, and biochemical research. Guanine is an organic base widely existing in animals and plants, is also an important base for forming nucleic acid, and is often applied to preparation of intermediates of antiviral drug acyclovir, preparation of drugs such as caffeine and the like, and biochemical research.
Adenine and guanine can be synthesized by human body, and food contains a large amount of purine compounds. However, the more purines that are absorbed by the body is not as good. Purine is catabolized in the human body to uric acid, which is excreted with urine. If too much purine is taken in human body, purine metabolic disorder and uric acid excretion disorder are often caused, so that hyperuricemia, namely gout which we often say, is caused. Therefore, gout patients should especially control the intake of high purine, and simple determination of purine content in food and medicine and the like which they are exposed to is important.
The nano-gold colorimetric method is a rapid and convenient detection method based on the phenomenon which can be observed by naked eyes. When the gold nano particles are in a dispersion state in the solution, the solution is wine red, and when the gold nano particles are in an agglomeration state in the solution, the solution is blue purple, so that the nano gold colorimetric method can be conveniently applied to the qualitative analysis of a sample. Meanwhile, the color change can bring the displacement of the maximum absorption peak on the ultraviolet visible absorption spectrum (from about 525nm to about 650 nm), and the absorbance ratio of the sample near 650nm to about 525nm is in direct proportion to the concentration of the substance to be detected in the sample in a certain range, so that a foundation is provided for the quantitative analysis of the sample by using a nanogold colorimetric method. However, the application of the nanogold colorimetric method usually requires that some recognition groups capable of interacting with the substance to be detected are connected on the surface of the nanogold, and this process usually brings about a complex modification process and waste of raw materials.
Disclosure of Invention
In view of the above problems, the present invention aims to provide a method for detecting adenine and guanine in a solution by using boron cluster nanogold, and aims to solve the technical problems of complicated operation, high detection cost, high raw material loss, high instrument requirement and the like of the existing detection method.
The method comprises the following steps:
step S1: preparing boron cluster reduced nanogold according to a conventional process, and diluting the boron cluster reduced nanogold to obtain a diluted solution;
step S2: adjusting the pH value of the diluted solution by using acid, and adding a sample aqueous solution to realize the specific recognition of adenine;
step S3: if no adenine exists in the system, acid is continuously added to adjust the pH value of the solution, so that the specific recognition of guanine can be realized;
step S4: performing ultraviolet visible absorption spectrum scanning on the obtained nano-gold sample solution according to nano-gold A650/A525The signal of (2) is strong or weak, and the concentration of adenine or guanine can be measured.
Preferably, in step S1, Cs is used2B12H12As reducing agent and stabilizer, Cs is added in a molar ratio of 1:12B12H12Adding into chloroauric acid solution, stirring at 25 deg.C for 30min to obtain boron cluster reduced nanogold.
Preferably, Cs2B12H12And chloroauric acid in the solution at a concentration of 0.4mM each, and the solvent was water.
Preferably, in step S2, the pH of the solution is in the range of 2.0-2.5, and the concentration of the sample aqueous solution is 0-200. mu.M.
Preferably, in step S3, the pH of the solution is in the range of 1.8-2.0.
The invention provides a method for detecting adenine and guanine in a solution by using boron cluster nanogold, which has the following advantages:
1. the nano-alloy is formed by adopting a one-step method, the synthesis process is simple, and the loss of raw materials is less;
2. the preparation work in the early detection stage is simple, and qualitative detection can be carried out after the solution is prepared;
3. the qualitative detection process is simple and convenient, the phenomenon is visible to naked eyes, and the observation is easy;
4. the quantitative detection has low requirements on instruments, and can be finished by a conventional ultraviolet visible spectrophotometer;
5. the detection of two purines can realize step-by-step detection, complex pretreatment operation is not needed, and only the pH value of the solution is adjusted and the color change time is recorded.
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FIG. 1 shows UV-VIS absorption spectra A of 0min and 10min at pH 2.0 in example I of the present invention650/A525Histogram for base type (base concentration 200. mu.M);
FIG. 2 shows UV-VIS absorption spectra A of 0h and 0.5h at pH 2.0 in example II of the present invention650/A525Graph against salt concentration (base concentration 40. mu.M);
FIG. 3 is a UV-VIS absorption spectrum A of 0h at pH 2.0 in example III of the present invention650/A525Graph for adenine concentration (base concentration 20. mu.M);
FIG. 4 is a graph showing the UV-VIS absorption spectrum A at pH 2.0 for 1h in example III of the present invention650/A525Graph against guanine concentration (base concentration 20. mu.M);
fig. 5 is a graph of the uv-vis absorption spectra of different base systems at 0h under pH 2.0 in example four of the present invention (the base concentrations are all 20 μ M);
FIG. 6 is a graph showing UV-VIS absorption spectra of different base systems (base concentrations of 20. mu.M) at pH 2.0 for 1h in example IV of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
A method for detecting adenine and guanine in a solution by using boron cluster nanogold comprises the following steps:
step S1: preparing boron cluster reduced nanogold according to a conventional process, and diluting the boron cluster reduced nanogold to obtain a diluted solution.
In the step, Cs is adopted when preparing the nano-gold reduced by the boron cluster2B12H12As reducing agent and stabilizer, Cs is added in a molar ratio of 1:12B12H12Adding into chloroauric acid solution, stirring at 25 deg.C for 30min to obtain boron cluster reduced nanogold. Wherein Cs2B12H12The concentration of chloroauric acid in the system is 0.4mM, and the solvent is water; when the nano-gold is diluted, the nano-gold is diluted by 2 to 10 times according to the type of a detection instrument, and preferably, the nano-gold is diluted by 4 to 5 times in the experiment.
Step S2: and (3) adjusting the pH value of the diluted solution by using acid, and adding a sample aqueous solution to realize the specific recognition of adenine.
In the step, the pH range of the solution is 2.0-2.5, only acid can be used for adjusting the pH of the solution, too high concentration buffer solution can bring too high salt concentration, so that the stability of the nanogold is influenced, and the concentration of the sample aqueous solution is 0-200 mu M.
Step S3: if no adenine exists in the system, acid is continuously added to adjust the pH of the solution, and the specific recognition of guanine can be realized.
In the step, the pH range of the solution is 1.8-2.0, and only acid can be used for adjusting the pH of the solution.
When adenine and guanine are detected, the pH value of the solution is not fixed, the detection can be completed in a certain pH range, the detection time is shorter as the pH value is lower, but the interference of other bases is larger at the time. For example, at pH 2.0, adenine can be detected quickly by mixing and measuring, whereas guanine detection requires 10min or even more than 1h depending on the concentration of guanine; and when the pH value is less than 1.8, guanine can realize rapid detection, but cytosine, thymine and uracil can change the color of the solution successively after a period of time.
Step S4: performing ultraviolet visible absorption spectrum scanning on the obtained nano-gold sample solution according to nano-gold A650/A525Signal strength ofThe concentration of adenine or guanine can be determined.
In the step, the obtained nano gold sample solution is subjected to ultraviolet visible absorption spectrum scanning, and the absorbance ratio at 650nm and 525nm is in direct proportion to the concentration of adenine or guanine in a certain range.
In order to explain the technical means of the present invention, the following description will be given by way of specific examples.
The first embodiment is as follows:
mixing Cs2B12H12The reduction-stable nanogold was diluted 4-fold and the pH of the solution was adjusted to 2.5, 2.0 and 1.8 with concentrated hydrochloric acid, the volume of concentrated hydrochloric acid used being recorded.
Taking another Cs2B12H12The reduction-stable nanogold was diluted 3 times, concentrated hydrochloric acid was added in the same volume ratio as above, and 1 volume of an aqueous solution of adenine, guanine, cytosine, thymine, and uracil (final base concentration: 200. mu.M) was added to the original nanogold solution, and the color change time of nanogold was recorded, and the results are shown in the following table.
The system was subjected to uv-vis absorption spectroscopy and the results are shown in fig. 1.
Example two:
mixing Cs2B12H12And (3) diluting the stable reduced nanogold by 2 times, adding concentrated hydrochloric acid with the same volume ratio and the NaCl aqueous solution with different volumes, which are adjusted to have the pH value of 2.0 in example 1, and fixing the volume of the solution to 3 times of the total volume of the original nanogold solution by using deionized water. Finally, deionized water or adenine water solution and guanine water solution with the volume of 1 time of the original nano gold solution are added. Wherein at salt concentrations below 70mM, no significant discoloration of the system occurs without the addition of purine, whereas the color changes occur after different times with the addition of adenine or guanine. The system was subjected to uv-vis absorption spectroscopy and the results are shown in fig. 2.
Example three:
mixing Cs2B12H12Diluting the stable nanogold by 3 times, adding concentrated hydrochloric acid and NaCl aqueous solution and adenine or guanine aqueous solution with different volumes, and fixing the volume by using deionized water to ensure that the nanogold in the final system is diluted by 4 times, wherein the NaCl concentration is 70mM, the pH value of the system is 2.0, the adenine concentration is 0-10 mu M, and the guanine concentration is 0-30 mu M. And scanning the system at 0h and 1h through ultraviolet-visible absorption spectrum. Wherein at 0h, the ultraviolet visible absorption spectrum A is in the range of 0.01-1 mu M650/A525Is proportional to the concentration of adenine; at 1h, in the range of 0.01-5 μ M, the ultraviolet visible absorption spectrum A650/A525Is proportional to the concentration of guanine. The results are shown in FIGS. 3 and 4.
Example four:
mixing Cs2B12H12Diluting the stable nanogold by 3 times, adding concentrated hydrochloric acid and NaCl aqueous solution and various base aqueous solutions with certain volume, and fixing the volume by using deionized water to ensure that the nanogold in the final system is diluted by 4 times, wherein the NaCl concentration is 70mM, the pH value of the system is 2.0, and the concentration of various base is 20 mu M. Wherein at 0h, the system added with 4 bases without adenine is red, and the system added with 5 bases is blue-violet; at 1h, the system with 3 bases without guanine appeared red, and the system with 4 bases appeared bluish-purple. The UV-VIS absorption spectrum scan was performed on the system at 0h and 1h, and the results are shown in FIGS. 5 and 6.
In the embodiment of the invention, the nano gold is agglomerated by utilizing the electrostatic interaction between two permanent negative charges of the boron cluster and N protonated by purine under an acidic condition, so that the color of the solution is changed, and the aims of quickly and conveniently identifying, detecting and quantifying are fulfilled. Meanwhile, because the strength of the action of adenine, guanine and boron clusters is inconsistent, the color changing time of the nanogold under different acidic conditions is inconsistent, and the stepwise detection of the two purines can be realized. Compared with the traditional detection method, the method has the advantages of convenience, rapidness, low detection cost, less raw material loss, low instrument requirement and the like.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (5)
1. A method for detecting adenine and guanine in a solution by using boron cluster nanogold is characterized by comprising the following steps:
step S1: preparing boron cluster reduced nanogold according to a conventional process, and diluting the boron cluster reduced nanogold to obtain a diluted solution;
step S2: adjusting the pH value of the diluted solution by using acid, and adding a sample aqueous solution to realize the specific recognition of adenine;
step S3: if no adenine exists in the system, acid is continuously added to adjust the pH value of the solution, so that the specific recognition of guanine can be realized;
step S4: performing ultraviolet visible absorption spectrum scanning on the obtained nano-gold sample solution according to nano-gold A650/A525The signal of (2) is strong or weak, and the concentration of adenine or guanine can be measured.
2. The method for detecting adenine and guanine in solution with boron cluster nano-gold as claimed in claim 1 wherein in step S1, Cs is used2B12H12As reducing agent and stabilizer, Cs is added in a molar ratio of 1:12B12H12Adding into chloroauric acid solution, stirring at 25 deg.C for 30min to obtain boron cluster reduced nanogold.
3. The method of claim 2, wherein Cs is adenine and guanine in the boron cluster nanogold detection solution2B12H12And chloroauric acid in the solution at a concentration of 0.4mM each, and the solvent was water.
4. The method for detecting adenine and guanine in a solution according to claim 1 wherein in step S2, the pH of the solution is in the range of 2.0 to 2.5 and the concentration of the aqueous sample solution is in the range of 0 to 200 μ M.
5. The method for detecting adenine and guanine in a solution according to claim 1 wherein in step S3, the pH of the solution is in the range of 1.8 to 2.0.
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Application publication date: 20200814 |