CN112730377A - Method for detecting serum potassium ions based on surface enhanced Raman spectroscopy - Google Patents
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Abstract
The invention discloses a method for detecting serum potassium ions based on surface enhanced Raman spectroscopy, which comprises the following steps: diluting a sample to be detected with deionized water to obtain a diluted sample to be detected; placing 0.3mL of diluted sample to be tested in a sample tube, adding 0.2-1% by mass of sodium tetraphenylborate aqueous solution, oscillating, mixing uniformly, standing for reaction, centrifuging, and pouring out supernatant; adding 0.5-1mL of deionized water into the sample tube, shaking and mixing uniformly, centrifuging, pouring out supernate, and repeating the operation once; adding 0.5-1mL of dilute hydrochloric acid aqueous solution with the concentration of 0.05-0.5mol/L into the sample tube, uniformly mixing by oscillation, and standing to obtain a solution to be detected; mixing the nano gold sol and the solution to be detected to obtain a mixed solution; and performing Raman spectrum detection on the mixed solution to obtain a Raman spectrum curve, and comparing the Raman spectrum curve with a potassium ion Raman spectrum working curve to determine the concentration of potassium ions in the sample to be detected.
Description
Technical Field
The invention belongs to the technical field of analysis and test, and particularly relates to a method for detecting serum potassium ions based on surface enhanced Raman spectroscopy.
Background
Potassium is a main cation for maintaining cell physiological activities, and has important effects in maintaining normal osmotic pressure and acid-base balance of organism, participating in sugar and protein metabolism, and ensuring normal function of neuromuscular. The normal concentration of potassium in serum is 3.5-5.5mmol/L, and low potassium (lower than 3.5mmol/L) and high potassium (higher than 5.5mmol/L) bring great harm to human body. Hypokalemia mainly compromises the digestive system, neuromuscular system, and renal function; hyperkalemia mainly causes damage to cardiac function, easily causes cardiac arrest, and simultaneously affects bones and a digestive system. At present, the clinical detection method for the blood potassium mainly comprises a flame photometry, an ion selective electrode method (ISE), a dry chemical method and a spectrophotometry:
the basic principle of flame photometry is: when the serum sample is atomized into fine mist by the atomizing device and sent into the flame for combustion, K is used+After the energy is obtained, a spectrum with special wavelength is emitted, and the light passes through the optical filter and is received by the optical detector. The radiation light energy is measured by a photoelectric system, and the known high and low value analytes are calibrated to obtain the potassium content; the flame photometry has the main problems that the sensitivity is influenced by a plurality of factors (including gas pressure, combustion-supporting gas pressure, sample injection speed, the sample is fully mixed after being added with distilled water, and the like), and the used gas propane has high danger and causes potential safety hazards to detection laboratories and personnel.
The ion selective electrode method is a quantitative analysis method based on measuring the potential of the battery. From K+The activity of the electrode is proportional to the ion concentration, so that the ion selective electrode method can quantitatively analyze the potassium ion concentration in the sample; the ion selective electrode method (ISE) has the main problems that the instrument needs to be calibrated continuously, the electrode needs to be replaced once in half a year, the maintenance cost is high, and the cost of a matched detection reagent is high.
The dry chemical method uses dry chemical slide which is coated with high-purity chemical reagent by gelatin binder by high-precision color Bola manufacturing process. The chemical slide contains K+Magnetic cipher for test, electronic computer measuring resultConverting into concentration and displaying; the multilayer film test dry sheet used by the dry chemical method is disposable, has high detection cost, and cannot be widely applied to clinical routine detection.
The spectrophotometry is mainly based on a macrocyclic chromophore method and an enzyme method, and adopts related substances and K+The principle of specific color development, the principle of measuring absorbance value and quantitative detection of K+And (4) concentration. In the spectrophotometry, the macrocyclic chromophore method cannot avoid the interference of bilirubin, and the enzyme method has expensive reagents, high detection cost and more influenced factors.
In conclusion, the methods generally have the defects of long detection time, high professional requirements for detection, high reagent detection cost and the like.
Disclosure of Invention
The purpose of the invention is as follows: in order to solve the problems in the prior art, the invention provides a method for detecting serum potassium ions based on surface enhanced Raman spectroscopy, and whether potassium ions exist in a sample can be accurately judged from a molecular level by means of the Raman spectroscopy principle.
The technical scheme adopted by the invention is as follows: a method for detecting serum potassium ions based on surface enhanced Raman spectroscopy comprises the following steps:
s100: taking a sample to be tested, and diluting the sample by n times with deionized water, wherein n is 1,2,3 … to obtain a diluted sample to be tested;
s200: placing the diluted sample to be detected in a sample tube, adding a sodium tetraphenylborate aqueous solution with the mass fraction of 0.2-1%, uniformly mixing by oscillation, standing for reaction, centrifuging, and pouring out supernatant; the volume ratio of the sample to be detected to the sodium tetraphenylborate aqueous solution is as follows: (12-18): 1;
s300: adding 0.5-1mL of deionized water into the sample tube, shaking and mixing uniformly, centrifuging and pouring out a supernatant;
s400: repeating the step S300 once;
s500: adding 0.5-1mL of dilute hydrochloric acid aqueous solution with the concentration of 0.05-0.5mol/L into the sample tube, uniformly mixing by oscillation, and standing to obtain a solution to be detected;
s600: mixing the nano gold sol and the solution to be detected to obtain a mixed solution, wherein the volume ratio of the mixed solution is as follows: nano gold sol: test solution ═ (1-5): 1;
s700: and performing Raman spectrum detection on the mixed solution to obtain a Raman spectrum curve, and comparing the Raman spectrum curve with a potassium ion Raman spectrum working curve to determine the concentration of potassium ions in the sample to be detected.
Further, the drawing step of the potassium ion Raman spectrum working curve comprises the following steps:
preparing a potassium ion standard solution with the concentration of 1 mmol/L;
diluting the potassium ion standard solution to different concentrations by water gradient to obtain potassium ion standard working solution;
taking 0.3mL of potassium ion standard working solutions with different concentrations, placing the solutions into respective sample tubes, and executing the following steps:
adding 0.2-1% by mass of sodium tetraphenylborate aqueous solution into the sample tube, oscillating, mixing uniformly, standing for reaction, centrifuging, and pouring out supernatant; the volume ratio of the potassium ion standard working solution to the sodium tetraphenylborate aqueous solution is as follows: (12-18): 1;
adding 0.5-1mL of deionized water into the sample tube, shaking and mixing uniformly, centrifuging and pouring out a supernatant; this step is repeatedly performed once;
adding 0.5-1mL of dilute hydrochloric acid aqueous solution with the concentration of 0.05-0.5mol/L into the sample tube, uniformly mixing by oscillation, and standing to obtain a solution to be detected;
mixing the nano gold sol and the solution to be detected to obtain a mixed solution, wherein the volume ratio of the nano gold sol to the solution to be detected is as follows: test solution ═ (1-5): 1;
performing Raman spectrum detection on the mixed solution to obtain a Raman spectrum curve;
995cm is selected-1Performing linear fitting on the intensity of the Raman characteristic peak and the potassium ion concentration, and calculating the corresponding relation as follows:
Cto be measured＝(0.159x10-4ITo be measured-0.0489) × (the dilution factor of the standard solution of potassium ion)
Wherein C isTo be measuredI.e. the concentration of potassium ions in the sample to be measured, ITo be measuredIndicating that the liquid to be detected is at 995cm-1Corresponding to the intensity of the Raman spectrum;
and forming a potassium ion Raman spectrum working curve by Raman spectrum curves with different concentrations.
Further, the preparation step of the nanogold sol in the step S600 includes: putting chloroauric acid aqueous solution with the mass fraction of 0.01-0.1% into a round-bottom flask, stirring and heating until boiling;
adding a sodium citrate solution with the mass fraction of 0.2-1% into the chloroauric acid aqueous solution, wherein the volume ratio of the chloroauric acid aqueous solution to the citric acid solution is 100 (1-0.7), stirring and heating until the liquid turns red, and cooling at room temperature to obtain the nano gold sol.
Further, the S700 may be replaced by the following steps:
performing Raman spectrum detection on the mixed solution to obtain a Raman spectrum curve;
obtaining the mixed solution at 995cm-1Corresponding to the intensity of the Raman spectrum;
determining the potassium ion concentration in the sample to be tested according to the following formula:
Cto be measured＝(0.159x10-4ITo be measured-0.0489)×n
Wherein C isTo be measuredI.e. the concentration of potassium ions in the sample to be measured, ITo be measuredIndicating that the liquid to be detected is at 995cm-1Corresponding to the intensity of the Raman spectrum, wherein n is the diluted multiple of the sample to be detected in S100.
Has the advantages that: the invention accurately judges the content of potassium ions in the sample from the molecular level by means of the Raman spectrum principle, only needs 5 minutes in the whole detection process of a single sample, has the advantages of simple and convenient sample treatment, less serum usage, low detection cost, quick detection, high detection precision, lower requirement on detection personnel, no dependence on large-scale detection equipment and the like, and is suitable for household or personal daily detection and monitoring.
Drawings
FIG. 1 is a spectrum of the change of potassium ion content corresponding to Raman spectrum.
Detailed Description
The invention is further illustrated below with reference to the figures and examples.
Example 1:
the method for detecting serum potassium ions based on surface enhanced raman spectroscopy of the embodiment comprises the following steps:
step 1: taking a proper amount of chloroauric acid aqueous solution with the mass fraction of 0.1 percent, stirring and heating the chloroauric acid aqueous solution in a round-bottom flask to boil; adding a sodium citrate solution with the mass fraction of 1% into a chloroauric acid aqueous solution, wherein the volume ratio of the sodium citrate solution is as follows: aqueous chloroauric acid solution: citric acid solution 100: 0.7; stirring and heating until the liquid changes into red, cooling at room temperature to obtain nano gold sol serving as a reinforcing reagent, and storing at normal temperature in a dark place;
step 2: preparing a dilute hydrochloric acid aqueous solution with the concentration of 0.1mol/L, and storing at normal temperature in a dark place;
and step 3: preparing 0.2 mass percent sodium tetraphenylborate aqueous solution, and refrigerating at 4 ℃ for later use;
and 4, step 4: accurately weighing 0.7455g of potassium chloride, diluting to a constant volume of 10mL with water to obtain a potassium ion standard solution of 1mmol/L, and refrigerating at 4 ℃ for later use;
and 5: diluting the potassium ion standard solution prepared in the step 4 to 0.5, 0.2, 0.1 and 0.02mmol/L by using water in a gradient manner to obtain a potassium ion standard working solution;
step 6: putting 0.3mL of potassium ion standard working solution into a sample tube, adding 20 mu L of the sodium tetraphenylborate aqueous solution prepared in the step 3 into the sample tube, uniformly mixing the solution by oscillation, standing the mixture for reaction for 2min, centrifuging the mixture, and pouring out supernatant; the sodium tetraphenylborate and potassium ions generate a complex reaction to generate a precipitate, and Raman pure detection of the potassium ions has no Raman response, so that the sodium tetraphenylborate plays a role in enriching the potassium ions and serving as a potassium ion detection marker.
And 7: adding 0.5mL of deionized water into the sample tube in the step 6, oscillating and uniformly mixing, pouring out supernate after centrifuging, and repeating the step once; this step is repeated for the purpose of: the sodium tetraphenylborate solution remained on the surface of the precipitate is cleaned, and the effect can be achieved by repeating once from the viewpoints of detection time and sample loss without performing twice or more times;
and 8: adding 0.5mL of the dilute hydrochloric acid aqueous solution prepared in the step 2 into the sample tube in the step 7, uniformly mixing by oscillation, and standing for 10s to obtain a solution to be detected; the solid precipitate generated in the step 6 can be dissolved in a dilute hydrochloric acid solution, and the solution state is relatively uniform and stable, thereby being beneficial to the acquisition of Raman signals.
And step 9: mixing 200 mu L of the nano gold sol prepared in the step 1 with 200 mu L of a solution to be detected, adding the nano gold sol into the solution to be detected to amplify the Raman signal of the object to be detected by about million times, and detecting the mixed solution on a machine to obtain a Raman spectrogram;
step 10: 995cm is selected-1Performing linear fitting on the intensity of the Raman characteristic peak and the potassium ion concentration, and calculating the corresponding relation as follows:
Cto be measured＝(0.159x10-4ITo be measured-0.0489) × (the dilution ratio of the standard solution of potassium ion in step 5)
Wherein, CTo be measuredI.e. the concentration of potassium ions in the sample to be measured, ITo be measuredShows that the sample is at 995cm-1Corresponding to the raman spectral intensity.
Step 11: and forming a potassium ion Raman spectrum working curve by Raman spectrum curves with different concentrations.
Step 12: placing 0.1 ml of a serum sample to be detected in a proper container, and adding a proper amount of deionized water to dilute the serum sample to obtain a diluted serum sample;
step 13: placing 0.3mL diluted serum sample in a sample tube, adding 20 μ L sodium tetraphenylborate aqueous solution with mass fraction of 0.2-1%, shaking, mixing, standing for 2min, centrifuging, and removing supernatant;
step 14: adding 0.5mL of deionized water into the sample tube, oscillating and uniformly mixing, pouring out a supernatant after centrifugation, and repeating the step once;
step 15: adding 0.5mL of dilute hydrochloric acid aqueous solution with the concentration of 0.1mol/L into the sample tube, uniformly mixing by oscillation, and standing for 10s to obtain a solution to be detected;
step 16: mixing 200 mu L of nano gold sol with 200 mu L of the solution to be detected obtained in the step 15, and detecting the mixed solution on a machine to obtain a Raman spectrogram; and comparing the Raman spectrum curve with a potassium ion Raman spectrum working curve to determine the concentration of potassium ions in the sample to be detected.
Example 2:
the method of this embodiment can be used when the conditions such as the amount of reagent and the batch of reagent are determined, and the method for detecting serum potassium ions based on surface enhanced raman spectroscopy of this embodiment includes the following steps:
step 1: placing 0.1 ml of a serum sample to be detected in a proper container, and adding a proper amount of deionized water to dilute the serum sample to obtain a diluted serum sample;
step 2: placing 0.3mL diluted serum sample in a sample tube, adding 20 μ L sodium tetraphenylborate aqueous solution with mass fraction of 0.2%, shaking, mixing, standing for 2min, centrifuging, and removing supernatant;
and step 3: adding 0.5mL of deionized water into the sample tube, oscillating and uniformly mixing, pouring out a supernatant after centrifugation, and repeating the step once;
and 4, step 4: adding 0.5mL of dilute hydrochloric acid aqueous solution with the concentration of 0.1mol/L into the sample tube, uniformly mixing by oscillation, and standing for 10s to obtain a solution to be detected;
and 5: mixing 250 mu L of nano gold sol and 50 mu L of solution to be detected to obtain mixed solution; detecting the mixed solution on a machine to obtain a Raman spectrogram; according to the Raman spectrogram, the solution to be detected containing potassium ions is 736cm-1、995cm-1、1272cm-1An obvious Raman spectrum peak is formed, and the intensity of the peak is in positive correlation with the concentration of potassium ions;
step 6: obtaining the mixed solution at 995cm-1Determining the concentration of potassium ions in the sample to be detected according to the following formula:
Cto be measured＝(0.159x10-4ITo be measured-0.0489) × (multiple of dilution of serum sample to be tested in step 1)
Wherein C isTo be measuredI.e. the concentration of potassium ions in the sample to be measured, ITo be measuredIndicating that the liquid to be detected is at 995cm-1Corresponding to the raman spectral intensity.
Claims (4)
1. A method for detecting serum potassium ions based on surface enhanced Raman spectroscopy is characterized in that: the method comprises the following steps:
s100: taking a sample to be tested, and diluting the sample by n times with deionized water, wherein n is 1,2,3 … to obtain a diluted sample to be tested;
s200: placing the diluted sample to be detected in a sample tube, adding a sodium tetraphenylborate aqueous solution with the mass fraction of 0.2-1%, uniformly mixing by oscillation, standing for reaction, centrifuging, and pouring out supernatant; the volume ratio of the sample to be detected to the sodium tetraphenylborate aqueous solution is (12-18) to 1;
s300: adding 0.5-1mL of deionized water into the sample tube, shaking and mixing uniformly, centrifuging and pouring out a supernatant;
s400: repeating the step S300 once;
s500: adding 0.5-1mL of dilute hydrochloric acid aqueous solution with the concentration of 0.05-0.5mol/L into the sample tube, uniformly mixing by oscillation, and standing to obtain a solution to be detected;
s600: mixing the nano gold sol and the solution to be detected to obtain a mixed solution, wherein the volume ratio of the mixed solution is as follows: nano gold sol: test solution ═ (1-5): 1;
s700: and performing Raman spectrum detection on the mixed solution to obtain a Raman spectrum curve, and comparing the Raman spectrum curve with a potassium ion Raman spectrum working curve to determine the concentration of potassium ions in the sample to be detected.
2. The method for detecting the potassium ions in the serum based on the surface enhanced Raman spectroscopy according to claim 1, wherein the method comprises the following steps: the drawing step of the potassium ion Raman spectrum working curve comprises the following steps:
preparing a potassium ion standard solution with the concentration of 1 mmol/L;
diluting the potassium ion standard solution to different concentrations by water gradient to obtain potassium ion standard working solution;
taking 0.3mL of potassium ion standard working solutions with different concentrations, placing the solutions into respective sample tubes, and executing the following steps:
adding 0.2-1% by mass of sodium tetraphenylborate aqueous solution into the sample tube, oscillating, mixing uniformly, standing for reaction, centrifuging, and pouring out supernatant; the volume ratio of the potassium ion standard working solution to the sodium tetraphenylborate aqueous solution is (12-18): 1;
adding 0.5-1mL of deionized water into the sample tube, shaking and mixing uniformly, centrifuging and pouring out a supernatant; this step is repeatedly performed once;
adding 0.5-1mL of dilute hydrochloric acid aqueous solution with the concentration of 0.05-0.5mol/L into the sample tube, uniformly mixing by oscillation, and standing to obtain a solution to be detected;
mixing the solution to be detected to obtain a mixed solution, wherein the volume ratio of the mixed solution is nano gold sol: test solution ═ (1-5): 1;
performing Raman spectrum detection on the mixed solution to obtain a Raman spectrum curve;
995cm is selected-1Performing linear fitting on the intensity of the Raman characteristic peak and the potassium ion concentration, and calculating the corresponding relation as follows:
Cto be measured＝(0.159x10-4ITo be measured-0.0489) × (the dilution factor of the standard solution of potassium ion)
Wherein C isTo be measuredI.e. the concentration of potassium ions in the sample to be measured, ITo be measuredIndicating that the liquid to be detected is at 995cm-1Corresponding to the intensity of the Raman spectrum;
and forming a potassium ion Raman spectrum working curve by Raman spectrum curves with different concentrations.
3. The method for detecting serum potassium ions based on surface enhanced Raman spectroscopy according to claim 1, wherein: the preparation method of the nano gold sol in S600 comprises the following steps: putting chloroauric acid aqueous solution with the mass fraction of 0.01-0.1% into a round-bottom flask, stirring and heating until boiling;
adding a sodium citrate solution with the mass fraction of 0.2-1% into the chloroauric acid aqueous solution, wherein the volume ratio of the chloroauric acid aqueous solution to the citric acid solution is 100 (1-0.7), stirring and heating until the liquid turns red, and cooling at room temperature to obtain the nano gold sol.
4. The method for detecting serum potassium ions based on surface enhanced Raman spectroscopy according to claim 1, wherein: the S700 may be replaced by the following steps:
performing Raman spectrum detection on the mixed solution to obtain a Raman spectrum curve;
obtaining the mixed solution at 995cm-1Corresponding to the intensity of the Raman spectrum;
determining the potassium ion concentration in the sample to be tested according to the following formula:
Cto be measured＝(0.159x10-4ITo be measured-0.0489)×n
Wherein, CTo be measuredI.e. the concentration of potassium ions in the sample to be measured, ITo be measuredIndicating that the liquid to be detected is at 995cm-1Corresponding to the intensity of the Raman spectrum, wherein n is the diluted multiple of the sample to be detected in S100.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104597035A (en) * | 2015-02-04 | 2015-05-06 | 吉林大学 | Method for detecting potassium ions by using SERS technology |
| CN107389658A (en) * | 2017-08-16 | 2017-11-24 | 广西师范大学 | One kind tetraphenylboron sodium part regulation and control stannic oxide/graphene nano band catalytically active surface enhancing Raman spectroscopy K+Method |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104597035A (en) * | 2015-02-04 | 2015-05-06 | 吉林大学 | Method for detecting potassium ions by using SERS technology |
| CN107389658A (en) * | 2017-08-16 | 2017-11-24 | 广西师范大学 | One kind tetraphenylboron sodium part regulation and control stannic oxide/graphene nano band catalytically active surface enhancing Raman spectroscopy K+Method |
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Application publication date: 20210430 |