Reagent and method for detecting fluorine ions in blood

Abstract

The invention belongs to the technical field of blood fluorine chemical analysis, and discloses a reagent and a method for detecting fluorine ions in blood. The invention discloses a fluorine ion detection reagent of an organic complex containing ferric ions, and the organic complex is selected from one of polyaniline, polydopamine, indocyanine green, Prussian blue, gallic acid, catechin and 1, 2, 3-pyrogallol. The method comprises the following steps: respectively reacting a series of standard solutions with a fluorine ion detection reagent, measuring the temperature change under the irradiation of near-infrared light to obtain a linear equation of the fluorine ion concentration and the temperature change, reacting a sample to be detected with the fluorine ion detection reagent under the same condition, measuring the temperature change, and substituting the measured temperature change into the linear equation to obtain the blood fluorine concentration in the sample to be detected. The method is small in repeated detection variability, high in accuracy and stability, low in detection cost and easy to popularize, and all methodological indexes can meet the requirement for detecting the concentration of the fluorine ions in the blood.

Description

Technical Field

[0001] The present invention relates to the technical field of blood fluoride chemical analysis, and more particularly to a reagent and method for detecting fluoride ions in blood. Background Art

[0002] Although fluoride is beneficial to the human body, it is not necessarily better to have more. Excessive fluoride intake may cause fluoride poisoning, leading to skeletal fluorosis and dental fluorosis; it may cause systemic poisoning in humans, and all tissues and organs in the body will be damaged to a certain extent, and fluoride is not easily excreted from the body through metabolism. Traditional methods for detecting fluoride ions include ion selective electrode method and ion chromatography method. These classic detection methods have relatively ideal detection sensitivity, and their practicality has been proven by time. However, the above two methods also have obvious disadvantages. On the one hand, these methods require expensive large instruments and professional operators, and are difficult to promote on a large scale to achieve rapid and real-time detection; on the other hand, the above methods are also susceptible to other ions (such as OH). - ) interference and suboptimal selectivity. In recent years, reported detection methods primarily include electrochemical methods, nuclear magnetic resonance methods, and fluorescence sensors. Most analytical methods suffer from low sensitivity, high cost, and complex sample handling and detection processes, limiting their practicality. Therefore, in many important applications, there is an urgent need for rapid, accurate, low-cost, and selective methods for analyzing and detecting these fluoride ions.

[0003] In view of this, developing a method that can detect the presence and content of fluoride ions plays an extremely important role in the treatment of clinical diseases, environmental protection and academic research. Summary of the Invention

[0004] In response to the above problems, the present invention provides a reagent and method for detecting fluoride ions in blood. One of the purposes is to provide a fluoride ion detection reagent with simple synthesis, mild reaction conditions and low cost; the second purpose is to provide a method with good selectivity and high sensitivity, which can quickly, sensitively and quantitatively detect fluoride ions in blood in aqueous solution.

[0005] One of the purposes of the present invention is to provide a fluoride ion detection reagent, which is an aqueous solution of an organic complex containing trivalent iron ions; the organic complex is selected from at least one of polyaniline, polydopamine, indocyanine green, Prussian blue, gallic acid, catechin, and 1,2,3-pyrogallol.

[0006] Furthermore, the trivalent iron ion is selected from Fe(NO)3 or Fe(Cl)3.

[0007] Furthermore, the mass fraction of the aqueous organic complex solution containing trivalent iron ions is 10-40%, preferably, the mass fraction is 30%.

[0008] Furthermore, the molar ratio of the trivalent iron ion to the organic complex is 1-2:5-8, preferably, the molar ratio is 2:7.

[0009] A second object of the present invention is to provide a method for detecting fluoride ions in blood, comprising the following steps:

[0010] (1) Prepare fluoride ion detection reagent;

[0011] (2) reacting the test solution with the fluoride ion detection reagent with a series of standard concentrations under the same near-infrared light irradiation, detecting the temperature change at the same time point, and obtaining a linear equation relationship between the temperature change and the fluoride ion concentration;

[0012] (3) Take the blood sample to be tested, centrifuge it at a centrifugal speed of 3000-5000 rpm for 10 min, and collect the supernatant to obtain serum or plasma;

[0013] (4) Under the same near-infrared light irradiation as step (2), the serum or plasma is reacted with the fluoride ion detection reagent to obtain a mixed solution, and the temperature change of the mixed solution at the same time point as step (2) is measured, and the blood fluoride concentration in the sample to be tested is obtained according to the linear equation relationship.

[0014] Furthermore, in step (1), the preparation is as follows: slowly adding an equal volume of trivalent iron ion aqueous solution to an organic complex aqueous solution, stirring the reaction at room temperature for 60-150 minutes, collecting the solid after reduced pressure distillation, and dissolving it in water to obtain the fluoride ion detection reagent.

[0015] Preferably, the volume ratio of the aqueous solution of trivalent iron ions to the aqueous solution of the organic complex is 1:1.

[0016] Furthermore, in step (2), the wavelength range of the near-infrared light is 750-1000 nm.

[0017] Furthermore, in step (2), the reaction temperature is 15-35°C and the reaction time is 2-5 minutes. Preferably, the reaction temperature is 25°C and the reaction time is 4 minutes.

[0018] Furthermore, in steps (2) and (3), the volume ratio of the test solution, the serum or plasma, and the fluoride ion detection reagent is 1:2.

[0019] The present invention uses the change of the thermal signal of the fluoride ion detection reagent to achieve rapid, sensitive and portable quantitative detection of the test component. Specifically, the linear graph (regression coefficient R) of the temperature value and concentration of a series of solutions of known concentrations of the test component is obtained. 2≥0.999), and a linear equation is obtained between temperature change (y) and fluoride ion concentration (x). Then, the temperature change of the unknown concentration of the component to be measured is measured and substituted into the linear equation to obtain the result.

[0020] It can be seen from the above technical solutions that, compared with the prior art, the present invention has the following beneficial effects:

[0021] 1. The blood sample of the present invention is only centrifuged, and plasma or serum is directly sampled, which reduces the error caused by pre-treatment and human operation error, improves the accuracy, repeatability and recovery rate of quantitative results, greatly shortens the detection time, makes the detection process simple and fast, reduces the experimental cost, and is more conducive to the detection of fluoride ion concentration in patients during clinical treatment.

[0022] 2. The method of the present invention can rapidly and sensitively perform quantitative analysis on the components to be tested in the test liquid, providing a new analytical testing method; the materials used in the method are simple, the operation time is short, and low-cost rapid, sensitive, and portable quantitative analysis can be achieved. DETAILED DESCRIPTION

[0023] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0024] The temperature change of the present invention is detected by a thermal imaging analyzer, the model of which is FLIR-E40.

[0025] Example 1

[0026] Provide a fluoride ion detection reagent:

[0027] 20 mM Fe(NO)3 solution was added to 70 mM indocyanine green aqueous solution, mixed in equal volumes, stirred and reacted at room temperature for 150 minutes, and the solid was collected after vacuum distillation and dissolved in water to obtain a fluoride ion detection reagent with a mass fraction of 30%.

[0028] Comparative Example 1

[0029] A fluoride ion detection reagent is provided: the difference between the fluoride ion detection reagent and the embodiment 1 is that the molar ratio of the Fe(NO)3 solution to the indocyanine green aqueous solution is 0.5:7, and a comparative reagent is obtained.

[0030] Comparative Example 2

[0031] A fluoride ion detection reagent is provided: the difference between the fluoride ion detection reagent and the embodiment 1 is that the molar ratio of the Fe(NO)3 solution to the indocyanine green aqueous solution is 2:9, and a comparative reagent is obtained.

[0032] Example 2

[0033] Provide a fluoride ion detection reagent:

[0034] 20 mM Fe(Cl)3 solution was added to 70 mM indocyanine green aqueous solution, mixed in equal volumes, stirred and reacted at room temperature for 150 min, and the solid was collected after vacuum distillation and dissolved in water to obtain a fluoride ion detection reagent with a mass fraction of 30%.

[0035] Example 3

[0036] Provide a fluoride ion detection reagent:

[0037] 20 mM Fe(NO)3 solution was added to 70 mM indocyanine green aqueous solution, mixed in equal volumes, stirred and reacted at room temperature for 150 minutes, and the solid was collected after vacuum distillation and dissolved in water to obtain a fluoride ion detection reagent with a mass fraction of 40%.

[0038] Example 4

[0039] Provide a fluoride ion detection reagent:

[0040] 20 mM Fe(NO)3 solution was added to 70 mM polydopamine aqueous solution, mixed in equal volumes, stirred and reacted at room temperature for 150 minutes, and the solid was collected after vacuum distillation and dissolved in water to obtain a fluoride ion detection reagent with a mass fraction of 40%.

[0041] Example 5

[0042] Provide a fluoride ion detection reagent:

[0043] A 20 mM Fe(NO)3 solution was added to a 70 mM 1,2,3-pyrogallol aqueous solution, mixed in equal volumes, and stirred for reaction at room temperature for 150 minutes. The solid was collected after vacuum distillation and dissolved in water to obtain a fluoride ion detection reagent with a mass fraction of 40%.

[0044] Example 6

[0045] A method for detecting fluoride ions in blood comprises the following steps:

[0046] Under 880 nm wavelength light, 0, 10, 20, 40, 60, 80, and 90 μmol / L fluoride ion standard solutions were mixed with fluoride ion detection reagents 1-5 and comparison reagents 1-2 at a volume ratio of 1:2, respectively. The mixture was reacted at 25°C for 4 minutes. The temperature change was detected, and the linear equation relationship between the temperature change (y) and the fluoride ion concentration (x) was obtained, as shown in Table 1 below.

[0047] Table 1

[0048] project Linear equations <![CDATA[R 2 ]]> Example 1 y=-0.06691x+7.3518 <![CDATA[R 2 =0.99991]]> Example 2 y=-0.09435x+4.4318 <![CDATA[R 2 =0.9997]]> Example 3 y=-0.05866x+6.2745 <![CDATA[R 2 =0.9991]]> Example 4 y=-0.0235x+6.5494 <![CDATA[R 2 =0.9992]]> Example 5 y=-0.03563x+7.6555 <![CDATA[R 2 =0.9992]]> Comparative Example 1 y=-0.28239x+4.4756 <![CDATA[R 2 =0.9695]]> Comparative Example 2 y=-0.25628x+2.1587 <![CDATA[R 2 =0.9148]]>

[0049] Under the conditions of the method of the present invention, the correlation coefficient R 2 It can reach 0.999, with good linearity, which can ensure the accuracy of the sample results measured by this method.

[0050] Example 7

[0051] A method for detecting fluoride ions in blood comprises the following steps:

[0052] Five blood samples to be tested were randomly selected from the sample library and centrifuged at 3000 rpm for 10 minutes. The supernatant was collected to obtain serum. Under the same conditions as in Example 1, the temperature change of the serum at the same time point as in Example 1 was measured. The blood fluoride concentration in the test sample was obtained according to the linear equation y = -0.06691x + 7.3518.

[0053] Quality control requirements:

[0054] Each sample is accompanied by a standard curve and six quality control samples (two each at high, medium, and low concentrations). The deviation of the quality control sample results should be less than 10%. A maximum of one-third of the quality control sample results are allowed to exceed this limit, but this cannot occur in the same quality control sample concentration. If the quality control sample results do not meet the above requirements, the test results of that batch of samples will be invalidated and retested. Substitute the temperature changes measured by the quality control samples into the above standard curve equation to calculate the fluoride ion concentration of the quality control samples.

[0055] The recovery rate of the control sample is calculated as follows: Recovery (%) = measured concentration / labeled concentration × 100. The recovery (%) should be within the range of 100 ± 10%. The final recovery rate measured in this embodiment can reach 96.8%.

[0056] In summary, under the conditions of the method of the present invention, the correlation coefficient of the standard curve of fluoride ion concentration and the recovery rate of the quality control product are in compliance with the regulations, proving the accuracy of the measured sample results.

[0057] The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

Claims

1. A fluoride ion detection reagent, characterized in that: It is an aqueous solution of an organic complex containing trivalent iron ions; The organic complex is selected from at least one of polyaniline, polydopamine, indocyanine green, Prussian blue, gallic acid, catechin, and 1,2,3-pyrogallol.

2. The method for detecting fluoride ions in blood according to claim 1, wherein: The trivalent iron ion is selected from Fe(NO)3 or Fe(Cl)3.

3. A fluoride ion detection reagent according to claim 1, characterized in that, The mass fraction of the organic complex aqueous solution containing trivalent iron ions is 10-40%.

4. A fluoride ion detection reagent according to claim 1, characterized in that, The molar ratio of the trivalent iron ion to the organic complex is 1-2:5-8.

5. A method for detecting fluoride ions in blood, characterized in that: The following steps are involved: (1) preparing the fluoride ion detection reagent according to any one of claims 1 to 4; (2) reacting the test solution with the fluoride ion detection reagent with a series of standard concentrations under the same near-infrared light irradiation, detecting the temperature change at the same time point, and obtaining a linear equation relationship between the temperature change and the fluoride ion concentration; (3) Take the blood sample to be tested, centrifuge it, and take the supernatant to obtain serum or plasma; (4) Under the same near-infrared light irradiation as step (2), the serum or plasma is reacted with the fluoride ion detection reagent to obtain a mixed solution, and the temperature change of the mixed solution at the same time point as step (2) is measured, and the fluoride ion concentration in the sample to be tested is obtained according to the linear equation.

6. The method for detecting fluoride ions in blood according to claim 5, characterized in that: In step (1), the preparation is as follows: slowly adding the aqueous solution of trivalent iron ions to the aqueous solution of the organic complex, stirring for reaction, collecting the solid after reduced pressure distillation, and adding water to dissolve it to obtain the fluoride ion detection reagent.

7. The method for detecting fluoride ions in blood according to claim 6, wherein: The volume ratio of the aqueous solution of trivalent iron ions to the aqueous solution of the organic complex is 1:

1.

8. The method for detecting fluoride ions in blood according to claim 5, wherein: In step (2), the wavelength range of the near-infrared light is 750-1000 nm.

9. The method for detecting fluoride ions in blood according to claim 5, wherein: In step (2), the reaction temperature is 15-35° C. and the reaction time is 2-5 min.

10. The method for detecting fluoride ions in blood according to claim 5, characterized in that: In steps (2) and (3), the volume ratio of the test solution, the serum or plasma, and the fluoride ion detection reagent is 1:2.