CN110779972A - Electrochemical detection method of trace vitamin B9 - Google Patents
Electrochemical detection method of trace vitamin B9 Download PDFInfo
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- CN110779972A CN110779972A CN201910905339.9A CN201910905339A CN110779972A CN 110779972 A CN110779972 A CN 110779972A CN 201910905339 A CN201910905339 A CN 201910905339A CN 110779972 A CN110779972 A CN 110779972A
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Abstract
The invention relates to an electrochemical detection method of trace vitamin B9, which belongs to the technical field of vitamin analysis and detection, wherein a specific sample treatment solution is matched with a sensor probe of a vitamin detector modified by a calixarene acetone saturated solution for use, and the electrochemical detection conditions are further optimized, so that trace vitamin B9 can be detected more sensitively, quickly and accurately finally, the application range is wide, and the method can be used for quickly detecting trace vitamin B9 in samples such as blood samples, medicines, foods and the like. The method is simple and easy to operate, has short detection time, does not need special technical personnel for operation, and is easy to popularize.
Description
Technical Field
The invention belongs to the technical field of vitamin analysis and detection, particularly relates to an electrochemical detection method of trace vitamin B9, and particularly relates to a method for detecting trace vitamin B9 in samples such as blood samples, medicines, foods and the like.
Background
Vitamin B9 is a trace organic substance that the body must take in food to maintain normal physiological functions, plays an important role in the growth and development of fetus, and the lack of B9 may cause serious development disorders of the central nervous system, such as no brain or spinal column protrusion or spina bifida, and megaloblastic anemia may occur.
At present, instruments and detection methods for detecting the content of vitamin B9 in a human body are different, and mainly comprise an ultraviolet spectrophotometry, a fluorescence analysis method, a high performance liquid chromatography and the like. The pretreatment technology in the high performance liquid chromatography is tedious and time-consuming, needs more organic solvents, and has larger serum requirement. The current instruments used for vitamin detection are: a vitamin analyzer was developed by Younglin corporation of korea to detect various vitamins in foods and medicines by using a high performance liquid chromatography method; germany Bayer R-Biopharm company produces an instrument for vitamin detection by means of fluorescence. Instruments such as a high performance liquid chromatograph, a fluorescence analyzer and the like are expensive, and special technical personnel are required for operation, so that the detection method is complicated, the detection time is long, and the popularization is difficult.
Disclosure of Invention
In view of the above, the present invention is directed to a method for electrochemically detecting trace vitamin B9.
In order to achieve the purpose, the invention provides the following technical scheme:
1. an electrochemical detection method of trace vitamin B9, which comprises the following steps:
(1) respectively and uniformly mixing vitamin B9 standard samples with different volume concentrations with BR buffer solution, then respectively detecting current signals generated by the vitamin B9 standard samples in an oxidation-reduction reaction on a sensor probe of a vitamin detector modified by a calixarene acetone saturated solution by using a differential pulse method, obtaining current signal values of the vitamin B9 standard samples with different concentrations, and making a standard curve;
(2) and (2) uniformly mixing a sample to be detected with BR buffer solution, detecting a current signal generated by the sample to be detected when the sample to be detected is subjected to an oxidation-reduction reaction on a sensor probe of the vitamin detector modified by the calixarene acetone saturated solution by using a differential pulse method to obtain a current signal value generated by the sample to be detected, and finally obtaining the content of vitamin B9 in the sample to be detected according to the current signal value generated by the sample to be detected and the standard curve formulated in the step (1).
Preferably, in the step (1) and the step (2), the concentration of the BR buffer solution is 0.001-2.0 mol/L, and the pH value is 1.0-9.0.
Preferably, in the step (1) and the step (2), the preparation method of the sensor probe of the vitamin detector modified by the calixarene acetone saturated solution is as follows: and (3) dripping 10-50 mu L of calixarene acetone saturated solution on the surface of a glassy carbon electrode which is polished by 0.05-1 mu m of alumina powder in a sensor probe of a vitamin detector, and completely volatilizing and drying the acetone to obtain the calixarene acetone-based composite electrode.
Preferably, in the step (1) and the step (2), the preparation method of the sensor probe of the vitamin detector modified by the calixarene acetone saturated solution is as follows: and dripping 30 mu L of calixarene acetone saturated solution on the surface of a glassy carbon electrode which is polished by 0.03 mu m of alumina powder in a sensor probe of a vitamin detector, and completely volatilizing and drying the acetone to obtain the calixarene acetone saturated solution.
Preferably, in the step (1) and the step (2), the enrichment electrodeposition potential of the vitamin detector is-600-300 mV, the enrichment electrodeposition time is 0-360 s, the initial potential is-500-100 mV, the termination potential is 100-1000 mV, the scanning speed is 10-500 mV/s, the sampling interval is 4-20 mV, the rest time is 10-60 s, the rest potential is 200-600 mV, the rest time is 10-60 s, and the measuring range is 0.0001-50 mA.
Preferably, in the step (1) and the step (2), the enrichment electrodeposition potential of the vitamin detector is 200mV, the enrichment electrodeposition time is 240s, the initial potential is 100mV, the termination potential is 1000mV, the scanning speed is 40mV/s, the sampling interval is 10mV, the rest time is 20s, the rest potential is 300mV, the rest time is 30s, and the measuring range is 0.0001-50 mA.
The invention has the beneficial effects that: the invention provides an electrochemical detection method of trace vitamin B9, wherein a specific sample treatment solution is matched with a vitamin detector sensor probe modified by a calixarene acetone saturated solution for use, detection conditions are further optimized, and finally, the method can detect trace vitamin B9 more sensitively, quickly and accurately, has a wide application range, and can be used for quickly detecting trace vitamin B9 in samples such as blood samples, medicines, foods and the like. The sensor probe of the vitamin detector (namely the glassy carbon electrode modified by the calixarene acetone saturated solution) modified by the calixarene acetone saturated solution can detect vitamin B9 more sensitively and accurately, and the sensitivity and the accuracy of vitamin B9 detection can be improved because the calixarene is specifically combined with substances with corresponding sizes; by optimizing the concentration and the pH value of the sample treatment solution, the current in the detection process can be reduced, higher sensitivity can be used for detection, lower concentration detection is facilitated, and the accuracy is higher; the optimized detection condition can stably detect the sample, so that the detection range is wider and the detection concentration is lower. The method is simple and easy to operate, has short detection time, does not need special technical personnel for operation, and is easy to popularize.
Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention. The objectives and other advantages of the invention may be realized and attained by the means of the instrumentalities and combinations particularly pointed out hereinafter.
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For the purposes of promoting a better understanding of the objects, aspects and advantages of the invention, reference will now be made to the following detailed description taken in conjunction with the accompanying drawings in which:
FIG. 1 is a standard curve chart prepared by the current signal values of standard samples of vitamin B9 at different concentrations in example 1;
FIG. 2 is a standard curve chart prepared by the current signal values of the vitamin B9 standard samples with different concentrations in example 2;
FIG. 3 is a standard curve chart prepared by the current signal values of the vitamin B9 standard samples with different concentrations in example 3; .
Detailed Description
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.
Example 1
Detecting the content of vitamin B9 in the blood sample
(1) Manufacturing of sensor probe of vitamin detector modified by calixarene acetone saturated solution
Dripping 30 mu L of calixarene acetone saturated solution on the surface of a glassy carbon electrode which is polished by 0.03 mu m of alumina powder in a sensor probe of a vitamin detector, and completely volatilizing and drying acetone;
(2) uniformly mixing vitamin B9 standard samples with the volume of 80uL and the concentration of 5nmol/L, 10nmol/L, 20nmol/L, 40nmol/L and 80nmol/L with BR buffer solution with the concentration of 1000uL of 0.05mol/L and the pH value of 6.5, detecting current signals generated in the redox reaction of each vitamin B9 standard sample on a sensor probe of a vitamin detector modified by a calixarene acetone saturated solution by using a differential pulse method to obtain current signal values of vitamin B9 standard samples with different concentrations, establishing a standard curve, and obtaining a linear regression equation of y 3.6275x +0.1037 as shown in figure 1, wherein R is y 3.6275x +0.1037 as shown in figure 1
2The method has the lowest detection concentration (final concentration) of 5nM/L, the detection range of 5-1600000nM/L and the accuracy of 2.68%; wherein the vitamin detector has an enrichment electrodeposition potential of 200mV, an enrichment electrodeposition time of 240s, an initial potential of 100mV, a termination potential of 1000mV, and a scanning speed of40mV/s, sampling interval of 10mV, rest time of 20s, rest potential of 300mV, rest time of 30s and measuring range of 0.0001-50 mA;
(3) uniformly mixing 80uL of blood sample to be detected with 1000uL of BR buffer solution with the concentration of 0.05mol/L and the pH value of 6.5, detecting a current signal generated by the blood sample to be detected in an oxidation-reduction reaction on a sensor probe of a vitamin detector modified by a calixarene acetone saturated solution by using a differential pulse method to obtain a current signal value generated by the blood sample to be detected, and finally obtaining the content of vitamin B9 in the blood sample to be detected as 24.56nmol/L according to the current signal value generated by the blood sample to be detected and the standard curve formulated in the step (2); wherein the enrichment electrodeposition potential of the vitamin detector is 200mV, the enrichment electrodeposition time is 240s, the initial potential is 100mV, the termination potential is 1000mV, the scanning speed is 40mV/s, the sampling interval is 10mV, the rest time is 20s, the rest potential is 300mV, the rest time is 30s, and the measuring range is 0.0001-50 mA.
Example 2
Detecting the content of vitamin B9 in the blood sample
(1) Manufacturing of sensor probe of vitamin detector modified by calixarene acetone saturated solution
Dripping 10 mu L of calixarene acetone saturated solution on the surface of a glassy carbon electrode which is polished by 0.05 mu m of alumina powder in a sensor probe of a vitamin detector, and volatilizing and drying the acetone completely;
(2) uniformly mixing vitamin B9 standard samples with the volume of 80uL and the concentration of 8nmol/L, 16nmol/L, 32nmol/L, 64nmol/L and 128nmol/L with BR buffer solution with the concentration of 1000uL of 0.1mol/L and the pH value of 4, detecting current signals generated in an oxidation-reduction reaction of each vitamin B9 standard sample on a sensor probe of a vitamin detector modified by a calixarene acetone saturated solution by using a differential pulse method to obtain current signal values of vitamin B9 standard samples with different concentrations, and establishing a standard curve, wherein as shown in figure 2, a linear regression equation is y 4.7511x +2.5732, and R is Y4.7511 x +2.5732
2The method has the minimum detection concentration (final concentration) of 8nM/L, the detection range of 8-16000000nM/L and the accuracy of 3.15%; wherein, the vitamin detector is provided with an enrichment electrodeThe deposition potential is 180mV, the enrichment electrodeposition time is 300s, the initial potential is-200 mV, the termination potential is 600mV, the scanning speed is 30mV/s, the sampling interval is 20mV, the rest time is 25s, the rest potential is 400mV, the rest time is 60s, and the measuring range is 0.0001-50 mA;
(3) uniformly mixing 80uL of blood sample to be detected with 1000uL of BR buffer solution with the concentration of 0.1mol/L and the pH value of 4, detecting a current signal generated by the blood sample to be detected in an oxidation-reduction reaction on a sensor probe of a vitamin detector modified by a calixarene acetone saturated solution by using a differential pulse method to obtain a current signal value generated by the blood sample to be detected, and finally obtaining the content of vitamin B9 in the blood sample to be detected to be 18.95nmol/L according to the current signal value generated by the blood sample to be detected and the standard curve formulated in the step (2); wherein the enrichment electrodeposition potential of the vitamin detector is 180mV, the enrichment electrodeposition time is 300s, the initial potential is-200 mV, the termination potential is 600mV, the scanning speed is 30mV/s, the sampling interval is 20mV, the rest time is 25s, the rest potential is 400mV, the rest time is 60s, and the measuring range is 0.0001-50 mA.
Example 3
Detecting the content of vitamin B9 in the blood sample
(1) Manufacturing of sensor probe of vitamin detector modified by calixarene acetone saturated solution
Dripping 50 mu L of calixarene acetone saturated solution on the surface of a glassy carbon electrode which is polished by 1.0 mu m of alumina powder in a sensor probe of a vitamin detector, and completely volatilizing and drying acetone;
(2) uniformly mixing vitamin B9 standard samples with the volume of 80uL and the concentration of 6nmol/L, 12nmol/L, 24nmol/L, 48nmol/L and 96nmol/L with BR buffer solution with the concentration of 1000uL of 1.5mol/L and the pH value of 8, detecting current signals generated in the redox reaction of each vitamin B9 standard sample on a sensor probe of a vitamin detector modified by a calixarene acetone saturated solution by using a differential pulse method to obtain current signal values of vitamin B9 standard samples with different concentrations, and establishing a standard curve, wherein as shown in figure 3, a linear regression equation is y 4.6833x +0.5356, and R is R4.6833 x +0.5356
20.9987, the lowest detected concentration (end) of the methodConcentration) of 6nM/L, detection range of 6-16000000nM/L, accuracy of 4.28%; wherein the enrichment electrodeposition potential of the vitamin detector is-120 mV, the enrichment electrodeposition time is 120s, the initial potential is 50mV, the termination potential is 800mV, the scanning speed is 100mV/s, the sampling interval is 5mV, the rest time is 45s, the rest potential is 600mV, the rest time is 10s, and the measuring range is 0.0001-50 mA;
(3) uniformly mixing 80uL of blood sample to be detected with 1000uL of BR buffer solution with the concentration of 1.5mol/L and the pH value of 8, detecting a current signal generated by the blood sample to be detected in an oxidation-reduction reaction on a sensor probe of a vitamin detector modified by a calixarene acetone saturated solution by using a differential pulse method to obtain a current signal value generated by the blood sample to be detected, and finally obtaining the content of vitamin B9 in the blood sample to be detected as 31.2nmol/L according to the current signal value generated by the blood sample to be detected and the standard curve formulated in the step (2); wherein the enrichment electrodeposition potential of the vitamin detector is-120 mV, the enrichment electrodeposition time is 120s, the initial potential is 50mV, the termination potential is 800mV, the scanning speed is 100mV/s, the sampling interval is 5mV, the rest time is 45s, the rest potential is 600mV, the rest time is 10s, and the measuring range is 0.0001-50 mA.
Finally, the above embodiments are only intended to illustrate the technical solutions of the present invention and not to limit the present invention, and although the present invention has been described in detail with reference to the preferred embodiments, it will be understood by those skilled in the art that modifications or equivalent substitutions may be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions, and all of them should be covered by the claims of the present invention.
Claims (6)
1. An electrochemical detection method of trace vitamin B9, which is characterized by comprising the following steps:
(1) respectively and uniformly mixing vitamin B9 standard samples with different volume concentrations with BR buffer solution, then respectively detecting current signals generated by the vitamin B9 standard samples in an oxidation-reduction reaction on a sensor probe of a vitamin detector modified by a calixarene acetone saturated solution by using a differential pulse method, obtaining current signal values of the vitamin B9 standard samples with different concentrations, and making a standard curve;
(2) and (2) uniformly mixing a sample to be detected with BR buffer solution, detecting a current signal generated by the sample to be detected when the sample to be detected is subjected to an oxidation-reduction reaction on a sensor probe of the vitamin detector modified by the calixarene acetone saturated solution by using a differential pulse method to obtain a current signal value generated by the sample to be detected, and finally obtaining the content of vitamin B9 in the sample to be detected according to the current signal value generated by the sample to be detected and the standard curve formulated in the step (1).
2. The method according to claim 1, wherein in the step (1) and the step (2), the concentration of the BR buffer is 0.001-2.0 mol/L, and the pH value is 1.0-9.0.
3. The method of claim 1, wherein in step (1) and step (2), the calixarene acetone saturated solution modified vitamin detector sensor probe is prepared by the following method: and (3) dripping 10-50 mu L of calixarene acetone saturated solution on the surface of a glassy carbon electrode which is polished by 0.05-1 mu m of alumina powder in a sensor probe of a vitamin detector, and completely volatilizing and drying the acetone to obtain the calixarene acetone-based composite electrode.
4. The method of claim 3, wherein in step (1) and step (2), the calixarene acetone saturated solution modified vitamin detector sensor probe is prepared by the following method: and dripping 30 mu L of calixarene acetone saturated solution on the surface of a glassy carbon electrode which is polished by 0.03 mu m of alumina powder in a sensor probe of a vitamin detector, and completely volatilizing and drying the acetone to obtain the calixarene acetone saturated solution.
5. The method of claim 1, wherein in the step (1) and the step (2), the vitamin detector has an enrichment electrodeposition potential of-600 to 300mV, an enrichment electrodeposition time of 0 to 360s, an initial potential of-500 to 100mV, an end potential of 100 to 1000mV, a scanning speed of 10 to 500mV/s, a sampling interval of 4 to 20mV, a rest time of 10 to 60s, a rest potential of 200 to 600mV, a rest time of 10 to 60s, and a range of 0.0001 to 50 mA.
6. The method of claim 5, wherein in step (1) and step (2), the vitamin detector has an enrichment electrodeposition potential of 200mV, an enrichment electrodeposition time of 240s, an initial potential of 100mV, an end potential of 1000mV, a scanning speed of 40mV/s, a sampling interval of 10mV, a rest time of 20s, a rest potential of 300mV, a rest time of 30s, and a measuring range of 0.0001-50 mA.
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| CN108490048A (en) * | 2018-03-16 | 2018-09-04 | 常州大学 | A kind of preparation method of the chiral sensor of CTAB self assembly calixarenes for electrochemical recognition amino acid enantiomer |
| CN109342529A (en) * | 2018-09-19 | 2019-02-15 | 上海大学 | Non-enzymatic catalysis sensor of glucose and preparation method thereof |
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- 2019-09-24 CN CN201910905339.9A patent/CN110779972A/en active Pending
Patent Citations (4)
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|---|---|---|---|---|
| JPH08145943A (en) * | 1994-11-17 | 1996-06-07 | Tokuyama Corp | Measuring method for cation concentration |
| CN102192932A (en) * | 2011-03-23 | 2011-09-21 | 天津市兰标电子科技发展有限公司 | Normal pulse stripping method for detecting content of vitamin in blood samples |
| CN108490048A (en) * | 2018-03-16 | 2018-09-04 | 常州大学 | A kind of preparation method of the chiral sensor of CTAB self assembly calixarenes for electrochemical recognition amino acid enantiomer |
| CN109342529A (en) * | 2018-09-19 | 2019-02-15 | 上海大学 | Non-enzymatic catalysis sensor of glucose and preparation method thereof |
Non-Patent Citations (1)
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| VISHWANATH D. VAZE等: "Electrochemical behavior of folic acid at calixarene based chemically modified electrodes and its determination by adsorptive stripping voltammetry", 《ELECTROCHIMICA ACTA》 * |
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Application publication date: 20200211 |