CN110208355B - Method for measuring interaction efficiency of quinone substances and carboxymethyl lysine in solution - Google Patents

Method for measuring interaction efficiency of quinone substances and carboxymethyl lysine in solution Download PDF

Info

Publication number
CN110208355B
CN110208355B CN201910406744.6A CN201910406744A CN110208355B CN 110208355 B CN110208355 B CN 110208355B CN 201910406744 A CN201910406744 A CN 201910406744A CN 110208355 B CN110208355 B CN 110208355B
Authority
CN
China
Prior art keywords
solution
cml
substances
quinone
catechol
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201910406744.6A
Other languages
Chinese (zh)
Other versions
CN110208355A (en
Inventor
李玉婷
李琳
李冰
朱杰
余祥英
刘宇佳
张霞
张振辉
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Dongguan University of Technology
Original Assignee
Dongguan University of Technology
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Dongguan University of Technology filed Critical Dongguan University of Technology
Priority to CN201910406744.6A priority Critical patent/CN110208355B/en
Publication of CN110208355A publication Critical patent/CN110208355A/en
Application granted granted Critical
Publication of CN110208355B publication Critical patent/CN110208355B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems
    • G01N27/48—Systems using polarography, i.e. measuring changes in current under a slowly-varying voltage

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Electrochemistry (AREA)
  • Molecular Biology (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

The invention discloses a method for measuring interaction efficiency of quinone substances and carboxymethyl lysine (CML) in a solution. The method comprises the steps of measuring cyclic voltammetry curves of catechol substance solutions before and after CML addition, respectively recording peak current and voltage characteristic values of the catechol substance solutions before and after CML addition, and then calculating the reduction rate of reduction peak current to obtain the interaction efficiency of quinone substances and CML in the solutions. In the method, after the phenolic substances are oxidized to form the quinone substances, the quinone substances can quickly interact with CML in a system, so that the side reaction of the quinone substances is reduced; the adhesion amount of the substance on the surface of the electrode is small, and the detection accuracy is high; the interference of factors such as the adhesion of polyphenol and oxidation products thereof on a working electrode, the related side reaction of quinone substances and the like on the interaction of the quinone substances and CML in the prior art is reduced; meanwhile, the determination method is simple, short in analysis time, good in repeatability and convenience, low in use cost and wide in application.

Description

Method for measuring interaction efficiency of quinone substances and carboxymethyl lysine in solution
Technical Field
The invention relates to the technical field of food safety detection and analytical chemistry, in particular to a method for determining interaction efficiency of quinone substances and carboxymethyl lysine (CML) in a solution.
Background
CML (carboxymethyl lysine) is currently the most widely studied typical end-product of Advanced Glycation (AGEs), which is widely found in various food products. CML is digested and absorbed and then accumulates in tissues, and existing researches prove that the CML has potential harm to human health. Therefore, the potential harm of the CML to the human health can be effectively reduced by researching a means for reducing the CML content.
The catechol substance can be oxidized to form corresponding catechol substances while playing an anti-oxidation role, and the catechol substances serving as strong electrophiles are easy to generate Michael addition reaction with amino on CML, so that the aim of eliminating the CML of the system is fulfilled. At present, the method for determining the interaction efficiency between the o-phenylenedioquinone substances and the CML mainly comprises the steps of oxidizing the o-phenylenedioquinone substances in a solution by using a potential controlled electrolytic coulometry method to obtain corresponding o-phenylenedioquinone substances, and then determining the interaction dynamics between the o-phenylenedioquinone substances and the CML by using a stopped-flow spectrum. The preparation of the o-phenylenedione substances by the potential electrolytic coulometry method requires that the cyclic voltammetry behavior of the selected o-phenylenedione substances on the working electrode is reversible and has no obvious adsorbability. At present, 4-methyl catechol is only successfully used for preparing corresponding quinone substances (4-methyl catechol), and the interaction condition of 4-methyl catechol and CML is measured in detail by using a stopped flow spectrum.
Most polyphenols containing catechol structure, for example: catechin, rutin, quercetin and the like are easy to be adsorbed on the surface of the working electrode, and quinone substances formed by oxidation of the catechin, rutin, quercetin and the like are easy to generate side reactions such as polymerization, rearrangement and the like, so that the possibility of preparing the corresponding quinone substances by using a controlled potential electrolytic coulometry method is not provided. Therefore, it is impossible to measure the interaction between the quinone-like substance obtained by oxidizing the polyphenol-like substance in the solution and CML by the conventional means.
Therefore, in view of the above-mentioned drawbacks of the detection method, it is necessary to provide a detection method with a wider application range.
Disclosure of Invention
The invention aims to provide a method for measuring the interaction efficiency of quinone substances and CML in a solution, aiming at the defects that in the prior art, the interaction efficiency of the quinone substances and the CML is detected by a potential electrolysis coulomb-stop flow spectrum combined method, the applied variety of catechol substances is less, and the application range is limited. The method utilizes the electrochemical activity of the catechol substance, measures the reduction peak current of the catechol substance before and after adding CML by cyclic voltammetry, and calculates the reduction rate of the reduction peak current, thereby obtaining the interaction efficiency of the quinone substance and the CML in the solution; the method is suitable for most of the catechol substances, is simple, convenient and fast in determination method, accurate and reliable in detection result, has stability and repeatability, and has wide application prospect in detection and evaluation of interaction efficiency of quinone substances and CML in food.
The above object of the present invention is achieved by the following scheme:
a method for determining the content of carboxymethyl lysine (CML) in a solution, comprising the steps of:
s1, dissolving a catechol substance in a buffer solution to obtain a solution A; dissolving CML in a buffer solution to obtain a solution B; dissolving catechol substances and CML in a buffer solution to obtain a solution C;
s2, taking a glassy carbon electrode as a working electrode, a platinum wire as a counter electrode, silver/saturated silver chloride as a reference electrode, connecting each electrode with an electrochemical workstation, constructing a three-electrode system, respectively detecting a solution A, a solution B and a solution C by using a cyclic voltammetry method to obtain a cyclic voltammetry curve of each solution, and respectively recording a peak current and a voltage characteristic value of each solution;
s3, according to the peak current and voltage characteristic values measured in the step S2, calculating according to the following formula, and then measuring the interaction efficiency of the quinone substances and the CML in the solution:
efficiency of interaction (reduction peak current reduction rate,%) (I)pC1 (phenol)-IpC1 (phenol + CML))/IpC1 (phenol)；
Wherein said IpC1 (phenol)Is the reduction peak-to-peak current of the solution A; said IpC1 (phenol + CML)Is the reduction peak-to-peak current of the solution C.
Because the structure of the catechol substance has electrochemical activity, in cyclic voltammetry scanning, when voltage is scanned in a positive direction, the catechol substance is oxidized on the surface of an electrode to form a corresponding quinone substance, and in a subsequent negative scanning process, the quinone substance is reduced to the original phenol substance on the surface of the electrode to form a pair of redox peaks; when the CML in the solution system reacts with the quinones to cause consumption of the quinones, the reduction peak current of the solution C is obviously reduced compared with that of the solution A in the negative scanning process, and the interaction efficiency of the quinones and the CML can be calculated by measuring the reduction rate of the reduction peak current in the cyclic voltammetry curve of the phenols before and after the addition of the CML.
When the original control potential electrolytic coulomb method is used for detection, a certain voltage is applied to completely oxidize phenol in the system to form quinone after a period of time (about 40 minutes), and then the quinone reacts with CML to determine the interaction efficiency; because the oxidation of the phenolic substances into the quinone substances takes a long time, the generated quinone substances are required to be very stable, otherwise, when the quinone substances generate a series of side reactions or are attached to the electrode to obstruct the subsequent preparation process, the accuracy of the detection result is affected, and when the quinone substances are attached to the electrode for a long time, the irreversible damage of the reticular glassy carbon electrode is caused. The whole scanning process of the cyclic voltammetry adopted by the invention is not more than 3 minutes, once the phenolic substances are oxidized on the electrode to form the quinone substances, the quinone substances can react with the CML in which the system coexists, the possibility of side reaction is reduced, and the quinone substances can not be attached to the surface of the electrode for a long time, so that the damage to the electrode is reduced.
Preferably, the catechol substance is one or more of catechol, 4-methyl catechol, protocatechuic acid ethyl ester, caffeic acid, chlorogenic acid, rosmarinic acid, rutin, (+) -catechin, (-) -epicatechin, luteolin or quercetin. The detection method provided by the invention is applicable to various catechol substances, and has a wide application range.
Preferably, in the cyclic voltammetry test process, the scanning potential range of the electrochemical workstation is-0.8-1.2V; the scanning speed is 0.01-0.10V/s; sensitivity of 10-5A/V; the initial scanning direction is the forward direction; the number of scanning segments is 2; the temperature was 25. + -. 1 ℃.
Preferably, before the solution A, the solution B and the solution C are detected, oxygen in the solution needs to be removed.
More preferably, before the solution A, the solution B and the solution C are detected, nitrogen is respectively introduced into each solution to remove oxygen.
More preferably, the nitrogen is introduced for 5-10 min.
Preferably, the glassy carbon working electrode has a diameter of 3 mm; before each scanning, the glassy carbon working electrode needs to be polished on a chamois leather dropwise added with an alumina suspension liquid with the particle size of 0.05 mu m, and then washed by distilled water. Before the test, the glass carbon electrode is polished to effectively remove attachments on the surface of the electrode, so that the surface of the electrode is clean before the test, and the detection accuracy is improved.
Preferably, the buffer solution is prepared by phosphate.
Preferably, the pH of the buffer solution is 5.0-8.0, wherein the concentration of phosphate is 0.2M.
Preferably, when the solute in the solution a is luteolin or quercetin, the solvent is a mixed solution of 0.2M phosphate buffer and ethanol, and the volume ratio is 4: 1.
Preferably, the concentration of the catechol in the solutions A and C is 0.5mM.
Preferably, the concentration of CML in the solution B and the solution C is 0.5-7.5 mM.
Compared with the prior art, the invention has the following beneficial effects:
(1) according to the invention, by using cyclic voltammetry, after phenolic substances are oxidized to form quinone substances, the quinone substances can quickly interact with CML in a system, so that side reactions of the quinone substances are reduced; the attachment on the surface of the electrode can be effectively removed by polishing the glassy carbon electrode before each test, so that the detection accuracy is improved; the interference of factors such as polyphenol and oxidation products thereof attached to a working electrode, quinone related side reactions and the like on the interaction of the quinone and the CML in the prior art is reduced.
(2) The method disclosed by the invention is simple, short in analysis time, good in repeatability and convenience, low in use cost, and good in application prospect in the field of interaction research of quinone substances and CML, and overcomes the defect that only the interaction of 4-methylphthaloquinone and CML can be measured by means of the prior art.
Drawings
FIG. 1 is a cyclic voltammogram (pH 7.0, scan rate 0.01V/s) of catechol interaction with CML.
FIG. 2 is a cyclic voltammogram of the interaction of 4-methylcatechol with CML (pH 7.0, scan rate 0.01V/s).
FIG. 3 is a cyclic voltammogram of protocatechuic acid interaction with CML (pH 7.0, scan rate 0.01V/s).
FIG. 4 is a cyclic voltammogram of protocatechuic acid interaction with CML (pH 7.0, scan rate 0.05V/s).
FIG. 5 is a cyclic voltammogram of the interaction of protocatechuic acid ethyl ester with CML (pH 7.0, scan rate 0.05V/s).
FIG. 6 is a cyclic voltammogram (pH 7.0, scan rate 0.01V/s) of caffeic acid interacting with CML.
FIG. 7 is a cyclic voltammogram (pH 7.0, scan rate 0.01V/s) of interaction of chlorogenic acid with CML.
FIG. 8 is a cyclic voltammogram of the interaction of rosmarinic acid with CML (pH 7.0, scan rate 0.01V/s).
FIG. 9 is a cyclic voltammogram of rutin interaction with CML (pH 7.0, scan rate 0.05V/s).
FIG. 10 is a cyclic voltammogram of the interaction of (+) -catechin with CML (pH 7.0, scan rate 0.05V/s).
FIG. 11 is a cyclic voltammogram of the interaction of (-) -epicatechin with CML (pH 7.0, scan rate 0.05V/s).
FIG. 12 is a cyclic voltammogram of luteolin interaction with CML (pH 7.0, scan rate 0.05V/s).
FIG. 13 is a cyclic voltammogram (pH 7.0, scan rate 0.05V/s) of the interaction of quercetin with CML.
Detailed Description
The present invention is further described in detail below with reference to specific examples, which are provided for illustration only and are not intended to limit the scope of the present invention. The test methods used in the following examples are all conventional methods unless otherwise specified; the materials, reagents and the like used are, unless otherwise specified, commercially available reagents and materials.
Example 1
A method for determining the interaction efficiency of quinone substances and CML in a solution comprises the following steps:
s1, dissolving catechol substance in a buffer solution to obtain a solution A; dissolving CML in a buffer solution to obtain a solution B; dissolving catechol substances and CML in a buffer solution to obtain a solution C;
s2, taking a glassy carbon electrode (with the diameter of 3mm) as a working electrode, a platinum wire as a counter electrode, silver/saturated silver chloride as a reference electrode, connecting each electrode with an electrochemical workstation, constructing a three-electrode system, detecting the solution A, the solution B and the solution C by using a cyclic voltammetry method to obtain a cyclic voltammetry curve of each solution, and recording the peak current and voltage characteristic value of each solution respectively; before testing, introducing high-purity nitrogen into each part of solution to be tested for 5-10 min to remove oxygen; before scanning each time, the glassy carbon working electrode needs to be polished on chamois leather dropwise added with alumina suspension liquid with the particle size of 0.05 mu m, and then washed by distilled water; the scanning potential range of the electrochemical workstation is-0.8-1.2V; the scanning speed is 0.01-0.10V/s; sensitivity of 10-5A/V; the initial scanning direction is the forward direction; the number of scanning segments is 2; the temperature is 25 +/-1 ℃;
s3, according to the peak current and voltage characteristic values measured in the step S2, calculating according to the following formula, and measuring the interaction efficiency of the quinone substances and the CML as follows:
interaction efficiency (reduction peak current reduction rate,%) is (I)pC1 (phenol)-IpC1 (phenol + CML))/IpC1 (phenol)；
Wherein said IpC1 (phenol)Is the reduction peak-to-peak current of the solution A; said IpC1 (phenol + CML)Is the reduction peak-to-peak current of the solution C.
1. The influence of the pH value, the scanning rate and the CML concentration of the buffer solution on the detection result is explored
Following the procedure of step S1 above, NaH is used2PO4、Na2HPO4Respectively preparing 0.2M phosphate buffer solutions with pH values of 5.0, 7.0 and 8.0, and then dissolving a certain amount of catechol or 4-methylcatechol to prepare a solution A, wherein the concentration of the catechol or 4-methylcatechol is 0.5 mM;
dissolving a certain amount of CML in 0.2M phosphoric acid buffer solution to prepare solution B, wherein the concentration of the CML is 0.5-7.5 mM;
dissolving a certain amount of catechol (or 4-methylcatechol) and CML in a buffer solution, and uniformly mixing by magnetic stirring to obtain a solution C, wherein the concentration of the catechol (or 4-methylcatechol) in the solution C is ensured to be 0.5mM, and the concentration of the CML is ensured to be 0.5-7.5 mM.
The measurement was performed by changing the scanning rates (0.01, 0.05 and 0.10V/S) according to the measurement procedure in the above step S2, and the measurement results are shown in Table 1 and Table 2, wherein Table 1 shows the measurement results of the interaction efficiency between phthaloquinone and CML (5.0mM) under different conditions, Table 2 shows the measurement results of the interaction efficiency between 4-methylphthalquinone and CML (5.0mM) under different conditions, and Table 3 shows the measurement results of the interaction efficiency between 4-methylphthalquinone and CML (0.5-7.5mM) at different concentrations at a scanning rate of 0.01V/S at pH 7.0. Wherein a greater reduction in reduction current indicates a greater degree of reaction, i.e., a greater efficiency of reaction of the quinone with the CML.
TABLE 1 interaction efficiency of o-benzoquinone with CML under different conditions
Figure BDA0002061477460000061
TABLE 2 efficiency of the interaction of 4-methylphthaloquinone with CML under different conditions
Figure BDA0002061477460000062
As can be seen from tables 1 and 2, the method of the present invention has good stability under the conditions of pH7.0 and 8.0, the RSD value is between 0.00 and 9.22, and the stability is good; at pH 5.0, the process stability is poor due to the low efficiency of the interaction between the quinone and CML, and the RSD value is high. Therefore, the method is more suitable for detecting the interaction efficiency of the quinone substances and the CML in a solution system with the pH of 7.0 and 8.0.
The invention judges the interaction between the CML and the quinone substances by measuring the reduction rate of the reduction peak current in the cyclic voltammetry curve of the phenol substances before and after the addition of the CML, and the interaction efficiency is higher when the reduction rate is higher. The high and low order of the interaction efficiency of the o-phenylenediquinone or 4-methyl o-phenylenediquinone and the CML under the conditions of different pH values and the same scanning rate is as follows: pH8.0> pH7.0 > pH 5.0; compared with the existing method for preparing 4-methyl o-phenylenediquinone by using a controlled potential electrolytic coulometry method, and then measuring the interaction efficiency of the 4-methyl o-phenylenediquinone substance and CML under different pH conditions by using a stopped flow spectrum (Food Chemistry,2018,244:25-28), namely, pH8.0 is more than pH7.0 is more than pH 5.0, so the method for measuring the interaction efficiency of the quinone substance and the CML in the solution has higher accuracy.
TABLE 3 interaction efficiency of 4-methylphthaloquinone with CML of different concentrations at pH7.0 and scan rate 0.01V/s
Figure BDA0002061477460000071
As can be seen from Table 3, the stability of the interaction efficiency of 4-methylphthaloquinone and CML with different concentrations measured by the method is better at pH7.0 and at a scanning rate of 0.01V/s, and the RSD value range is 0.35-5.62. With the increase of the CML content of the system, the interaction efficiency of the 4-methylphthalquinone and the CML is remarkably increased, and the theoretical expectation is met.
2. The influence of different substituents on the benzene ring on the interaction efficiency of the quinone substances and the CML is explored
The test procedure was as described in example 1, except that the solution A was prepared by dissolving catechol, 4-methylcatechol or protocatechuic acid in a phosphate buffer (0.2M, pH7.0) to give a polyphenol substance concentration of 0.5 mM; the solution C is prepared by dissolving catechol, 4-methyl catechol or protocatechuic acid and CML in phosphate buffer solution (0.2M, pH7.0), wherein the concentration of polyphenols is 0.5mM, and the concentration of CML is 5.0 mM. During the test, the operating parameters of the electrochemical workstation were: scanning potential range, -0.8-1.2V; scanning rate, 0.01V/s; sensitivity, 10-5A/V; initial scan direction, forward; number of scan segments, 2. Cyclic voltammograms of each solution were obtained and the peak current and voltage characteristics of each solution were recorded separately and calculated according to the formula in example 1. The calculation results are shown in table 4.
TABLE 4 efficiency of interaction of different quinones with CML
Figure BDA0002061477460000072
As can be seen from Table 4, under the same conditions, CML interacted most efficiently with protocatechuic acid quinone formed by oxidation of protocatechuic acid, intermediate with catechol formed by oxidation of catechol, and least efficiently with 4-methylcatechol formed by oxidation of 4-methylcatechol. The methyl group on the benzene ring has an electron donating effect, so that the electron cloud density of the quinone substances is increased, and the electrophilic reaction activity is reduced; the carboxyl group on the benzene ring has an electron withdrawing effect to reduce the electron cloud density of the quinone substances, so that the electrophilic reaction activity of the quinone substances is enhanced. In conclusion, the test result of the invention accords with the theoretical expectation and has higher stability (the RSD value ranges from 0.00 to 9.22).
3. Reaction degree of protocatechuic acid and protocatechuic acid carboxyl ethyl esterification oxidation to corresponding quinones and CML
The test procedure was as described in example 1, except that the solution A was prepared by dissolving protocatechuic acid or protocatechuic acid ethyl ester in a phosphate buffer (0.2M, pH7.0), respectively, and the concentration of polyphenols was 0.5 mM; the solution C is protocatechuic acid or protocatechuic acid ethyl ester and CML respectively dissolved in phosphate buffer (0.2M, pH7.0) to obtain solution C, wherein the concentration of polyphenols is 0.5mM, and the concentration of CML is 5.0 mM. During the test, the operating parameters of the electrochemical workstation were: scanning potential range, -0.8-1.2V; scanning rate, 0.05V/s; sensitivity, 10-5A/V; initial scan direction, forward; number of scan segments, 2. Cyclic voltammograms of each solution were obtained and the peak current and voltage characteristics of each solution were recorded separately and calculated according to the formula in example 1. The calculation results are shown in table 5.
TABLE 5 efficiency of interaction of various catechol quinones with CML
Figure BDA0002061477460000081
As can be seen from Table 5, under the same conditions, the interaction efficiency of CML with protocatechuic acid quinone formed by oxidation of protocatechuic acid is higher than that of protocatechuic acid ethyl ester quinone formed by oxidation of protocatechuic acid ethyl ester, the electron withdrawing effect of carboxyl group is weakened by the esterification of carboxyl group, and thus the electrophilic reaction capability is reduced, and the test result of the invention is in line with the theoretical expectation and has higher stability (RSD value is in the range of 0.39-6.22).
4. Reaction degree of quinone substances formed by oxidation of different phenolic acid substances with CML
The test procedure was as described in example 1, except that each of the solutions A was prepared by dissolving caffeic acid, chlorogenic acid or rosmarinic acid in a phosphate buffer (0.2M, pH7.0) to give a solution containing polyphenols in a concentration of 0.5 mM; the solution C is prepared by dissolving caffeic acid, chlorogenic acid or rosmarinic acid and CML in phosphate buffer (0.2M, pH7.0), wherein the concentration of polyphenols is 0.5mM, and the concentration of CML is 5.0 mM. During the test, the operating parameters of the electrochemical workstation were: scanning potential range, -0.8-1.2V; scanning rate, 0.01V/s; sensitivity, 10-5A/V; initial scan direction, forward; number of scan segments, 2. Cyclic voltammograms of each solution were obtained and the peak current and voltage characteristics of each solution were recorded separately and calculated according to the formula in example 1. The calculation results are shown in table 6.
TABLE 6 reaction degree of Quinones with CML formed by oxidation of various phenolic acids
Figure BDA0002061477460000091
As can be seen from Table 6, under the same conditions, the interaction efficiency of CML and caffeic acid quinone formed by caffeic acid oxidation is higher than that of quinines oxidized by chlorogenic acid or rosmarinic acid, and the steric hindrance effect brought by large substituents on benzene rings of chlorogenic acid quinone and rosmarinic acid quinone weakens the interaction efficiency of CML and the test result of the invention is in line with theoretical expectation and has higher stability (RSD value is in the range of 2.80-7.27).
5. Reaction degree of quinone substances formed by oxidation of flavonoids with CML
The test procedure was as described in example 1, except that the solution A was prepared by dissolving rutin, (+) -catechin or (-) -epicatechin in phosphate buffer (0.2M, pH7.0), or luteolin or quercetin in a mixture of 0.2M phosphate buffer and ethanol (volume ratio: 4:1, pH7.0), and the concentration of polyphenols was 0.5 mM; the solution C is prepared by dissolving rutin, (+) -catechin or (-) -epicatechin and CML in phosphate buffer solution (0.2M, pH7.0), or dissolving luteolin or a mixed solution (volume ratio: 4:1, pH7.0) of quercetin and CML in 0.2M phosphate buffer solution and ethanol, wherein the concentration of polyphenols is 0.5mM, and the concentration of CML is 5.0 mM. During the test, the operating parameters of the electrochemical workstation were: scanning potential range, -0.8-1.2V; scanning rate, 0.05V/s; sensitivity, 10-5A/V; initial scan direction, forward; number of scan segments, 2. Cyclic voltammograms of each solution were obtained and the peak current and voltage characteristics of each solution were recorded separately and calculated according to the formula in example 1. The calculation results are shown in table 7.
TABLE 7 interaction efficiency of quinone with CML by oxidation of flavonoids
Figure BDA0002061477460000092
As can be seen from Table 7, under the same conditions, the interaction efficiency of CML and luteolin quinone formed by oxidizing luteolin is the highest among the selected flavones, which is related to the lower density of benzene ring electron cloud, the test result of the invention is in line with the theoretical expectation, the RSD value range is 5.48-42.73, and the test stability of partial flavonoids is not good.
In conclusion, the method is suitable for detecting the reaction efficiency of the CML and the quinone substances formed by oxidizing various different phenolic substances, accords with theoretical expectation, shows that the detection result is accurate, and simultaneously shows that the RSD value of the detection result is small, and shows that the detection method and the detection result have stability.
It should be finally noted that the above examples are only intended to illustrate the technical solutions of the present invention, and not to limit the scope of the present invention, and that other variations and modifications based on the above description and thought may be made by those skilled in the art, and that all embodiments need not be exhaustive. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the claims of the present invention.

Claims (4)

1. A method for measuring the interaction efficiency of quinone substances and carboxymethyl lysine in a solution is characterized by comprising the following steps:
s1, dissolving a catechol substance in a buffer solution to obtain a solution A; dissolving CML in a buffer solution to obtain a solution B; dissolving catechol substances and CML in a buffer solution to obtain a solution C;
s2, taking a glassy carbon electrode as a working electrode, a platinum wire as a counter electrode, silver/saturated silver chloride as a reference electrode, connecting each electrode with an electrochemical workstation, constructing a three-electrode system, respectively detecting a solution A, a solution B and a solution C by using a cyclic voltammetry method to obtain a cyclic voltammetry curve of each solution, and respectively recording a peak current and a voltage characteristic value of each solution;
s3, according to the peak current and voltage characteristic values measured in the step S2, calculating according to the following formula, and then measuring the interaction efficiency of the quinone substances and the CML in the solution:
efficiency of interaction/% (I)pC1 (phenol)-IpC1 (phenol + CML))/IpC1 (phenol)；
Wherein said IpC1 (phenol)The reduction peak current of the structural unit of the catechol which is a phenolic substance in the solution A; said IpC1 (phenol + CML)The reduction peak-to-peak current of the structural unit of catechol which is a phenolic substance in the solution C;
the catechol substance is one or more of catechol, protocatechuic acid ethyl ester, caffeic acid, chlorogenic acid, rosmarinic acid, rutin, (+) -catechin, (-) -epicatechin, luteolin or quercetin;
in the cyclic voltammetry test process, the scanning potential range of the electrochemical workstation is-0.8-1.2V; the scanning speed is 0.01-0.10V/s; sensitivity of 10-5A/V; the initial scanning direction is the forward direction; the number of scanning segments is 2; the temperature is 25 +/-1 ℃;
the buffer solution is prepared from phosphate; the pH value of the buffer solution is 7.0-8.0, and the concentration of phosphate is 0.2M;
in the solution A and the solution C, the concentration of the catechol substance is 0.5 mM; in the solutions B and C, the concentration of CML is 0.5-7.5 mM;
the diameter of the glassy carbon working electrode is 3 mm; before each scanning, the glassy carbon working electrode needs to be polished on a chamois leather dropwise added with alumina suspension liquid with the particle size of 0.05 mu m, and then is cleaned by distilled water;
when the solute in the solution A is luteolin or quercetin, the solvent is a mixed solution of 0.2M phosphate buffer solution and ethanol, and the volume ratio is 4: 1.
2. The method for determining the interaction efficiency of quinone with CML in a solution as claimed in claim 1, wherein said solution A, solution B and solution C are subjected to a detection process by removing oxygen from the solution.
3. The method for determining the interaction efficiency of quinone substances with CML in solution as claimed in claim 2, wherein nitrogen is introduced into each solution to remove oxygen before the buffer solution A, the buffer solution B and the mixed solution C are tested.
4. The method for determining the interaction efficiency of quinone substances and CML in a solution according to claim 3, wherein the nitrogen is introduced for 5-10 min.
CN201910406744.6A 2019-05-16 2019-05-16 Method for measuring interaction efficiency of quinone substances and carboxymethyl lysine in solution Active CN110208355B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201910406744.6A CN110208355B (en) 2019-05-16 2019-05-16 Method for measuring interaction efficiency of quinone substances and carboxymethyl lysine in solution

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201910406744.6A CN110208355B (en) 2019-05-16 2019-05-16 Method for measuring interaction efficiency of quinone substances and carboxymethyl lysine in solution

Publications (2)

Publication Number Publication Date
CN110208355A CN110208355A (en) 2019-09-06
CN110208355B true CN110208355B (en) 2022-07-05

Family

ID=67787472

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201910406744.6A Active CN110208355B (en) 2019-05-16 2019-05-16 Method for measuring interaction efficiency of quinone substances and carboxymethyl lysine in solution

Country Status (1)

Country Link
CN (1) CN110208355B (en)

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101349673B (en) * 2008-09-04 2012-12-19 南京大学 Electrochemical method for non-eletricity enriching and measuring trace amount tervalence inorganic arsenic in water system
CN102288669B (en) * 2011-05-04 2013-06-26 青岛大学 Electrochemical method based on graphene-modified electrode for simultaneous determination of rutin and quercetin
CN102809592B (en) * 2012-07-13 2014-03-12 江苏大学 Electro-polymerization preparation method of electrochemical sensor for quickly detecting EGCG (Epigallocatechin-3-Gallate)
CN105918772B (en) * 2016-05-30 2020-01-14 华南理工大学 Carboxymethyl lysine eliminating agent and application thereof

Also Published As

Publication number Publication date
CN110208355A (en) 2019-09-06

Similar Documents

Publication Publication Date Title
Huang et al. Electrochemical behavior and voltammetric determination of tryptophan based on 4-aminobenzoic acid polymer film modified glassy carbon electrode
Gholivand et al. Electrooxidation behavior of warfarin in Fe3O4 nanoparticles modified carbon paste electrode and its determination in real samples
Rice et al. Simultaneous voltammetric and chemical monitoring of dopamine release in situ
Zhou et al. Selective determination of dopamine and uric acid using electrochemical sensor based on poly (alizarin yellow R) film-modified electrode
Kong et al. Palygorskite–expanded graphite electrodes for catalytic electro-oxidation of phenol
Hua et al. Electrochemical behavior of 5-fluorouracil on a glassy carbon electrode modified with bromothymol blue and multi-walled carbon nanotubes
CN102141538B (en) Method for determining concentration of phosphite by cyclic voltammetry
CN104569121B (en) Method for detecting lead ion concentration in solution
CN110208355B (en) Method for measuring interaction efficiency of quinone substances and carboxymethyl lysine in solution
Alghamdi Voltammetric analysis of montelukast sodium in commercial tablet and biological samples using the hanging mercury drop electrode
CN109374708A (en) With the method for hydroxylated multi-walled carbon nanotubes the@cubic mesoporous measurement of carbon composite membrane electrochemical sensor trace olaquindox and carbadox
Cao et al. Determination of active ingredients of Apocynum venetum by capillary electrophoresis with electrochemical detection
CN107102052B (en) Based on the uric acid electrochemical sensor containing active copper carbon dots and its application
JPH08511625A (en) Gas concentration measurement
El-Shahawi et al. Determination of trace levels of diosmin in a pharmaceutical preparation by adsorptive stripping voltammetry at a glassy carbon electrode
CN106990150A (en) A kind of preparation method of novel molecular engram polymer rutin electrochemical sensor
Wu et al. Voltammetric Investigation of Vitamin B~ 6 at a Glassy Carbon Electrode and Its Application in Determination
CN101893595A (en) Chemically modified electrode, preparation thereof and method for rapid determination of acid value of plant oil
CN108426939A (en) A method of detection Antioxidants In Food content
Tuzhi et al. Voltammetric measurement of haloperidol following adsorptive accumulation at glassy-carbon electrodes
CN110702770B (en) Method for identifying reaction product of 4-methylphthaloquinone and amino compound in solution
CN111551595B (en) Composite material modified acetylene black carbon paste electrode for measuring content of triclosan and preparation method and application thereof
CN104458859A (en) La0.88Sr0.12MnO3 nano-fiber modified carbon paste electrode and preparation method and application thereof
Qing-Cui et al. Determination of effective ingredients in compound sulfamethoxazole tablets by capillary electrophoresis with amperometric detection
CN109060907A (en) Application of the poly- complex film modified glass-carbon electrode of gold/acetylene black in detection hydroquinone

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant