CN116660338A - Method for quantitatively detecting vitamin K3 - Google Patents
Method for quantitatively detecting vitamin K3 Download PDFInfo
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- CN116660338A CN116660338A CN202310586274.2A CN202310586274A CN116660338A CN 116660338 A CN116660338 A CN 116660338A CN 202310586274 A CN202310586274 A CN 202310586274A CN 116660338 A CN116660338 A CN 116660338A
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- MJVAVZPDRWSRRC-UHFFFAOYSA-N Menadione Chemical compound C1=CC=C2C(=O)C(C)=CC(=O)C2=C1 MJVAVZPDRWSRRC-UHFFFAOYSA-N 0.000 title claims abstract description 205
- 235000012711 vitamin K3 Nutrition 0.000 title claims abstract description 104
- 239000011652 vitamin K3 Substances 0.000 title claims abstract description 104
- 238000000034 method Methods 0.000 title abstract description 10
- 238000001514 detection method Methods 0.000 claims abstract description 62
- 230000006698 induction Effects 0.000 claims abstract description 46
- 238000006243 chemical reaction Methods 0.000 claims abstract description 14
- 239000000523 sample Substances 0.000 claims description 19
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims description 15
- 239000012488 sample solution Substances 0.000 claims description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 10
- 239000002904 solvent Substances 0.000 claims description 5
- 238000004445 quantitative analysis Methods 0.000 abstract description 3
- 239000000243 solution Substances 0.000 description 50
- 239000000126 substance Substances 0.000 description 8
- 239000012085 test solution Substances 0.000 description 8
- 239000012153 distilled water Substances 0.000 description 6
- 150000003716 vitamin K3 derivatives Chemical class 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- XUFQPHANEAPEMJ-UHFFFAOYSA-N famotidine Chemical compound NC(N)=NC1=NC(CSCCC(N)=NS(N)(=O)=O)=CS1 XUFQPHANEAPEMJ-UHFFFAOYSA-N 0.000 description 3
- 239000011259 mixed solution Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 201000000839 Vitamin K Deficiency Bleeding Diseases 0.000 description 2
- 206010047634 Vitamin K deficiency Diseases 0.000 description 2
- 208000016794 vitamin K deficiency hemorrhagic disease Diseases 0.000 description 2
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 1
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 1
- 208000032843 Hemorrhage Diseases 0.000 description 1
- 206010025476 Malabsorption Diseases 0.000 description 1
- 229930003448 Vitamin K Natural products 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- 239000003114 blood coagulation factor Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002848 electrochemical method Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 208000031169 hemorrhagic disease Diseases 0.000 description 1
- 238000012625 in-situ measurement Methods 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- SHUZOJHMOBOZST-UHFFFAOYSA-N phylloquinone Natural products CC(C)CCCCC(C)CCC(C)CCCC(=CCC1=C(C)C(=O)c2ccccc2C1=O)C SHUZOJHMOBOZST-UHFFFAOYSA-N 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000003805 procoagulant Substances 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 235000019168 vitamin K Nutrition 0.000 description 1
- 239000011712 vitamin K Substances 0.000 description 1
- 150000003721 vitamin K derivatives Chemical class 0.000 description 1
- 229940046010 vitamin k Drugs 0.000 description 1
Classifications
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- 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
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/302—Electrodes, e.g. test electrodes; Half-cells pH sensitive, e.g. quinhydron, antimony or hydrogen electrodes
-
- 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
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 Analysing Biological Materials (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The invention relates to a method for quantitatively detecting vitamin K3, which is characterized by comprising the following steps of: application of HCHO-NaHSO 3 ‑Na 2 SO 3 The pH clock reaction system is used as a detection solution, and quantitative analysis of the vitamin K3 is realized according to different responses of the system to different concentrations of the vitamin K3, namely different induction times. The quantitative analysis method for vitamin K3 has the characteristics of high accuracy, easiness in operation, convenience, quickness and the like.
Description
Technical Field
The invention relates to an analysis and detection method, in particular to a method for establishing HCHO- NaHSO 3 - Na 2 SO 3 The method for quantitatively analyzing the vitamin K3 is realized according to different responses of the system to the vitamin K3 with different concentrations, namely different induction time, and belongs to the field of analytical chemistry.
Background
Vitamin K3 with molecular formula of C 11 H 8 O 2 Is an analytical reagent in chemical analysis. Vitamin K3 is mainly a procoagulant, and can be used for treating hemorrhagic diseases caused by vitamin K deficiency, such as neonatal hemorrhage, vitamin K deficiency caused by intestinal malabsorption, and hypoprothrombinia. Vitamin K3 is artificially synthesized vitamin K, which is used as the necessary substance for human physiological coagulation, and participates in the synthesis of coagulation factors to maintain blood coagulation in human bodyIs a physiological process of (a).
The current detection methods for vitamin K3 comprise liquid chromatography, spectrophotometry, electrochemical methods and the like. However, such detection methods often require large equipment and are expensive to test and are not suitable for in-situ measurement. Therefore, it is necessary to find a detection and analysis method which has good detection effect and is simple and convenient and quick to operate. The structure of the vitamin K3 is shown as a structural formula (I).
Vitamin K3 structural formula (I)
Disclosure of Invention
The invention aims to provide a novel quantitative detection method for vitamin K3, namely, HCHO is adopted- NaHSO 3 - Na 2 SO 3 The pH clock system is a method for quantitatively detecting the vitamin K3 by using a detection solution, and the method is a standard curve (working curve) method developed based on the sensitive response of the pH clock system to the vitamin K3. Specifically, "HCHO" is applied- NaHSO 3 - Na 2 SO 3 The pH clock reaction system is used as a detection solution, and a map of pH change along with time is recorded; when the pH clock reaction starts, adding the same volume of a series of vitamin K3 sample solutions to be detected with different concentrations into the pH clock system, and realizing quantitative detection of the vitamin K3 sample to be detected according to different induction time generated by the system when the concentrations of the solutions to be detected in the pH clock system are different.
Establishing a working curve according to the relation between the concentration of vitamin K3 in a pH clock system and the induction time; wherein the abscissa is the concentration of vitamin K3 in the pH clock system, the ordinate is the induction time t, and the concentration of vitamin K3 in the system is 7.25X10 -5 mol/L to 5.08X10 -4 When the mol/L ratio is between the two mol/L ratios, the induction time t and the concentration of the vitamin K3 form a linear relation, so that the quantitative detection of the vitamin K3 in the sample can be realized.
The quantitative detection method is different from the prior art in that the invention applies HCHO- NaHSO 3 - Na 2 SO 3 The pH clock system is used as a detection solution, and the response of the system to different concentrations of vitamin K3 is different, namely, the induction time is different, so that the quantitative analysis of the vitamin K3 is realized.
The concentration of vitamin K3 detected in the detection solution (pH clock system) is 7.25X10 -5 -5.08×10 -4 mol/L。
When vitamin K3 is detected in the detection solution (pH clock system), the temperature of the pH clock system is controlled to be any specific temperature in the range of 10-18 ℃.
With the above pH clock system, the concentration range in which vitamin K3 can be detected is the optimum concentration range determined experimentally. In the concentration range, the induction time has good response to the change of the concentration of vitamin K3, and the linear correlation coefficient is large. In addition, the concentration ranges of the components in the detection solution (pH clock system) are shown in table 1, and the optimal concentrations of the detection solution (pH clock system) obtained through a plurality of experiments are shown in table 2:
table 1: concentration of each component in a pH clock system
| HCHO(mol/ L) | NaHSO 3 (mol/L) | Na 2 SO 3 (mol/L) |
| 0.043-0.0635 | 0.035-0.0625 | 0.0035-0.00625 |
Table 2: optimum concentration of each component in pH clock system
| HCHO(mol/ L) | NaHSO 3 (mol/L) | Na 2 SO 3 (mol/L) |
| 0.0501 | 0.0499 | 0.00499 |
The specific experimental steps are as follows:
1. preparing 40mL of detection solution (pH clock system) according to the concentration range specified in Table 1, wherein the temperature is controlled to be a specific temperature value between 10 ℃ and 18 ℃ and is kept unchanged; the prepared working electrode (pH composite electrode, lei Ci, E-331) was inserted into the solution, the other end of the working electrode was connected to a computer through a potential/temperature/pH integrated tester (ZHFX-595, jiaxing Disheng electronic technologies Co., ltd.), and after the chemical signal acquisition and analysis program in the computer was opened to set the acquisition time and sampling speed, the start key was clicked rapidly to monitor the pH of the solution. The computer records the acquired pH profile, i.e., pH clock profile, over time. When the substance needs to be detected, the substance to be detected is rapidly added at the same time when the reaction of the pH clock system is started, and the pH clock pattern of the pH change along with time is recorded in the same way.
Basic parameters of the pH clock profile include:
induction time: the time required from the start of the reaction of the pH clock system to the pH jump.
pH jump range: the pH corresponding to the beginning of the pH jump is changed to the pH corresponding to the end of the pH jump.
Establishing a working curve for detecting the relation between the concentration of vitamin K3 in a solution and the pH induction time
Preparing a vitamin K3 solution with the concentration of 0.0725mol/L to 0.508mol/L as a sample solution by using ethanol as a solvent, and respectively adding 40 mu L of sample solutions with different concentrations into a pH clock system of 40mL by using a pipette while the reaction of the pH clock system is started, so that the concentration of the vitamin K3 in the system is 7.25 multiplied by 10 -5 mol/L to 5.08X10 -4 mol/L; the response variable of the pH clock system is the induction time and is marked as t; when the concentration of vitamin K3 in the system is different, the induction time t of the pH clock system is also different; plotting with the concentration of vitamin K3 in the system as an abscissa and t as an ordinate; when the concentration of vitamin K3 in the system is 7.25X10 -5 mol/L to 5.08X10 -4 And when the mol/L ratio is between the mol/L ratio, the pH clock system induction time t and the concentration of the vitamin K3 form a linear relation to obtain a working curve.
Quantitative detection of vitamin K3
The method comprises the steps of adding a sample to be detected with unknown concentration into a pH clock system of a detection solution when the pH clock system starts to react, measuring the induction time (t) of the corresponding pH clock system, and according to the corresponding relation between t and the unknown vitamin K3 concentration on a working curve, obtaining the vitamin K3 concentration in the detection system, and further calculating the vitamin K3 concentration in the sample to be detected.
Drawings
FIG. 1 is a graph showing the pH of a detection solution (pH clock system) with time when a sample to be detected is not added in example 1.
FIG. 2 is a graph of example 1, incorporating 7.25X10 -5 After the mol/L vitamin K3, the pH value of the solution (pH clock system) is detected to change with time.
FIG. 3 is a schematic diagram of example 1, incorporating 1.45X10 -4 After the mol/L vitamin K3, the pH value of the solution (pH clock system) is detected to change with time.
FIG. 4 is a graph showing the working curve between pH induction time t and vitamin K3 concentration in example 1.
FIG. 5 is a graph showing the pH of the test solution (pH clock system) with time when no sample to be tested was added in example 2.
FIG. 6 is a diagram of example 2 incorporating 2.9X10 -4 After the mol/L vitamin K3, the pH value of the solution (pH clock system) is detected to change with time.
FIG. 7 is a sample of example 2, to which 4.35X 10 is added -4 After the mol/L vitamin K3, the pH value of the solution (pH clock system) is detected to change with time.
FIG. 8 is a graph showing the working curve between pH induction time t and vitamin K3 concentration in example 2.
FIG. 9 is a graph showing the pH of the test solution (pH clock system) with time when no sample to be tested was added in example 3.
FIG. 10 is a sample of example 3, incorporating 4.35×10 -4 After the mol/L vitamin K3, the pH value of the solution (pH clock system) is detected to change with time.
FIG. 11 is a diagram of example 3, incorporating 5.08X10 s -4 After the mol/L vitamin K3, the pH value of the solution (pH clock system) is detected to change with time.
FIG. 12 is a graph showing the working curve between pH induction time t and vitamin K3 concentration in example 3.
Description of the embodiments
Examples
Application to HCHO- NaHSO 3 - Na 2 SO 3 The pH clock system serving as a substrate is used as a detection solution to quantitatively analyze the vitamin K3. And adding vitamin K3 sample solution with no concentration into the pH clock system in an equal volume, establishing a working curve (such as a linear relation) related between the concentration of vitamin K3 in the detection system and the induction time, achieving the purpose of detecting the vitamin K3 in the pH clock system, and further calculating the concentration of the vitamin K3 in the sample to be detected.
(1) Preparing a detection solution
Firstly, distilled water is used for preparing HCHO solution with the concentration of 0.12mol/L and NaHSO with the concentration of 0.15mol/L respectively 3 And 0.015mol/L Na 2 SO 3 Is a mixed solution of (a) and (b). To a 50mL small beaker was added 10.0mL of distilled water solution, 13.3mL of NaHSO in sequence 3 - Na 2 SO 3 Mix solution, 16.7mL of 0.12mol/L HCHO solution to ensure "HCHO- NaHSO 3 - Na 2 SO 3 "the concentration of each component in the pH clock system is HCHO 0.0501mol/L, naHSO 3 0.0499mol/L、Na 2 SO 3 0.00499mol/L, total volume 40mL, temperature was controlled at 12 ℃.
Simultaneously, ethanol is used as a solvent to prepare a series of vitamin K3 sample solutions with different concentrations.
(2) Obtaining a pH clock pattern
The profile of the pH of the prepared test solution over time was recorded by a computer equipped with a chemical signal acquisition analysis program (no test sample added). As shown in fig. 1. The pH induction time was 230s for blank control. Two sets of detection solutions with the same concentration of each component as the detection solution are additionally prepared. For one of the groups, at the same time as the reaction started, 40. Mu.L of a 0.0725mol/L sample solution of vitamin K3 was added to a pH clock system of 40mL so that the concentration of vitamin K3 in the detection solution was 7.25X10 -5 mol/L, the added vitamin K3 shortens the induction time to 206s as shown in figure 2; for the other group, 40. Mu.L of a 0.145mol/L sample solution of vitamin K3 was added to a pH clock system of 40mL at the same time as the start of the reaction, so that the concentration of vitamin K3 in the detection solution was 1.45X10 -4 mol/L, vitamin K3 was added so that the induction time became 187s as shown in FIG. 3. Figures 2 and 3 demonstrate that different concentrations of vitamin K3 in the test solutions lead to different induction times for the appearance of the pH clock system. When the concentration of vitamin K3 in the detection system is 7.25X10 -5 mol/L to 5.08X10 -4 The results of different induction times of the pH clock system due to different concentrations between mol/L can be observed.
(3) Quantitative detection
An operating curve is established based on the relationship between the concentration of vitamin K3 in the detection system and the induction time, as shown in FIG. 4, wherein the abscissa is the concentration of vitamin K3 in the pH clock system, and the ordinate is the induction time t, when the concentration of vitamin K3 in the detection system is 7.25X10 -5 mol/L to 5.08X10 -4 When the mol/L is in a mol/L range, the induction time and the concentration of vitamin K3 are in a linear relation, and the linear equation is t= -244032c #Vitamin K3) +222.57, R 2 =0.997. Therefore, the quantitative detection of the vitamin K3 in the sample can be realized.
Examples
(1) Preparing a detection solution
Firstly, distilled water is used for preparing HCHO solution with the concentration of 0.12mol/L and NaHSO with the concentration of 0.15mol/L respectively 3 And 0.015mol/L Na 2 SO 3 Is a mixed solution of (a) and (b). To a 50mL small beaker was added 10mL of distilled water solution, 14mL of NaHSO in sequence 3 - Na 2 SO 3 Mix solution, 16mL of 0.12mol/L HCHO solution to ensure "HCHO- NaHSO 3 - Na 2 SO 3 "the concentration of each component in the pH clock system is HCHO 0.048mol/L, naHSO 3 0.0525mol/L、Na 2 SO 3 0.00525mol/L, total volume 40mL, temperature was controlled at 12 ℃.
Simultaneously, ethanol is used as a solvent to prepare a series of vitamin K3 sample solutions with different concentrations.
(2) Obtaining a pH clock pattern
The profile of the pH of the prepared test solution over time was recorded by a computer equipped with a chemical signal acquisition analysis program (no test sample added), as shown in FIG. 5. The pH induction time was 231s for blank control. Two sets of detection solutions with the same concentration of each component as the detection solution are additionally prepared. For one of the groups, a 40. Mu.L 0.29mol/L sample solution of vitamin K3 was added to a pH clock system of 40mL at the same time as the reaction was started, so that the concentration of vitamin K3 in the detection solution was 2.9X10 -4 mol/L, vitamin K3 was added to shorten the induction time to 151s as shown in FIG. 6; for the other group, a sample solution of vitamin K3 of 40. Mu.L of 0.435mol/L was added to a pH clock system of 40mL at the same time as the reaction was started, so that the concentration of vitamin K3 in the detection solution was 4.35X10 -4 mol/L, vitamin K3 was added so that the induction time became 113s as shown in FIG. 7. Fig. 6 and 7 demonstrate that different concentrations of vitamin K3 in the test solutions lead to different induction times for the appearance of the pH clock system. When the concentration of vitamin K3 in the detection system is 7.25X10 -5 mol/L to 5.08X10 -4 The concentration of the solution is different to lead the pH clock system to be outThe results of the different induction times can be observed.
(3) Quantitative detection
An operating curve is established based on the relationship between the concentration of vitamin K3 in the detection system and the induction time, as shown in FIG. 8, wherein the abscissa is the concentration of vitamin K3 in the pH clock system, and the ordinate is the induction time t, when the concentration of vitamin K3 in the detection system is 7.25X10 -5 mol/L to 5.08X10 -4 Between mol/L, the induction time and the concentration of vitamin K3 are in a linear relation, and the linear equation is t= -240894c (vitamin K3) +221.05, R 2 = 0.9951. Therefore, the quantitative detection of the vitamin K3 in the sample can be realized.
Examples
(1) Preparing a detection solution
Firstly, distilled water is used for preparing HCHO solution with the concentration of 0.12mol/L and NaHSO with the concentration of 0.15mol/L respectively 3 And 0.015mol/L Na 2 SO 3 Is a mixed solution of (a) and (b). To a 50mL small beaker was added 10.2mL of distilled water solution, 13.2mL of NaHSO in sequence 3 - Na 2 SO 3 Mix solution, 16.6mL of 0.12mol/L HCHO solution to ensure "HCHO- NaHSO 3 - Na 2 SO 3 "the concentration of each component in the pH clock system is HCHO 0.0498mol/L, naHSO 3 0.0495mol/L、Na 2 SO 3 0.00495mol/L, total volume 40mL, temperature was controlled at 12 ℃.
Simultaneously, ethanol is used as a solvent to prepare a series of vitamin K3 sample solutions with different concentrations.
(2) Obtaining a pH clock pattern
The profile of the pH of the prepared test solution over time was recorded by a computer equipped with a chemical signal acquisition analysis program (no test sample added). As shown in fig. 9. The pH induction time was 230.6s for the blank. Two sets of detection solutions with the same concentration of each component as the detection solution are additionally prepared. For one of the groups, 40. Mu.L of a 0.435mol/L sample solution of vitamin K3 was added to a pH clock system of 40mL at the same time as the reaction was started, so that the concentration of vitamin K3 in the detection solution was 4.35X10 -4 mol/L, vitamin K3 is added to makeThe induction time was reduced to 113s as shown in FIG. 10; for the other group, 40. Mu.L of a 0.508mol/L sample solution of vitamin K3 was added to a pH clock system of 40mL at the same time as the reaction was started, so that the concentration of vitamin K3 in the detection solution was 5.08X10 -4 mol/L, vitamin K3 was added so that the induction time became 102s as shown in FIG. 11. Fig. 10 and 11 demonstrate that different concentrations of vitamin K3 in the test solutions lead to different induction times for the appearance of the pH clock system. When the concentration of vitamin K3 in the detection system is 7.25X10 -5 mol/L to 5.08X10 - 4 The results of different induction times of the pH clock system due to different concentrations between mol/L can be observed.
(3) Quantitative detection
An operating curve is established based on the relationship between the concentration of vitamin K3 in the detection system and the induction time, as shown in FIG. 12, wherein the abscissa is the concentration of vitamin K3 in the pH clock system, and the ordinate is the induction time t, when the concentration of vitamin K3 in the detection system is 7.25X10 -5 mol/L to 5.08X10 -4 Between mol/L, the induction time and the concentration of vitamin K3 are in a linear relation, and the linear equation is t= -242971c (vitamin K3) +222.7, R 2 = 0.9963. Therefore, the quantitative detection of the vitamin K3 in the sample can be realized.
Claims (4)
1. A quantitative detection method of vitamin K3 is characterized in that:
ethanol is used as a solvent to prepare a solution of a sample to be detected;
application of HCHO-NaHSO 3 - Na 2 SO 3 The pH clock reaction system is used as a detection solution, and a map of pH change along with time is recorded; the temperature of the pH clock system is controlled at any specific temperature within the range of 10-18 ℃, when the pH clock reaction starts, the equal volumes of a series of sample solutions to be detected with different concentrations are respectively added into the pH clock system, and quantitative detection of the samples to be detected is realized according to different induction time generated by the system when the concentrations of the solutions to be detected in the pH clock system are different;
detecting the molar concentration of each component in the solutionThe range is as follows: HCHO 0.043-0.0635mol/L, naHSO 3 0.035-0.0625mol/L、Na 2 SO 3 0.0035-0.00625mol/L；
The sample to be detected is vitamin K3 solution.
2. The quantitative detection method according to claim 1, wherein: establishing a working curve according to the relation between the concentration of the solution to be detected in the pH clock system and the induction time; wherein the abscissa is the concentration of vitamin K3 in the solution to be detected in the pH clock system, and the ordinate is the induction time t; when the concentration of vitamin K3 in the system is 7.25X10 -5 mol/L to 5.08X10 -4 When the mol/L ratio is between the two, the induction time t and the concentration of the vitamin K3 form a linear relation, so that the quantitative detection of the vitamin K3 in the sample is realized.
3. The quantitative detection method according to claim 1 or 2, characterized in that: the molar concentration of each component in the detection solution is HCHO 0.0501mol/L, naHSO 3 0.0499mol/L、Na 2 SO 3 0.00499mol/L。
4. The quantitative detection method according to claim 1 or 2, characterized in that: the temperature of the pH clock system was controlled at 12℃when the vitamin K3 solution was tested.
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