CN1786693A - Process for determining content of chlorine ion and kit for diagnosing chlorine ion therefor - Google Patents
Process for determining content of chlorine ion and kit for diagnosing chlorine ion therefor Download PDFInfo
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Abstract
The invention relates to measuring method for chloride ion content and diagnosing kit. The process includes the following steps: utilizing alpha-amylase to hydrolyze numerous poly- glucose into few poly- glucose, coupling into glucosaccharase, or glucose oxidase and take reaction with peroxidase, oxidizing the colorless reduction type chromogen combined system into pigmented quinone imide chromogen or amine chromogen dye, by detecting the variance of 400- 600nm absorbance between reaction period, and the content of chloride ion in sample would be quantitatively responsed. The method has high specificity, has no pollution from inside and outside material, accurate test result. The diagnose kit is made up into bi-agent or tri-agent that could reduce cross influence from each component and keeps stability of the agent that is convenient for long-term storage. The method could quickly test on commonness ultraviolet/ visible light, analyzer or semi-transfer/ full automatically biochemical study instrument, and doesn't need particularity or extra instrument. The testing cost is cheap, and is convenient for popularization.
Description
Technical Field
The invention relates to a method for measuring the content of chloride ions in a sample and a diagnostic kit for measuring the content of chloride ions, belonging to the technical field of medical examination and measurement.
Background
Chlorine element exists in the form of ions in body fluid, almost increases and decreases in balance with sodium ions, and the determination of the content of the chlorine ions has very important significance in the clinical and analytical fields.
In the prior art, the method for measuring the content of chloride ions mainly comprises the following steps: chemical determination method-mercuric nitrate titration method, mercuric thiocyanate colorimetric method, coulometry method and ion selective electrode method. The mercuric chloride titration method needs skilled operation, is difficult to judge the titration end point, has variable depth and intensity of the end point due to different protein contents of different samples, and is interfered by bilirubin, hemoglobin and blood fat, so the method has obviously poor precision and low working efficiency. Mercury thiocyanate colorimetry also reacts with other elements in samples such as serum, e.g., F, Br and I, when measured, but is negligible due to amounts less than 1mmol/l, but is more influential when certain drugs and bilirubin are involved. The colorimetric method eliminates the influence of subjective factors of human beings, and can carry out batch measurement in an automatic biochemical analyzer, so that the colorimetric method is a method commonly used in clinic.
The electric quantity analysis method belongs to a physical method and has the following principle: placing silver electrode in specimen under constant current, releasing Ag from the electrode+With Cl-The reaction begins to generate an undissociated AgCl precipitate when Cl is present-All with Ag+Free Ag after the action is finished+When the solution conductivity is obviously increased, a sensor and a timer are designed, the current is cut off immediately when the solution conductivity is increased, and Cl in the sample is calculated according to the time and the current intensity used for titration-And (4) content. The method is simple, convenient and rapid, and is a better determination method under the condition of well controlling current, and Br in a sample-And I-Has certain interference on the measurement result, and can be ignored due to little quantity.
The ion selective electrode method is the best method for measuring the chloride ion content at present. Because of the determination of chloride ionsAll electrodes of (1) are equal to K+、Na+The electrodes are matched, and only 100 mul of whole blood is needed to measure K in the sample+、Na+And Cl-The content of (a). The method is rapid, accurate and simple to operate, and is widely used clinically at present. However, the service life of the chloride ion electrode is short, the general validity period is only 4-6 months, and the measurement results of instruments of different types have certain errors.
In recent years, enzymatic detection techniques have been developed. The method is popular due to the advantages of high specificity, good stability, high detection speed, accurate and reliable result and the like. Similar to other substances, the technology of enzyme method for detecting the content of chloride ions in various samples such as serum and plasma has been one direction of research.
Disclosure of Invention
The purpose of the invention is: provides a method for determining the content of chloride ions by an enzyme method and a chloride ion diagnostic kit prepared by applying the method. By adopting the reagent in the kit, the content of the chloride ions can be measured by utilizing an ultraviolet/visible light analyzer or a semi-automatic/full-automatic biochemical analyzer, the measuring speed is high, the accuracy is high, and a technical support is provided for popularizing the technology for detecting the content of the chloride ions by an enzyme method and replacing the existing chloride ion detection method in the field of medical detection.
In order to achieve one of the purposes of the invention, the method for measuring the content of the chloride ions comprises the following steps:
first, a sample to be measured is mixed with a reagent containing a combination of "inactivated α -amylase", "polyglucose- (glucose) n", glucosidase, glucose oxidase, peroxidase, and reduced chromogen, and the following reactions are carried out:
① the chloride ions in the sample activated the "inactivated α -amylase",
② the active α -amylase "hydrolyzes" polysaccharidic-n (glucose) to form a plurality of "oligosaccharidic-m" (glucose),
③ oligo-glucose- (D-glucosyl) -m is hydrolyzed into glucose under the action of glucosidase,
④ the oxidation of glucose to gluconolactone and hydrogen peroxide by the action of glucose oxidase,
⑤ the hydrogen peroxide reacts with peroxidase and reduced chromogen to generate indamine chromogen or quinoneimine chromogen and water,
the reaction process is represented by a principle reaction formula as follows:
wherein, when m is 1, 2 or 3{ m, etc. 1, (glucose) m does not participate in the ③ th reaction }, and n is a natural number greater than or equal to 4;
and then, placing the reaction mixture under an ultraviolet/visible light analyzer or a semi-automatic/full-automatic biochemical analyzer, and detecting the rising speed of the absorbance with the main wavelength of 400-600 nm, thereby measuring and calculating the content of the chloride ions in the sample.
In the method for determining the content of chloride ions, the reduced chromogen combination is composed of the reduced chromogen A and another component B. The method specifically comprises the following steps:
reduced chromogen A is: "3-methyl-2-benzothiazolinone hydrazone" (abbreviated as MBTH in English, and called 3-methyl-2-benzothiazolinone-hydrazone in total) or "4-amino arsenopyrine " (abbreviated as 4-aminoantipyrine in total);
the other component B is one of the following 15 substances:
① carbolic acid (phenol),
② N-ethyl-N- (3-thiopropyl) -m-thiadiazine (N-ethyl-N- (3-sulfopropyl) -m-anisidine, ESPAS for short),
③ N, N-Diethyl-m-toluidine (N, N-Diethyl-m-toluidine),
④ 2, 4-bischlorocarbonic acid (2, 4-Dichloriphenol),
⑤ 2, 4, 6-triple bromo-3-hydroxy-benzenesulfonic acid (2, 4, 6-Tribromo-3-hydroxy-benzosulfonic acid, TBHB for short),
⑥ 3, 5-Dichlorocarbonic acid sulfonic acid (3, 5-Dichlorophenolsulfonic acid),
⑦ 3, 5-Dichloro-2-hydroxy-benzenesulfonic acid (3, 5-dichoro-2-hydroxy-benzosulfonic acid, DHBS for short),
⑧ N-ethyl-N- (2-hydroxy-3-thiopropyl) -m-toluidine sodium salt (N-ethyl-N- (2-hydroxy-3-sulfopropyl) -m-toluidine sodium, TOOS for short),
⑨ Tribromohydroxybenzoic acid (Tribromohydroxybenzoic acid),
⑩ Dimethylaniline (DMA),
(11) N-Ethyl-N- (2-hydroxy-3-thiopropyl) -m-toluidine (EHSPT for short, which is called N-Ethyl-N- (2-hydroxy-3-sulfopropyl) -m-tolulidine),
(12)2, 2 '-biazonitrogen-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (2, 2' -azino-bis- (3-ethylben-zazoline-6-sulfonic acid), ABTS for short),
(13)2, 2 '-azino-bis (3-ethylbenzothiopyrroline-6-sulfonic acid), having the name 2, 2' -azino-bis- (3-ethyllbenztbizoline-6-sulfonic acid),
(14) 4-hydroxy-3-methoxybenzoic acid (4-hydroxy-3-methoxybenzoic acid, HMB for short),
(15) 3-methyl-ethyl-hydroxyaniline (3-methyl-ethyl-hydroxyaniline, MEHA for short).
In the measuring process, the using ratio of the sample to be measured to the reagent is controlled to be 1: 10-1: 500 according to the volume, the reaction temperature is controlled to be 20-50 ℃, the reaction time is controlled to be 2-30 minutes, and the secondary wavelength is set to be more than 500-700 nm during the detection.
The chloride ion diagnostic kit for realizing another purpose of the invention can be a single agent consisting of the following components:
40 to 200mmol/l of a buffer solution,
(glucose) n is 1 to 30mmol/l,
500-50000U/l of inactivated α -amylase,
500-50000U/l of glucosidase,
500-50000U/l of glucose oxidase,
500-50000U/l of peroxidase,
0.1-20 mmol/l of reduced chromogen composition,
the total volume of the stabilizer/reagent is 10-80%.
The meaning of the reduced chromogen combination is as described above, and the concentration of the two components A and B is 0.1-20 mmol/l.
Thecomponents in the single preparation can also be combined to prepare double preparations so as to be beneficial to eliminating internal and external glucose pollution, such as:
wherein, the reduced chromogen combination (I) can be: 3-methyl-2-benzothiazolinone hydrazone or 4-amino arsenopyridine ; the reduced chromogen combination (II) may be one of the 15 substances described above.
The formula of the double-agent is not limited to the formula, the reduced chromogen combination (I) and (II) can be exchanged, and the components in the reagent I, namely deactivated α -amylase, glucosidase, glucose oxidase, peroxidase and the like, can be placed in the reagent II, and the component in the reagent II, namely (glucose) n, can be placed in the reagent I, so that various formulas can be formed, not only one.
The reagent can be prepared into the following three reagents, which is not only favorable for eliminating the pollution of glucose at internal and external sources, but also favorable for the stability of the reagent. Such as:
similar to the double dose, the reduced chromogen combination (one) in the three doses can be 3-methyl-2-benzothiazolinone hydrazone or 4-amino-arsenic , and the combination (two) can be one of the 15 substances.
The formula of the three agents is not limited to the above formula, the reduced chromogen combination (I) and (II) can be separately placed at any position of the reagent I, II or III, the components in the reagent II, namely inactivated α -amylase, glucosidase, glucose oxidase, peroxidase and the like, can be placed in the reagent I or the reagent III, and the component in the reagent III, namely (glucose) n, can also be placed in the reagent I or the reagent II, so that a plurality of formulas can be formed, and the three formulas are not listed.
Among the components of the diagnostic kit for determining the chloride ion content, the basic requirement for selecting the buffer solution is that the pH value is within the range of 6.0-11.0, and can be as follows: the buffer solution may be at least one of a "disodium hydrogen Phosphate-citric acid buffer solution", "Glycylglycine (glycine) buffer solution", "citric acid-sodium citrate buffer solution", "sodium carbonate-sodium bicarbonate buffer solution", "boric acid-borax buffer solution", "glycine-sodium hydroxide buffer solution", "borax-sodium hydroxide buffer solution", or "phosphoric acid (phosphonate) buffer solution", but the selection range of the buffer solution is not limited by these lists.
In addition, in order to reduce the cross effect among the components of the reagents and maintain the stability of the reagents for long-term storage, a stabilizer is usually added into the single-dose, double-dose reagent I/reagent II and three-dose reagent I/reagent II/reagent III, and the amount of the stabilizer accounts for about 10-80% (or the concentration is within the range of 10-50 mmol/l) of the total volume of the reagents. Substances used as stabilizers may be: at least one of ethylene glycol, propylene glycol, glycerol, ammonium sulfate, thioglycolic acid, adenosine diphosphate, bovine serum albumin, carbonate, cholate, dextran, ethylenediamine tetraacetic acid, flavin adenine dinucleotide, flavin mononucleotide, glutamate, reduced glutathione, lactose, mannitol, or succinate.
Research shows that the diagnosis kit is ideal and also a preferred scheme of the invention in the following formula component relationship no matter single, double or triple dose in consideration of the accuracy of the determination result and the economy of the preparation cost:
80 to 120mmol/l of a buffer solution,
(glucose) n is 10 to 20mmol/l,
10000-40000U/l of inactivated α -amylase,
5000-10000U/l of glucosidase,
20000-30000U/l of glucose oxidase,
20000-40000U/l of peroxidase,
0.1-10 mmol/l of reduced chromogen composition,
the total volume of the stabilizer/reagent is 20-50%.
According to the method, α -amylase with lost activity is activated by chloride ions, after the activity is regained, polyglucose is hydrolyzed into oligo-glucose, the oligo-glucose is further hydrolyzed into glucose monosaccharide under the action of glucosidase, then glucose oxidase is coupled to react to generate gluconolactone and hydrogen peroxide, and when the hydrogen peroxide and peroxidase are subjected to enzyme coupling reaction again, a colorless reduced Chromogen (Chromogen) combination system is oxidized into a colored quinoneimine Chromogen (Quioniimine) or Indamine Chromogen (Indamine) dye.
The outstanding substantive features and remarkable progress of the technical scheme of the invention are mainly shown in the following steps:
(1) the method completely utilizes an enzymology method, the enzymolysis reaction has the characteristic of high specificity, and the content of chloride ions in a tested sample is quantitatively reflected by oxidizing colorless reduced chromogens into colored quinonimine chromogens or indamine chromogens dye, so that the test result is accurate;
(2) the components participating in the enzyme coupling reaction are externally added, so that the enzyme coupling reaction is not polluted byinternal and external substances, and the accuracy of the test process is high;
(3) the method is simple and easy to operate, can quickly obtain a detection result, and the reaction is carried out under the condition of a buffer solution, so that the environment is not polluted;
(4) the method can be used for rapid detection on a common ultraviolet/visible light analyzer or a semi-automatic/full-automatic biochemical analyzer, does not need special or additional instruments, has low test cost, is convenient to popularize and apply, and is expected to replace other methods in clinical and chemical analysis to become a conventional analysis method;
(5) the determination method provided by the invention can be used for preparing liquid reagents, dry powder reagents, dry reagents and other reagents in various forms, and is used for determining the content of chloride ions in various samples;
(6) the liquid chlorine ion content diagnosis kit provided by the invention has good stability, the three-dimensional space structures of various enzymes in the kit are kept complete, and the application test effect is well ensured. After the double-agent or the triple-agent is prepared, the cross influence among various components can be further reduced, the detection result is more reliable, the reagent is more stable, and the reagent can be stored for a long time.
Detailed Description
The technical solution of the present invention is further illustrated by the following specific examples. These examples are merely exemplary applications and are not to be construed as limiting the scope of the invention as claimed.
EXAMPLE one (Single dose)
Preparing a chloride ion diagnostic kit according to the following components and dosage:
the buffer solution of glycylglycine is 80mmol/l,
(glucose) n is 10mmol/l,
10000U/l of inactivated α -amylase,
the content of the glucosidase is 5000U/l,
20000U/l of glucose oxidase,
the peroxidase is 20000U/l,
4-amino arsenic trioxide 2mmol/l,
10mmol/l of carbolic acid is added,
ethylene glycol 50% (based on the total volume of the reagents).
Setting on a full-automatic biochemical analyzer (Hitachi-7080): the temperature is 37 ℃, the reaction time is 10 minutes, the test main wavelength is 495-505 nm, the test auxiliary wavelength is more than 600nm, the volume ratio of the tested chloride ion sample to the reagent is 1: 25, and the reaction direction is positive reaction (the absorbance is increased, the same below).
And adding the sample and the prepared single agent, automatically and uniformly mixing the sample and the prepared single agent in an analyzer, and detecting and recording the rising condition of the absorbance at 495-505 nm. And (4) according to methods such as a rate method, an end point method and the like, measuring and calculating the content of the chloride ions in the sample by contrasting the corresponding standard curves. The process is repeated for the same batch of samples for many times, and the test result has good repeatability and high accuracy.
EXAMPLE two (two-agent)
Preparing a chloride ion diagnostic kit according to the following components and dosage:
reagent I-
100mmol/l of buffer solution of citric acid and sodium citrate,
the inactivated α -amylase 25000U/l,
the content of the glucosidase is 7500U/l,
the glucose oxidase 25000U/l,
the peroxidase (30000U/l),
1mmol/l of 4-amino arsenicum,
50% of glycerol (based on the total volume of the reagent I);
reagent II- (III) -
100mmol/l of buffer solution of citric acid and sodium citrate,
(glucose) n 15mmol/l,
1mmol/l of 2, 4, 6-triple bromo-3-hydroxy-benzenesulfonic acid,
ethylene glycol 20 mmol/l.
Setting on a full-automatic biochemical analyzer (Hitachi-7080): the temperature is 30 ℃, the reaction time is 15 minutes, the test main wavelength is 546nm, the test auxiliary wavelength is more than 630nm, the ratio of the total volume of the tested chloride ion sample to the reagent I and the reagent II is 1: 25, the ratio of the dosage of the reagent I to the reagent II is 4: 1, and the reaction direction is positive and negative.
Adding the sample and the reagent I, adding the reagent II after 5 minutes, automatically mixing the sample and the reagent I in an analyzer, and detecting and recording the increase of the absorbance at 546 nm. And (4) according to methods such as a rate method, an end point method and the like, measuring and calculating the content of the chloride ions in the sample by contrasting the corresponding standard curves. The process is repeated for the same batch of samples for many times, and the test result has good repeatability and high accuracy.
EXAMPLE three (three doses)
Preparing achloride ion diagnostic kit according to the following components and dosage:
reagent I-
Glycine-sodium hydroxide buffer solution 120mmol/l,
inactivated α -amylase 40000U/l,
2mmol/l of 3-methyl-2-benzothiazolone hydrazone,
propylene glycol 50% (based on the total volume of reagent I);
reagent II- (III) -
Glycine-sodium hydroxide buffer solution 120mmol/l,
the content of the glucosidase is 10000U/l,
glucose oxidase 30000U/l,
the content of the peroxidase is 40000U/l,
propylene glycol 50% (based on the total volume of reagent II);
reagent III-
Glycine-sodium hydroxide buffer solution 120mmol/l,
(glucose) n is 20mmol/l,
2mmol/l of bis-methylaniline,
ethylene glycol 20 mmol/l.
Setting on a full-automatic biochemical analyzer (Hitachi-7080): the temperature is 25 ℃, the reaction time is 20 minutes, the test main wavelength is 578nm, the test auxiliary wavelength is more than 700nm, the ratio of the tested sample to the total volume of the reagent I, the reagent II and the reagent III is 1: 25, the dosage of the reagent I, the reagent II and the reagent III is 8: 1, and the reaction direction is positive and negative.
Adding the sample, the reagent I and the reagent II, adding the reagent III after 5 minutes, automatically and uniformly mixing the sample, the reagent I and the reagent II in an analyzer, and detecting and recording the rising condition of the absorbance at 578 nm. And (4) according to methods such as a rate method, an end point method and the like, measuring and calculating the content of the chloride ions in the sample by contrasting the corresponding standard curves. The process is repeated for the same batch of samples for many times, and the test result has good repeatability and high accuracy.
EXAMPLE four (preferred double dose)
Preparing a chloride ion diagnostic kit according to the following components and dosage:
reagent I-
100mmol/l of buffer solution of disodium hydrogen phosphate to citric acid,
the glucose oxidase enzyme has a ratio of 10000U/I,
20000U/l of glucose oxidase,
the peroxidase is 20000U/l,
1mmol/l of 4-amino arsenicum,
50% of glycerol (based on the total volume of the reagent I);
reagent II- (III) -
100mmol/l of buffer solution of disodium hydrogen phosphate to citric acid,
20000U/l of inactivated α -amylase,
(glucose) n is 10mmol/l,
2mmol/l of bis-methylaniline,
glycerol 50% (based on total volume of reagent II).
Setting on a full-automatic biochemical analyzer (Hitachi-7080): the temperature is 30 ℃, the reaction time is 10 minutes, the test main wavelength is 578nm, the test auxiliary wavelength is more than 700nm, the ratio of the total volume of the tested sample to the reagent I and the reagent II is 1: 25, the ratio of the dosage of the reagent I to the reagent II is 4: 1, and the reaction direction is positive and negative.
Firstly adding a sample and a reagent I, adding a reagent II after 5 minutes, automatically mixing the three in an analyzer, and carrying out the following principle reaction:
the increase in absorbance at 578nm was detected and recorded. And (4) according to methods such as a rate method, an end point method and the like, measuring and calculating the content of the chloride ions in the sample by contrasting the corresponding standard curves. The process is repeated for the same batch of samples for many times, and the test result has good repeatability and high accuracy.
The invention has the obvious characteristic of eliminating the pollution of internal and external glucose. The elimination of the internal and external glucose occurs during the first half of the entire reaction time, the contaminated internal and external glucose is consumed during the latter half, and the glucose required for testing the chloride ion content is derived from the action of the chloride ions in the sample during the latter half.
In a word, experiments prove that the measuring method of the invention can completely measure the content of chloride ions in various samples such as plasma, serum and the like by an ultraviolet/visible light analyzer or a semi-automatic/full-automatic biochemical analyzer, has high detection speed, high sensitivity and good accuracy, and is not polluted by internal and external substances. Moreover, the chloride ion diagnostic kit provided by the invention has good stability, and can accurately detect the content of chloride ions in various types of samples after being stored for a long time.
Claims (10)
1. A method of determining chloride ion content, comprising the steps of:
(1) mixing a sample to be tested with a reagent containing deactivated α -amylase, polysaccharidase- (glucose) -n- (glucosyl) n, glucosidase, glucose oxidase, peroxidase and reduced chromogen combination, and carrying out the following reactions:
① the chloride ions in the sample activated the "inactivated α -amylase",
② the active α -amylase "hydrolyzes" polysaccharidic-n (glucose) to form a plurality of "oligosaccharidic-m" (glucose),
③ oligo-glucose- (D-glucosyl) -m is hydrolyzed into glucose under the action of glucosidase,
④ the oxidation of glucose to gluconolactone and hydrogen peroxide by the action of glucose oxidase,
⑤ the hydrogen peroxide reacts with peroxidase and reduced chromogen to generate indamine chromogen or quinoneimine chromogen and water,
the reaction process is represented by a principle reaction formula as follows:
wherein m is 1, 2 or 3, and n is a natural number greater than or equal to 4;
(2) and (3) placing the reaction mixture under an ultraviolet/visible light analyzer or a semi-automatic/full-automatic biochemical analyzer, detecting the rising speed of the absorbance with the main wavelength of 400-600 nm, and measuring and calculating the content of the chloride ions in the sample.
2. The method for determining chloride ion content according to claim 1, wherein: the reduced chromogen combination is composed of a reduced chromogen A and another component B, wherein the reduced chromogen A is 3-methyl-2-benzothiazolone hydrazone or 4-amino arsenic , and the other component B is one of the following 15 substances: carbolic acid, N-ethyl-N- (3-thiopropyl) -m-thiadineamine, N, N-diethyl-m-toluidine, 2, 4-bischlorocarbolic acid, 2, 4, 6-tribromo-3-hydroxy-benzenesulfonic acid, 3, 5-bischlorocarbolic acid sulfonic acid, 3, 5-bischloro-2-hydroxy-benzenesulfonic acid, N-ethyl-N- (2-hydroxy-3-thiopropyl) -m-toluidine sodium salt, tribromohydroxybenzoic acid, bismethylaniline, N-ethyl-N- (2-hydroxy-3-thiopropyl) -m-toluidine, 2, 2' -biazoxy-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, 2, 2' -azino-bis (3-ethylbenzothiopyrroline-6-sulfonic acid), 4-hydroxy-3-methoxybenzoic acid, 3-methyl-ethyl-hydroxyaniline.
3. The method for determining chloride ion content according to claim 1 or 2, wherein: the using ratio of the sample to be detected to the reagent is controlled to be 1: 10-1: 500 according to the volume.
4. The method for determining chloride ion content according to claim 1 or 2, wherein: the reaction temperature is controlled to be 20-50 ℃, the reaction time is controlled to be 2-30 minutes, and the secondary wavelength is set to be more than 500-700 nm during detection.
5. A chloride ion diagnostic kit, wherein the kit comprises the following components:
40 to 200mmol/l of a buffer solution,
(glucose) n is 1 to 30mmol/l,
500-50000U/l of inactivated α -amylase,
500-50000U/l of glucosidase,
500-50000U/l of glucose oxidase,
500-50000U/l of peroxidase,
0.1-20 mmol/l of reduced chromogen composition,
the total volume of the stabilizer/reagent is 10-80%.
6. The chloride ion diagnostic kit according to claim 5, characterized in that:
the reduced chromogen combination is composed of a reduced chromogen A and another component B, wherein the concentration of A and B is 0.1-20 mmol/l
The reduced chromogen A is 3-methyl-2-benzothiazolinone hydrazone or 4-amino arsenic ;
the other component B is one of the following 15 substances: carbolic acid, N-ethyl-N- (3-thiopropyl) -m-thiadineamine, N, N-diethyl-m-toluidine, 2, 4-bischlorocarbolic acid, 2, 4, 6-tribromo-3-hydroxy-benzenesulfonic acid, 3, 5-bischlorocarbolic acid sulfonic acid, 3, 5-bischloro-2-hydroxy-benzenesulfonic acid, N-ethyl-N- (2-hydroxy-3-thiopropyl) -m-toluidine sodium salt, tribromohydroxybenzoic acid, bismethylaniline, N-ethyl-N- (2-hydroxy-3-thiopropyl) -m-toluidine, 2, 2' -biazoxy-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, 2, 2' -azino-bis (3-ethylbenzothiopyrroline-6-sulfonicacid), 4-hydroxy-3-methoxybenzoic acid, 3-methyl-ethyl-hydroxyaniline.
7. The chloride ion diagnostic kit according to claim 5 or 6, characterized in that: the pH range of the buffer solution is 6.0-11.0.
8. The chloride ion diagnostic kit according to claim 7, characterized in that: the buffer solution is at least one of disodium hydrogen phosphate-citric acid buffer solution, glycylglycine buffer solution, citric acid-sodium citrate buffer solution, sodium carbonate-sodium bicarbonate buffer solution, boric acid-borax buffer solution, glycine-sodium hydroxide buffer solution, borax-sodium hydroxide buffer solution or phosphoric acid buffer solution.
9. The chloride ion diagnostic kit according to claim 5 or 6, characterized in that: the stabilizer is at least one of ethylene glycol, propylene glycol, glycerol, ammonium sulfate, sulfenyl ethanol, adenosine diphosphate, bovine serum albumin, carbonate, cholate, dextran, ethylene diamine tetraacetic acid, flavin adenine dinucleotide, flavin mononucleotide, glutamate, reduced glutathione, lactose, mannitol or succinate.
10. The chloride ion diagnostic kit according to any one of claims 5 to 9, characterized in that: the reagent is prepared into single, double or triple dose.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108181252A (en) * | 2018-02-09 | 2018-06-19 | 广东贝特涞生物科技有限公司 | Disinfectant detection device, adding set and adding method |
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| CN108181252A (en) * | 2018-02-09 | 2018-06-19 | 广东贝特涞生物科技有限公司 | Disinfectant detection device, adding set and adding method |
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