CN105158248A - Preparation method of single hydroxyl phenolic urine metabolite detection reagent - Google Patents
Preparation method of single hydroxyl phenolic urine metabolite detection reagent Download PDFInfo
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- CN105158248A CN105158248A CN201510536672.9A CN201510536672A CN105158248A CN 105158248 A CN105158248 A CN 105158248A CN 201510536672 A CN201510536672 A CN 201510536672A CN 105158248 A CN105158248 A CN 105158248A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/82—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a precipitate or turbidity
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
Abstract
The invention relates to the field of detection reagents, in particular to a single hydroxyl phenolic urine metabolite detection reagent. The preparation method includes the following steps of preparing 0.2-1g/ml of a ferric sulphate solution serving as a solution A; preparing 0.1-1g/ml of a mercury sulfate solution serving as a solution B; preparing 0.15-1g/ml of a copper nitrate solution serving as a solution C; preparing 0.15-1g/ml of a ferric nitrate solution as a solution D; adding a preservative after mixing the solution A, the solution B, the solution C and the solution D in the mass ratio of 1:(1-2):(1.5-2):1 with uniform stirring to obtain the light-green single hydroxyl phenolic urine metabolite detection reagent, wherein the temperature of the solution A, the solution B, the solution C and the solution C and mixed liquor of the solution A, the solution B, the solution C and the solution D ranges from 30 DEG C to 35DEG C. The detection reagent is prepared from the solution A, the solution B, the solution C and the solution D with the optimal mass ratio and the best mixed concentration, so that the reagent is high in flexibility, low in false positive rate and false negative rate, easy to operate and low in cost.
Description
Technical field
The present invention relates to and detect reagent field, be specifically related to a kind of preparation method of urine monohydroxyphenyl metabolites detection reagent.
Background technology
At present, a lot of for the method for diagnosing tumour both at home and abroad, commonly detect various tumor markers as blood PSA, AFP, CEA, ferritin, CA125 and lipoprotein etc., although the recall rate of these marks is higher, but be all have abrasive detection mostly, and complicated operation, expensive, be unsuitable for large-scale crowd examination.Detect tumour by urine to avoid blood drawing this easily causes the link of infection, also have both at home and abroad many methods as Urine by HPLC polyamines, ELISA method measures ferritinuria and the TYR measured in urine and metabolin and protein chip thereof and detects tumor markers etc.Its method cuts both ways, and as HPLC method measures urinary polyamine, although highly sensitive, selectivity is good, and complicated operation is not easily carried out clinical.
Summary of the invention
The present invention is directed to the deficiencies in the prior art, provide a kind of highly sensitive, simple to operate, accuracy is high, the preparation method of lower-cost urine monohydroxyphenyl metabolites detection reagent.
The present invention is by the following technical solutions: a kind of preparation method of urine monohydroxyphenyl metabolites detection reagent, is characterized in that, comprises the steps:
(1) ferrum sulfuricum oxydatum solutum of 0.2-1g/ml is prepared, as solution A;
(2) the mercuric sulfate solution of 0.1-1g/ml is prepared, as B solution;
(3) copper nitrate solution of 0.15-1g/ml is prepared, as C solution;
(4) iron nitrate solution of 0.15-1g/ml is prepared, as solution D;
(5) by solution A, B solution, C solution, solution D with mass ratio be 1:(1-2): (1.5-2): 1
After mixing, add antiseptic, stir and make absinthe-green urine monohydroxyphenyl metabolites detection reagent;
Wherein, solution A, B solution, C solution, solution D and solution A, B solution, C solution and solution D
The temperature of mixed liquor be between 30-35 degree Celsius.
The preparation process of described solution A is as follows: be dissolved in by 20--25g iron in the sulfuric acid solution of the 2-3M of 245-250ml, after iron dissolves completely, make solution A.
The preparation process of described B solution is as follows: be dissolved in by 10--25g mercury in the sulfuric acid solution of the 2-3M of 245-250ml, after mercury dissolves completely, make B solution.
The preparation process of described C solution is as follows: be dissolved in by 15--25g copper in the salpeter solution of the 2-3M of 245-250ml, after copper dissolves completely, make C solution.
The preparation process of described solution D is as follows: be dissolved in by 15--25g iron in the salpeter solution of the 2-3M of 245-250ml, after iron dissolves completely, make solution D.
In step (5), antiseptic is one or more of Sodium Benzoate, potassium sorbate, dehydro sodium acetate, calcium propionate, sodium Diacetate, sodium lactate or propylparaben.
In step (5), solution A, B solution, C solution, solution D are that 1:1-2:1.5:1 mixes with mass ratio.
The present invention adopts solution A, B solution, C solution, solution D with optimum quality ratio and optium concentration mixing to make detection reagent, highly sensitive, false positive rate and false negative rate low, and cost simple to operate is low.
Embodiment
Below in conjunction with specific embodiment, the present invention is further described in detail.
Preparation embodiment 1
(1) ferrum sulfuricum oxydatum solutum is prepared, as solution A; 20g iron is dissolved in the sulfuric acid solution of the 2M of 250ml, after iron dissolves completely, makes solution A
(2) mercuric sulfate solution is prepared, as B solution; 15g mercury is dissolved in the sulfuric acid solution of the 2M of 250ml, after mercury dissolves completely, makes B solution;
(3) copper nitrate solution is prepared, as C solution; 15g copper is dissolved in the salpeter solution of the 2M of 250ml, after copper dissolves completely, makes C solution.
(4) iron nitrate solution is prepared, as solution D; 20g iron is dissolved in the salpeter solution of the 2M of 250ml, after iron dissolves completely, makes solution D.
(5) be after 1:2:1.5:1 mixing with mass ratio by solution A, B solution, C solution, solution D, add antiseptic, stir and make absinthe-green urine monohydroxyphenyl metabolites detection reagent;
The temperature of the mixed liquor of solution A, B solution, C solution, solution D and solution A, B solution, C solution and solution D be 30 ?between 35 degrees Celsius.
Preparation embodiment 2
(1) ferrum sulfuricum oxydatum solutum is prepared, as solution A; 25g iron is dissolved in the sulfuric acid solution of the 3M of 250ml, after iron dissolves completely, makes solution A
(2) mercuric sulfate solution is prepared, as B solution; 20g mercury is dissolved in the sulfuric acid solution of the 3M of 250ml, after mercury dissolves completely, makes B solution;
(3) copper nitrate solution is prepared, as C solution; 20g copper is dissolved in the salpeter solution of the 2M of 250ml, after copper dissolves completely, makes C solution.
(4) iron nitrate solution is prepared, as solution D; 20g iron is dissolved in the salpeter solution of the 2M of 250ml, after iron dissolves completely, makes solution D.
(5) be after 1:1.5:2:1 mixing with mass ratio by solution A, B solution, C solution, solution D, add antiseptic, stir and make absinthe-green urine monohydroxyphenyl metabolites detection reagent;
The temperature of the mixed liquor of solution A, B solution, C solution, solution D and solution A, B solution, C solution and solution D be 30 ?between 35 degrees Celsius.
Preparation embodiment 3
(1) ferrum sulfuricum oxydatum solutum is prepared, as solution A; 20g iron is dissolved in the sulfuric acid solution of the 2M of 250ml, after iron dissolves completely, makes solution A
(2) mercuric sulfate solution is prepared, as B solution; 15g mercury is dissolved in the sulfuric acid solution of the 2M of 250ml, after mercury dissolves completely, makes B solution;
(3) copper nitrate solution is prepared, as C solution; 25g copper is dissolved in the salpeter solution of the 2M of 250ml, after copper dissolves completely, makes C solution.
(4) iron nitrate solution is prepared, as solution D; 25g iron is dissolved in the salpeter solution of the 2M of 250ml, after iron dissolves completely, makes solution D.
(5) be after 1:2:1.5:1 mixing with mass ratio by solution A, B solution, C solution, solution D, add antiseptic, stir and make absinthe-green urine monohydroxyphenyl metabolites detection reagent;
The temperature of the mixed liquor of solution A, B solution, C solution, solution D and solution A, B solution, C solution and solution D be 30 ?between 35 degrees Celsius.
Comparative example 1
(1) mercuric sulfate solution is prepared, as solution A; 15g mercury is dissolved in the sulfuric acid solution of the 2M of 250ml, after mercury dissolves completely, makes solution A;
(2) nickel nitrate solution is prepared, as B solution; 25g copper is dissolved in the salpeter solution of the 2M of 250ml, after copper dissolves completely, makes B solution.
(3) mercuric nitrate solution is prepared, as C solution; 25g iron is dissolved in the salpeter solution of the 2M of 250ml, after iron dissolves completely, makes C solution.
(4) be after 1:2:1.5:1 mixing with mass ratio by solution A, B solution, C solution, add antiseptic, stir and make absinthe-green urine monohydroxyphenyl metabolites detection reagent;
The temperature of the mixed liquor of solution A, B solution, C solution and solution A, B solution, C solution be 30 ?between 35 degrees Celsius.
Comparative example 2
(1) ferrum sulfuricum oxydatum solutum is prepared, as solution A; 20g iron is dissolved in the sulfuric acid solution of the 2M of 250ml, after iron dissolves completely, makes solution A
(2) mercuric sulfate solution is prepared, as B solution; 15g mercury is dissolved in the sulfuric acid solution of the 2M of 250ml, after mercury dissolves completely, makes B solution;
(3) copper nitrate solution is prepared, as C solution; 25g copper is dissolved in the salpeter solution of the 2M of 250ml, after copper dissolves completely, makes C solution.
(4) iron nitrate solution is prepared, as solution D; 25g iron is dissolved in the salpeter solution of the 2M of 250ml, after iron dissolves completely, makes solution D.
(5) be after 1:2:1.5:1 mixing with mass ratio by solution A, B solution, C solution, solution D, add antiseptic, stir and make absinthe-green urine monohydroxyphenyl metabolites detection reagent;
The temperature of the mixed liquor of solution A, B solution, C solution, solution D and solution A, B solution, C solution and solution D is 25 degrees Celsius.
Get 100 hepatocarcinoma patient urine specimens made a definite diagnosis and 10 without cancer sample (i.e. negative sample), carry out urine monohydroxyphenyl metabolites detection with preparation embodiment 1 ?3 and comparative example 1 ?2 respectively, testing result is as following table 1 and table 2.
Detection method is: get examinee urine 3ml, in the detection reagent of instillation 0.5ml, shake up, leave standstill 3min, examination criteria is: occurring white/yellow precipitation or do not occur precipitation person, is feminine gender, occur that grey/black precipitate person is positive, can arrange color board to by grey to the different colours of black, color darker instruction book hydroxyl phenolic metabolism thing content is more, and tumour may be more serious.Wherein color grade 0 is negative findings, and all the other color grades are grey or black.
The testing result of table 1 pair hepatocarcinoma patient
The testing result of table 2 pair negative sample
As shown in Table 1, detection reagent of the present invention is about 95% to the recall rate of hepatocarcinoma patient, higher than existing detection reagent.As shown in Table 2, detection reagent of the present invention can not cause false positive, is also better than existing detection reagent.
It should be noted that; the foregoing is only embodiments of the invention; not thereby the scope of the claims of the present invention is limited; every utilize instructions of the present invention to do equivalent structure or equivalent flow process conversion; or be directly or indirectly used in other correlative technology fields, be all in like manner included in scope of patent protection of the present invention.
Claims (7)
1. a preparation method for urine monohydroxyphenyl metabolites detection reagent, is characterized in that, comprises the steps:
(1) ferrum sulfuricum oxydatum solutum of 0.2-1g/ml is prepared, as solution A;
(2) the mercuric sulfate solution of 0.1-1g/ml is prepared, as B solution;
(3) copper nitrate solution of 0.15-1g/ml is prepared, as C solution;
(4) iron nitrate solution of 0.15-1g/ml is prepared, as solution D;
(5) by solution A, B solution, C solution, solution D with mass ratio be 1:(1-2): (1.5-2): after 1 mixing, add antiseptic, stir and make absinthe-green urine monohydroxyphenyl metabolites detection reagent;
Wherein, the temperature of the mixed liquor of solution A, B solution, C solution, solution D and solution A, B solution, C solution and solution D is between 30-35 degree Celsius.
2. the preparation method of urine monohydroxyphenyl metabolites detection reagent according to claim 1, it is characterized in that, the preparation process of described solution A is as follows: be dissolved in by 20--25g iron in the sulfuric acid solution of the 2-3M of 245-250ml, after iron dissolves completely, make solution A.
3. the preparation method of urine monohydroxyphenyl metabolites detection reagent according to claim 1, it is characterized in that, the preparation process of described B solution is as follows: be dissolved in by 10--25g mercury in the sulfuric acid solution of the 2-3M of 245-250ml, after mercury dissolves completely, make B solution.
4. the preparation method of urine monohydroxyphenyl metabolites detection reagent according to claim 1, it is characterized in that, the preparation process of described C solution is as follows: be dissolved in by 15--25g copper in the salpeter solution of the 2-3M of 245-250ml, after copper dissolves completely, make C solution.
5. the preparation method of urine monohydroxyphenyl metabolites detection reagent according to claim 1, it is characterized in that, the preparation process of described solution D is as follows: be dissolved in by 15--25g iron in the salpeter solution of the 2-3M of 245-250ml, after iron dissolves completely, make solution D.
6. the preparation method of urine monohydroxyphenyl metabolites detection reagent according to claim 1, it is characterized in that, in step (5), antiseptic is one or more of Sodium Benzoate, potassium sorbate, dehydro sodium acetate, calcium propionate, sodium Diacetate, sodium lactate or propylparaben.
7. the preparation method of urine monohydroxyphenyl metabolites detection reagent according to claim 1, is characterized in that, in step (5), solution A, B solution, C solution, solution D are that 1:1-2:1.5:1 mixes with mass ratio.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510536672.9A CN105158248A (en) | 2015-08-28 | 2015-08-28 | Preparation method of single hydroxyl phenolic urine metabolite detection reagent |
| PCT/CN2015/090540 WO2017035883A1 (en) | 2015-08-28 | 2015-09-24 | Method for preparing urine monohydroxy phenol metabolite detection reagent |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106645120A (en) * | 2016-11-21 | 2017-05-10 | 江苏华鸣生物科技有限公司 | Kit for detecting monohydroxyphenol metabolite in urine and preparation method of kit |
| CN107024472A (en) * | 2017-02-20 | 2017-08-08 | 北京众生合生物科技有限公司 | The single hydroxyl amphyl of urine determines the method for preparation and use of reagent |
| CN107356589A (en) * | 2017-07-11 | 2017-11-17 | 青岛浩铂生物科技有限公司 | A kind of preparation method of reagent thereof that single hydroxyl phenol derivatives are detected using sulfonating reaction |
| CN108287160A (en) * | 2017-01-10 | 2018-07-17 | 成都健浩生物科技有限公司 | The preparation method of the single hydroxyl amphyl measure reagent of urine |
| CN111201224A (en) * | 2017-10-16 | 2020-05-26 | 新生命医药科技有限公司 | Uropolyamines as biomarkers for prostate cancer detection |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR900002747B1 (en) * | 1988-03-11 | 1990-04-28 | 삼일제약 주식회사 | Improvemented millon reagent for using diagnosis in cancer screening |
| CN1192532A (en) * | 1997-03-05 | 1998-09-09 | 王恩铭 | Reagent for analysis of urine from tumour patient |
| CN104535565B (en) * | 2014-12-25 | 2016-03-02 | 江苏东博生物医药有限公司 | Single hydroxyl phenolic metabolism thing urine detection reagent and preparation method thereof |
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- 2015-08-28 CN CN201510536672.9A patent/CN105158248A/en active Pending
- 2015-09-24 WO PCT/CN2015/090540 patent/WO2017035883A1/en active Application Filing
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106645120A (en) * | 2016-11-21 | 2017-05-10 | 江苏华鸣生物科技有限公司 | Kit for detecting monohydroxyphenol metabolite in urine and preparation method of kit |
| CN108287160A (en) * | 2017-01-10 | 2018-07-17 | 成都健浩生物科技有限公司 | The preparation method of the single hydroxyl amphyl measure reagent of urine |
| CN107024472A (en) * | 2017-02-20 | 2017-08-08 | 北京众生合生物科技有限公司 | The single hydroxyl amphyl of urine determines the method for preparation and use of reagent |
| CN107356589A (en) * | 2017-07-11 | 2017-11-17 | 青岛浩铂生物科技有限公司 | A kind of preparation method of reagent thereof that single hydroxyl phenol derivatives are detected using sulfonating reaction |
| CN111201224A (en) * | 2017-10-16 | 2020-05-26 | 新生命医药科技有限公司 | Uropolyamines as biomarkers for prostate cancer detection |
| CN111201224B (en) * | 2017-10-16 | 2023-04-25 | 新生命医药科技有限公司 | Urinary polyamine as biomarker for prostate cancer detection |
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| WO2017035883A1 (en) | 2017-03-09 |
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Application publication date: 20151216 |