CN108627654B - Composition for eliminating interference of calcium dobesilate medicine on creatinine enzymatic detection - Google Patents
Composition for eliminating interference of calcium dobesilate medicine on creatinine enzymatic detection Download PDFInfo
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- CN108627654B CN108627654B CN201810663778.9A CN201810663778A CN108627654B CN 108627654 B CN108627654 B CN 108627654B CN 201810663778 A CN201810663778 A CN 201810663778A CN 108627654 B CN108627654 B CN 108627654B
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- G—PHYSICS
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/70—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving creatine or creatinine
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
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- G—PHYSICS
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- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/34—Genitourinary disorders
- G01N2800/347—Renal failures; Glomerular diseases; Tubulointerstitial diseases, e.g. nephritic syndrome, glomerulonephritis; Renovascular diseases, e.g. renal artery occlusion, nephropathy
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Abstract
The invention relates to a composition for eliminating interference of calcium dobesilate drugs on creatinine enzyme method detection, which comprises the following components: r1 reagent, R2 reagent, and polymeric oxidizer; the high molecular oxidant is one or more of high molecular peroxyacid, high molecular selenium oxide, high molecular chlorosulphide, N-imide, water-soluble tetrazole and dessimutane oxidant. The high molecular oxidant in the composition disclosed by the invention can react with calcium dobesilate in a sample to be detected, so that the calcium dobesilate loses the effect of interfering the detection of the creatinine in an enzyme method, and the detection accuracy of the creatinine oxidase method is improved, thereby solving the problem of serious negative interference of high-concentration calcium dobesilate in blood of a patient on the detection result of the creatinine item in the enzyme method during the taking of a calcium dobesilate medicament by the patient.
Description
Technical Field
The invention belongs to the field of medical inspection, and particularly relates to a composition for eliminating interference of calcium dobesilate drugs on creatinine enzymatic detection.
Background
Creatinine, an end product of the metabolism of creatine and phosphocreatine, is produced primarily by the nonenzymatic reaction of phosphocreatine in muscle. The degree of impaired renal function can be known from the measurement of creatinine in blood, and a higher creatinine measurement indicates a more serious impairment of renal function.
The alkaline picric acid method is a traditional creatinine detection method, is well used for a long time, but has poor stability and is gradually eliminated by various clinical laboratories; the creatinine enzyme method is a creatinine detection method appearing in recent years, and has become a main creatinine item detection method in clinical departments due to good stability and precision and high linear range.
Calcium dobesilate is a blood vessel protective drug and is mainly used for treating diabetic nephropathy, chronic renal insufficiency, chronic arteriovenous insufficiency and the like. Recently, there are many reports in the literature that calcium dobesilate has severe negative interference on the enzyme assay of creatinine, and Xiuzhi Guo, Li' an Hou, Yicong Yin, Jie Wu, Fang Zhao, Lianggyu Xia, Xinqi Cheng, Qian Liu, Li Liu, Ermu Xu, Ling Qiu, etc. have studied the specific mechanism of interference of calcium dobesilate on the enzyme assay of creatinine, and have found that the hydroquinone ring of calcium dobesilate can consume H2O2Reagents that lead to numerous coupled trinder's reactions are negatively biased to varying degrees.
Disclosure of Invention
In view of the defects of the prior art, the invention aims to provide a composition for eliminating the interference of a calcium dobesilate medicament on the detection of a creatinine enzyme method, and aims to solve the problem of the interference of the calcium dobesilate medicament on the detection of the creatinine enzyme method.
The technical scheme of the invention is as follows:
a composition for eliminating interference of calcium dobesilate drugs on enzymatic creatinine assay, comprising: r1 reagent, R2 reagent, and polymeric oxidizer;
the high molecular oxidant is one or more of high molecular peroxyacid, high molecular selenium oxide, high molecular chlorosulphide, N-imide, water-soluble tetrazole and dessimutane oxidant;
the R1 reagent comprises the following components in percentage by concentration:
the R2 reagent comprises the following components in percentage by concentration:
the composition, wherein the polymeric peroxyacid comprises peroxyacetic acid and peroxypropionic acid.
The composition, wherein the macromolecular selenium oxide comprises selenium oxychloride and bis (4-methoxyphenyl) selenium oxide.
The composition, wherein the polymer chlorinated sulfide comprises chloromethyl methyl sulfide and chloromethyl diphenyl sulfide.
The composition, wherein the N-substituted imide comprises chlorobutyl diacetylimide, N-ethyl maleimide and N-methyl maleimide.
The composition comprises water-soluble tetrazolium, water-soluble tetrazolium salt-1, water-soluble tetrazolium salt-2, water-soluble tetrazolium salt-3, water-soluble tetrazolium salt-4, water-soluble tetrazolium salt-5, water-soluble tetrazolium salt-6, water-soluble tetrazolium salt-7 and water-soluble tetrazolium salt-8.
The composition, wherein the dessimutan oxidant comprises 1,1,1- (triacetoxy) -1, 1-dihydro-1, 2-phenyliodoyl-3 (1H) -one, dessimutan.
The composition, wherein the concentration of the polymeric oxidant is 0.01-10 mmol/L.
The composition, wherein the buffer is a tris buffer, and the pH is 8.2; the interference removing agent is potassium ferrocyanide; the preservative is Pc 300.
Has the advantages that: the high molecular oxidant in the composition disclosed by the invention can react with calcium dobesilate, so that the calcium dobesilate loses the effect of interfering the creatinine enzyme method detection, and the problem of serious negative interference of high-concentration calcium dobesilate in blood of a patient on the detection result of an enzyme method creatinine project during the period that the patient takes a calcium dobesilate drug is solved.
Drawings
FIG. 1 is a graph of test results for the examples provided in the present invention.
Detailed Description
The invention provides a composition for eliminating interference of calcium dobesilate drugs on creatinine enzymatic detection, and the invention is further described in detail below in order to make the purpose, technical scheme and effect of the invention clearer and more clear. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The test methods described below are all conventional methods unless otherwise specified, and the test materials used are readily available from commercial companies unless otherwise specified.
A composition for eliminating interference of calcium dobesilate drugs on enzymatic creatinine assay detection, comprising: r1 reagent, R2 reagent, and polymeric oxidizer;
the high molecular oxidant is one or more of high molecular peroxyacid, high molecular selenium oxide, high molecular chlorosulphide, N-imide, water-soluble tetrazole and dessimutane oxidant;
the R1 reagent comprises the following components in percentage by concentration:
the R2 reagent comprises the following components in percentage by concentration:
in one embodiment of the present invention, the polymeric peroxyacid in the polymeric oxidizer is a polymeric compound having a general molecular formula of R-CO-O-OH, wherein the R group in the general formula is a group unrelated to the reaction and may be an alkyl group, a hydrocarbon group, or the like; the polymer selenium oxide in the polymer oxidizing agent is a polymer compound containing a-SeO-group, and examples thereof include selenium oxychloride, bis (4-methoxyphenyl) selenium oxide, and the like.
Further, the concentration of the macromolecular oxidant in the composition is 0.01mmol/L to 10 mmol/L; preferably 0.01mmol/L to 5mmol/L, more preferably 0.05mmol/L to 1 mmol/L. When the addition concentration of the polymeric oxidizing agent is less than 0.05mmol/L, a large amount of the composition is required, which is not favorable for detection and analysis, and when the addition concentration of the polymeric oxidizing agent is more than 1mmol/L, the amount of the composition required for detection is relatively small, which is inconvenient for operation.
The composition provided by the invention can be applied to the detection of creatinine by an enzyme method, and can also be applied to other clinical biochemical projects based on the reaction principle of trinder's (coupled end-point colorimetry), such as Uric Acid (UA), free fatty acid (NEFA), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and the like.
The present invention is further illustrated by the following specific examples, which employ the sarcosine oxidase method.
1. The specific reagent components are as follows:
r1 reagent:
| RI reagent material | Concentration of | Sources of materials |
| Tris buffer | 100mm/L(PH8.2) | Chinese medicine |
| Potassium ferrocyanide | 0.1g/L | Chinese medicine |
| TX-100 | 0.001 | sigma |
| TODB chromogen | 2mm/L | Xibao (medicine for health) |
| Catalase enzyme | 200KU/L | Roche of Roche |
| Ascorbic acid oxidase | 20KU/L | Roche of Roche |
| Sarcosine oxidase | 30KU/L | Novel enzyme in vast sea |
| Creatine dehydrogenase | 30KU/L | Novel enzyme in vast sea |
| Preservative Pc300 | 0.005 | sigma |
R2 reagent:
2. the specific implementation scheme is as follows:
the additive, i.e., the polymeric oxidizer, may be added to the R1 reagent or the R2 reagent, and is not limited thereto.
3. The specific operation steps are as follows:
20 parts of normal human serum without turbidity, jaundice and hemolysis is collected to prepare mixed serum. Taking 6 1.5ml EP tubes, respectively adding 0.9ml of mixed serum and numbering according to 1-6; preparing a 640 mu g/ml calcium dobesilate solution, and diluting and releasing 320 mu g/ml, 160 mu g/ml, 80 mu g/ml and 40 mu g/ml calcium dobesilate solution at equal ratio; 0.1ml of purified water is added into No. 1 mixed serum, 0.1ml of 40 mu g/ml calcium dobesilate solution is added into No. 2 mixed serum, 0.1ml of 80 mu g/ml calcium dobesilate solution is added into No. 3 mixed serum, 0.1ml of 160 mu g/ml calcium dobesilate solution is added into No. 4 mixed serum, 0.1ml of 320 mu g/ml calcium dobesilate solution is added into No. 5 mixed serum, 0.1ml of 640 mu g/ml calcium dobesilate solution is added into No. 6 mixed serum, and the uniformly mixed samples are uniformly mixed by using a vortex instrument to obtain 6 samples with the same creatinine concentration and the calcium dobesilate content of 0 mu g/ml, 4 mu g/ml and 8 mu g/ml … … 64 mu g/ml.
The reagents of each example were scaled on a Hitachi 7170 fully automatic biochemical analyzer using Landao composite calibrators according to the following on-machine parameters:
| dominant wavelength | Sub-wavelength | S | R1 | R2 | Detection method | Calibration mode | Light measuring point |
| 546nm | 660nm | 4μL | 180μL | 60μL | Two point end point | Two point linearity | 15,31 |
Then, the prepared samples are respectively detected, each sample is respectively detected for 3 times, the average value is taken and recorded as a result, and the detection results are as follows:
the maximum limit concentration of calcium dobesilate in the serum of patients taking the medicine is usually 60 mug/ml, and in the above operation steps, the maximum concentration of 64 mug/ml is selected to cover the range. For example, 320. mu.g/ml, 160. mu.g/ml, 80. mu.g/ml and 40. mu.g/ml calcium dobesilate solutions are released by equal dilution, and the test can be accurately and quickly carried out by equal dilution. Of course, other concentrations in the range of 0-64. mu.g/ml are possible.
Referring to fig. 1, which is a graph showing the test results of the above embodiments, reference numeral 1 indicates embodiment 1, reference numeral 2 indicates embodiment 2, and reference numeral … 10 indicates embodiment 10. As can be seen from the figure, the deviation of the results of example 7 is minimal, and the deviation of the results of example 7 is much smaller than that of example 1 (without adding the polymeric oxidizing agent), especially at high concentrations.
In conclusion, the high molecular oxidant in the composition disclosed by the invention can react with calcium dobesilate in a sample to be detected, so that the calcium dobesilate loses the effect of interfering the detection of the creatinine by the enzyme method, and the detection accuracy of the creatine oxidase method is improved, thereby solving the problem of serious negative interference of high-concentration calcium dobesilate in blood of a patient on the detection result of the creatinine item by the enzyme method during the taking of a calcium dobesilate medicament by the patient.
It is to be understood that the invention is not limited to the examples described above, but that modifications and variations may be effected thereto by those of ordinary skill in the art in light of the foregoing description, and that all such modifications and variations are intended to be within the scope of the invention as defined by the appended claims.
Claims (8)
1. A composition for eliminating interference of calcium dobesilate drugs on enzymatic creatinine assay detection is characterized by comprising: r1 reagent, R2 reagent, and polymeric oxidizer;
the high molecular oxidant is one or more of high molecular peroxyacid, high molecular selenium oxide, high molecular chlorosulphide, N-imide, water-soluble tetrazole and dessimutane oxidant;
the R1 reagent comprises the following components in percentage by concentration:
the R2 reagent comprises the following components in percentage by concentration:
the preservative is Pc 300; the concentration of the macromolecular oxidant is 0.01-10 mmol/L.
2. The composition of claim 1, wherein the polymeric peroxyacid is peroxyacetic acid or peroxypropionic acid.
3. The composition of claim 1, wherein the polymeric selenium oxide comprises selenium oxychloride, bis (4-methoxyphenyl) selenium oxide.
4. The composition as claimed in claim 1, wherein the high molecular weight chlorinated sulfide comprises chloromethyl methyl sulfide, chloromethyl diphenyl sulfide.
5. The composition of claim 1, wherein the N-substituted imide comprises chlorobutyldiacetimide, N-ethylmaleimide, N-methylmaleimide.
6. The composition of claim 1, wherein the water-soluble tetrazolium comprises water-soluble tetrazolium salt-1, water-soluble tetrazolium salt-2, water-soluble tetrazolium salt-3, water-soluble tetrazolium salt-4, water-soluble tetrazolium salt-5, water-soluble tetrazolium salt-6, water-soluble tetrazolium salt-7 and water-soluble tetrazolium salt-8.
7. The composition as claimed in claim 1, wherein said dessimutan oxidant comprises 1,1,1- (triacetoxy) -1, 1-dihydro-1, 2-benziodox-3 (1H) -one, dessimutan.
8. The composition according to claim 1, characterized in that the buffer is a tris buffer, pH 8.2; the interference removing agent is potassium ferrocyanide.
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| WO2020113521A1 (en) * | 2018-12-06 | 2020-06-11 | 深圳迈瑞生物医疗电子股份有限公司 | Method for eliminating doxorubicin interference in immunoassay and immunoassay kit |
| CN111443201A (en) * | 2020-02-25 | 2020-07-24 | 深圳市安帝宝科技有限公司 | Aspirin drug resistance detection kit |
| CN111394424B (en) * | 2020-03-11 | 2023-09-29 | 海丰生物科技（北京）有限公司 | Human serum creatinine content detection reagent and method for resisting calcium dobesilate interference |
| CN111766234B (en) * | 2020-08-07 | 2022-11-29 | 武汉瀚海新酶生物科技有限公司 | Creatinine detection reagent for resisting interference of calcium dobesilate and other medicines |
| CN112626170A (en) * | 2020-08-15 | 2021-04-09 | 中山标佳生物科技有限公司 | Uric acid kit for quickly and simply eliminating drug interference and preparation method thereof |
| CN111733208B (en) * | 2020-08-15 | 2021-02-02 | 中山标佳生物科技有限公司 | Creatinine kit capable of eliminating calcium dobesilate and etamsylate and preparation method thereof |
| CN112029820B (en) * | 2020-09-07 | 2023-08-18 | 上海科华生物工程股份有限公司 | Creatinine content detection kit for resisting calcium dobesilate interference and application thereof |
| CN113817742A (en) * | 2021-08-23 | 2021-12-21 | 湖北擎科生物科技有限公司 | Aptamer combined with catalase and application |
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Application publication date: 20181009 Assignee: Yichang Hanzhi Biotechnology Co.,Ltd. Assignor: WUHAN NEW BIOCALYSIS SOLUTION Co.,Ltd. Contract record no.: X2022420000012 Denomination of invention: Composition for eliminating interference of calcium dobesilate drug on creatinine enzymatic detection Granted publication date: 20210316 License type: Common License Record date: 20220128 |
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