CN105241831A - Stable good-interference resistance serum zinc detection reagent and detection method - Google Patents
Stable good-interference resistance serum zinc detection reagent and detection method Download PDFInfo
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- CN105241831A CN105241831A CN201510656657.8A CN201510656657A CN105241831A CN 105241831 A CN105241831 A CN 105241831A CN 201510656657 A CN201510656657 A CN 201510656657A CN 105241831 A CN105241831 A CN 105241831A
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- reagent
- serum zinc
- detects
- damping fluid
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Abstract
The invention relates to the technical field of serum zinc detection and especially relates to a serum zinc detection reagent. A reagent R1 contains a buffer, salicylaldoxime, dimethylglyoximate, a fluorocarbon surfactant FSO-300, 2, 9-dimethyl-1, 10-phenanthroline, ascorbic acid, potassium ferrocyanide, polyethylene glycol 6000, glycol, trisodium citrate, NaCl and an antiseptic. A reagent R2 contains a buffer, dimethyl sulfoxide, nitro-PAPS, a fluorocarbon surfactant FSO-300, ascorbic acid, potassium ferrocyanide, polyethylene glycol 6000, glycol, trisodium citrate, NaCl and an antiseptic. The Tris buffer, the stabilizing agent and a plurality of anti-interference agents are used, can effectively eliminate interference caused by copper, iron, calcium and magnesium ions and can substantially improve reagent stability. The used novel non-ionic fluorocarbon surfactant FSO-300 can promote serum zinc cracking, prevent reaction system turbidity, improve chromogenic substance stability, prevent a pretreatment process such as protein removal and substantially improve reagent sensitivity and stability.
Description
Technical field
The present invention relates to serum zinc detection technique field, particularly a kind of serum zinc detects reagent, also relates to the detection method using this to detect reagent.
Background technology
Zinc is that the mankind sustain life one of necessary trace element, and it is combined with albumin and enters blood after enteron aisle is absorbed by intestinal mucosa cells.Zinc is discharged primarily of in ight soil.The important physiological function of zinc is the co-factor maintaining multiple enzyme in human body, about has the metalloenzyme of more than 90 kinds must have the normal function of zinc competence exertion, such as erythrocytic carbonic anhydrase, alkaline phosphatase and synthesize relevant multiple enzyme etc. with DNA, RNA.Zinc plays an important role to the structure maintaining enzyme, and after being combined with memebrane protein, the structure of Absorbable organic halogens film, prevents film lipid peroxidation.
Zinc in food especially in fish, eggs and cheese content very abundant.Zinc is the nutrient that body weight for humans is wanted, and its english abbreviation is: Zn.Zn content relative constancy in normal human blood, content is about 0.6 ~ 1.2mg/L.Serum zinc reduction sees acute myocardial infarction AMI, acute organ burn, hepatopathy, kidney trouble, diabetes etc.Serum zinc raises and sees hyperthyroidism, hypophysis and hypoadrenocorticism, polycythemia vera, eosinophilia, hyperpietic.
At present, the detection method that serum zinc is conventional has compleximetry, fluorimetry, colourimetry, atomic absorption spectrophotometry etc., and compleximetry sample consumption is large, and waste time and energy, specificity is low.Although fluorimetry is simple to operate, need specific instrument-fluorophotometer.And
Atomic absorption spectrophotometry specificity is high, simple to operate, but instrument is very expensive, is difficult to universal at basic hospital.
Given this, the present invention adopts colourimetry, and this method is simple to operate, quick, highly sensitive, is applicable to automated analysis.By optimizing reaction system, the isoionic interference of iron, copper, calcium, magnesium effectively can be avoided.Add the stability that stabilizing agent Macrogol 6000, ethylene glycol, NaCl etc. effectively can improve reagent; And preferred nonionic fluorocarbon surfactant FSO-300 adds the cracking that can promote serum zinc, reaction system can be prevented muddy, strengthen the stability of substance that show color, avoid the pre-treatment step such as removing protein.Significantly improve the sensitivity of reagent, fast easy and simple to handle, being applicable to automated analysis, is a kind of more stable, serum zinc (Zn) reagent that antijamming capability is strong.
Summary of the invention
The object of this invention is to provide one for detecting the reagent of serum zinc (Zn) and using this reagent to detect the method for serum zinc content.This kit adopts colourimetry, effectively can detect the content of serum zinc, and antijamming capability is strong, the advantages such as good stability.
Ultimate principle:
Nitro-PAPS reacts with Zn in alkaline solution, and raw empurpled compound, has maximum absorption peak at 570nm place.The interference of copper ion can by adding preferred neocuproine-2, and 9-dimethyl-1,10-phenanthroline is removed; The interference of ferric ion can by adding the elimination such as ascorbic acid and potassium ferrocyanide; And the earth alkali metal such as calcium, magnesium can eliminate its interference by adding trisodium citrate etc.
The present invention is obtained by following steps:
A kind of serum zinc detects reagent, and comprise reagent R1 and reagent R2, described reagent R1's and reagent R2 is composed as follows:
The component of reagent R1 is:
Damping fluid
200mmol/L,
Salicylaldoxime
5mmol/L,
Dimethylglyoximate
2mmol/L,
Fluorocarbon surfactant FSO-300
10ml/L,
2,9-dimethyl-1,10-phenanthroline
3mmol/L,
Ascorbic acid
5mmol/L,
Potassium ferrocyanide
1mmol/L,
Macrogol 6000
10g/L,
Ethylene glycol
5ml/L,
Trisodium citrate
12g/L,
NaCl
···················································································9g/L，
Antiseptic
0.5g/L;
The component of reagent R2 is:
Damping fluid
200mmol/L,
Dimethyl sulfoxide (DMSO)
5ml/L,
Nitro-PAPS
0.5mmol/L,
Fluorocarbon surfactant FSO-300
10ml/L,
Ascorbic acid
5mmol/L,
Potassium ferrocyanide
1mmol/L,
Macrogol 6000
10g/L,
Ethylene glycol
5ml/L,
Trisodium citrate
12g/L,
NaCl
···················································································9g/L，
Antiseptic
0.5g/L.
Described serum zinc detects reagent, and in reagent R1, damping fluid is 25 DEG C, and pH is the Tris damping fluid of 8.2.
Described serum zinc detects reagent, and in reagent R2, damping fluid is 25 DEG C, and pH is the Tris damping fluid of 8.2.
Described serum zinc detects reagent, and described antiseptic is NaN
3.
Described serum zinc detects the detection method that reagent detects serum zinc content, and use automatic clinical chemistry analyzer to utilize end-point method to measure, detection predominant wavelength is 570nm.
Described detection method, the ratio of R1 reagent and R2 reagent is 4:1.
Beneficial effect of the present invention:
1) adopt new buffer system and stabilizing agent and add multiple anti-interference agent, effectively can remove copper, iron and calcium, the isoionic interference of magnesium, significantly improve the stability of reagent;
2) adopt new non-ionic type fluorocarbon surfactant FSO-300, the cracking of serum zinc can be promoted, reaction system can be prevented muddy, strengthen the stability of substance that show color, avoid the pre-treatment step such as removing protein, significantly improve sensitivity and the stability of reagent;
3) reagent accuracy and have good stability, low price, easy to use, can meet clinical needs completely.
Accompanying drawing explanation
Fig. 1 is the correlation curve figure of two kinds of reagent;
Fig. 2 is two kinds of reagent effect phase stability curve figure;
Fig. 3 embodiment 1 reagent test method;
Fig. 4 embodiment 1 reagent interference free performance compares;
Fig. 5 embodiment 1 reagent and market is common and the Determination of Zn in Serum kit got the nod contrasts testing result.
Embodiment
Below in conjunction with specific embodiment, the present invention is further described.
embodiment 1
The detection reagent of serum zinc, bag reagent R1 and reagent R2:
1) the consisting of of its R1:
Tris(trishydroxymethylaminomethane) damping fluid (pH=8.2,25 DEG C)
100mmol/L
Salicylaldoxime
5mmol/L,
Dimethylglyoximate
2mmol/L,
Fluorocarbon surfactant FSO-300
10ml/L,
2,9-dimethyl-1,10-phenanthroline
3mmol/L,
Ascorbic acid
5mmol/L,
Potassium ferrocyanide
1mmol/L,
Macrogol 6000
10g/L,
Ethylene glycol
5ml/L,
Trisodium citrate
12g/L,
NaCl
··················································································································9g/L，
Liquid BPF aN
30.5g/L
2) component of reagent R2 is:
Tris(trishydroxymethylaminomethane) damping fluid (pH=8.2,25 DEG C)
100mmol/L
Dimethyl sulfoxide (DMSO)
5ml/L,
Nitro-PAPS
0.5mmol/L,
Fluorocarbon surfactant FSO-300
10ml/L,
Ascorbic acid
5mmol/L,
Potassium ferrocyanide
1mmol/L,
Macrogol 6000
10g/L,
Ethylene glycol
5ml/L,
Trisodium citrate
12g/L,
NaCl
··············································································································9g/L，
Antiseptic
0.5g/L,
Liquid BPF aN
30.5g/L.
3) using method of the present embodiment reagent:
The serum zinc that the present embodiment describes detects reagent, adopts the automatic clinical chemistry analyzer with double reagent function in use, as Hitachi 7180 fully-automatic analyzer etc., utilizes end-point method to measure.Be placed on corresponding reagent position according to the ratio of 4:1 by R1 and R2, place distilled water, standard items and sample at the correspondence position of sample disc, operation is as Fig. 3:
calculate: serum zinc content (μ
mol/L)=(A measures ÷ A standard) × C standard.
embodiment 2
Interference is tested: get fresh mix serum, be divided into 2 equal portions, then every equal portions are divided into 7 equal portions again, add different interfering materials, makes its concentration in serum reach the requirement of Fig. 4.Then use embodiment 1 gained reagent respectively, and the serum zinc approved (Zn) reagent common with market is the content of Zn in comparative determination serum simultaneously, control group measurement result with add disturbance material after the measurement result respectively organized see Fig. 4.Mensuration average × 100% of relative deviation (%)=(the mensuration average of the mensuration average-check sample of interference sample)/check sample.
As seen from Figure 4, embodiment 1 reagent is at triglyceride≤1000mg/dL, cholerythrin≤50mg/dL, Cu
2+≤ 60 μm of ol/L, Fe
3+≤ 60 μm of ol/L, Ca
2+≤ 5mmol/L, Mg
2+≤ 4mmol/L does not obviously disturb test result.And control group reagent is when above-mentioned concentration interfering material exists, be subject to obvious interference, this illustrates that the interference free performance of embodiment 1 reagent significantly improves, and is far superior to contrast agent by after optimization reaction buffer system, interpolation anti-interference agent and nonionic fluorocarbon surfactant FSO-300.
embodiment 3
Correlativity is tested: utilize embodiment 1 formulated reagent, the serum zinc kit of certain company that the State Food and Drug Administration common with market is approved carries out control test, and have detected 20 clinical serum samples, testing result as shown in Figure 5 simultaneously.And obtain the correlation curve (as shown in Figure 1) of two kinds of reagent, shown by testing result, the related coefficient of two kits is 0.9994, and describing both has great correlativity.
embodiment 4
The stability contrast test of reagent: to the reagent in embodiment 1, even packing 13 groups, the amount of reagent often organized is R1 be 20mL, R2 is 5mL; And serum zinc (Zn) kit getting certain company of the common State Food and Drug Administration's accreditation in 13 groups of market compares.Be placed in 2-8 DEG C of refrigerator, a taking-up on the same day group reagent monthly detects Zn quality-control product, and (target value is 19.5
μm ol/L), as shown in Figure 2, it is more stable that the serum zinc (Zn) more common than market under 2-8 DEG C of condition of storage of embodiment 1 reagent measures kit to testing result.
By checking, it is good that this reagent and similar detection reagent contrast correlativity, and clinical detection sample results is consistent, can reach the application requirement of market to product, and good in anti-interference performance is that a kind of more stable, good serum zinc (Zn) detects reagent.
Claims (7)
1. serum zinc detects a reagent, and it is characterized in that comprising reagent R1 and reagent R2, described reagent R1's and reagent R2 is composed as follows:
The component of reagent R1 is:
Damping fluid
200mmol/L,
Salicylaldoxime
5mmol/L,
Dimethylglyoximate
2mmol/L,
Fluorocarbon surfactant FSO-300
10ml/L,
2,9-dimethyl-1,10-phenanthroline
3mmol/L,
Ascorbic acid
5mmol/L,
Potassium ferrocyanide
1mmol/L,
Macrogol 6000
10g/L,
Ethylene glycol
5ml/L,
Trisodium citrate
12g/L,
NaCl
···················································································9g/L，
Antiseptic
0.5g/L;
The component of reagent R2 is:
Damping fluid
200mmol/L,
Dimethyl sulfoxide (DMSO)
5ml/L,
Nitro-PAPS
0.5mmol/L,
Fluorocarbon surfactant FSO-300
10ml/L,
Ascorbic acid
5mmol/L,
Potassium ferrocyanide
1mmol/L,
Macrogol 6000
10g/L,
Ethylene glycol
5ml/L,
Trisodium citrate
12g/L,
NaCl
···················································································9g/L，
Antiseptic
0.5g/L.
2. serum zinc according to claim 1 detects reagent, and it is characterized in that in reagent R1, damping fluid is 25 DEG C, pH is the Tris damping fluid of 8.2.
3. serum zinc according to claim 1 detects reagent, and it is characterized in that in reagent R1, damping fluid is 25 DEG C, pH is the Tris damping fluid of 8.2.
4. serum zinc according to claim 1 detects reagent, it is characterized in that in reagent R1 and R2, surfactant is fluorocarbon surfactant FSO-300.
5. serum zinc according to claim 1 detects reagent, it is characterized in that described antiseptic is NaN
3.
6. the serum zinc according to any one of claim 1-5 detects the detection method that reagent detects serum zinc, it is characterized in that using automatic clinical chemistry analyzer to utilize end-point method to measure, and detection predominant wavelength is 570nm.
7. detection method according to claim 6, is characterized in that the ratio of R1 reagent and R2 reagent is 4:1.
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Cited By (7)
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| CN106370862A (en) * | 2016-08-30 | 2017-02-01 | 山东博科生物产业有限公司 | Stable and sensitive fibronectin detection reagent |
| CN107290522A (en) * | 2017-06-15 | 2017-10-24 | 中国农业大学 | A kind of kit of zinc ion content quick detection |
| CN108169224A (en) * | 2017-12-27 | 2018-06-15 | 山东博科生物产业有限公司 | A kind of serum magnesium detection reagent and preparation |
| CN108444805A (en) * | 2018-02-13 | 2018-08-24 | 广州聚佰生物科技有限公司 | A kind of purifying tablets for rapid detection for pesticide residue pre-treatment |
| CN109374884A (en) * | 2018-12-24 | 2019-02-22 | 四川沃文特生物技术有限公司 | A kind of PCT concentration detection kit and preparation method thereof |
| CN112858422A (en) * | 2019-11-12 | 2021-05-28 | 北京怡成生物电子技术股份有限公司 | Electrochemical uric acid test strip for removing interference of ascorbic acid and manufacturing and application thereof |
| CN112986223A (en) * | 2021-02-25 | 2021-06-18 | 吉林大学第一医院 | Reagent strip for rapid detection of seminal plasma biochemical zinc ions |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106370862A (en) * | 2016-08-30 | 2017-02-01 | 山东博科生物产业有限公司 | Stable and sensitive fibronectin detection reagent |
| CN106370862B (en) * | 2016-08-30 | 2018-07-20 | 山东博科诊断科技有限公司 | A kind of stabilization, sensitive fibronectin detection reagent |
| CN107290522A (en) * | 2017-06-15 | 2017-10-24 | 中国农业大学 | A kind of kit of zinc ion content quick detection |
| CN108169224A (en) * | 2017-12-27 | 2018-06-15 | 山东博科生物产业有限公司 | A kind of serum magnesium detection reagent and preparation |
| CN108444805A (en) * | 2018-02-13 | 2018-08-24 | 广州聚佰生物科技有限公司 | A kind of purifying tablets for rapid detection for pesticide residue pre-treatment |
| CN108444805B (en) * | 2018-02-13 | 2020-10-02 | 广州聚佰生物科技有限公司 | Purifying sheet for quick pesticide residue detection pretreatment |
| CN109374884A (en) * | 2018-12-24 | 2019-02-22 | 四川沃文特生物技术有限公司 | A kind of PCT concentration detection kit and preparation method thereof |
| CN112858422A (en) * | 2019-11-12 | 2021-05-28 | 北京怡成生物电子技术股份有限公司 | Electrochemical uric acid test strip for removing interference of ascorbic acid and manufacturing and application thereof |
| CN112858422B (en) * | 2019-11-12 | 2022-12-20 | 北京怡成生物电子技术股份有限公司 | Electrochemical uric acid test strip for removing interference of ascorbic acid and manufacturing and application thereof |
| CN112986223A (en) * | 2021-02-25 | 2021-06-18 | 吉林大学第一医院 | Reagent strip for rapid detection of seminal plasma biochemical zinc ions |
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