CN114705640A - High-sensitivity serum zinc determination reagent with strong anti-interference capability and preparation method thereof - Google Patents
High-sensitivity serum zinc determination reagent with strong anti-interference capability and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a high-sensitivity serum zinc determination reagent with strong anti-interference capability and a preparation method thereof.A first raw material and a second raw material are weighed according to a ratio and are respectively added into corresponding mixing containers for stirring, dissolving and mixing to obtain a first mixed solution; adjusting the pH value of the first mixed solution to 8.0-9.0, quantifying by using purified water to a configured amount, and then filtering and standing to obtain a first reagent; weighing a third raw material and a fourth raw material according to a ratio, and respectively adding the third raw material and the fourth raw material into corresponding mixing containers to stir, dissolve and mix to obtain a second mixed solution; and adjusting the pH value of the second mixed solution to 8.0-9.0, quantifying by using purified water to a configured amount, and then filtering and standing to obtain the second reagent for improving the using effect of the reagent.
Description
Technical Field
The invention relates to the technical field of medical detection, in particular to a high-sensitivity serum zinc determination reagent with strong anti-interference capability and a preparation method thereof.
Background
Serum zinc ion detection principle (pyridylazo multi-metal complex method): under the existence of anti-protein, organic interference agent and metal masking agent with specific concentration, the pyridine azo complex is reacted with zinc in alkaline solution to produce purple compound with maximum absorption peak at 500-600 nm wavelength. The concentration of the zinc ions in the sample can be calculated by measuring the photometric value of the purple compound generated by the zinc ions with standard concentration and establishing a standard curve of absorbance change and concentration. When the human serum zinc ions are detected, the reagent has weak anti-interference capability on endogenous metal ion interference substances such as copper ions, iron ions, calcium ions, magnesium ions and the like, so that the test result is inaccurate. Meanwhile, the anti-interference agent is selected improperly, so that the combination reaction of the pyridine azo multi-element metal complex to zinc ions is seriously interfered, and the sensitivity of the reagent is not high and the detection result is inaccurate. And the performance of the reagent is unstable, and the requirement that the general in vitro diagnostic reagent has to reach the expiration date of 12-18 months cannot be met, so that the using effect of the reagent is poor.
Disclosure of Invention
The invention aims to provide a high-sensitivity serum zinc determination reagent with strong anti-interference capability and a preparation method thereof, and the use effect of the reagent is improved.
In order to achieve the above object, in a first aspect, the present invention provides a method for preparing a high-sensitivity serum zinc assay reagent with strong interference resistance, comprising the following steps:
weighing a first raw material and a second raw material according to a ratio, and respectively adding the first raw material and the second raw material into corresponding mixing containers to stir, dissolve and mix to obtain a first mixed solution;
adjusting the pH value of the first mixed solution to 8.0-9.0, quantifying by using purified water to a configured amount, and then filtering and standing to obtain a first reagent;
weighing a third raw material and a fourth raw material according to a ratio, and respectively adding the third raw material and the fourth raw material into corresponding mixing containers to stir, dissolve and mix to obtain a second mixed solution;
and adjusting the pH value of the second mixed solution to 8.0-9.0, quantifying by using purified water to a configured amount, and then filtering and standing to obtain a second reagent.
Wherein, weigh first raw materials and second raw materials according to the ratio, add respectively and stir in the mixing vessel who corresponds and dissolve and mix, obtain first mixed solution, include:
weighing a first raw material according to a ratio, adding the first raw material into a mixing container containing 75-85% of purified water by mass ratio, and stirring until the first raw material is dissolved;
weighing a second raw material according to a ratio, and adding the second raw material into the other mixing container in two batches for stirring and dissolving;
and mixing the first raw material solution and the second raw material solution to obtain a first mixed solution.
Adjusting the pH value of the first mixed solution to 8.0-9.0, quantifying by using purified water to a configured amount, and then filtering and standing to obtain a first reagent, wherein the first reagent comprises:
judging whether the pH value of the first mixed solution is 8.0-9.0 or not;
if not, adjusting by using a hydrochloric acid solution or a sodium hydroxide solution;
and (4) quantifying the purified water to a configured amount, and then filtering and standing to obtain a first reagent.
Wherein, utilize purified water ration to carry out the filtration after the configuration volume and stand, obtain first reagent, include:
quantifying to a configured amount by using purified water, uniformly stirring, and standing for 15 minutes or more;
filtering with a nylon filter membrane with the pore size of 0.4-1.0 micron to obtain the first reagent.
In a second aspect, the present invention provides a high-sensitivity serum zinc determination reagent with strong anti-interference capability, and the preparation method of the high-sensitivity serum zinc determination reagent with strong anti-interference capability according to the first aspect is applied to the high-sensitivity serum zinc determination reagent with strong anti-interference capability,
the high-sensitivity serum zinc determination reagent with strong anti-interference capability comprises a first reagent and a second reagent, wherein the mass ratio of the first reagent to the second reagent is 4: 1;
the first reagent comprises a first raw material and a second raw material, wherein the first raw material comprises 20-200 mmol/L of buffer solution with the pH value of 8.0-9.0, 4-10 g/L of sodium potassium tartrate, 0.05-0.1 g/L of ferrotitanium reagent, 0.05-0.15 g/L of pyrrolidine dithioamino-benzoic acid ammonium and 0.1-0.5 g/L of preservative, and the second raw material comprises 0.1-0.5 g/L of dimethylglyoxime, 0.1-0.5 g/L of salicylaldoxime, 5-10 ml/L of absolute ethyl alcohol and 1-5 g/L of nonionic surfactant;
the second reagent comprises a third raw material and a fourth raw material, wherein the third raw material comprises 20-200 mmol/L of buffer solution with the pH value of 8.0-9.0 and 0.1-0.5 g/L of preservative, and the fourth raw material comprises 0.15-0.5 g/L of pyridine azo multi-metal complex, 5-10 g/L of nonionic surfactant and 5-10 ml/L of absolute ethyl alcohol.
Wherein the buffer solution comprises one or more of N-tris (hydroxymethyl) methyl-3-aminopropane sulfonic acid, tris (hydroxymethyl) methylglycine, N-di-hydroxyethylglycine, N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid-sodium hydroxide, piperazine-1, 4-diethylsulfonic acid-sodium hydroxide.
Wherein the nonionic surfactant comprises one or two of fatty alcohol-polyoxyethylene ether and alkylphenol polyoxyethylene ether.
Wherein, the preservative can be one or two of methyl chloroisothiazolinone and methyl isothiazolinone.
The high-sensitivity serum zinc determination reagent with strong anti-interference capability comprises a first reagent and a second reagent, wherein the mass ratio of the first reagent to the second reagent is 4:1, the first raw material and the second raw material are weighed according to a ratio and are respectively added into corresponding mixing containers for stirring, dissolving and mixing to obtain a first mixed solution; adjusting the pH value of the first mixed solution to 8.0-9.0, quantifying by using purified water to a configured amount, and then filtering and standing to obtain a first reagent; weighing a third raw material and a fourth raw material according to a ratio, and respectively adding the third raw material and the fourth raw material into corresponding mixing containers to stir, dissolve and mix to obtain a second mixed solution; and adjusting the pH value of the second mixed solution to 8.0-9.0, quantifying by using purified water to a configured amount, and then filtering and standing to obtain the second reagent for improving the using effect of the reagent.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a schematic step diagram of a preparation method of a high-sensitivity serum zinc determination reagent with strong anti-interference capability provided by the invention.
FIG. 2 is a first embodiment of the present invention for correlation analysis of clinical results with the same type of detection reagents.
Detailed Description
Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the accompanying drawings are illustrative and intended to explain the present invention and should not be construed as limiting the present invention.
In the description of the present invention, "a plurality" means two or more unless specifically defined otherwise.
Referring to fig. 1, the present invention provides a method for preparing a high-sensitivity serum zinc determination reagent with strong interference resistance, comprising the following steps:
s101, weighing the first raw material and the second raw material according to the proportion, and respectively adding the first raw material and the second raw material into corresponding mixing containers to be stirred, dissolved and mixed to obtain a first mixed solution.
Specifically, a first raw material is weighed according to a ratio, all ingredients of the first raw material are sequentially added into a mixing container containing 75-85% of purified water (accounting for 75-85% of the total amount required by the preparation of the first reagent) by mass ratio, and the mixture is stirred one by one until the ingredients are completely dissolved; weighing a second raw material according to a ratio, dividing the second raw material into two batches, adding the two batches of the second raw material into the other mixing container, stirring and dissolving, namely stirring the second raw material of the first batch until the second raw material is fully and uniformly mixed, and then adding the second raw material of the second batch one by one until the second raw material is completely dissolved; and then, fully mixing the first raw material solution and the second raw material solution to obtain a first mixed solution.
S102, adjusting the pH value of the first mixed solution to 8.0-9.0, quantifying by using purified water to a configured amount, and then filtering and standing to obtain a first reagent.
Specifically, it is tested whether the first mixed solution has a pH value of 8.0 to 9.0, otherwise adjusted to this range with a hydrochloric acid solution or a sodium hydroxide solution.
And quantifying with purified water to a preparation amount, uniformly stirring, standing and balancing the solution for at least 15 minutes, and filtering insoluble substances with a 0.4-1.0 micron aperture nylon filter membrane to obtain the first reagent.
S103, weighing the third raw material and the fourth raw material according to the proportion, and respectively adding the weighed third raw material and the fourth raw material into corresponding mixing containers to be stirred, dissolved and mixed to obtain a second mixed solution.
Specifically, a third raw material is weighed according to a ratio, all ingredients of the third raw material are sequentially added into a mixing container containing 75-85% of purified water (accounting for 75-85% of the total amount required by the preparation of the third reagent) by mass ratio, and the mixture is stirred one by one until the ingredients are completely dissolved; weighing a fourth raw material according to a ratio, adding the fourth raw material into another mixing container for stirring and dissolving in two batches, namely stirring the fourth raw material of the first batch until the fourth raw material is fully and uniformly mixed, and then adding the fourth raw material of the second batch one by one until the fourth raw material is completely dissolved; and then, fully mixing the third raw material solution and the fourth raw material solution to obtain a second mixed solution.
And S104, adjusting the pH value of the second mixed solution to 8.0-9.0, quantifying by using purified water to a configured amount, and then filtering and standing to obtain a second reagent.
Specifically, it is tested whether the pH of the second mixed solution is 8.0 to 9.0, otherwise it is adjusted to this range with a hydrochloric acid solution or a sodium hydroxide solution.
And (3) quantifying to a preparation amount by using purified water, uniformly stirring, standing and balancing the solution for at least 15 minutes, and filtering insoluble substances by using a nylon filter membrane with the aperture of 0.4-1.0 micrometer to obtain a second reagent.
The invention provides a high-sensitivity serum zinc determination reagent with strong anti-interference capability, and the preparation method of the high-sensitivity serum zinc determination reagent with strong anti-interference capability is applied to the high-sensitivity serum zinc determination reagent with strong anti-interference capability,
the high-sensitivity serum zinc determination reagent with strong anti-interference capability comprises a first reagent and a second reagent, wherein the mass ratio of the first reagent to the second reagent is 4: 1;
the first reagent comprises a first raw material and a second raw material, wherein the first raw material comprises 20-200 mmol/L of buffer solution with the pH value of 8.0-9.0, 4-10 g/L of sodium potassium tartrate, 0.05-0.1 g/L of ferrotitanium reagent, 0.05-0.15 g/L of pyrrolidine dithioamino-benzoic acid ammonium and 0.1-0.5 g/L of preservative, and the second raw material comprises 0.1-0.5 g/L of dimethylglyoxime, 0.1-0.5 g/L of salicylaldoxime, 5-10 ml/L of absolute ethyl alcohol and 1-5 g/L of nonionic surfactant; the second reagent comprises a third raw material and a fourth raw material, wherein the third raw material comprises 20-200 mmol/L of buffer solution with the pH value of 8.0-9.0 and 0.1-0.5 g/L of preservative, and the fourth raw material comprises 0.15-0.5 g/L of pyridine azo multi-metal complex, 5-10 g/L of nonionic surfactant and 5-10 ml/L of absolute ethyl alcohol.
In the embodiment, the first reagent is used for masking endogenous interfering substances (metal, protein and organic substances) and adjusting the pH value in the reaction process, the second reagent is a pyridine-azo multi-element metal complex solution which forms a complete detection reagent in the detection process, and the pH value of the reaction system is 8.0-9.0. The reaction ratio of the first reagent to the second reagent is 4: 1. the first reagent comprises a first raw material and a second raw material, wherein the first raw material comprises 20-200 mmol/L of buffer solution with the pH value of 8.0-9.0, 4-10 g/L of sodium potassium tartrate, 0.05-0.1 g/L of ferrotitanium reagent, 0.05-0.15 g/L of pyrrolidine dithioamino-benzoic acid ammonium and 0.1-0.5 g/L of preservative, and the second raw material comprises 0.1-0.5 g/L of dimethylglyoxime, 0.1-0.5 g/L of salicylaldoxime, 5-10 ml/L of absolute ethyl alcohol and 1-5 g/L of nonionic surfactant; the second reagent comprises a third raw material and a fourth raw material, wherein the third raw material comprises 20-200 mmol/L of buffer solution with the pH value of 8.0-9.0 and 0.1-0.5 g/L of preservative, and the fourth raw material comprises 0.15-0.5 g/L of pyridine azo multi-metal complex, 5-10 g/L of nonionic surfactant and 5-10 ml/L of absolute ethyl alcohol.
The buffer solution can be GOOD' S biological buffer solution with the PH ranging from 8.0 to 9.0, such as one or more of N-tris (hydroxymethyl) methyl-3-aminopropanesulfonic acid (TAPS), tris (hydroxymethyl) methylglycine (Tricine), N-bis-hydroxyethylglycine (Bicine), N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid-sodium hydroxide (HEPES-NaOH), piperazine-1, 4-diethylsulfonic acid-sodium hydroxide (PIPES-NaOH) buffer solution; the nonionic surfactant can be one or more of fatty alcohol polyoxyethylene ether and alkylphenol polyoxyethylene ether as protein dissociation agent and solubilizer, such as fatty alcohol polyoxyethylene (7) ether (AEO7), triton X-100, or their mixture. The preservative can be one or a mixture of methyl Chloroisothiazolinone (CMIT) and Methyl Isothiazolinone (MIT), such as Bodha PC-300 bacteriostatic agent, ProClin300 preservative. The pyridine azo multi-metal complex can be 2- (5-bromo-2-pyridylazo) -5- [ N-propyl-N- (3-sulfopropyl) amino ] phenol disodium salt (5-Br-PAPS), 2- (5-Nitro-2-pyridylazo) -5- (N-propyl-N-sulfopropylamino) phenol disodium salt (Nitro-PAPS), 2- (5-bromo-2-pyridylazo) -5- (diethylamino) phenol (5-Br-PADAP), 2- (3, 5-dibromo-2-pyridylazo) -5-dimethylaminophenol (3, 5-diBr-DMPAP).
First embodiment
High-sensitivity serum zinc determination reagent with strong anti-interference capability
A first reagent:
reagent composition
And a second reagent:
reagent composition
The preparation method comprises the following steps:
preparation of reagent I
(1) Adding purified water accounting for about 80% of the total amount required by the preparation of the reagent I into a mixing container, respectively and accurately weighing a whole batch of TAPS, potassium sodium tartrate, titanium iron reagent, pyrrolidine ammonium dithiobenzoate and Boehmeria PC-300 bacteriostatic agent, sequentially adding into the mixing container, and stirring one by one until the bacteriostatic agent is completely dissolved.
(2) And (2) accurately weighing the absolute ethyl alcohol and the triton X-100 in full batch respectively in another mixing container, fully mixing uniformly, adding and dissolving the dimethylglyoxime and the salicylaldoxime in full batch one by one, and fully mixing with the solution prepared in the step (1) after fully stirring and dissolving.
(3) It is tested whether the pH is 8.5. + -. 0.05 or not, otherwise adjusted to this range with 6mol/L hydrochloric acid solution or 10mol/L sodium hydroxide solution.
(4) Quantifying with purified water to a certain amount, stirring, standing for 15min until the solution is balanced, and filtering insoluble substances with 0.8 micrometer aperture nylon filter membrane.
Preparation of reagent two
(1) Purified water accounting for about 80% of the total amount required by the preparation of the reagent II is added into the mixing container, and the whole batch of the TAPS and Boehringer PC-300 bacteriostatic agent is accurately weighed and stirred until the bacteriostatic agent is completely dissolved.
(2) And (2) accurately weighing the anhydrous ethanol and the triton X-100 in full batch respectively in another mixing container, fully mixing uniformly, adding the 5-Br-PAPS in full batch, fully stirring for dissolving, and fully mixing with the solution prepared in the step (1).
(3) It is tested whether the pH is 8.4. + -. 0.05 or not, otherwise adjusted to this range with 6mol/L hydrochloric acid solution or 10mol/L sodium hydroxide solution.
(4) Quantifying with purified water to a certain amount, stirring, standing for 15min until the solution is balanced, and filtering insoluble substances with 0.8 micrometer aperture nylon filter membrane.
Second embodiment
High-sensitivity serum zinc determination reagent with strong anti-interference capability
A first reagent:
reagent composition
And a second reagent:
reagent composition
Preparation method
Preparation of reagent I
(1) Adding purified water accounting for about 80% of the total amount required by the preparation of the reagent I into a mixing container, accurately weighing the whole batch of HEPES, NaOH, potassium sodium tartrate, ferrotitanium reagent, ammonium pyrrolidine dithioaminobenzoate and ProClin300 preservative respectively, sequentially adding into the mixing container, and stirring one by one until the preservative is completely dissolved.
(2) And (2) accurately weighing the anhydrous ethanol and the AEO7 in full batches in another mixing container respectively, fully mixing uniformly, adding and dissolving the dimethylglyoxime and the salicylaldoxime in full batches one by one, and fully mixing with the solution prepared in the step (1) after fully stirring and dissolving.
(3) It is tested whether the pH is 8.0. + -. 0.05 or not, otherwise adjusted to this range with 6mol/L hydrochloric acid solution or 10mol/L sodium hydroxide solution.
(4) Quantifying with purified water to a certain amount, stirring, standing for 15min until the solution is balanced, and filtering insoluble substances with 0.8 micrometer aperture nylon filter membrane.
Preparation of reagent two
(1) Purified water accounting for about 80% of the total amount required by the preparation of the reagent II is added into a mixing container, and a full batch of HEPES, NaOH and ProClin300 preservatives are accurately weighed and stirred until the preservatives are completely dissolved.
(2) And (2) accurately weighing the anhydrous ethanol and the AEO7 in the whole batch respectively in another mixing container, fully and uniformly mixing, adding the Nitro-PAPS in the whole batch, fully stirring and dissolving, and fully mixing with the solution prepared in the step (1).
(3) It is tested whether the pH value is 8.4. + -. 0.05, otherwise it is adjusted to this range with 6mol/L hydrochloric acid solution or 10mol/L sodium hydroxide solution.
(4) Quantifying with purified water to a certain amount, stirring, standing for 15min until the solution is balanced, and filtering insoluble substances with 0.8 micrometer aperture nylon filter membrane.
Third embodiment
A first reagent:
reagent components
And a second reagent:
reagent composition
Preparation method
Preparation of reagent I
(1) Adding purified water accounting for about 80% of the total amount required by the preparation of the reagent I into a mixing container, accurately weighing the Tricine, the potassium sodium tartrate, the ferrotitanium reagent, the ammonium pyrrolidine dithioaminobenzoate and the ProClin300 preservative in full batches respectively, sequentially adding into the mixing container, and stirring one by one until the materials are completely dissolved.
(2) And (2) accurately weighing the anhydrous ethanol, the AEO7 and the triton X-100 in a whole batch respectively in another mixing container, fully mixing uniformly, adding and dissolving the dimethylglyoxime and the salicylaldoxime in the whole batch one by one, and fully mixing with the solution prepared in the step (1) after fully stirring and dissolving.
(3) It was tested whether the pH was 8.7. + -. 0.05, otherwise adjusted to this range with 6mol/L hydrochloric acid solution or 10mol/L sodium hydroxide solution.
(4) Quantifying with purified water to a certain amount, stirring, standing for 15min until the solution is balanced, and filtering insoluble substances with 0.8 micrometer aperture nylon filter membrane.
Preparation of reagent two
(1) Purified water accounting for about 80% of the total amount required by the preparation of the reagent II is added into a mixing container, and the full batch of Tricine and ProClin300 preservatives are accurately weighed and stirred until the preservatives are completely dissolved.
(2) And (2) accurately weighing the anhydrous ethanol and the AEO7 in the whole batch respectively in another mixing container, fully and uniformly mixing, adding the Nitro-PAPS in the whole batch, fully stirring and dissolving, and fully mixing with the solution prepared in the step (1).
(3) It was tested whether the pH was 8.7. + -. 0.05, otherwise adjusted to this range with 6mol/L hydrochloric acid solution or 10mol/L sodium hydroxide solution.
(4) Quantifying with purified water to a certain amount, stirring, standing for 15min until the solution is balanced, and filtering insoluble substances with 0.8 micrometer aperture nylon filter membrane.
The principle is as follows: the pyridine azo complex is reacted with zinc in alkaline solution to produce purple complex with maximum absorption peak at 500-600 nm wavelength. The concentration of the zinc ions in the sample can be calculated by measuring the photometric value of the purple compound generated by the zinc ions with standard concentration and establishing a standard curve of absorbance change and concentration.
In a stability load experiment in which the first reagent and the second reagent are placed at 37 ℃ for 7 days and a storage stability experiment in which the first reagent and the second reagent are stored at 4-8 ℃ for 12 months, the appearance of the reagents has no obvious change, when the detection is carried out according to the detection method disclosed by the invention, the upper limit of the concentration of the zinc ions measured by the reagents can reach 160 mu mol/L, and if a sample contains the following interferents, the detection result is not influenced: bilirubin is less than or equal to 100 mu mol/L, hemoglobin is less than or equal to 1g/L, heparin sodium is less than or equal to 5IU/mL, CU2+≤150μmol/L、Fe3+≤150μmol/L、Ca2+≤10mmol/L、Mg2+Less than or equal to 6 mmol/L. The reagent has no obvious change in appearance, sensitivity, accuracy and linear range, and is in full accordance with the clinical detection requirement.
The reagent can adopt a manual testing method and can also detect a sample through an analysis instrument such as a full-automatic biochemical analyzer, and the like, and the method specifically adopts the following steps:
TABLE 1 Zinc assay reagent parameters table
The calculation formula is as follows:
in order to ensure the reliability of the test result, the reagent detection needs to be calibrated by a zinc standard or standard serum before the sample detection.
Interference experiments: the agents were tested for interference resistance using the additive method. The interferents were selected according to the items and concentrations indicated in table 2, the interferent concentrate was prepared 10 times the amount indicated by the interferent concentration, 1 part of the concentrate was aspirated, 9 parts of the sample serum was added and mixed well, the control sample was replaced with the interferent concentrate with the same volume of physiological saline, each sample was tested 3 times, and the relative deviation of the mean was calculated. And (3) judging: no interference was found if the test results deviate by no more than ± 10% from the blank.
TABLE 2 anti-interference verification of zinc assay reagents
| Interfering species | Zn mean value (mu mol/L) | Relative deviation of |
| Blank control group (non-interfering substance) | 17.9 | 0 |
| Bilirubin (100. mu. mol/L) | 17.5 | -2.23％ |
| Hemoglobin (1g/L) | 18.2 | 1.68％ |
| Heparin sodium (5IU/mL) | 18.0 | 0.56％ |
| Calcium ion (10mmol/L) | 18.3 | 2.23％ |
| Copper ion (150. mu. mol/L) | 18.1 | 1.12％ |
| Iron ion (150. mu. mol/L) | 18.2 | 1.68％ |
| Magnesium ion (6mmol/L) | 18.0 | 0.56％ |
And (3) correlation verification: the reagent is prepared by using the formula of the embodiment 1, the contrast detection is carried out on the reagent and a serum zinc determination kit sold in a certain company approved by the national drug administration in the market, 100 clinical sample test samples are simultaneously detected on a Hitachi 7180 biochemical analyzer, and correlation and matching T test analysis is carried out on two groups of test results. The result shows that as shown in FIG. 2, the linear correlation coefficient R of the two groups of data is greater than 0.9750, and the P of the T test analysis is greater than 0.05, which indicates that the reagent has good correlation with the similar detection reagent in the clinical test result and the result has no obvious difference
The invention has the beneficial effects that:
1) five metal complexing agents of potassium sodium tartrate, dimethylglyoxime, salicylaldoxime, a ferrotitanium reagent and pyrrolidine ammonium dithioaminobenzoate are combined to form an anti-interference agent system of the zinc ion detection reagent, and compared with the traditional formula, the anti-interference capability of the whole zinc ion detection reagent is greatly improved. Wherein the pyrrolidine ammonium dithioamino benzoate and salicylaldoxime mask copper ions, potassium sodium tartrate masks calcium and magnesium ions, a ferrotitanium reagent masks iron ions, the pyrrolidine ammonium dithioamino benzoate masks copper ions, and dimethylglyoxime masks nickel ions; the detection system can ensure that the main endogenous interferent CU in serum2+≤150μmol/L、Fe3+≤150μmol/L、Ca2+≤10mmol/L、Mg2+No more than 6mmol/L has no obvious interference on the test result.
2) The method avoids the problems that the detection sensitivity of the reagent is reduced and the test is inaccurate because the competitive inhibition of a strong metal complexing agent such as a copper reagent and the like and a pyridine-azo multi-element metal complex is generated in the zinc ion detection process.
The high-sensitivity serum zinc determination reagent with strong anti-interference capability comprises a first reagent and a second reagent, wherein the mass ratio of the first reagent to the second reagent is 4:1, the first raw material and the second raw material are weighed according to a ratio and are respectively added into corresponding mixing containers for stirring, dissolving and mixing to obtain a first mixed solution; adjusting the pH value of the first mixed solution to 8.0-9.0, quantifying by using purified water to a configured amount, and then filtering and standing to obtain a first reagent; weighing a third raw material and a fourth raw material according to a ratio, and respectively adding the third raw material and the fourth raw material into corresponding mixing containers to stir, dissolve and mix to obtain a second mixed solution; and adjusting the pH value of the second mixed solution to 8.0-9.0, quantifying by using purified water to a configured amount, and then filtering and standing to obtain the second reagent for improving the using effect of the reagent.
While the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (8)
1. A preparation method of a high-sensitivity serum zinc determination reagent with strong anti-interference capability is characterized by comprising the following steps:
weighing a first raw material and a second raw material according to a ratio, and respectively adding the first raw material and the second raw material into corresponding mixing containers to stir, dissolve and mix to obtain a first mixed solution;
adjusting the pH value of the first mixed solution to 8.0-9.0, quantifying by using purified water to a configured amount, and then filtering and standing to obtain a first reagent;
weighing a third raw material and a fourth raw material according to a ratio, and respectively adding the third raw material and the fourth raw material into corresponding mixing containers to stir, dissolve and mix to obtain a second mixed solution;
and adjusting the pH value of the second mixed solution to 8.0-9.0, quantifying by using purified water to a configured amount, and then filtering and standing to obtain a second reagent.
2. The method for preparing the reagent for measuring serum zinc with strong interference resistance and high sensitivity according to claim 1, wherein the first raw material and the second raw material are weighed according to the mixture ratio and are respectively added into the corresponding mixing containers to be stirred, dissolved and mixed to obtain a first mixed solution, and the method comprises the following steps:
weighing a first raw material according to a ratio, adding the first raw material into a mixing container containing 75-85% of purified water by mass ratio, and stirring until the first raw material is dissolved;
weighing a second raw material according to a ratio, and adding the second raw material into the other mixing container in two batches for stirring and dissolving;
and mixing the first raw material solution and the second raw material solution to obtain a first mixed solution.
3. The method for preparing a reagent for measuring serum zinc with high sensitivity and high interference rejection according to claim 1, wherein the step of adjusting the pH of the first mixed solution to 8.0 to 9.0, quantifying the pH to a predetermined amount with purified water, and then filtering and standing the solution to obtain the first reagent comprises:
judging whether the pH value of the first mixed solution is 8.0-9.0 or not;
if not, adjusting by using a hydrochloric acid solution or a sodium hydroxide solution;
and (4) quantifying the purified water to a configured amount, and then filtering and standing to obtain a first reagent.
4. The method for preparing a reagent for measuring serum zinc with high sensitivity and high interference rejection according to claim 3, wherein the step of obtaining the first reagent by filtering and standing after quantifying the purified water to a predetermined amount comprises:
quantifying to a configured amount by using purified water, uniformly stirring, and standing for 15 minutes or more;
filtering with a nylon filter membrane with the pore size of 0.4-1.0 micron to obtain the first reagent.
5. A high-sensitivity serum zinc determination reagent with strong interference resistance, the preparation method of the high-sensitivity serum zinc determination reagent with strong interference resistance according to any one of claim 1 to claim 4 is applied to the high-sensitivity serum zinc determination reagent with strong interference resistance, and the preparation method is characterized in that,
the high-sensitivity serum zinc determination reagent with strong anti-interference capability comprises a first reagent and a second reagent, wherein the mass ratio of the first reagent to the second reagent is 4: 1;
the first reagent comprises a first raw material and a second raw material, wherein the first raw material comprises 20-200 mmol/L of buffer solution with the pH value of 8.0-9.0, 4-10 g/L of sodium potassium tartrate, 0.05-0.1 g/L of ferrotitanium reagent, 0.05-0.15 g/L of pyrrolidine dithioamino-benzoic acid ammonium and 0.1-0.5 g/L of preservative, and the second raw material comprises 0.1-0.5 g/L of dimethylglyoxime, 0.1-0.5 g/L of salicylaldoxime, 5-10 ml/L of absolute ethyl alcohol and 1-5 g/L of nonionic surfactant;
the second reagent comprises a third raw material and a fourth raw material, wherein the third raw material comprises 20-200 mmol/L of buffer solution with the pH value of 8.0-9.0 and 0.1-0.5 g/L of preservative, and the fourth raw material comprises 0.15-0.5 g/L of pyridine azo multi-metal complex, 5-10 g/L of nonionic surfactant and 5-10 ml/L of absolute ethyl alcohol.
6. The reagent for measuring serum zinc with high sensitivity and high interference rejection of claim 5,
the buffer solution comprises one or more of N-tris (hydroxymethyl) methyl-3-aminopropane sulfonic acid, tris (hydroxymethyl) methylglycine, N-di-hydroxyethylglycine, N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid-sodium hydroxide, piperazine-1, 4-diethylsulfonic acid-sodium hydroxide.
7. The reagent for measuring serum zinc with high sensitivity and high interference rejection of claim 5,
the nonionic surfactant comprises one or two of fatty alcohol-polyoxyethylene ether and alkylphenol polyoxyethylene.
8. The reagent for measuring serum zinc with high sensitivity and high interference rejection of claim 5,
the preservative can be one or two of methyl chloro isothiazolinone and methyl isothiazolinone.
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