A semi-quantitative urine zinc test strip, its preparation method and application

Abstract

The present invention relates to a semi-quantitative urine zinc test strip, a preparation method and application thereof, and belongs to the field of analysis and detection. A semi-quantitative urine zinc test strip is provided, which includes a zinc test strip; the zinc test strip is obtained by soaking and drying the test strip in a substrate impregnation liquid, and the substrate impregnation liquid includes: 0.01% by mass of 2-(5-bromo-2-pyridyl azo)-5-[N-propyl-N-(3-sulfopropyl) amino] phenol disodium salt, 0.5-1M N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid, 0.1%-1% by mass of polyethylene glycol monooctylphenyl ether, 1%-5% by mass of a masking agent, and the solvent is deionized water; wherein the masking agent includes sodium hexametaphosphate and N'-(sodium toluenesulfonate) thiourea. This urine zinc semi-quantitative test strip can realize visual distinction and interpretation of low concentration zinc ions, and has strong anti-interference ability and high accuracy, making it suitable for home use.

Description

Technical Field

[0001] The invention relates to a urine zinc semi-quantitative detection test strip, a preparation method and application thereof, and belongs to the technical field of analysis and detection. Background Art

[0002] Urine zinc testing can detect the dynamic changes of human zinc at an early stage and more accurately reflect the corresponding zinc metabolism level in the body, which is of great significance.

[0003] The invention patent with publication number CN1766585A measures the reagent and rapid detection method of the zinc content in urine. However, its detection method comprises the following steps: adding 0.5ml urine to a test tube, adding 0.2ml strong acid, mixing, then adding 0.05ml sodium thiocyanate solution, mixing, and then adding 0.05ml crystal violet solution. It can be seen that the detection method of the patent is a liquid colorimetric method, colorimetric in liquid, color development is not obvious, and the interpretation is not accurate enough; it is also necessary to use a test tube, and urine, strong acid (such as sulfuric acid or hydrochloric acid), sodium thiocyanate solution, crystal violet solution are quantitatively, mixed and other operations, not only the steps are cumbersome, but also the use of corrosive strong acid, the risk factor is high, the operator's operation standardization requirements are high, and it is not suitable for home use.

[0004] The invention patent with publication number CN110672598A discloses a dry chemical reagent strip for detecting calcium, magnesium and zinc in breast milk, wherein the formula of the zinc ion detection reagent is 15-40g of 2-(N-morpholine)ethanesulfonic acid, 0.05-3g of 4-(2-pyridyl azo)resorcinol sodium salt, 1-10ml of ethanol, 0.3-24g of sucrose, 2-10ml of glycerol, 0.1-4g of triethanolamine, 0.5-3g of sodium citrate, 0.05-0.1g of sodium hydroxide, and 200-800ml of purified water or distilled water; visual comparison with a standard colorimetric card is performed, and the interpretation results are divided into the following three levels: when the zinc concentration is <400ug / dl, it is low; when the zinc concentration is 400-800ug / dl, it is normal; when the zinc concentration is >800ug / dl, it is high. As shown in Comparative Example 1 of the present application, the method of the patent is not suitable for visually distinguishing and judging the zinc ion concentration (0 mg / L, 0.15 mg / L, 0.5 mg / L, 1 mg / L) in urine.

[0005] The invention patent with publication number CN109580600A discloses a rapid detection reagent strip for measuring biochemical zinc ions in seminal plasma. The reagent strip is firstly coated with a coating liquid including a buffer, a color developer, a film former, a surfactant, a masking agent and a preservative on a transparent base film, and a color developing layer is formed after drying; and then a polymer film is pasted on the color developing layer to prepare a reagent strip. Its embodiment specifically discloses a detection method using 4-(2-pyridyl azo)-resorcinol sodium salt as a color developer, but the establishment of its standard curve still needs to analyze the absorbance with the aid of a chemical analyzer, and the zinc concentration in seminal plasma is relatively high, which can reach 30 mg / L. As shown in Comparative Example 2 of the present application, the method of the patent is not suitable for visually distinguishing and judging the zinc ion concentration (0 mg / L, 0.15 mg / L, 0.5 mg / L, 1 mg / L) in urine.

[0006] A zinc ion detection test paper and its preparation method and application are disclosed in the invention patent with publication number CN104502342A, and the impregnation solution formula is: 0.05g diphenylcarbazone, 0.05g hydroxylamine hydrochloride, 100mL ethanol, 0.2g sodium thiosulfate, 100mL distilled water. Its detection method comprises the following steps: after the zinc ion detection test paper is immersed in the sample solution, it is taken out and then the color developing auxiliary agent is added dropwise to develop color. It can be seen that its detection step is more complicated. And the patent does not provide any experimental data to confirm its technical effect, particularly the corresponding relationship between zinc ion concentration and color. As shown in the comparative example 3 of the present application, the method of the patent is not suitable for visual distinction and judgment of zinc ion concentration (0mg / L, 0.15mg / L, 0.5mg / L, 1mg / L) in urine.

[0007] The invention patent with publication number CN1584552A discloses a seminal plasma zinc detection reagent, which determines the zinc ion content in seminal plasma through the synergistic effect of thiourea or thiourea derivatives, fluoride, and the color developer 5-Br-PAPS. An enzyme marker is required for colorimetric analysis, which has high requirements on the operator's operational standardization and is not suitable for home use. In addition, the zinc concentration in seminal plasma is relatively high, reaching 30 mg / L. As shown in Comparative Example 4 of the present application, the method of the patent is also not suitable for visual distinction and judgment of zinc ion concentration in urine (0 mg / L, 0.15 mg / L, 0.5 mg / L, 1 mg / L).

[0008] It can be seen that there is an urgent need to develop a semi-quantitative urine zinc test strip that can visually distinguish and interpret low-concentration zinc ions, and has strong anti-interference ability and high accuracy. Summary of the invention

[0009] [Technical issues]

[0010] Provided is a urine zinc semi-quantitative detection test strip, which can realize visual distinction and interpretation of low-concentration zinc ions, has strong anti-interference ability, high accuracy, and is suitable for home use.

[0011] [Technical solution]

[0012] On the one hand, a semi-quantitative urine zinc test strip is provided; the semi-quantitative urine zinc test strip comprises a zinc test strip; the zinc test strip is obtained by soaking a substrate in a substrate impregnation liquid and drying it, the substrate impregnation liquid comprises: 0.01% by mass of 2-(5-bromo-2-pyridyl azo)-5-[N-propyl-N-(3-sulfopropyl) amino] phenol disodium salt, 0.5-1M N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid, 0.1%-1% by mass of polyethylene glycol monooctylphenyl ether, 1%-5% by mass of a masking agent, and the solvent is deionized water; wherein the masking agent comprises sodium hexametaphosphate and N′-(sodium p-toluenesulfonate) thiourea.

[0013] In some embodiments, the substrate impregnation solution further includes: a pigment with a mass fraction of 0.005-0.05%; the pigment includes: one of brilliant blue, erythrosine, and green pigment.

[0014] In some embodiments, the substrate impregnation liquid includes: 0.01% by mass of 2-(5-bromo-2-pyridyl azo)-5-[N-propyl-N-(3-sulfopropyl) amino] phenol disodium salt, 0.5M N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid, 0.1% by mass of polyethylene glycol monooctylphenyl ether, 1% by mass of sodium hexametaphosphate, 1% by mass of N′-(sodium p-toluenesulfonate) thiourea, 0.005% by mass of brilliant blue, and the solvent is deionized water.

[0015] In some embodiments, the semi-quantitative urine zinc test strip also includes a shell; the shell is configured to accommodate and fix the zinc test strip; the shell is provided with a dripping hole; the dripping hole is located directly above the zinc test strip, and is used for dripping the urine sample onto the zinc test strip and observing the color development result.

[0016] In some embodiments, the dripping hole is circular in shape.

[0017] On the other hand, a method for preparing the aforementioned urine zinc semi-quantitative test strip is provided, which comprises the following steps:

[0018] (1) Preparation of zinc detection test strips: soak the test paper in the substrate impregnation solution for 5 to 10 minutes, and then dry it at 55° C. for 0.5 to 2 hours; take it out and cut the test paper into test strips to obtain zinc detection test strips;

[0019] (2) Assembling the zinc detection test strip and the shell to obtain a urine zinc semi-quantitative detection test strip.

[0020] In some embodiments, the method for preparing a semi-quantitative urine zinc test strip comprises the following steps:

[0021] The urine to be tested is dripped onto the zinc detection test paper strip, and after complete color development, it is compared with the standard colorimetric card to visually determine which of the following ranges the zinc content in the urine is in: zinc content is less than or equal to 0.15 mg / L, zinc content is greater than 0.15 mg / L and less than 1 mg / L, or zinc content is greater than or equal to 1 mg / L; wherein, the standard colorimetric card is obtained by dripping a zinc ion standard solution of known concentration onto the zinc detection test paper strip, collecting the color development result when the color development is complete, and is prepared according to the corresponding relationship between the concentration of the zinc ion standard solution and the color development result.

[0022] In some embodiments, when the aforementioned urine zinc semi-quantitative test strip is used, the visual judgment method is as follows:

[0023] When the zinc test strip is brown-yellow, the zinc content in the urine is less than or equal to 0.15 mg / L, indicating severe zinc deficiency; when the zinc test strip is crimson-purple, the zinc content in the urine is greater than 0.15 mg / L and less than 1 mg / L, indicating normal zinc; when the zinc test strip is purple, the zinc content in the urine is greater than or equal to 1 mg / L, indicating that the zinc level is too high.

[0024] [Beneficial Effects]

[0025] (1) In view of the technical difficulty that the existing zinc test strips cannot visually distinguish the key concentrations of 0 mg / L, 0.15 mg / L, 0.5 mg / L and 1 mg / L for urine zinc concentration detection without the aid of additional equipment, the present invention provides a urine zinc semi-quantitative detection test strip, which includes a zinc detection test strip; the zinc detection test strip is obtained by soaking the test strip in a substrate impregnation liquid and drying it, the substrate impregnation liquid includes: 0.01% by mass of 2-(5-bromo-2-pyridyl azo)-5-[N-propyl-N-(3-sulfopropyl) amino] phenol disodium salt, 0.5-1M N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid, 0.1%-1% by mass of polyethylene glycol monooctylphenyl ether, 1%-5% by mass of a masking agent, and the solvent is deionized water; wherein the masking agent includes sodium hexametaphosphate and N′-(sodium p-toluenesulfonate) thiourea. The urine zinc semi-quantitative detection test strip of the present invention can realize visual distinction and interpretation of low-concentration zinc ions, and has strong anti-interference ability and high accuracy. When detecting urine samples, the urine zinc semi-quantitative detection test strip of the present invention only needs to be dripped and then visually compared in color, which is simple to operate and is particularly suitable for home use.

[0026] (2) The present invention has found that the formulation of the substrate impregnation solution of the present invention is crucial for visually distinguishing the key concentrations of 0 mg / L, 0.15 mg / L, 0.5 mg / L, and 1 mg / L for urine zinc concentration detection. Based on the formulation of the present invention, changing the type or concentration range of the color developer, changing the type of masking agent, and changing the type of pigment will have a significant impact on the color development effect.

[0027] (3) The present invention determines the type of developer that can significantly change color with low-concentration zinc by screening, and optimizes the concentration of the developer. In view of the problem that calcium, magnesium, copper, iron and other ions with higher concentrations in urine will affect the detection of zinc, the present invention screens the masking agent, which can specifically bind to the interfering substances in the sample so that it does not affect the detection of zinc, and does not need to pre-treat the sample again, effectively shielding the interfering substances. In view of the problem that the color displayed after the urine zinc is combined with the developer is light and not easy to judge, the present invention adjusts the color of the test paper by adding a coloring pigment, and enhances the color change without affecting the determination, thereby improving the visual easiness of judgment and the accuracy of the results. The developer of the present invention uses 2-(5-bromo-2-pyridyl azo)-5-[N-propyl-N-(3-sulfopropyl) amino] phenol disodium salt with different positive and negative colors, and cooperates with the pigment to make the developer clear after combining with zinc, which is convenient for distinguishing urine zinc samples of different concentrations. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 A physical picture of the urine zinc semi-quantitative detection test strip prepared by the present invention;

[0029] Figure 2 The actual color development pictures of the test strips prepared in Example 5-1 and Comparative Examples 1 to 4 of the present invention. DETAILED DESCRIPTION

[0030] Example 1: A semi-quantitative urine zinc test strip and its preparation method

[0031] The urine zinc semi-quantitative test strip includes an upper shell, a zinc test strip and a lower shell from top to bottom. Figure 1 The lower shell is configured as a supporting structure for the zinc test strip; the upper shell and the lower shell are adapted in shape, and the zinc test strip is fixed between the upper shell and the lower shell; for example, a slot is provided on the lower shell to fix the zinc test strip. The upper shell is provided with a sample drop hole, which is located directly above the zinc test strip and is used for dropping urine samples and observing results; the shape of the sample drop hole includes but is not limited to a circle; the material of the upper shell and / or the lower shell includes but is not limited to polystyrene.

[0032] The preparation process of the zinc test strip comprises the following steps:

[0033] The substrate (white cotton fiber test paper) was soaked in the substrate impregnation solution for 8 minutes, and then placed in a constant temperature drying oven at 55°C for 30 minutes; taken out, and the test paper was cut into 10mm*10mm test strips using a chopper to obtain zinc detection test strips; wherein the composition of the substrate impregnation solution is: 0.01% by mass of 2-(5-bromo-2-pyridyl azo)-5-[N-propyl-N-(3-sulfopropyl) amino] phenol disodium salt (color developer), 0.5M N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid (buffer), 0.1% by mass of polyethylene glycol monooctylphenyl ether (dispersant), 1% by mass of sodium hexametaphosphate (masking agent 1), 1% by mass of N′-(sodium p-toluenesulfonate) thiourea (masking agent 2), and the remainder is deionized water, as shown in Table 1.

[0034] The zinc detection test strip, the upper shell and the lower shell are assembled to obtain a high-sensitivity urine zinc semi-quantitative detection test strip.

[0035] Example 2: Effect of Urine Zinc Semi-Quantitative Test Strips Using Different Color Developing Agents on the Detection of Urine Zinc Concentration

[0036] The method for preparing the semi-quantitative urine zinc test strip is similar to that of Example 1, except that the type of the color developer in the substrate impregnation solution is changed to one of 4-(2-pyridylazo)resorcinol sodium salt, 1-(2-pyridylazo)-2-naphthol, and dithizone, and the other parameters and conditions are the same as those of Example 1. The comparison of the substrate impregnation solutions of Example 1 and Example 2 is shown in Table 1.

[0037] Zinc ion standard solutions of different concentrations (0 mg / L, 0.15 mg / L, 0.5 mg / L, 1 mg / L) were respectively added dropwise to the zinc detection test strips of the urine zinc semi-quantitative detection test strips of Example 1 and Example 2 through the droplet hole, and the color change was observed after 1 minute.

[0038] Table 1

[0039]

[0040] The results are shown in Table 2. Only the urine zinc semi-quantitative test strip using 2-(5-bromo-2-pyridyl azo)-5-[N-propyl-N-(3-sulfopropyl) amino] phenol disodium salt as the color developer in Example 1 can successfully distinguish 0 mg / L, 0.15 mg / L, 0.5 mg / L, and 1 mg / L zinc ion samples, while other color developers (4-(2-pyridyl azo) resorcinol sodium salt, dithizone, 1-(2-pyridyl azo) resorcinol sodium salt, It can be seen that the use of 2-(5-bromo-2-pyridyl azo)-5-[N-propyl-N-(3-sulfopropyl) amino] phenol disodium salt as a color developer in the present invention is one of the key technical means to successfully distinguish 0 mg / L, 0.15 mg / L, 0.5 mg / L, and 1 mg / L zinc ion samples.

[0041] Table 2

[0042] 0mg / L 0.15 mg / L 0.5mg / L 1mg / L Example 1 yellow orange color red Magenta Example 2-1 orange color orange color orange color orange color Example 2-2 green green green green Example 2-3 Orange Red Orange Red Orange Red Orange Red

[0043] Example 3: Effect of using different concentrations of 2-(5-bromo-2-pyridyl azo)-5-[N-propyl-N-(3-sulfopropyl)amino]phenol disodium salt urine zinc semi-quantitative test strips on the detection of urine zinc concentration

[0044] The method for preparing the semi-quantitative urine zinc test strip is based on Example 1, except that the concentration of the color developer 2-(5-bromo-2-pyridyl azo)-5-[N-propyl-N-(3-sulfopropyl) amino] phenol disodium salt in the substrate impregnation solution is changed to 0.005wt%, 0.03wt%, 0.05wt%, and 0.1wt%, respectively, and the other parameters and conditions are the same as those in Example 1. The comparison of the substrate impregnation solutions of Example 1 and Example 3 is shown in Table 3.

[0045] Zinc ion standard solutions of different concentrations (0 mg / L, 0.15 mg / L, 0.5 mg / L, 1 mg / L) were respectively aspirated and added dropwise to the zinc detection test strips of the urine zinc semi-quantitative detection test strips of Example 1 and Example 3 through the droplet hole, and the color change was observed after 1 minute.

[0046] Table 3

[0047]

[0048] The results are shown in Table 4. Only the urine zinc semi-quantitative test strip of 2-(5-bromo-2-pyridyl azo)-5-[N-propyl-N-(3-sulfopropyl) amino] phenol disodium salt with a concentration of 0.01wt% in Example 1 can successfully distinguish 0 mg / L, 0.15 mg / L, 0.5 mg / L, and 1 mg / L zinc ion samples, while the urine zinc semi-quantitative test strip of 2-(5-bromo-2-pyridyl azo)-5-[N-propyl-N-(3-sulfopropyl) amino] phenol disodium salt with a concentration of 0.005wt%, 0.03wt%, 0.05wt%, and 0.1wt% in Example 3 cannot successfully distinguish 0 mg / L, 0.15 mg / L, 0.5 mg / L, and 1 It can be seen that the concentration of 2-(5-bromo-2-pyridyl azo)-5-[N-propyl-N-(3-sulfopropyl) amino] set to 0.01wt% in the present invention is one of the key technical means to successfully distinguish 0 mg / L, 0.15 mg / L, 0.5 mg / L, and 1 mg / L zinc ion samples.

[0049] Table 4

[0050] 0 mg / L 0.15 mg / L 0.5 mg / L 1 mg / L Example 1 yellow orange color red Magenta Example 3-1 yellow red Magenta Magenta Example 3-2 yellow orange color red red Example 3-3 orange color orange color orange color red Embodiment 3-4 orange color orange color orange color orange color

[0051] Example 4: Effect of using different masking agents on the detection of urine zinc concentration using semi-quantitative urine zinc test strips

[0052] The method for preparing a semi-quantitative urine zinc test strip is based on Example 1, except that the type of masking agent in the substrate impregnation solution is changed to select one or more of sodium hexametaphosphate, sodium tartrate, sodium fluoride, and N'-(sodium p-toluenesulfonate) thiourea, or the masking agent is omitted (blank control), and the other parameters and conditions are the same as those in Example 1; a comparison of the substrate impregnation solutions of Example 1 and Example 4 is shown in Table 5.

[0053] Prepare sample 1: Add single metal ions as interferents to zinc ion standard solutions of different concentrations (0 mg / L, 0.15 mg / L, 0.5 mg / L, 1 mg / L), including: 1 mg / L copper ion, 0.5 mg / L iron ion, 200 mg / L calcium ion and 160 mg / L magnesium ion.

[0054] Prepare sample 2: Add mixed metal ions as interferents to zinc ion standard solutions of different concentrations (0 mg / L, 0.15 mg / L, 0.5 mg / L, 1 mg / L), including: 1 mg / L copper ion, 0.5 mg / L iron ion, 200 mg / L calcium ion and 160 mg / L magnesium ion.

[0055] The zinc ion solutions of different concentrations (0 mg / L, 0.15 mg / L, 0.5 mg / L, 1 mg / L) containing metal ion interfering substances were respectively aspirated and dropped onto the zinc detection test strips of the semi-quantitative urine zinc detection test strips of Example 1 and Example 4 through the droplet hole, and the color change was observed after 1 minute.

[0056] Table 5

[0057]

[0058] Results: Sodium hexametaphosphate and N'-(sodium p-benzenesulfonate)thiourea could mask the interfering substances in urine.

[0059] The results are shown in Table 6. The urine zinc semi-quantitative detection test strip using sodium hexametaphosphate as a masking agent in Example 4-1 can successfully distinguish between zinc ion samples of 0 mg / L, 0.15 mg / L, 0.5 mg / L, and 1 mg / L respectively containing interferences of 0.5 mg / L iron ions, 200 mg / L calcium ions, and 160 mg / L magnesium ions; the urine zinc semi-quantitative detection test strip using N'-(sodium p-benzenesulfonate) thiourea as a masking agent in Example 4-4 can successfully distinguish between zinc ion samples of 0 mg / L, 0.15 mg / L, 0.5 mg / L, and 1 mg / L containing interferences of 1 mg / L copper ions; while the urine zinc semi-quantitative detection test strip using sodium tartrate as a masking agent in Example 4-2 and sodium fluoride as a masking agent in Example 4-3 cannot simultaneously distinguish between zinc ion samples of 0.5 mg / L and 1 mg / L containing interferences of 0.5 mg / L iron ions, 200 mg / L calcium ions, and 160 mg / L magnesium ions. It can be seen that the use of ammonium hexametaphosphate and N'-(sodium p-toluenesulfonate) thiourea as a masking agent in the present invention is one of the key technical means for successfully distinguishing 0mg / L, 0.15mg / L, 0.5mg / L, 1mg / L zinc ion samples containing 1mg / L copper ions, 0.5mg / L iron ions, 200mg / L calcium ions and 160mg / L magnesium ions. In addition, the experimental results of mixed ion interference samples confirmed that the technical solution of Example 1 has excellent resistance to interference from mixed metal ions (1mg / L copper ions, 0.5mg / L iron ions, 200mg / L calcium ions and 160mg / L magnesium ions).

[0060] Table 6

[0061]

[0062]

[0063] Example 5: Effect of using different pigments of urine zinc semi-quantitative test strips on the detection of urine zinc concentration

[0064] The preparation method of the semi-quantitative urine zinc test strip is based on Example 1, with the only difference being that different pigments are added to the substrate impregnation solution, and one of brilliant blue, erythrosine, and green pigment is selected, and the concentration of the pigment is 0.005wt%. Other parameters and conditions are the same as those in Example 1; a comparison of the substrate impregnation solutions of Example 1 and Example 5 is shown in Table 7.

[0065] Zinc ion standard solutions of different concentrations (0 mg / L, 0.15 mg / L, 0.5 mg / L, 1 mg / L) were respectively aspirated and added dropwise to the zinc detection test strips of the urine zinc semi-quantitative detection test strips of Example 1 and Example 5 through the droplet hole, and the color change was observed after 1 minute.

[0066] Table 7

[0067]

[0068] The results are shown in Table 8 and Figure 2 As shown, Example 1, Example 5-1, Example 5-2, and Example 5-3 can all successfully distinguish zinc ion samples of 0 mg / L, 0.15 mg / L, 0.5 mg / L, and 1 mg / L. However, the color of the test paper of Example 1 after color change is lighter. Comparing the color development effects of different pigments and no pigment added, it can be seen that after adding the pigment Brilliant Blue to Example 5-1, zinc ion samples of 0 mg / L, 0.15 mg / L, 0.5 mg / L, and 1 mg / L can be more clearly distinguished.

[0069] Table 8

[0070] 0mg / L 0.15 mg / L 0.5mg / L 1mg / L Example 1 yellow orange color red Magenta Example 5-1 Yellow Green Brown Purple Purple Example 5-2 orange color Dark Orange red Rose red Example 5-3 Grass Green green tan Tan

[0071] Example 6: Real urine sample test

[0072] This embodiment uses the urine zinc semi-quantitative test strip of embodiment 5-1 as an example to test the real urine sample. Take a human urine sample and drip it onto the zinc test strip of the urine zinc semi-quantitative test strip of embodiment 5-1, and observe the color change of the zinc test strip after 1 minute. After a lot of research, it is found that the method for judging the urine zinc content of the urine zinc semi-quantitative test strip of embodiment 5-1 is as follows: when the zinc test strip is brownish yellow, the zinc content in the urine is less than or equal to 0.15 mg / L, indicating severe zinc deficiency; when the zinc test strip is purple, the zinc content in the urine is greater than 0.15 mg / L and less than 1 mg / L, indicating normal zinc; when the zinc test strip is purple, the zinc content in the urine is greater than or equal to 1 mg / L, indicating that zinc is too high. The evaluation standard of urine zinc is set according to the reference Li Qing, Wang Keling. Application evaluation of urine zinc colorimetric detection reagent [J]. China Medical Herald, 2011, 8 (7): 81-82.

[0073] The results obtained according to the above method were compared with the quantitative results of zinc content in urine by PAN colorimetry, and the results are shown in Table 9.

[0074] Table 9

[0075]

[0076] As shown in Table 9, the determination results of zinc ions in 40 urine samples detected by the urine zinc semi-quantitative test strip of the present invention are consistent with the quantitative detection results of the PAN method by up to 100%, which proves that the method of the present invention has high accuracy. Compared with the PAN method, which requires instrument matching for detection and is inconvenient to operate, the urine zinc semi-quantitative test strip of the present invention can complete the detection without the aid of additional instruments and equipment, and is simple to operate and has good practicality.

[0077] Comparative Example 1

[0078] The zinc test paper was prepared according to the preparation method of the test paper in patent CN 110672598A, wherein the reaction layer used whatman filter paper as a carrier, the pore size was 5um, and the thickness was 50um. A whatman filter paper with a length and width of 20*20cm was immersed in the zinc test paper reagent, and immediately taken out after being immersed for 2 minutes and placed in an oven at 60°C for drying. After 90 minutes, it was taken out and cut into 5mm*5mm square pieces with a spacing of 2mm, aligned and bonded together by double-sided tape, and placed in an aluminum foil bag for sealing.

[0079] Among them, the formula of the zinc test paper reagent is 15-40g of 2-(N-morpholine)ethanesulfonic acid, 0.05-3g of 4-(2-pyridyl azo)resorcinol sodium salt, 1-10ml of ethanol, 0.3-24g of sucrose, 2-10ml of glycerol, 0.1-4g of triethanolamine, 0.5-3g of sodium citrate, 0.05-0.1g of sodium hydroxide, and 200-800mL of purified water or distilled water. This comparative example 1 is an experiment based on the scheme recorded in Example 1 of CN 110672598A.

[0080] Comparative Example 2

[0081] A zinc test strip was prepared according to the preparation method of the test strip in Example 2 of patent CN 109580600A. Solution A was first evenly coated on a plastic base film and dried for 3 hours to form a color development layer. Solution B was then evenly coated on the color development layer. Solution B was dried for 15 minutes to form a shielding film. A nitrocellulose membrane was laid on the nitrocellulose membrane to form a polymer film. The film was dried in a low-humidity environment. Finally, a hydrophilic nylon membrane was laid on the nitrocellulose membrane and cut into a specific size to obtain a reagent strip.

[0082] Wherein, the formula of solution A is: 5g 4-(2-pyridyl azo)-resorcinol sodium salt, 100g gelatin, 12g polyethylene glycol, 13g sodium citrate, 8g sodium azide, Tris buffer (pH=9.0) is fixed to 1000mL;

[0083] The formula of solution B is as follows: 90 g gelatin, Tris buffer (pH=9.0) is fixed to 1000 mL.

[0084] Comparative Example 3

[0085] The zinc test paper is prepared according to the preparation method of the test paper in Example 1 of patent CN104502342A. The color test paper is prepared by soaking a quantitative filter paper in an immersion solution, taking it out and drying it; when testing, the zinc test paper needs to be immersed in the sample solution for 1 to 2 seconds, then taken out and 1 drop of color development aid (0.5M acetic acid and 0.5M sodium acetate are mixed in a ratio of 1:1) is added;

[0086] The impregnation solution formula is: 0.05g diphenylcarbazone, 0.05g hydroxylamine hydrochloride, 100mL ethanol, 0.2g sodium thiosulfate, and 100mL distilled water.

[0087] Comparative Example 4

[0088] Referring to the method for preparing the semi-quantitative urine zinc test strip of Example 1, the only difference is that the formula of the substrate impregnation solution is adjusted according to Example 3 of CN1584552A, that is, color developing solution A: phosphate buffer solution with a thiourea content of 2%, a sodium fluoride content of 0.2%, a sodium pyrophosphate content of 0.05%, and a 5-Br-PAPS content of 0.15%; color developing solution B: an anhydrous ethanol solution with a diacetyl oxime content of 0.5%; color developing solution A and color developing solution B are mixed in a volume ratio of 15:4 to obtain a substrate impregnation solution.

[0089] The zinc semi-quantitative test strips of Comparative Examples 1 to 4 were used to test the zinc ion standard solutions of different concentrations (0 mg / L, 0.15 mg / L, 0.5 mg / L, 1 mg / L). After the color development was complete, the color development was recorded. The results are shown in Tables 10 and Figure 2 shown.

[0090] Table 10

[0091]

[0092] From Table 10 and Figure 2It can be seen that the zinc test strips of Comparative Examples 1 to 4 are unable to distinguish between 0 mg / L, 0.15 mg / L, 0.5 mg / L, and 1 mg / L zinc ion standard solutions, and are unable to distinguish whether the zinc concentration in urine is in the interval of less than or equal to 0.15 mg / L, greater than 0.15 mg / L and less than 1 mg / L, and greater than or equal to 1 mg / L, and are thus unable to judge whether the zinc concentration in urine is low, normal, or high.

[0093] The embodiments provided above are not intended to limit the scope of the present invention, and the steps described are not intended to limit the execution order thereof. Those skilled in the art may make obvious improvements to the present invention in combination with existing common knowledge, which also fall within the scope of protection defined by the claims of the present invention.

Claims

1. A semi-quantitative urine zinc test strip, characterized in that: The urine zinc semi-quantitative test strips include zinc test strips; The zinc detection test paper strip is obtained by soaking a substrate in a substrate impregnation liquid and drying the substrate, wherein the substrate impregnation liquid comprises: 0.01% by mass of 2-(5-bromo-2-pyridyl azo)-5-[N-propyl-N-(3-sulfopropyl) amino] phenol disodium salt, 0.5-1 M N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid, 0.1%-1% by mass of polyethylene glycol monooctylphenyl ether, 1%-5% by mass of a masking agent, and the solvent is deionized water; wherein sodium hexametaphosphate and N'-(sodium p-toluenesulfonate) thiourea are masking agents.

2. The urine zinc semi-quantitative test strip according to claim 1, characterized in that: The substrate impregnation solution also includes: a pigment with a mass fraction of 0.005-0.05%; the pigment includes: one of brilliant blue, erythrosine, and green pigment.

3. The urine zinc semi-quantitative test strip according to claim 1, characterized in that: The substrate impregnation liquid comprises: 0.01% by mass of 2-(5-bromo-2-pyridyl azo)-5-[N-propyl-N-(3-sulfopropyl) amino] phenol disodium salt, 0.5 M N-2-hydroxyethylpiperazine-N´-2-ethanesulfonic acid, 0.1% by mass of polyethylene glycol monooctylphenyl ether, 1% by mass of sodium hexametaphosphate, 1% by mass of N'-(sodium p-toluenesulfonate) thiourea, 0.005% by mass of brilliant blue, and the solvent is deionized water.

4. The urine zinc semi-quantitative test strip according to claim 1, characterized in that: The semi-quantitative urine zinc test strip also includes a shell; the shell is configured to accommodate and fix the zinc test strip; the shell is provided with a sample dropping hole; the sample dropping hole is located directly above the zinc test strip and is used for dropping a urine sample onto the zinc test strip and observing the color development result.

5. The urine zinc semi-quantitative test strip according to claim 4, characterized in that: The shape of the sample dropping hole is circular.

6. The method for preparing the urine zinc semi-quantitative test strip according to any one of claims 1 to 5, characterized in that: The steps include: (1) Preparation of zinc detection test strips: soak the test paper in the substrate impregnation solution for 5-10 min, then dry it at 55 °C for 0.5-2 h; take it out and cut the test paper into test strips to obtain zinc detection test strips; (2) Assembling the zinc detection test strip and the shell to obtain a urine zinc semi-quantitative detection test strip.

7. Use of the urine zinc semi-quantitative detection test strip according to any one of claims 1 to 5 in the urine zinc semi-quantitative detection, characterized in that: The steps include: The urine to be tested is dripped onto the zinc test paper strip. After complete color development, it is compared with the standard colorimetric card to visually determine whether the zinc content in the urine is within the following ranges: zinc content is less than or equal to 0.15 mg / L, zinc content is greater than 0.15 mg / L and less than 1 mg / L, or zinc content is greater than or equal to 1 mg / L.

8. The use according to claim 7, characterized in that: When the urine zinc semi-quantitative test strip according to claim 3 is used, the visual judgment method is as follows: When the zinc test strip is brown-yellow, the zinc content in the urine is less than or equal to 0.15 mg / L, indicating severe zinc deficiency; when the zinc test strip is crimson-purple, the zinc content in the urine is greater than 0.15 mg / L and less than 1 mg / L, indicating normal zinc; when the zinc test strip is purple, the zinc content in the urine is greater than or equal to 1 mg / L, indicating that the zinc level is too high.

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