Dry urine spot acquisition card and application thereof

Abstract

The invention provides a dry urine spot collection card and application thereof, the dry urine spot collection card comprises a first outer surface area, a sample collection area and a second outer surface area which are sequentially connected along the transverse direction, and the sample collection area is vertically connected with an information collection area. The acquisition card provided by the invention can effectively protect the sample, improve the stability of the sample to be detected and improve the detection accuracy.

Description

Technical Field

[0001] The present invention belongs to the technical field of medical sampling devices, and particularly relates to a dry urine spot collection card and its application, and more particularly to a dry urine spot collection card with high sample stability and its application. Background Art

[0002] Catecholamine substances in urine mainly include dopamine (DA), epinephrine (E), norepinephrine (NE), metanephrine (MN), normetanephrine (NMN), 3-methoxytyramine (3-MT), homovanillic acid (HVA) and vanillylmandelic acid (VMA). The contents of these several compounds are related to the clinical diagnostic criteria for pheochromocytoma. Currently, 24-hour urine is mainly used clinically to detect this sample. Since the sample is only stable in an acidic environment, concentrated hydrochloric acid needs to be added to the urine in advance to maintain urine acidification, so as to ensure the stability of these 8 compounds. This has thus caused the sample collection to be cumbersome, and at the same time, the addition of concentrated hydrochloric acid has also made the sample preservation and protection have certain difficulties and risks.

[0003] CN214621846U discloses a dry blood spot and dry urine spot collection card. The collection card includes a protective layer, a sample collection area and a collection information area connected in sequence along the horizontal direction. The protective layer can be folded along the connection line between the protective layer and the sample collection area to cover the sample collection area. The sample collection area includes two separated sampling areas. The protective layer provided on the collection card of this utility model can effectively protect the dry blood spots and dry urine spots in the sample collection area, protect trace substance molecules with detection significance, and improve the accuracy of qualitative and quantitative analysis results. This collection card is convenient to use, has no pollution to the environment, and has a low cost. It can be used for the situation where urine and blood samples need to be collected and preserved for the detection of trace substances in dry urine spots and dry blood spots.

[0004] CN217520789U discloses a urine specimen collection card. The collection card is set in three groups. The collection card includes a double-layer hard paper sheet and also includes a urine filter paper sheet connected to the double-layer hard paper sheet. The double-layer hard paper sheet is in a rectangular shape with a rounded top corner. The top of the urine filter paper sheet is clamped by the double-layer hard paper sheet and fixedly connected by pasting. A horizontal folding line is provided at the junction of the double-layer hard paper sheet and the urine filter paper sheet. A vertical folding line is provided between the collection cards. The top length of the urine filter paper sheet is less than the bottom length of the double-layer hard paper sheet. Thin films are symmetrically arranged on both sides of the double-layer hard paper sheet, and protective paper is arranged on the inner side of the thin films. The top of the thin films is pasted to the bottom side of the collection card. This utility model has a novel structure and is convenient to use. During preservation and detection, external impurities are avoided by covering with a thin film, providing the accuracy of detection.

[0005] CN209117426U discloses a urine sample collection card, which includes a waterproof sheet. One end of the waterproof sheet has a hand-held end, and the other end is embedded with a water-absorbing sheet. At least one side surface of the water-absorbing sheet is exposed on the outer surface of the urine sample collection card; a sampling and storage set includes the urine sample collection card and a storage container. The storage container includes a container body and a container cover that can be opened and closed relative to each other. The container body and the container cover are closed to form a sealed cavity, and the sealed cavity is used to seal and accommodate the urine sample collection card and dry the urine sample collection card. The urine sample collection card and the sampling and storage set provided by this utility model can conveniently and time-savingly realize the daily collection, reliable storage and transportation of urine samples, meet the needs of remote collection and transportation, and have the advantages of safety, reliability, convenient operation and strong timeliness.

[0006] However, none of the above-mentioned collection cards involve any content related to sample protection, which affects the test results. Therefore, how to provide a collection card that can effectively protect samples has become an urgent problem to be solved. Summary of the Invention

[0007] Aiming at the deficiencies of the prior art, the purpose of the present invention is to provide a dry urine spot collection card and its application, especially a dry urine spot collection card with high sample stability and its application. The collection card provided by the present invention can effectively protect samples, improve the stability of samples to be tested, and improve the accuracy of detection.

[0008] To achieve the purpose of this invention, the following technical solutions are adopted:

[0009] On the one hand, the present invention provides a dry urine spot collection card, which includes a first outer surface area, a sample collection area, and a second outer surface area connected in sequence along the horizontal direction. The sample collection area is vertically connected to the information collection area;

[0010] The first outer surface area can be folded along the connection line between the first outer surface area and the sample collection area to cover the sample collection area from the front;

[0011] The second outer surface area can be folded along the connection line between the second outer surface area and the sample collection area to cover the sample collection area from the back;

[0012] The sample collection area is composed of two layers of cardboard and a filter paper sandwiched between the two layers of cardboard. The two layers of cardboard are open at least at two opposite positions to expose the filter paper, forming at least two sampling areas.

[0013] The above-mentioned specific collection card can effectively protect samples, improve the stability of samples to be tested, and improve the accuracy of detection.

[0014] Preferably, a light-shielding layer is provided on the surface of the first outer surface area and the second outer surface area away from the sample collection area after folding, and the light-shielding layer is an aluminum foil paper layer, a tin foil paper layer or a dark ink layer.

[0015] The above setting can effectively protect the sample from being stored in a light-shielded environment, improve the stability of the sample to be tested, and improve the accuracy of detection.

[0016] Preferably, a deoxidizer packet is provided on the surface of the second outer surface area away from the sample collection area after folding, and the deoxidizer packet contains a deoxidizer.

[0017] By setting the deoxidizer packet, the above collection card can make the sample in a deoxidized environment after collection and being put into the sealed bag, improve the stability of the sample to be tested, and improve the accuracy of detection.

[0018] Preferably, the deoxidizer includes any one or a combination of at least two of iron powder, dithionite, glucose oxidase, vitamin C or oleic acid.

[0019] Preferably, the shape of the sampling area is circular, square, rectangular, rhombic, fan-shaped, trapezoidal or irregular.

[0020] Preferably, the filter paper is soaked in a protective agent solution.

[0021] The above process of treating with the protective agent solution can greatly improve the protection effect of the collection card on the sample to be tested, effectively improve the stability of the sample to be tested, and further improve the accuracy of detection.

[0022] Preferably, the protective agent includes any one or a combination of at least two of acetic acid, sodium metabisulfite, potassium metabisulfite, vitamin C, trifluoroacetic acid, perchloric acid or formic acid.

[0023] Preferably, the protective agent is a combination of acetic acid and sodium metabisulfite.

[0024] The above specific combination is compounded by acetic acid and sodium metabisulfite, and through their synergistic effect, it can effectively improve the protection effect of the collection card.

[0025] Preferably, in the protective agent solution, the mass fraction of acetic acid is 0.5 - 1.5%, and the mass fraction of sodium metabisulfite is 0.01 - 0.05%. Among them, the mass fraction of acetic acid can be 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4% or 1.5%, etc., and the mass fraction of sodium metabisulfite can be 0.01%, 0.015%, 0.02%, 0.025%, 0.03%, 0.035%, 0.04%, 0.045% or 0.05%, etc., but is not limited to the above-listed values, and other unlisted values within the above numerical range are equally applicable.

[0026] On the other hand, the present invention also provides the application of the dry urine spot collection card as described above in clinical urine detection.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] The present invention provides a dry urine spot collection card, which can effectively protect the sample, improve the stability of the sample to be detected, and improve the accuracy of detection; and through specific structural settings and processing procedures, the stability of the sample to be detected is further improved, and the accuracy of detection is improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 is a schematic diagram of the front unfolded structure of the dry urine spot collection card in Example 1;

[0030] Figure 2 is a schematic diagram of the back unfolded structure of the dry urine spot collection card in Example 1;

[0031] Where 1 - the first outer surface area, 2 - the sample collection area, 3 - the second outer surface area, 4 - the information collection area. DETAILED DESCRIPTION OF THE INVENTION

[0032] The technical solutions of the present invention will be further described below through specific embodiments. Those skilled in the art should understand that the embodiments are only for helping to understand the present invention and should not be regarded as specific limitations to the present invention.

[0033] Example 1

[0034] This example provides a dry urine spot collection card, the front unfolded structure of which is as Figure 1 , and the back unfolded structure is as Figure 2 shown; where 1 is the first outer surface area (7×5 cm), 2 is the sample collection area (10×5 cm), 3 is the second outer surface area (7×5 cm), 4 is the information collection area, the sample collection area is composed of two layers of cardboard (each with a thickness of 1 mm) and a filter paper sandwiched between the two layers of cardboard, and the two layers of cardboard are opened at two opposite positions to expose the filter paper, forming two circular sampling areas with a diameter of 2 cm; the back of the first outer surface area and the front of the second outer surface area are respectively printed with black ink to form a light-shielding layer, and a deoxidizer package containing sodium dithionite, Ca(OH)2 and activated carbon is pasted on the light-shielding layer of the second outer surface area. The filter paper is pre-soaked in a protective agent solution (aqueous solution, where the mass fraction of acetic acid is 1% and the mass fraction of sodium metabisulfite is 0.025%), and then dried in a vacuum drying oven at 60°C.

[0035] During use, after the sample is collected, fold the first outer surface area along the connection line towards the front, fold the second outer surface area along the connection line towards the back, and then put it into a bag to achieve sample collection and preservation. The test sample is protected by a triple protection of a protective agent, a light-shielding layer, and a deoxidizer, effectively improving the stability of the sample and thus the accuracy of the detection.

[0036] Use the above-mentioned collection card to sample the urine sample to be tested, and then test. The test reagents and methods are as follows:

[0037] (1) Detection of catecholamine compounds:

[0038] Use the sample extraction solution (urinary catecholamine) kit (Shanghai RunDa RongJia Biotechnology Co., Ltd., Shanghai Medical Device Preparation 20230104) to test dopamine - DA, epinephrine - E, norepinephrine - NE, metanephrine - MN, normetanephrine - NMN, 3 - methoxytyramine - 3 - MT, homovanillic acid - HVA, vanillylmandelic acid - VMA. After extraction and direct dilution treatment using an Agilent Boyaa Auto M32 fully automatic extractor, inject the sample. Experimental conditions:

[0039]

[0040]

[0041] The S1 - S6 standard curve (artificial urine) samples and third - party quality controls are respectively labeled as follows:

[0042]

[0043] Sample treatment method:

[0044] Take 400 μL of urine sample, drop it on the dry urine spot collection card, and dry it in a well - ventilated place away from light at 20 °C for 4 h. Then use a punch to take 2 circular pieces with a diameter of 0.3 cm, add 400 μL of water, and extract with 16 mL of internal standard solution by ultrasonic for 15 min.

[0045] A) Magnetic filler treatment: Activate with 1 mL of methanol and equilibrate with 1 mL of water;

[0046] B) Eluent: Dilute according to the ratio of EUC - pH regulator 3: methanol = 1:49;

[0047] C) Take 0.38 mL of the sample into a centrifuge tube, add 0.38 mL of pH regulator 1 dilution solution, add pH regulator 2 dilution solution to adjust the pH to 7, and then take 1 mL and add it to the magnetic filler in step A).

[0048] D) Take 1 mL of water and 0.5 mL of methanol to wash the magnetic packing material successively. Finally, elute it with 0.5 mL of the eluent prepared in step B), collect the eluent and dry it with nitrogen. The residue is redissolved with 200 μL of the LEUC - complex solution and vortexed for 30 s. For determination.

[0049] The experimental results are as follows:

[0050] Linearity:

[0051]

[0052]

[0053] Accuracy (using the third - party quality control of the Masscheck brand produced by Chromsystem):

[0054]

[0055]

[0056] Conclusion: This collection card can be applied to the detection of dopamine, epinephrine, norepinephrine, metanephrine, normetanephrine and 3 - methoxytyramine in dried urine spots.

[0057] (2) Detection of homovanillic acid and vanillylmandelic acid

[0058] Experimental conditions:

[0059]

[0060]

[0061] Linear concentration of the samples in the kit:

[0062]

[0063] Sample treatment method:

[0064] Take 400 μL of urine sample and drop it on the dried urine spot collection card. Air - dry it in a well - ventilated place at 20 °C in the dark for 4 h. Then use a hole - puncher to take 2 circular pieces with a diameter of 0.3 cm, add 400 μL of water and 16 mL of internal standard solution, and extract by ultrasonic for 15 min.

[0065] Take 10 μL of the extract, add 90 μL of the diluent in the kit to dilute it, centrifuge at 14800 rpm for 5 min, and take the supernatant for direct injection, thus obtained.

[0066] The results are as follows:

[0067] Linearity:

[0068]

[0069]

[0070] Accuracy (using a third-party quality control of the Masscheck brand produced by Chromsystem):

[0071]

[0072] Conclusion: This acquisition card can be applied to the detection of vanillylmandelic acid and homovanillic acid in dried urine spots. Comparison of preservatives:

[0073] (1) Based on Example 1, the filter paper of the acquisition card was replaced respectively to test different preservatives. Except for replacing the preservative, the rest was the same as Example 1. The test filter papers included ordinary quantitative filter paper without treatment with the preservative solution (M1), quantitative filter paper with a 1.025% acetic acid aqueous solution as the preservative solution (M2), quantitative filter paper with a 1% acetic acid + 0.025% sodium metabisulfite aqueous solution as the preservative solution (M3, i.e., Example 1), quantitative filter paper with a 1% acetic acid + 0.025% vitamin C aqueous solution as the preservative solution (M4), quantitative filter paper with a 1.025% vitamin C aqueous solution as the preservative solution (M5), and quantitative filter paper with a 1.025% sodium metabisulfite aqueous solution as the preservative solution (M6).

[0074] Take fresh urine samples, drop 400 μL of urine samples on the dried urine spot acquisition card in sequence, air dry in the dark for 4 h, prepare 6 replicates for each condition, and process them according to the catecholamine compound detection method in Example 1. Take out one replicate for processing on the 0th, 1st, 3rd, 5th, and 7th days respectively. Taking the detection data on the 0th day of each as 100%, compare the detection results of the dried urine spots under each condition within 7 days. The results are as follows:

[0075] Epinephrine:

[0076] Number of days M1 M2 M3 M4 M5 M6 0 100％ 100％ 100％ 100％ 100％ 100％ 1 93.21％ 112.97％ 112.45％ 102.00％ 99.76％ 99.23％ 3 73.23％ 88.10％ 97.05％ 92.84％ 86.98％ 85.32％ 5 65.12％ 78.66％ 98.27％ 70.63％ 76.21％ 72.01％ 7 59.03％ 73.10％ 95.19％ 59.28％ 60.01％ 69.12％

[0077] Norepinephrine:

[0078] Number of days M1 M2 M3 M4 M5 M6 0 100％ 100％ 100％ 100％ 100％ 100％ 1 102.13％ 118.28％ 107.92％ 109.35％ 98.76％ 98.64％ 3 98.23％ 96.14％ 96.25％ 122.21％ 96.45％ 101.23％ 5 95.67％ 122.97％ 100.87％ 125.45％ 103.24％ 99.08％ 7 92.34％ 99.69％ 104.29％ 92.29％ 99.17％ 97.65％

[0079] Metanephrine:

[0080] Number of days M1 M2 M3 M4 M5 M6 0 100％ 100％ 100％ 100％ 100％ 100％ 1 92.16％ 117.96％ 109.91％ 97.96％ 99.66％ 98.12％ 3 82.62％ 99.18％ 96.27％ 93.78％ 93.34％ 90.23％ 5 75.43％ 99.19％ 100.93％ 82.94％ 80.53％ 86.17％ 7 67.22％ 94.92％ 93.69％ 75.85％ 64.32％ 82.34％

[0081] 3-Methoxytyramine:

[0082] Number of days M1 M2 M3 M4 M5 M6 0 100％ 100％ 100％ 100％ 100％ 100％ 1 101.25％ 115.22％ 102.50％ 96.45％ 92.14％ 101.88％ 3 92.17％ 94.69％ 89.07％ 91.20％ 85.62％ 99.43％ 5 88.65％ 96.38％ 95.52％ 88.38％ 80.18％ 84.58％ 7 85.13％ 95.64％ 88.46％ 79.74％ 70.97％ 86.86％

[0083] Dopamine:

[0084] Number of days M1 M2 M3 M4 M5 M6 0 100％ 100％ 100％ 100％ 100％ 100％ 1 88.67％ 115.17％ 97.75％ 99.00％ 92.40％ 96.45％ 3 65.43％ 93.31％ 89.15％ 95.44％ 87.65％ 90.14％ 5 56.89％ 90.85％ 96.09％ 86.70％ 81.17％ 88.56％ 7 48.19％ 78.65％ 87.75％ 72.17％ 70.02％ 80.08％

[0085] Metanephrine:

[0086]

[0087]

[0088] Conclusion: It can be seen that in the dry urine spot collection card without stabilizer, all 6 compounds showed obvious degradation within 7 days, among which the degradation rates of E and DA were the highest. While in the dry urine spot collection card with stabilizer, adding acetic acid and sodium metabisulfite had the best effect, which could ensure that the contents of all 6 compounds were stable within ±15%.

[0089] (2) Based on Example 1, the filter papers of the collection card were replaced respectively to test different protective agents. Except for replacing the protective agent, the rest were the same as in Example 1. The test filter papers included ordinary quantitative filter paper without treatment with protective agent solution (M0), quantitative filter paper with 1.025% acetic acid aqueous solution as the protective agent solution (M1), quantitative filter paper with 1% acetic acid + 0.0125% sodium metabisulfite aqueous solution as the protective agent solution (M2), quantitative filter paper with 1% acetic acid + 0.025% sodium metabisulfite aqueous solution as the protective agent solution (M3, i.e., Example 1), and quantitative filter paper with 1% acetic acid + 0.03125% sodium metabisulfite aqueous solution as the protective agent solution (M4).

[0090] Take fresh urine samples, and successively add 400 μL of urine samples dropwise onto the dry urine spot collection cards, air-dry in the dark for 4 h. Prepare 5 replicates for each condition, and process them according to the detection methods of homovanillic acid (HVA) and vanilmandelic acid (VMA) in Example 1. Take out one replicate for treatment on each of the 0th, 1st, 3rd, 5th, and 7th days respectively, and take the detection data on the 0th day of each as 100% to compare the detection results of dry urine spots under each condition within 7 days. The results are as follows:

[0091] Homovanillic acid:

[0092]

[0093]

[0094] Vanilmandelic acid:

[0095] Number of days M0 M1 M2 M3 M4 0 100.00％ 100.00％ 100.00％ 100.00％ 100.00％ 1 71.17％ 102.43％ 101.27％ 110.68％ 98.30％ 3 44.32％ 72.86％ 93.84％ 100.02％ 100.54％ 4 23.21％ 63.89％ 89.03％ 94.42％ 98.78％ 7 5.46％ 29.14％ 66.47％ 88.32％ 88.60％

[0096] Conclusion: It can be seen that in the dry urine spot collection card with a 1.025% glacial acetic acid formula, vanillylmandelic acid (VMA) is significantly degraded within 7 days, while the effect of adding 1% acetic acid and 0.025% sodium metabisulfite is the best, similar to the effect of adding 1% acetic acid and 0.03125% sodium metabisulfite, indicating that 1% acetic acid and 0.025% sodium metabisulfite are sufficient to ensure the stability of the compounds and can ensure that the contents of the two compounds are both stable within ±15%.

[0097] Example 2:

[0098] Taking the signal-to-noise ratio close to 10:1 as the standard, the samples were diluted with artificial urine, and the sensitivities of each compound were tested under the conditions of Example 1 as follows: (unit: μg / mL)

[0099]

[0100]

[0101] M0 M1 M2 M3 M4 Homovanillic acid 0.052 0.043 0.031 0.028 0.035 Vanillylmandelic acid 0.045 0.032 0.030 0.025 0.036

[0102] Example 3:

[0103] Two different concentrations of test samples, low and high, were added to the urine, and 400 μL of the urine sample was respectively dropped onto the dry urine spot collection card, air-dried in the dark for 4 h for the spike recovery test. 6 replicates were prepared in parallel for each sample, and the results are as follows:

[0104] It can be seen that for M3 with the compounding of acetic acid and sodium metabisulfite, the recovery rates of each compound can pass, indicating that the compounding of these two compounds can significantly improve

[0105]

[0106]

[0107] From the above tests, it can be found that the collection card provided by the present invention can effectively prevent the degradation of the detection items in the sample, improve the stability of the sample, and improve the accuracy and sensitivity of the detection; and the present invention further improves the effect of the collection card by selecting the compounding of acetic acid and sodium metabisulfite, having a synergistic effect and controlling their ratio.

[0108] The applicant declares that the present invention uses the above embodiments to illustrate the dry urine spot collection card and its application of the present invention, but the present invention is not limited to the above embodiments, that is, it does not mean that the present invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, the equivalent replacement of each raw material of the product of the present invention, the addition of auxiliary components, the selection of specific methods, etc., all fall within the protection scope and the disclosure scope of the present invention.

[0109] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solutions of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0110] In addition, it should be noted that, in the various specific technical features described in the above specific embodiments, without conflict, they can be combined in any appropriate manner. To avoid unnecessary repetition, the present invention will not separately describe various possible combination manners.

Claims

1. A dry urine spot collection card, characterized in that, The dry urine spot collection card includes a first outer surface area, a sample collection area, and a second outer surface area that are sequentially connected along the horizontal direction. The sample collection area is vertically connected to the information collection area; The first outer surface area can be folded along the connection line between the first outer surface area and the sample collection area to cover the sample collection area from the front; The second outer surface area can be folded along the connection line between the second outer surface area and the sample collection area to cover the sample collection area from the back; The sample collection area is composed of two layers of cardboard and filter paper sandwiched between the two layers of cardboard. The two layers of cardboard are opened at least at two opposite positions to expose the filter paper, forming at least two sampling areas.

2. The dry urine spot collection card according to claim 1, characterized in that, A light-shielding layer is provided on the surface of the first outer surface area and the second outer surface area away from the sample collection area after folding. The light-shielding layer is an aluminum foil paper layer, a tin foil paper layer, or a dark ink layer.

3. The dry urine spot collection card according to claim 1 or 2, characterized in that A deoxidizer packet is provided on the surface of the second outer surface area away from the sample collection area after folding. The deoxidizer packet contains a deoxidizer.

4. The dry urine spot collection card according to claim 3, characterized in that, The deoxidizer includes any one or a combination of at least two of iron powder, dithionite, glucose oxidase, vitamin C, or oleic acid.

5. The dry urine spot collection card according to any one of claims 1-4, characterized in that The shape of the sampling area is circular, square, rectangular, rhombic, fan-shaped, trapezoidal, or irregular.

6. The dry urine spot collection card according to any one of claims 1-5, characterized in that, The filter paper is soaked in a protective agent solution.

7. The dry urine spot collection card according to claim 6, wherein The protective agent includes any one or a combination of at least two of acetic acid, sodium metabisulfite, potassium metabisulfite, vitamin C, trifluoroacetic acid, perchloric acid, or formic acid.

8. The dry urine spot collection card according to claim 7, characterized in that, The protective agent is a combination of acetic acid and sodium metabisulfite.

9. The dry urine spot collection card according to claim 8, characterized in that, In the protective agent solution, the mass fraction of acetic acid is 0.5 - 1.5%, and the mass fraction of sodium metabisulfite is 0.01 - 0.05%.

10. Use of the dry urine spot collection card according to any one of claims 1 - 9 in clinical urine detection.

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