Urine protective agent and application thereof

Abstract

The invention relates to the technical field of urine preservation, in particular to a urine protective agent and application thereof. The urine protective agent is prepared from diazoalkyl urea, Proclin300 and EDTA (Ethylene Diamine Tetraacetic Acid) salt. According to the urine protective agent, the storage stability of citric acid, oxalic acid and cystine in urine can be considered under the conditions of high temperature and refrigeration temperature aiming at three important target monitoring substances including citric acid, oxalic acid and cystine in the urine, and reagents used by the urine protective agent are easy to obtain; the urine storage bag is easy to prepare, can be widely applied to any scene for storing urine or urine collection containers, assists the development of precise medical examination, and has a wide application prospect.

Description

Technical Field

[0001] The present invention relates to the technical field of urine preservation, and in particular to a urine protective agent and application thereof. Background Art

[0002] Urinary stones are a common disease with a high incidence and recurrence rate. Urinary stones are caused by a variety of factors. The Chinese expert consensus on metabolic assessment and recurrence prevention of urinary stones points out that metabolic assessment factors include: oxalic acid, uric acid, cystine, etc. Among them, citric acid, oxalic acid and cystine in urine are important detection indicators, which are closely related to the health status of the human body. These various factors have different physical and chemical factors, and their physical and chemical properties such as stability and solubility vary greatly. In particular, oxalic acid easily combines with calcium to form calcium oxalate precipitation, which affects the oxalic acid test results.

[0003] Some literature points out the use of 6 mol / L hydrochloric acid for urine protection, and some literature uses a mixed solution of boric acid, thymol and hydrochloric acid as a stabilizer. However, different stabilizers have different disadvantages. For example, although hydrochloric acid can acidify the sample, increase the solubility of oxalic acid, and reduce the crystallization of calcium oxalate, it is difficult to purchase hydrochloric acid and needs to be registered; and the increase in the amount of boric acid can easily cause the detection result of citric acid to be low.

[0004] Therefore, how to develop a urine protectant that can take into account the storage stability of citric acid, oxalic acid and cystine in urine under both high temperature and refrigerated temperature conditions has become a technical problem that urgently needs to be solved in this field. Summary of the invention

[0005] In order to solve the above technical problems, the present invention provides a urine protective agent, including diazolidinyl urea, Proclin 300 and EDTA salt.

[0006] The present invention obtains the above-mentioned urine protective agent by screening from a combination of numerous preservative ingredients and chelating agent ingredients through a large amount of compounding. The urine protective agent can take into account the storage stability of citric acid, oxalic acid and cystine in urine at both high temperature (e.g., 37°C) and refrigerated temperature (2-8°C), and can be stored for up to 3 days at high temperature and up to 14 days at refrigerated temperature.

[0007] Preferably, the EDTA salt is EDTA potassium salt or EDTA sodium salt.

[0008] More preferably, the potassium EDTA salt is at least one of dipotassium EDTA and tetrapotassium EDTA; and the sodium EDTA salt is at least one of disodium EDTA and tetrasodium EDTA.

[0009] Preferably, the mass ratio of diazolidinyl urea, Proclin 300 and EDTA salt is (2-4):1:(4-10).

[0010] More preferably, the mass ratio of diazolidinyl urea, Proclin 300 and EDTA salt is (2-4):1:(4-8).

[0011] Most preferably, the mass ratio of diazolidinyl urea, Proclin 300 and EDTA salt is (2-3):1:(4-5).

[0012] Furthermore, the present invention provides use of the urine protecting agent described in any of the above embodiments in urine storage.

[0013] Preferably, the urine protective agent containing 4-10 mg diazolidinyl urea, 1-4 mg Proclin 300 and 8-40 mg EDTA salt is added to every 2-4 mL of urine.

[0014] Furthermore, the present invention provides a urine storage tube, which contains the urine protective agent described in any of the above embodiments.

[0015] In the specific implementation process, the material of the urine storage tube includes but is not limited to PET.

[0016] In the specific implementation process, the capacity of the urine storage tube includes but is not limited to 4 mL.

[0017] In the specific implementation process, the specifications of the urine storage tube include but are not limited to 13*75mm.

[0018] Furthermore, the present invention provides a urine diagnostic reagent or kit, which contains the urine protective agent described in any of the above embodiments.

[0019] Compared with the prior art, the present invention has the following beneficial effects: The urine protectant of the present invention can monitor three important target substances in urine, namely citric acid, oxalic acid and cystine, and can take into account the storage stability of citric acid, oxalic acid and cystine in urine under high temperature (37°C) and refrigerated temperature (2~8°C). The reagents used in the urine protectant of the present invention are easily available, easy to prepare and can be widely used in any scene of urine storage or urine collection container, which helps the development of precision medical testing and has broad application prospects. DETAILED DESCRIPTION

[0020] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described clearly and completely below. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0021] If no specific techniques or conditions are specified in the examples, all are carried out according to conventional methods or the techniques or conditions described in the literature in the field, or according to the product instructions. If no manufacturer is specified for the reagents and instruments used, all are conventional products that can be purchased through regular channels.

[0022] Example 1

[0023] This example tests the degradation rates of citric acid, oxalic acid and cystine in urine under storage conditions of 37°C and 4°C with different urine protectants (stabilizers). The specific steps are as follows: The experiment used a human urine sample, and spiked its high and low points (low point concentrations were: citric acid concentration of 120μg / mL, oxalic acid concentration of 30μg / mL, cystine concentration of 12μg / mL; high point concentrations were: citric acid concentration of 700μg / mL, oxalic acid concentration of 125μg / mL, cystine concentration of 70μg / mL), and the sample was divided into several equal portions, each 4 mL, and placed in a centrifuge tube. Different urine protective agents were added to each sample to investigate the stability of citric acid, oxalic acid and cystine in human urine samples stored at 37 degrees for 1 day, 3 days, 5 days and 4 degrees Celsius for 1 day, 3 days, 7 days, and 14 days. The sample at each time point was measured three times, and the results were compared with the test value of the initial urine sample (0 day), and the deviation was calculated (deviation% = (test value at different time points-initial test value) / initial test value*100), and the deviation was qualified within ±15%. Oxalic acid, citric acid and cystine were tested by AB Sciex4500. The specific test conditions refer to Example 1 in CN117330684A.

[0024] The test results of citric acid degradation rates of different urine protectants at 37°C and 4°C are shown in Table 1.

[0025] Table 1 Degradation rate of citric acid by different urine protectants at 37℃ and 4℃

[0026] It can be seen that for citric acid, at 37°C, all urine protectants tested met the requirements after 3 days of storage, but did not meet the requirements after 5 days of storage. At 4°C, only the 4 mg diazolidinyl urea-2 mg Proclin300 and 4 mg diazolidinyl urea-2 mg Proclin300-8 mg EDTA disodium groups still met the requirements after 14 days of storage.

[0027] The test results of oxalic acid degradation rates of different urine protectants at 37°C and 4°C are shown in Table 2.

[0028] Table 2 Oxalic acid degradation rates of different urine protectants at 37℃ and 4℃

[0029] It can be seen that for oxalic acid, at 37°C, all urine protectants tested did not meet the requirements after 5 days of storage. After 3 days of storage, only four groups, 4 mg diazolidinyl urea, 2 mg Proclin300, 4 mg diazolidinyl urea-2 mg Proclin300-8 mg EDTA disodium, and 4 mg diazolidinyl urea-4 mg gentamicin-8 mg EDTA disodium, met the requirements. At 4°C, only 4 mg diazolidinyl urea-2 mg Proclin300-8 mg EDTA disodium still met the requirements after 14 days of storage.

[0030] The test results of cystine degradation rates of different urine protectants at 37°C and 4°C are shown in Table 3.

[0031] Table 3 Cystine degradation rates of different urine protective agents at 37℃ and 4℃

[0032] It can be seen that for cystine, at 37°C, all urine protectants tested did not meet the requirements after 5 days of storage, and only 4 mg diazolidinyl urea-2 mg Proclin300-8 mg EDTA disodium met the requirements after 3 days of storage. At 4°C, all urine protectants tested met the requirements after 14 days of storage.

[0033] In summary, 4 mg diazolidinyl urea-2 mg Proclin300-8 mg EDTA disodium as a urine protectant can enable urine to be stored at 37°C for 3 days and at 4°C for up to 14 days.

[0034] Example 2

[0035] In order to further investigate the effects of different chelating agents, this example used equal amounts of dipotassium EDTA, tetrasodium EDTA, tetrapotassium EDTA, EGTA (ethylene glycol bis-α-aminoethyl ether tetraacetic acid) and BAPTA tetrapotassium salt (1,2-bis(2-aminophenoxy)ethane-N,N,N`,N`-tetraacetic acid tetrapotassium) to replace disodium EDTA, that is, 4 mg diazolidinyl urea-2 mg Proclin300-8 mg dipotassium EDTA, 4 mg diazolidinyl urea-2 mg Proclin300-8 mg tetrasodium EDTA, 4 mg diazolidinyl urea-2 mg Proclin300-8 mg tetrapotassium EDTA, 4 mg diazolidinyl urea-2 mg Proclin300-8 mg EGTA, and 4 mg diazolidinyl urea-2 mg Proclin300-8 mg respectively. BAPTA tetrapotassium salt was used as a urine protectant. The degradation rates of citric acid, oxalic acid and cystine in urine of different urine protectants were tested under the detection method of Example 1 at 37° C. and 4° C. storage conditions.

[0036] The test results of citric acid degradation rates of different urine protectants at 37°C and 4°C are shown in Table 4.

[0037] Table 4 Degradation rate of citric acid by different urine protectants at 37℃ and 4℃

[0038] It can be seen that for citric acid, at 37°C, all urine protectants tested met the requirements after 3 days of storage, but did not meet the requirements after 5 days of storage. At 4°C, all urine protectants tested met the requirements after 14 days of storage.

[0039] The test results of oxalic acid degradation rates of different urine protectants at 37°C and 4°C are shown in Table 5.

[0040] Table 5 Oxalic acid degradation rates of different urine protective agents at 37℃ and 4℃

[0041] It can be seen that for oxalic acid, at 37°C, all urine protectants tested met the requirements after 3 days of storage, but did not meet the requirements after 5 days of storage. At 4°C, urine protectants containing different EDTA salts met the requirements after 14 days of storage, but did not meet the requirements after being replaced with EGTA and BAPTA tetrapotassium salts after 14 days of storage.

[0042] The test results of cystine degradation rates of different urine protectants at 37°C and 4°C are shown in Table 6.

[0043] Table 6 Cystine degradation rates of different urine protectants at 37℃ and 4℃

[0044] It can be seen that for cystine, at 37°C, all urine protectants tested met the requirements after 5 days of storage. At 4°C, all urine protectants tested met the requirements after 14 days of storage.

[0045] In summary, the three urine protective agents, 4 mg diazolidinyl urea-2 mg Proclin300-8 mg EDTA dipotassium, 4 mg diazolidinyl urea-2 mg Proclin300-8 mg EDTA tetrasodium, and 4 mg diazolidinyl urea-2 mg Proclin300-8 mg EDTA tetrapotassium, can achieve the same urine protective effect as 4 mg diazolidinyl urea-2 mg Proclin300-8 mg EDTA disodium in Example 1, and can be maintained for three days at 37°C and 14 days at 4°C.

[0046] Example 3

[0047] In this example, the urine protection of 4 mg diazolidinyl urea-2 mg Proclin 300-8 mg disodium EDTA in Example 1 and the urine protectant in Example 2 at -20°C was tested. The test method was the same as that in Example 1. The degradation rates of citric acid, oxalic acid and cystine in urine are shown in Tables 7 to 9, respectively.

[0048] Table 7 Degradation rate of citric acid of different urine protective agents at -20℃

[0049] It can be seen that for citric acid, all urine protectants tested met the requirements when stored at -20°C for 30 days.

[0050] Table 8 Oxalic acid degradation rate of different urine protective agents at -20℃

[0051] It can be seen that for oxalic acid, all urine protectants tested met the requirements when stored at -20°C for 30 days.

[0052] Table 9 Cystine degradation rate of different urine protective agents at -20℃

[0053] It can be seen that for cystine, all urine protectants tested met the requirements when stored at -20°C for 30 days.

[0054] In summary, all urine protectants tested met the detection requirements for citric acid, oxalic acid, and cystine when stored at -20°C for 30 days.

[0055] Example 4

[0056] This example takes diazolidinyl urea-Proclin 300-EDTA dipotassium as an example to explore the effects of different ratios on urine protection. The following schemes are set respectively: Regimen 1: 4 mg diazolidinyl urea-2 mg Proclin300-8 mg EDTA dipotassium (2:1:4); Regimen 2: 4 mg diazolidinyl urea-1 mg Proclin300-8 mg EDTA dipotassium (4:1:8); Plan 3: 10 mg diazolidinyl urea-4 mg Proclin300-40 mg EDTA dipotassium (2.5:1:10).

[0057] Plan 4: 4 mg diazolidinyl urea-1 mg Proclin300-10 mg EDTA dipotassium (4:1:10).

[0058] The test method is the same as that of Example 1. The degradation rates of citric acid, oxalic acid and cystine in urine are shown in Tables 10 to 12, respectively.

[0059] Table 10 Degradation rate of citric acid at 4°C with different ratios

[0060] Table 11 Oxalic acid degradation rate of different mixing ratios at 4℃

[0061] Table 12 Cystine degradation rate of different ratios at 4°C

[0062] It can be seen that the urine protectants with different ratios mentioned above can meet the detection requirements of citric acid, oxalic acid and cystine after being stored at 4°C for 14 days.

[0063] Comparative Example

[0064] In this comparative example, 4 mg imidazolidinyl urea-2 mg Proclin 300-8 mg EDTA disodium and 4 mg DMDMH hydantoin-2 mg Proclin 300-8 mg EDTA disodium were used as urine protective agents, and the test method of Example 1 was used for the test.

[0065] The degradation rate results of citric acid, oxalic acid and cystine in urine are shown in Tables 13 to 15, respectively.

[0066] Table 13 Citric acid degradation rates of different urine protective agents at 37°C and 4°C

[0067] It can be seen that for citric acid, the urine protectant containing imidazolidinyl urea can be stored for 3 days at 37°C and for 14 days at 4°C; while the urine protectant containing DMDMH hydantoin can barely be stored for 3 days at 37°C and for 14 days at 4°C.

[0068] Table 14 Oxalic acid degradation rate of different urine protective agents at 37℃ and 4℃

[0069] It can be seen that for oxalic acid, urine protectants containing imidazolidinyl urea and DMDMH hydantoin cannot be stored at 4°C for 14 days.

[0070] Table 15 Cystine degradation rates of different urine protectants at 37°C and 4°C

[0071] It can be seen that for cystine, the urine protectant containing imidazolidinyl urea and DMDMH hydantoin cannot be stored at 37°C for 3 days.

[0072] In summary, the urine protectant of this comparative example cannot simultaneously achieve the technical effects of storage at 37°C for 3 days and storage at 4°C for 14 days.

[0073] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A urine protective agent, characterized in that: Includes diazolidinyl urea, Proclin 300 and EDTA salts.

2. The urine protective agent according to claim 1, characterized in that: The EDTA salt is EDTA potassium salt or EDTA sodium salt.

3. The urine protective agent according to claim 2, characterized in that: The EDTA potassium salt is at least one of dipotassium EDTA and tetrapotassium EDTA; the EDTA sodium salt is at least one of disodium EDTA and tetrasodium EDTA.

4. The urine protective agent according to any one of claims 1 to 3, characterized in that The mass ratio of diazolidinyl urea, Proclin 300 and EDTA salt is (2~4):1:(4~10).

5. The urine protective agent according to claim 4, characterized in that: The mass ratio of diazolidinyl urea, Proclin 300 and EDTA salt is (2~4):1:(4~8).

6. The urine protective agent according to claim 5, characterized in that: The mass ratio of diazolidinyl urea, Proclin 300 and EDTA salt is (2~3):1:(4~5).

7. Use of the urine protecting agent according to any one of claims 1 to 6 in urine storage.

8. The use according to claim 7, characterized in that: The urine protectant containing 4-10 mg diazolidinyl urea, 1-4 mg Proclin 300 and 8-40 mg EDTA salt is added to every 2-4 mL of urine.

9. A urine storage tube, characterized in that: It contains the urine protecting agent according to any one of claims 1 to 6.

10. A urine diagnostic reagent or kit, characterized in that: It contains the urine protecting agent according to any one of claims 1 to 6.

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