Soft materials, containers, liquid calibrators, and test kits sealed with sealing agents

Abstract

This invention provides a soft material, container, liquid calibrator, and test kit sealed with a sealing agent. The sealing agent in the soft material is a steroid compound. This invention solves the problem in existing technologies where steroid hormones are easily adsorbed by the soft material of the container during storage, and is applicable to the field of soft materials.

Description

Technical Field

[0001] This invention relates to the field of soft materials, and more specifically, to a soft material, container, liquid calibrator, and test kit sealed with a sealing agent. Background Technology

[0002] Steroid hormones are a class of tetracyclic aliphatic hydrocarbon compounds with a cyclopentane-polyhydrophenanthrene core. They are synthesized by the gonads, placenta, and zona reticularis of the adrenal cortex in animals, and play a role in promoting the maturation of sexual organs, the development of secondary sexual characteristics, and maintaining sexual function. Modern clinical medical research shows that the detection and monitoring of steroid hormones can be an effective indicator for the diagnosis of endocrine and metabolic diseases, especially gonadal and adrenal diseases. Accurate quantitative determination of steroid hormones in the serum or plasma of healthy individuals can effectively provide early warning of endocrine and metabolic diseases. For patients with existing clinical symptoms of endocrine and metabolic diseases, accurate quantitative determination of these substances can help clinicians make rapid and accurate diagnoses.

[0003] For example, androstenedione is a 19-carbon steroid hormone secreted by the adrenal glands and gonads, serving as an intermediate product in the biochemical synthesis of the male hormone testosterone and the female hormone estradiol. Measuring the level of androstenedione in the blood can be used as an effective indicator for analyzing androgens. Elevated levels of androstenedione can be used to predict the presence of adrenal gland disease, adrenal tumors, or gonadal tumors. Measuring androstenedione levels can also be used to monitor the effectiveness of glucocorticoid treatment.

[0004] In androstenedione immunoassay, a certain concentration of androstenedione needs to be stored as a calibration standard. Existing androstenedione calibrators are typically stored in containers with some soft material (usually rubber or silicone rubber) to facilitate needle insertion for sampling. Because androstenedione is hydrophobic, it easily adsorbs into the pores of the inner wall of the similarly hydrophobic soft material, leading to a decrease in the concentration of the calibration standard and affecting the accuracy of the test results. Similarly, other steroid hormones, besides androstenedione, also have the problem of calibrators being easily adsorbed by soft materials during storage. Summary of the Invention

[0005] The main objective of this invention is to provide a soft material, container, liquid calibrator, and test kit sealed with a sealing agent, in order to solve the problem in the prior art that steroid hormones are easily adsorbed by the soft material of the container during storage.

[0006] To achieve the above objectives, according to a first aspect of the present invention, a soft material sealed with a sealing agent is provided, wherein the sealing agent in the soft material is a steroidal compound.

[0007] Further, the soft material is silicone material, including solid silicone or liquid silicone; preferably, the main material of the solid silicone is methyl vinyl silicone rubber; preferably, the solid silicone also includes a first reinforcing agent and a first crosslinking agent, the first reinforcing agent being silica and the first crosslinking agent being hydroxyl silicone oil; preferably, the main material of the liquid silicone is polyorganosiloxane; preferably, the liquid silicone also includes a second reinforcing agent and a second crosslinking agent, the second reinforcing agent being silica or vinyl MQ resin and the second crosslinking agent being hydroxyl silicone oil.

[0008] Furthermore, sealing can include physical sealing or chemical sealing.

[0009] Further, physical sealing includes: immersing the soft material in a solution of the steroidal compound for incubation, drying, and obtaining the soft material sealed with the sealing agent; preferably, the solution of the steroidal compound includes at least one of phosphate buffer, borate buffer, Tris hydrochloric acid buffer, or carbonate buffer; the concentration of the steroidal compound in the solution is 10-100 ng / mL, more preferably 20 ng / mL.

[0010] Further, chemical blocking includes: contacting a soft material and a steroid compound under acidic conditions to carry out an addition reaction to obtain a soft material blocked by a blocking agent; the steroid compound contains a carbonyl group in its structure; the acidic conditions include a buffer solution with a pH of 4.0-5.0, preferably including at least one of phosphate buffer, acetate buffer, and citrate buffer.

[0011] To achieve the above objective, according to a second aspect of the present invention, a method for preparing a soft material sealed with a sealing agent is provided, the method comprising: sealing the soft material with a sealing agent, wherein the sealing agent is a steroidal compound.

[0012] Furthermore, closure includes physical adsorption or chemical cross-linking.

[0013] Further, physical adsorption includes: immersing the soft material in a solution of the steroidal compound for incubation, drying, and obtaining a soft material blocked by a blocking agent; preferably, the solution of the steroidal compound includes at least one of phosphate buffer, acetate buffer, citrate buffer, borate buffer, Tris hydrochloric acid buffer, or carbonate buffer; the concentration of the steroidal compound in the solution is 10-100 ng / mL, more preferably 20 ng / mL; the pH of the solution of the steroidal compound is 6.5-7.5.

[0014] Furthermore, chemical cross-linking includes: contacting a soft material and a steroid compound under acidic conditions to carry out an addition reaction to obtain a soft material blocked by a blocking agent; the steroid compound contains a carbonyl group in its structure; the acidic conditions include a buffer solution with a pH of 4.0-5.0.

[0015] Further, the soft material is silicone material, including solid silicone or liquid silicone; preferably, the main material of the solid silicone is methyl vinyl silicone rubber; preferably, the solid silicone also includes a first reinforcing agent and a first crosslinking agent, the first reinforcing agent being silica and the first crosslinking agent being hydroxyl silicone oil; preferably, the main material of the liquid silicone is polyorganosiloxane; preferably, the liquid silicone also includes a second reinforcing agent and a second crosslinking agent, the second reinforcing agent being silica or vinyl MQ resin and the second crosslinking agent being hydroxyl silicone oil.

[0016] To achieve the above objectives, according to a third aspect of the present invention, a container is provided comprising the aforementioned soft material sealed with a sealant.

[0017] To achieve the above objectives, according to a fourth aspect of the present invention, a liquid calibrator is provided, which is stored in the aforementioned container, the liquid calibrator comprising a target steroid hormone, wherein the steroid compound acting as a blocking agent has a different chemical structure from the target steroid hormone.

[0018] Further, the target steroid hormones include sex hormones and / or corticosteroids; preferably, the sex hormones include one or more of androgens, estrogens, or progestins; preferably, the corticosteroids include glucocorticoids and / or mineralocorticoids.

[0019] Furthermore, the liquid calibrators also include one or more of the following: phosphate buffer, acetate buffer, citrate buffer, borate buffer, Tris hydrochloride buffer, or carbonate buffer.

[0020] To achieve the above objectives, according to a fifth aspect of the present invention, a sandwich assay kit for detecting a target steroid hormone is provided. The assay kit includes the aforementioned liquid calibrator and further includes: a first antibody for specifically binding to the target steroid hormone and a second antibody for specifically binding to a new epitope formed after the first antibody and the target steroid hormone bind.

[0021] By applying the technical solution of the present invention, the soft material sealed with the sealing agent contains steroidal compounds. Compared with conventional soft materials, the adsorption capacity for target steroid hormones is greatly reduced, which can be used for the preservation of target steroid hormones and reduce the amount of target steroid hormones adsorbed by the soft material of the container during the preservation process. Detailed Implementation

[0022] It should be noted that, unless otherwise specified, the embodiments and features described in this application can be combined with each other. The present invention will now be described in detail with reference to the embodiments.

[0023] Terminology Explanation:

[0024] Soft materials: Soft materials such as silicone are used for sealing in containers and come into contact with the samples stored in the containers, including but not limited to rubber stoppers, gaskets or rings.

[0025] Steroid hormones: compounds with the following basic cyclopentane-polyhydrophenanthrene skeleton, as shown in Formula I. R1, R2 and R3 are common substituent positions on the skeleton. Common substituents include, but are not limited to, methyl groups. Other substituent groups or olefin bonds may also be present at other positions on the skeleton, including, but not limited to, naturally occurring hormones or artificially synthesized steroid hormone derivatives.

[0026]

[0027] Target steroid hormones: In this application, steroid hormones to be preserved and prevented from being adsorbed by soft materials are included, including but not limited to naturally occurring hormones or synthetically produced steroid hormone derivatives. Naturally occurring hormones include sex hormones of the gonads and corticosteroids of the adrenal cortex. Sex hormones include androgens (dehydroepiandrosterone, androstenedione, testosterone, etc.), estrogens (estradiol, estradiol, etc.), and progestins (pregnenolone, progesterone, etc.). Corticosteroids include cortisol, aldosterone, etc., including but not limited to glucocorticoids or mineralocorticoids. Synthetically produced steroid hormone derivatives include but are not limited to metestosterone.

[0028] Steroidal compounds: In this application, steroidal hormones used to seal soft materials are included, but are not limited to naturally occurring hormones or artificially synthesized steroidal hormone derivatives.

[0029] As mentioned in the background section, steroid hormones are easily adsorbed by soft materials in containers during storage. In this application, the inventors attempted to develop a soft material that could reduce the adsorption of steroid hormones, and proposed a series of protection schemes for this application.

[0030] In a first typical embodiment of this application, a soft material sealed with a sealing agent, wherein the sealing agent is a steroidal compound, is provided.

[0031] In the aforementioned soft material, steroidal compounds are sealed. This sealing allows the steroidal compounds to adsorb into the easily adsorbed pores in the soft material in advance, reducing the adsorption problem of the soft material on the target steroid hormone to be preserved.

[0032] By using one or more steroidal compounds to block soft materials, further adsorption of target steroid hormones by the soft materials can be prevented. The reduction in steroidal compound adsorption is not limited to the specific type of steroidal compound used for blocking; that is, after blocking soft materials with one or more steroidal compounds, the adsorption capacity of the blocked soft material for all cyclopentane-polyhydrophenanthrene (steroidal nucleus) compounds is reduced.

[0033] To address the issue of steroid hormone adsorption by soft materials, a sealing agent is preferentially used to seal the soft material before adding the steroid hormone calibrator solution to the container. This prevents the soft material from adsorbing the stored steroid hormone (target steroid hormone), thus ensuring its stable preservation. Preferably, using a small molecule compound with a structure similar to the target steroid hormone as the sealing agent yields better results. Preferably, structural similarity includes differences in substituents on the basic steroid core skeleton or differences in the positions of double bonds in the multi-rings on the basic steroid core skeleton. Preferably, using a steroid compound with the same or similar structure as the target steroid compound for sealing results in a soft material with low adsorption capacity for the target steroid compound.

[0034] Taking androstenedione as an example, the soft material of the container (here, a silicone film for sealing the container) is first sealed using a compound containing cyclopentanoperhydrophenanthrene, which has a similar structure to androstenedione. The sealing soft material has pores that easily adsorb androstenedione. Preferably, the steroidal compounds involved in the sealing may include testosterone, estradiol, progesterone, aldosterone, and other steroidal compounds. Cyclopentanoperhydrophenanthrene is the basic structure of all steroid hormones; it is composed of three cyclohexanes and one cyclopentane. Different steroid hormones differ only in the number of carbon atoms and the substituents.

[0035] In a preferred embodiment, the steroid compound includes one or more of sex hormones or corticosteroids; preferably, the sex hormone includes one or more of androgens, estrogens, or progestins; preferably, the corticosteroid includes glucocorticoids and / or mineralocorticoids.

[0036] In a preferred embodiment, the soft material is silicone material, which includes solid silicone or liquid silicone; preferably, the main material of the solid silicone is methyl vinyl silicone rubber; preferably, the solid silicone also includes a first reinforcing agent and a first crosslinking agent, wherein the first reinforcing agent is silica and the first crosslinking agent is hydroxyl silicone oil; preferably, the main material of the liquid silicone is polyorganosiloxane; preferably, the liquid silicone also includes a second reinforcing agent and a second crosslinking agent, wherein the second reinforcing agent is silica or vinyl MQ resin (CAS NO: 68988-89-6) and the second crosslinking agent is hydroxyl silicone oil.

[0037] The main material of liquid silica gel is polyorganosiloxane. By reacting this main material with a crosslinking agent, soft polysiloxane-based materials are prepared. These materials have a low degree of polymerization, which reduces their adsorption capacity. This is because a lower degree of polymerization results in a smaller molecular weight, weaker intermolecular forces, a more porous structure, larger pore size, and a smaller specific surface area. Therefore, such materials have a lower adsorption capacity, reducing the adsorption of steroids or other types of compounds and exhibiting better stability. The aforementioned crosslinking agent can transform linear or slightly branched macromolecules into a three-dimensional network structure, thereby improving the strength, heat resistance, wear resistance, and solvent resistance of the soft materials.

[0038] In the aforementioned liquid silicone soft materials, reinforcing agents can further improve the strength of the soft material. For example, silica has strong chemical adsorption activity; its surface hydroxyl groups can combine with water through hydrogen bonds to form a multimolecular adsorption layer, and it can also adsorb many small organic molecules. Another example is the addition of vinyl MQ silicone resin to liquid silicone. Liquid rubber reinforced with vinyl MQ silicone resin exhibits lower hardness (hardness decreases linearly with decreasing specific surface area), lower specific surface area, and higher light transmittance. The smaller the specific surface area, the weaker the adsorption capacity. Simultaneously, with the increase of vinyl MQ resin content, the change in light transmittance is very small (within 10%). Lower light transmittance indicates lower density, a more porous structure, larger pore size, smaller specific surface area, and weaker adsorption capacity.

[0039] As a sealing material, polysiloxane soft material (i.e., liquid silica gel) is used as the sealing target. Based on the fact that the adsorption of steroidal compounds by the above-mentioned polysiloxane soft material is smaller than that of ordinary soft materials, the polysiloxane soft material can be further sealed with steroidal compounds, including by means of physical sealing or chemical sealing. This can obtain a soft material with a further reduced adsorption capacity for steroidal compounds. Using this kind of polysiloxane soft material to preserve steroidal compounds can minimize the loss of steroidal compounds.

[0040] In a preferred embodiment, the sealing includes physical sealing or chemical sealing.

[0041] In a preferred embodiment, physical sealing includes: immersing the soft material in a solution of a steroid compound for incubation, followed by drying to obtain a soft material sealed with a sealing agent; preferably, the solution of the steroid compound includes at least one of phosphate buffer, acetate buffer, citrate buffer, borate buffer, Tris hydrochloric acid buffer, or carbonate buffer; the concentration of the steroid compound in the solution is 10-100 ng / mL, more preferably 20 ng / mL. Silica gel has an open porous structure, is hydrophobic, and has strong adsorption capacity, enabling it to adsorb various substances. Steroid hormones are primarily hydrophobic and have a small molecular weight, making it easy for them to fill the pores of silica gel through hydrophobic interactions, preventing further adsorption of other substances. This hydrophobic interaction is relatively stable; therefore, utilizing the hydrophobic interaction of physical sealing can ensure the stable adsorption of steroid compounds in the soft material, meeting the needs of actual products.

[0042] In a preferred embodiment, chemical sealing includes: contacting a soft material and a steroid compound under acidic conditions to carry out an addition reaction to obtain a soft material sealed with a sealing agent, wherein the steroid compound contains a carbonyl group in its structure.

[0043] The acidic conditions mentioned above include buffer solutions with a pH of 4.0-5.0, and preferably the buffer solution includes at least one of phosphate buffer, acetate buffer, and citrate buffer.

[0044] Because there are a large number of residual silanol groups on the silica gel, and some steroidal compounds contain carbonyl groups, under acidic conditions, the carbonyl groups of the steroidal compounds react with the hydroxyl groups on the silica gel material. Alcohols and aldehydes react to form hemiacetals, which further generate acetals. This allows the steroidal compounds to be chemically bonded to the soft material, completing the above-mentioned chemical sealing, filling the pores of the silica gel, and preventing the silica gel from further adsorbing other substances.

[0045] The carbonyl structure described above is located on the substituents at positions 3, 17, and 17 of the steroid compound (skeletal designation as shown in Formula II) or other positions. The structure of a steroid compound may contain one or more carbonyl groups.

[0046]

[0047] Steroidal compounds containing carbonyl groups include, but are not limited to, pregnenolone, 17α-pregnenolone, dehydroepiandrosterone, progesterone, 17α-hydroxyprogesterone, androstenedione, testosterone, dihydrotestosterone, deoxycorticosterone, 11-deoxycortisol, estrogen, aldosterone, corticosterone, or cortisol. In a second typical embodiment of this application, a container is provided comprising the aforementioned soft material sealed with a sealing agent.

[0048] The aforementioned containers include, but are not limited to, vials, wide-mouth bottles, etc., and are sealed using a soft material. This soft material may come into contact with the stored sample. The soft material can take the form of, but is not limited to, rubber stoppers, gaskets, or rubber rings. Using this soft material, sealed with a sealing agent, as the sealing material in the aforementioned containers ensures that even if the soft material comes into contact with the sample in the container, it will not adsorb the sample, thus guaranteeing stable sample storage and content determination. Preferably, the aforementioned soft material allows a sampling needle to pass through for sampling.

[0049] In a third typical embodiment of this application, a liquid calibrator is provided, which is stored in the aforementioned container. The liquid calibrator includes a target steroid hormone, and the steroid compound used as a blocking agent has a different chemical structure from the target steroid hormone.

[0050] In the aforementioned liquid calibrators, containers containing a soft material sealed with steroidal compounds are used for storage to prevent the content of the target steroid hormone from being affected. Furthermore, the soft material does not release the steroidal compounds used for sealing, thus avoiding contamination of the stored sample (target steroid hormone).

[0051] The target steroid hormone used to preserve the sample is not the same compound as the steroid compound used as the sealing agent; they have different chemical formulas. For example, if the target steroid hormone is androstenedione, the steroid compound can be flexibly selected from other steroid compounds that are not androstenedione, including but not limited to testosterone, estradiol, progesterone, and aldosterone. Even if trace amounts of the sealing agent are released into the liquid sample, it will not affect the content of the target steroid hormone.

[0052] In a preferred embodiment, the target steroid hormone includes one or more of sex hormones or corticosteroids; preferably, the sex hormone includes one or more of androgens, estrogens, or progestins; preferably, the corticosteroid includes glucocorticoids and / or mineralocorticoids.

[0053] In a preferred embodiment, the liquid calibrator further includes one or more of phosphate buffer, acetate buffer, citrate buffer, borate buffer, Tris hydrochloride buffer, or carbonate buffer.

[0054] In a fourth typical embodiment of this application, a sandwich method for detecting a target steroid hormone is provided. The detection kit includes the liquid calibrator described above. The detection kit also includes: a first antibody for specifically binding to the target steroid hormone, and a second antibody for specifically binding to a new epitope formed after the first antibody and the target steroid hormone bind together.

[0055] The aforementioned sandwich-method detection kit for target steroid hormones includes a liquid calibrator, a first antibody, and a second antibody. This kit can be used to detect the target steroid hormone in the calibrator and in the sample. The first antibody specifically binds to the target steroid hormone. Furthermore, the second antibody binds to newly formed epitopes in the first binding product, forming a second binding product. Using existing techniques such as chemiluminescence, the content of the second binding product can be accurately detected to obtain the amount of the target steroid hormone.

[0056] Taking a chemiluminescence detection platform as an example, the detection principle and steps are as follows: the target steroid hormone in the liquid calibrator specifically binds to the first antibody coated with magnetic microspheres to form an immune complex; after incubation, unbound substances are removed by magnetic separation and washing; then a luminescent label is added for incubation, and the second antibody labeled with ABEI (N-(4-aminobutyl)-N-ethylisoluminol) specifically binds to the new epitope formed by the target steroid hormone and the first antibody on the magnetic microspheres, forming a ternary immune complex. After incubation, unbound substances are removed by magnetic separation and washing. Finally, the substrate solution for the fully automated immunoassay system is added to initiate the chemiluminescence reaction and generate a light signal. The relative light intensity (RLU) measured by a photomultiplier tube is proportional to the concentration of the target steroid hormone in the liquid calibrator. Since the second antibody only specifically binds to the new epitope formed by the binding of the first antibody and the steroid hormone, even if other steroid hormones are present in the liquid calibrator, the immune complex formed with the first antibody will not be bound by the second antibody. Therefore, using a sandwich method can further avoid the problem of inaccurate detection results.

[0057] The beneficial effects of this application will be explained in more detail below with reference to specific embodiments.

[0058] Example 1: Soft Materials and Sealant

[0059] The soft materials used in this embodiment shown in Table 1 were purchased externally. Material 1 is solid silicone, purchased from Maigao Fine Advanced Materials (Shenzhen) Co., Ltd.; Materials 2 and 3 are liquid silicone, purchased from Shin-Etsu Silicone International Trading (Shanghai) Co., Ltd.

[0060] Table 1

[0061] Main material (70wt%) Reinforcing agent (25wt%) Crosslinking agent (4 wt%) Material 1 Methyl vinyl silicone rubber silicon dioxide Hydroxy silicone oil Material 2 Polyorganosiloxane silicon dioxide Hydroxy silicone oil Material 3 Polyorganosiloxane Vinyl MQ resin Hydroxy silicone oil

[0062] Using a soft material as the container, the container was blocked according to Table 2 using a blocking solution (the buffer solution was 20 wt% Tris-HCl solution, pH 7.5) containing 20 ng / mL testosterone, estradiol, progesterone or aldosterone as the blocking agent.

[0063] Experimental groups 1-5 adopted physical sealing. The specific sealing steps were as follows: the soft material was immersed in the above sealing liquid and shaken at 37°C for 24 hours. Then the soft material was taken out and dried in an oven at 40°C for 12 hours.

[0064] Experimental groups 6-8 were chemically blocked. A 20 ng / mL progesterone chemical blocking solution was prepared (the buffer solution was 20 wt% phosphate buffer, pH 4.5). The solution was shaken at 37°C for 4 h, and then the soft material was taken out and dried in an oven at 40°C for 12 h.

[0065] Table 2

[0066]

[0067] Example 2: Concentration decrease rate of androstenedione calibrator

[0068] The adsorption effect of different experimental groups and control groups on androstenedione was verified by using liquid calibrators at three concentrations: low, medium, and high (0.29 ng / mL, 3.02 ng / mL, and 6.08 ng / mL).

[0069] For the physically sealed group, the adsorption was verified by storing at 2-8℃ for 1 month and at 37℃ for 3 days. For the chemically sealed group, the adsorption was verified by storing at 2-8℃ for 3 months and at 37℃ for 7 days. To simulate the storage and transportation process, the liquid calibrators stored in containers containing soft materials were placed on a shaker and shaken throughout the process.

[0070] The formula for calculating the decrease rate is: (original concentration - current concentration) / original concentration × 100%. The low-value decrease rate, medium-value decrease rate, and high-value decrease rate are the decrease rates of androstenedione concentration in low, medium, and high concentration samples, respectively. The lower the decrease rate, the less androstenedione in the liquid calibrator is adsorbed by the container.

[0071] The assay was performed using the MAGLUMI 4000P chemiluminescence immunoassay analyzer, manufactured in-house by New Industries Biotech, and the androstenedione sandwich assay kit.

[0072] The detection principle and steps are as follows: First, the calibrator solution, buffer solution, and magnetic microspheres are mixed and incubated together according to the instrument program. Androstenedione in the solution specifically binds to the primary antibody coated on the magnetic microspheres, forming an immune complex. After incubation, unbound substances are removed by magnetic separation washing. Second, a luminescent label is added and incubated. The ABEI-labeled secondary antibody specifically binds to the new epitope formed by the androstenedione and primary antibody on the magnetic microspheres, forming a ternary immune complex. After incubation, unbound substances are removed by magnetic separation washing. Finally, the substrate solution for the fully automated immunoassay system is added, initiating the chemiluminescent reaction and generating a light signal. The relative light intensity (RLU) measured by a photomultiplier tube is proportional to the androstenedione concentration in the sample. The decrease rate is calculated based on the detection results, as shown in Table 3.

[0073] Table 3

[0074]

[0075] The results showed that, compared with control group 1, the concentration of androstenedione in the liquid calibrators decreased to some extent after long-term and short-term storage at 2-8℃ and 37℃. Among them, experimental group 1, which used physical sealing, had better results than experimental groups 2, 3, 4 and 5. After storage at 2-8℃ for 1 month and at 37℃ for 3 days, the average decrease rate was only 5.91% and 5.16%, respectively.

[0076] Chemical sealing ensures a low rate of degradation during long-term storage. In experimental group 6, after storage at 2-8℃ for 3 months and at 37℃ for 7 days, the concentration degradation rate of androstenedione calibrator was only 9.44% and 7.34%, respectively.

[0077] Experimental group 7 used material 2 to preserve liquid calibrators, and the anti-adsorption effect was better than that of experimental group 6. After preservation at 2-8℃ for 3 months and at 37℃ for 7 days, the average reduction rate was 9.15% and 9.23%, respectively. After preservation at 2-8℃ for 1 month and at 37℃ for 3 days, the average reduction rate was 5.11% and 4.31%, respectively.

[0078] In Experimental Group 8, material 3 was used to preserve the liquid calibrator. The anti-adsorption effect was further superior to that of Experimental Groups 6 and 7. After storage at 2-8℃ for 3 months and at 37℃ for 7 days, the average degradation rate was only 4.63% and 4.15%, respectively. After storage at 2-8℃ for 1 month and at 37℃ for 3 days, the average degradation rate was only 2.88% and 1.60%, respectively.

[0079] Example 3: Concentration decrease rate of estradiol calibrator

[0080] The concentration of estradiol calibrator at a certain level was used to verify the rate of concentration decrease in experimental group 1 during the storage of estradiol calibrator.

[0081] Similar to Example 3, the MAGLUMI 4000P chemiluminescence immunoassay analyzer and estradiol sandwich assay kit, manufactured by New Industries Biotechnology, were used for instrumental testing.

[0082] The detection principle and steps are as follows: First, the calibrator solution, buffer solution, and magnetic microspheres are mixed and incubated according to the instrument program. Estradiol in the solution specifically binds to the primary antibody coated on the magnetic microspheres, forming an immune complex. After incubation, unbound substances are removed by magnetic separation washing. Second, a luminescent label is added and incubated. The ABEI-labeled secondary antibody specifically binds to the new epitope formed by the androstenedione on the magnetic microspheres and the primary antibody, forming a ternary immune complex. After incubation, unbound substances are removed by magnetic separation washing. Finally, the substrate solution for the fully automated immunoassay system is added, initiating the chemiluminescent reaction and generating a light signal. The relative light intensity (RLU) measured by a photomultiplier tube is proportional to the estradiol concentration in the sample, and the decrease rate is calculated based on the detection results.

[0083] The results showed that after storage at 2-8°C for 1 month and at 37°C for 3 days, the decrease rate of estradiol in the liquid calibrator was 6.18% and 6.12%, respectively. Therefore, the preservation method in this invention is applicable to the preservation of different steroid hormones.

[0084] As can be seen from the above description, the embodiments of the present invention achieve the following technical effects: the soft material sealed with the sealing agent has a significantly reduced adsorption capacity for steroidal compounds compared to conventional soft materials. Furthermore, the polysiloxane soft material itself has an even lower adsorption capacity for steroidal compounds compared to conventional soft materials in the prior art; after sealing with steroidal compounds, the adsorption capacity is further reduced. This soft material can be used for the preservation of steroidal compounds. Using a container containing this soft material to store the target steroid hormone reduces the amount of the target steroid hormone adsorbed by the container's soft material during preservation, and the soft material does not release the sealing steroidal compounds, thus avoiding any impact on the quantity and purity of the preserved steroidal compounds. The sample stored in the container includes the liquid standard, which maintains a stable concentration during storage, thereby ensuring the accuracy of the test in subsequent use.

[0085] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A liquid calibrant, characterized in that, The liquid calibrator comprises a container and a target steroid hormone preserved in the container, The container comprises a soft material sealed by a sealing agent; wherein, The target steroid hormone is androstenone, and the sealing agent is testosterone; The soft material is a silica gel material, and the silica gel is a solid silica gel; The main material of the solid silica gel is methyl vinyl silicone rubber; The sealing is physical sealing; The solid silica gel further comprises a first reinforcing agent and a first crosslinking agent, the first reinforcing agent is silica, and the first crosslinking agent is hydroxyl silicone oil.

2. The liquid calibrant of claim 1, wherein, The physical sealing comprises: incubating the soft material in a solution of the steroid compound, drying to obtain the soft material sealed by the sealing agent.

3. The liquid calibrant of claim 2, wherein, The solution of the steroid compound comprises at least one of a phosphate buffer, a borate buffer, a Tris hydrochloride buffer or a carbonate buffer; The concentration of the steroid compound in the solution of the steroid compound is 10-100 ng / mL; The pH of the solution of the steroid compound is 6.5-7.

5.

4. The liquid calibrant of claim 3, wherein, The concentration of the steroid compound in the solution of the steroid compound is 20 ng / mL.

5. The liquid calibrant according to any one of claims 1 to 4, characterized in that, The target steroid hormone comprises a sex hormone and / or a corticosteroid.

6. The liquid calibrant of claim 5, wherein, The sex hormone comprises one or more of an androgen, an estrogen or a progestogen.

7. The liquid calibrant of claim 5, wherein, The corticosteroid comprises a glucocorticoid and / or a mineralocorticoid.

8. The liquid calibrant of claim 1, wherein, The liquid calibrator further comprises one or more of a phosphate buffer, an acetate buffer, a citrate buffer, a borate buffer, a Tris hydrochloride buffer or a carbonate buffer.

9. A sandwich assay kit for detecting a target steroid hormone, characterized by, The detection kit comprises the liquid calibrator of any one of claims 1-8, The detection kit further comprises: a first antibody for specifically binding to the target steroid hormone, a second antibody for specifically binding to a neoepitope formed after the first antibody and the target steroid hormone bind together.

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