A saliva sampling sponge for low-absorption THC detection and its collection rod

Abstract

The present invention relates to a saliva sampling sponge for low-absorption THC detection and its collection rod. The sampling sponge is made by the following steps: (1) Sponge cleaning and impurity removal: Clean and remove impurities from the medical sponge; (2) SiO2 modification treatment: Modify the cleaned medical sponge with silicon dioxide; (3) Composite enzyme loading treatment: Spray and load the modified medical sponge in a composite enzyme solution, and the composite enzyme is a complex of protease and glucoamylase; (4) Low-temperature drying of the medical sponge after the composite enzyme loading treatment; (5) Clove oil loading treatment: Spray and load the medical sponge after the composite enzyme loading treatment with a clove oil solution, and store it sealed at low temperature. By subjecting the sponge to SiO2 loading treatment, composite enzyme and clove oil loading treatment, the present invention can effectively improve the collection of THC in the oral cavity, thereby improving the detection sensitivity.

Description

Technical Field

[0001] The invention relates to a saliva sampling sponge and a collection stick for low-absorption THC detection, and belongs to the field of THC detection. Background Art

[0002] Marijuana (THC) testing can reveal evidence of marijuana use. Blood and urine are widely used specimens for marijuana analysis. The primary target analyte in blood testing is THC-COOH, which can only detect small amounts of THC and can only provide information on short-term abuse, including behavioral impairment and the degree of intoxication. However, collection is invasive, making roadside monitoring difficult and requiring laboratory analysis. The primary target analyte in urine testing is THC-COOH, whose concentration is primarily affected by factors such as drug dosage, method of abuse, frequency of abuse, and individual differences in drug metabolism and excretion rates. Urine testing offers advantages such as a wide range, a wide detection window (within one week), and low cost. However, collection requires specialized facilities and personnel of the same gender, and is prone to fraud. Saliva is a newly developed specimen, widely used in traffic safety cases involving drunk driving and drugged driving. Its primary target analyte is THC, with relatively few metabolites. Compared with traditional methods of collecting samples (such as blood and urine tests), saliva testing is more sensitive and has the advantages of being easy to operate and fast. Compared with urine testing, saliva testing is easier for the testees to accept and is not restricted by venue and gender. However, the existing saliva THC test has a lower detection sensitivity than urine and blood tests. The main reasons are as follows: (1) Saliva collection has an important impact on saliva drug detection. Currently, medical-grade PVA sponges are generally used as collection carriers. Medical-grade PVA sponges are a polymer material with excellent liquid absorption performance and a very soft surface when in contact with the human body. They are completely naturally degradable and are very suitable for collecting saliva samples. However, the hydrophobic groups in PVA molecules, such as vinyl groups (-CH2-CH2-), do not contain polar functional groups and are relatively hydrophobic, which enables them to have good compatibility with oil molecules (usually non-polar). At the same time, the van der Waals force and hydrophobic force between vinyl groups help PVA interact with oil molecules, thereby promoting the absorption of oil substances. This makes it difficult to separate the THC adsorbed on the sponge, resulting in a low THC content in the collected saliva. (2) THC in the mouth is affected by the organic matter in the oral mucosa and saliva, and exists in large quantities in the oral mucosa and saliva organic matter, making it difficult for THC in the mouth to diffuse into the water in the saliva and be collected. Summary of the Invention

[0003] The present invention provides a saliva sampling sponge and a collection stick for low-absorption THC detection, which solve the problem of low detection sensitivity in existing THC saliva detection.

[0004] To solve the above technical problems, the technical solution adopted by the present invention is as follows:

[0005] A saliva sampling sponge for low-absorption THC detection, and the sampling sponge is made by the following steps:

[0006] (1) Sponge cleaning and impurity removal: Clean and remove impurities from the medical sponge;

[0007] (2) SiO2 modification treatment: Modify the cleaned medical sponge with silicon dioxide;

[0008] (3) Composite enzyme loading treatment: Spray and load the modified medical sponge with a composite enzyme solution, and the composite enzyme is a complex of protease and glucoamylase;

[0009] (4) Low-temperature drying of the medical sponge after the composite enzyme loading treatment;

[0010] (5) Clove oil loading treatment: Spray and load the medical sponge after the composite enzyme loading treatment with a clove oil solution, and seal and store. That's it.

[0011] Further, preferably: The specific sponge cleaning is as follows: Immerse the medical sponge in ethanol and ultrasonically wash it 1-2 times, 10 minutes each time, then rinse it with deionized water until there is no alcohol smell, and finally dry it for standby.

[0012] Further, preferably: The specific SiO2 modification treatment is to add nano-SiO2 to water, ultrasonically for 30 minutes to form a silicon dioxide dispersion liquid, then put the cleaned medical sponge into the above solution, slowly squeeze and dip it 1-3 times and then take it out, and dry it.

[0013] Further, preferably: The concentration of the silicon dioxide dispersion liquid is 3-5 mg / mL.

[0014] Further, preferably: The specific composite enzyme loading treatment is as follows: Dissolve protease and glucoamylase in deionized water to form a composite enzyme solution, then evenly spray the composite enzyme solution on the soaked surface of the sponge after SiO2 modification treatment, and dry it. That's it.

[0015] Further, preferably: The protease content in the composite enzyme solution is 30-50 mg / L, and the glucoamylase content is 3-5 mg / L.

[0016] Further, preferably: The composite enzyme solution is spray-treated in an amount of 30 mL / m2.

[0017] Further, preferably: The clove oil solution is prepared by adding clove oil to a 70% ethanol solution to form a solution with a mass concentration of 0.5-1%; the clove oil solution is spray-treated in an amount of 10 mL / m2.

[0018] The present invention also provides a saliva sampling swab for low-absorption THC detection, which includes a sampling head or a sampling piece, and the sampling head or the sampling piece uses the medical sponge of the present invention.

[0019] Further, preferably, it includes a liquid-absorbing sponge head and an indicating tube. A test strip is arranged in the indicating tube. The liquid-absorbing sponge head is connected to the indicating tube. A notch is arranged at the rear end of the liquid-absorbing sponge head, and the front end of the test strip can be inserted into the notch. The rear end of the indicating tube is connected with a handle. A convex ring is arranged around the connection part of the handle and the indicating tube, and the handle is an airbag.

[0020] The beneficial effects of the present invention:

[0021] By subjecting the sponge to SiO2 loading treatment, the present invention effectively reduces the adsorption of THC by the PVA sponge, enabling more THC to enter the saliva and be collected. At the same time, the sponge is subjected to loading treatment with a composite enzyme and clove oil. The composite enzyme can effectively decompose the organic substances in the oral cavity and saliva, helping the THC adsorbed by the organic substances to be released into the water, and clove oil effectively improves the permeability of the oral mucosa, enabling the THC remaining in the oral mucosa to be released into the saliva. Through the above treatment, the collection of THC in the oral cavity can be effectively improved, thereby improving the detection sensitivity. Description of the Drawings

[0022] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0023] Figure 1 It is a schematic structural diagram of the saliva collection swab of the present invention. Detailed Embodiments

[0024] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the drawings in the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts belong to the scope of protection of the present invention.

[0025] A saliva sampling sponge for low-absorption THC detection, and the sampling sponge is made by the following steps:

[0026] (1)Sponge cleaning and impurity removal: Soak the medical sponge in ethanol and ultrasonically wash it twice, 10 minutes each time. Ultrasonic temperature: 25 - 30 °C, power: 150 W. Then rinse it with deionized water until there is no alcohol smell, and finally dry it at 60 °C;

[0027] (2)SiO2 modification treatment: Add nano - SiO2 (A200) to water, ultrasonically for 30 minutes. Ultrasonic temperature: 25 - 30 °C, power: 150 W, to form a silica dispersion liquid. Then put the cleaned medical sponge into the above solution, slowly squeeze and dip it twice and then take it out, and dry it at 60 °C. The concentration of the silica dispersion liquid is 4 mg / mL;

[0028] (3)Composite enzyme loading treatment: Spray - load the modified medical sponge in the composite enzyme solution. The composite enzyme is a complex of protease and glucoamylase. Specifically: Dissolve protease (papain, 1.5 - 2 million u / g) and glucoamylase (50,000 u / g) in deionized water to form a composite enzyme solution. Then evenly spray the composite enzyme solution on the surface of the sponge soaked after SiO2 modification treatment, and dry it. The protease content in the composite enzyme solution is 30 mg / L, and the glucoamylase content is 3 mg / L; The composite enzyme solution is sprayed according to the amount of 30 mL / m 2 for spraying treatment;

[0029] (4)Dry the medical sponge with the composite enzyme loaded at 40 °C;

[0030] (5)Clove oil loading treatment: Spray - load the medical sponge with the composite enzyme loaded in the clove oil solution and store it sealed. Specifically, the clove oil solution is formed by adding clove oil to a 70% ethanol solution to form a solution with a mass concentration of 0.8%; The clove oil solution is sprayed according to the amount of 10 mL / m 2 for spraying treatment.

[0031] A saliva sampling sponge for low - absorption THC detection, and the sampling sponge is made by the following steps:

[0032] (1)Sponge cleaning and impurity removal: Soak the medical sponge in ethanol and ultrasonically wash it twice, 10 minutes each time. Ultrasonic temperature: 25 - 30 °C, power: 150 W. Then rinse it with deionized water until there is no alcohol smell, and finally dry it at 60 °C;

[0033] (2)SiO2 modification treatment: Add nano - SiO2 (A200) to water, ultrasonically for 30 minutes. Ultrasonic temperature: 25 - 30 °C, power: 150 W, to form a silica dispersion liquid. Then put the cleaned medical sponge into the above solution, slowly squeeze and dip it twice and then take it out, and dry it at 60 °C. The concentration of the silica dispersion liquid is 3 mg / mL;

[0034] (3)Composite enzyme loading treatment: The modified medical sponge is subjected to spray loading treatment in the composite enzyme solution. The composite enzyme is a complex of protease and glucoamylase. Specifically: Protease (papain, 1.5 - 2 million u / g) and glucoamylase (50,000 u / g) are dissolved in deionized water to form a composite enzyme solution. Then the composite enzyme solution is evenly sprayed on the soaked surface of the sponge after SiO2 modification treatment and dried. The protease content in the composite enzyme solution is 30 mg / L, and the glucoamylase content is 4 mg / L; The composite enzyme solution is sprayed at a rate of 30 mL / m2;

[0035] (4)The medical sponge after composite enzyme loading treatment is dried at 40 °C;

[0036] (5)Clove oil loading treatment: The medical sponge after composite enzyme loading treatment is subjected to spray loading treatment with clove oil solution and sealed for storage. Specifically, the clove oil solution is prepared by adding clove oil to a 70% ethanol solution to form a solution with a mass concentration of 0.5%; The clove oil solution is sprayed at a rate of 10 mL / m2.

[0037] A saliva sampling sponge for low-absorption THC detection, and the sampling sponge is made by the following steps:

[0038] (1)Sponge cleaning and impurity removal: The medical sponge is soaked in ethanol and ultrasonically washed 2 times, 10 minutes each time. Ultrasonic temperature: 25 - 30 °C, power: 150 W. Then it is rinsed with deionized water until there is no alcohol smell, and finally dried at 60 °C;

[0039] (2)SiO2 modification treatment: Nano-SiO2 (A200) is added to water and ultrasonically treated for 30 minutes. Ultrasonic temperature: 25 - 30 °C, power: 150 W to form a silica dispersion. Then the cleaned medical sponge is put into the above solution, slowly squeezed and dipped 2 times and taken out, and dried at 60 °C. The concentration of the silica dispersion is 5 mg / mL;

[0040] (3)Composite enzyme loading treatment: The modified medical sponge is subjected to spray loading treatment in the composite enzyme solution. The composite enzyme is a complex of protease and glucoamylase. Specifically: Protease (papain, 1.5 - 2 million u / g) and glucoamylase (50,000 u / g) are dissolved in deionized water to form a composite enzyme solution. Then the composite enzyme solution is evenly sprayed on the soaked surface of the sponge after SiO2 modification treatment and dried. The protease content in the composite enzyme solution is 40 mg / L, and the glucoamylase content is 4 mg / L; The composite enzyme solution is sprayed at a rate of 30 mL / m 2 2;

[0041] (4)Drying the medical sponge after the composite enzyme loading at 40°C;

[0042] (5)Clove oil loading treatment: Spraying and loading the medical sponge after the composite enzyme loading treatment with the clove oil solution, and storing it sealed. Specifically, the clove oil solution is prepared by adding clove oil to a 70% ethanol solution to form a solution with a mass concentration of 0.6%; the clove oil solution is sprayed at a rate of 10 mL / m 2 for spraying treatment.

[0043] A saliva sampling sponge for low-absorption THC detection, and the sampling sponge is made by the following steps:

[0044] (1)Sponge cleaning and impurity removal: Soaking the medical sponge in ethanol and ultrasonically washing it twice, 10 minutes each time, ultrasonic temperature: 25 - 30°C, power: 150W, then rinsing it with deionized water until there is no alcohol smell, and finally drying it at 60°C;

[0045] (2)SiO2 modification treatment: Adding nano-SiO2 (A200) to water, ultrasonically treating for 30 minutes, ultrasonic temperature: 25 - 30°C, power: 150W, to form a silica dispersion liquid, then putting the cleaned medical sponge into the above solution, slowly squeezing and dipping it twice and then taking it out, drying it at 60°C, and the concentration of the silica dispersion liquid is 5mg / mL;

[0046] (3)Composite enzyme loading treatment: Spraying and loading the modified medical sponge with the composite enzyme solution. The composite enzyme is a complex of protease and glucoamylase. Specifically: Dissolving protease (papain, 1.5 - 2 million u / g) and glucoamylase (50,000 u / g) in deionized water to form a composite enzyme solution, then evenly spraying the composite enzyme solution on the surface of the sponge soaked after the SiO2 modification treatment, and drying it. The protease content in the composite enzyme solution is 50mg / L, and the glucoamylase content is 5mg / L; the composite enzyme solution is sprayed at a rate of 30 mL / m 2 for spraying treatment;

[0047] (4)Drying the medical sponge after the composite enzyme loading at 40°C;

[0048] (5)Clove oil loading treatment: Spraying and loading the medical sponge after the composite enzyme loading treatment with the clove oil solution, and storing it sealed. Specifically, the clove oil solution is prepared by adding clove oil to a 70% ethanol solution to form a solution with a mass concentration of 1%; the clove oil solution is sprayed at a rate of 10 mL / m 2 for spraying treatment.

[0049] Such as Figure 1As shown in the figure, a saliva sampling stick for low-absorption THC detection includes a sampling head or a sampling piece, and the sampling head or the sampling piece uses the medical sponge of the present invention. The structure of the specific saliva collection stick can be designed according to actual needs. One structure provided by the present invention is as follows:

[0050] A saliva sampling stick for low-absorption THC detection includes a liquid-absorbing sponge head 1 and an indicating tube 2. A test strip 3 is arranged in the indicating tube 2. The liquid-absorbing sponge head 1 is connected to the indicating tube 2. A notch is arranged at the rear end of the liquid-absorbing sponge head 1. The front end of the test strip 3 can be inserted into the notch. The rear end of the indicating tube 2 is connected with a handle 5. A convex ring 4 is arranged around the connection part of the handle 5 and the indicating tube 2. The handle is an airbag.

[0051] The saliva sampler of the present invention uses the modified sponge as the washing liquid sponge head, which can effectively improve the detection sensitivity and is conducive to the popularization and use of saliva THC detection.

[0052] In addition, the method of adding a test strip to the rear end of the liquid-absorbing sponge head of the present invention is convenient for observing the amount of saliva taken during the sampling process, so as to achieve quantitative sampling and improve the disease testing efficiency.

[0053] Comparative Examples 1-6

[0054] It is basically the same as Example 1, except that different loadings are used for loading the medical PVA sponge to investigate the influence of different loadings on the THC detection sensitivity. The specific loadings are shown in Table 1.

[0055] Table 1 Loading ratios of different comparative examples

[0056] SiO2 / (mg / mL) Protease / (50 mg / L) Glucoamylase / (5 mg / L) Clove oil / (%) Example 1 4 35 3 0.8 Comparative Example 1 0 0 0 0 Comparative Example 2 4 0 0 0 Comparative Example 3 0 35 0 0 Comparative Example 4 0 35 5 0 Comparative Example 5 0 0 0 0.8 Comparative Example 6 4 35 5 0 Comparative Example 7 4 0 0 0.8

[0057] Water absorption rate:

[0058] At room temperature, the dry sponge samples (diameter 12 mm, height 10 mm) prepared by different examples are immersed in distilled water. After the samples are left standing and swollen for 15 min, the excess liquid on the surface is removed with filter paper and their weights are measured. The calculation formula for the water absorption rate of the sponge is as follows:

[0059] Water absorption rate (%) = (weight of the sponge after water absorption - weight of the sponge before water absorption) / weight of the sponge before water absorption × 100.

[0060] The specific results are shown in Table 2.

[0061] Table 2 Measurement results of the absorption rates of the sponges prepared by different examples

[0062] Example 1 Example 2 Example 3 Example 4 Water absorption rate / % 326 331 301 298 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Comparative Example 5 Comparative Example 6 Comparative Example 7 Water absorption rate / % 368 325 352 348 365 316 321

[0063] As can be seen from Table 1, by mixing nanoparticles of silicon dioxide with a sponge, a composite material is formed, making its pores smaller or more irregular, improving the hydrophobicity of the sponge and reducing the adsorption of oil molecules. However, the overall reduction is not significant. Compared with Comparative Example 1, the water absorption rate of the modified sponge decreased by 10.1% - 19.0%, which has little impact on the overall water absorption rate of the sponge.

[0064] THC detection sensitivity

[0065] Prepare a THC solution of 20 - 100 ng using artificial saliva (Saliva juice, pH 6.8), and then add 50 mg of porcine small intestine mucosal powder per 100 mL and mix evenly for standby. Preparation method of porcine small intestine mucosal powder: After the porcine small intestine is washed with distilled water, scrape the inner wall mucosa, blot the water with absorbent paper, and vacuum freeze-dry it (temperature: -35 to -50 °C, time: 24 h) to form a small intestine mucosal powder with a water content of less than 5%.

[0066] Immerse the sampling stick of the present invention (the absorbent sponge head has a diameter of 12.5 mm and a height of 10 mm) in 10 mL of the above solution for 5 min, then take it out, collect the liquid of the absorbent sponge head, and use a THC colloidal gold immunochromatographic test strip with a detection sensitivity of 20 ng for determination. Add 200 μL of the solution to the micropore of the THC colloidal gold immunochromatographic test strip and observe after reacting for 5 - 10 min. When the color of the T line is lighter than the G2 line or the T line does not appear, it is positive. See Table 3 for details.

[0067] Blank control: Prepare a 20 - 100 ng / mL THC standard working solution using a chromatography solution of 0.01 mol / L PBS solution (pH = 7.4) containing 10% TX-100.

[0068] Add 200 μL of the standard working solution to the micropore of the THC colloidal gold immunochromatographic test strip and observe after reacting for 5 - 10 min.

[0069] Table 3 Test strip detection results of sponges prepared in different examples

[0070] 20 ng / mL 30 ng / mL 40 ng / mL 50 ng / mL 60 ng / mL 70 ng / mL 80 ng / mL 90 ng / mL 100 ng / mL Blank control + + + + + + + + + Example 1 — + + + + + + + + Example 2 — + + + + + + + + Example 3 — + + + + + + + + Example 4 — — + + + + + + + Comparative Example 1 — — — — — — + + + Comparative Example 2 — — — — + + + + + Comparative Example 3 — — — — — + + + + Comparative Example 4 — — — — — + + + + Comparative Example 5 — — — + + + + + + Comparative Example 6 — — — + + + + + + Comparative Example 7 — — — + + + + + +

[0071] In the embodiments of the present invention, porcine small intestinal mucosa powder is used to simulate the oral mucosa, and artificial saliva (Salivajuice, pH 6.8) and porcine small intestinal mucosa powder are used to simulate the environment of THC in the oral cavity. As can be seen from Table 3, the present invention uses a composite enzyme to enzymatically hydrolyze the organic matter in the oral cavity, effectively reducing its adhesiveness, so that more of it is collected by the sponge. The use of clove oil can effectively improve the permeability of the oral mucosa, enabling more THC remaining in the oral mucosa to dissolve with saliva and be collected. The treatment with silica can effectively reduce the lipophilicity of the sponge, causing the THC collected by the sponge to be collected into the saliva with saliva. Through the above treatments, the collection of THC remaining in the oral cavity can be effectively improved, thereby improving the sensitivity of saliva THC detection. Compared with Comparative Example 1 without modification treatment, the detection sensitivity of the present invention is increased from 80 ng / mL to 30 ng / mL, greatly improving the sensitivity of saliva THC detection, which is conducive to the popularization and utilization of saliva THC detection.

[0072] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A saliva sampling sponge for low-absorption THC detection, characterized in that, The sampling sponge described above is made by the following steps: (1) Sponge cleaning and impurity removal: Clean and remove impurities from the medical sponge; (2) SiO2 modification treatment: Modify the cleaned medical sponge with silicon dioxide; (3) Composite enzyme loading treatment: Spray and load the modified medical sponge with a composite enzyme solution. The composite enzyme is a complex of protease and glucoamylase; (4) Low-temperature drying of the medical sponge after composite enzyme loading treatment; (5) Clove oil loading treatment: Spray and load the medical sponge after composite enzyme loading treatment with a clove oil solution, seal and store at low temperature, then it is ready; The specific SiO2 modification treatment is to add nano-SiO2 into water, ultrasonicate for 30 min to form a silicon dioxide dispersion liquid, then put the cleaned medical sponge into the above solution, slowly squeeze and dip it 1 - 3 times and then take it out and dry it; the concentration of the silicon dioxide dispersion liquid is 3 - 5 mg / mL; The specific composite enzyme loading treatment is: dissolve protease and glucoamylase in deionized water to form a composite enzyme solution, then evenly spray the composite enzyme solution on the soaked surface of the sponge after SiO2 modification treatment, and dry it, then it is ready.

2. The saliva sampling sponge for low-absorption THC detection according to claim 1, wherein: The specific sponge cleaning is: soak the medical sponge in ethanol and ultrasonically wash it 1 - 2 times, 10 min each time, then rinse it with deionized water until there is no alcohol smell, and dry it for standby.

3. A saliva sampling sponge for low-absorption THC detection according to claim 1, characterized in that: The protease content in the composite enzyme solution is 30 - 50 mg / L, and the glucoamylase content is 3 - 5 mg / L.

4. A saliva sampling sponge for low-absorption THC detection according to claim 3, characterized in that: The described complex enzyme solution is spray-treated in an amount of 30 mL / m 2 .

5. A saliva sampling sponge for low-absorption THC detection according to claim 1, characterized in that: The clove oil solution is prepared by adding clove oil to a 70% ethanol solution to form a solution with a mass concentration of 0.5 - 1%; the clove oil solution is spray-treated in an amount of 10 mL / m 2 of the amount.

6. A saliva sampling swab for low-absorption THC detection, characterized in that: It includes a sampling head or a sampling piece, and the sampling head or the sampling piece uses the medical sponge described in any one of claims 1 - 5.

7. The saliva sampling swab for low-absorption THC detection according to claim 6, characterized in that: The saliva sampling stick for low-absorption THC detection described above includes a liquid-absorbing sponge head and an indicating tube. A test strip is arranged in the indicating tube. The liquid-absorbing sponge head is connected to the indicating tube. A notch is arranged at the rear end of the liquid-absorbing sponge head. The front end of the test strip can be inserted into the notch; a handle is connected to the rear end of the indicating tube. A convex ring is arranged around the connection part of the handle and the indicating tube. The handle is an airbag.

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