Saliva sampling sponge for low-absorption THC detection and sampling rod of saliva sampling sponge

Abstract

The invention relates to a low-absorption THC detection saliva sampling sponge and a sampling rod thereof. The sampling sponge is prepared by the following steps: (1) sponge cleaning and impurity removal: cleaning a medical sponge to remove impurities; (2) SiO2 modification treatment: carrying out modification treatment on the cleaned medical sponge by adopting silicon dioxide; (3) carrying out compound enzyme loading treatment: carrying out spray loading treatment on the modified medical sponge in compound enzyme liquid, wherein the compound enzyme is a compound of protease and saccharifying enzyme; (4) performing low-temperature drying on the medical sponge subjected to the compound enzyme loading treatment; and (5) carrying out clove oil loading treatment: carrying out spray loading treatment on the medical sponge subjected to compound enzyme loading treatment by using a clove oil solution, and sealing and storing at low temperature. According to the method, SiO2 loading treatment is carried out on the sponge, and loading treatment is carried out on the sponge through the compound enzyme and the clove oil, so that the collection of THC in the oral cavity can be effectively improved, and the detection sensitivity is further improved.

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 detect evidence of marijuana use. Blood and urine are widely used test materials for marijuana analysis. The main target analyte of blood testing is THC-COOH, which can only detect a small amount of THC and can only provide short-term abuse information, including behavioral impairment and degree of intoxication, but the collection is invasive, difficult to monitor on the roadside, and requires laboratory analysis. The main target analyte of urine testing is THC-COOH, and its concentration is mainly affected by factors such as drug dosage, abuse method, frequency of abuse, and individual differences in drug metabolism and excretion rates. Urine testing has the advantages of a wide range, a wide detection window (within one week), and low testing costs, but collection requires special places and staff of the same gender, and is easy to falsify. Saliva is a new test material developed in recent years and has been widely used in the field of drunk driving and drug driving cases involving traffic safety. Its main target analyte is THC, and its metabolites are relatively small. Compared with traditional methods of collecting samples (such as blood and urine testing), 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 testee to accept and is not restricted by venue and gender. However, compared with urine and blood testing, the existing saliva THC test has a lower detection sensitivity. The main reasons are as follows: (1) The collection of saliva has an important impact on saliva drug detection. Currently, medical-grade PVA sponge is generally used as a collection carrier. Medical-grade PVA sponge is a polymer material with excellent liquid absorption performance and extremely soft surface when in contact with the human body. It can be completely naturally degraded and is very suitable for the collection of saliva samples. However, the hydrophobic groups in PVA molecules, such as vinyl (-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 the interaction between PVA and 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, making it difficult for THC in the mouth to diffuse into the moisture in the saliva and be collected. Summary of the invention

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

[0004] In order to solve the above technical problems, the technical solution adopted by the present invention is: A saliva sampling sponge for low-absorption THC detection, wherein the sampling sponge is prepared by the following steps: (1) Cleaning and removing impurities from the sponge: Clean and remove impurities from the medical sponge; (2) SiO2 modification: The cleaned medical sponge is modified with silicon dioxide; (3) Composite enzyme loading treatment: spraying the modified medical sponge with a composite enzyme solution, wherein the composite enzyme is a complex of protease and saccharifying enzyme; (4) Low-temperature drying of the medical sponge treated with complex enzyme loading; (5) Clove oil loading treatment: spray the medical sponge loaded with the complex enzyme with a clove oil solution and seal it for storage.

[0005] Further, preferably, the sponge cleaning is specifically performed by soaking the medical sponge in ethanol and ultrasonically washing it 1-2 times, each time for 10 minutes, then rinsing it with deionized water until there is no alcohol smell, and finally drying it for later use.

[0006] Further, preferably: the SiO2 modification treatment is specifically adding nano-SiO2 into water, ultrasonicating for 30 minutes to form a silicon dioxide dispersion, then placing the cleaned medical sponge into the above solution, slowly squeezing and dipping it 1-3 times, then taking it out and drying it.

[0007] Furthermore, preferably, the concentration of the silicon dioxide dispersion is 3-5 mg / mL.

[0008] Further, preferably: the complex enzyme loading treatment is specifically: dissolving the protease and saccharifying enzyme in deionized water to form a complex enzyme solution, and then evenly spraying the complex enzyme solution on the surface of the sponge soaked after SiO2 modification, and drying.

[0009] Furthermore, preferably, the protease content in the complex enzyme solution is 30-50 mg / L, and the saccharifying enzyme content is 3-5 mg / L.

[0010] Furthermore, preferably: the complex enzyme solution is sprayed at a volume of 30 mL / m2.

[0011] Further, preferably: the clove oil solution is prepared by adding clove oil to 70% ethanol solution to form a solution with a mass concentration of 0.5-1%; the clove oil solution is sprayed at an amount of 10 mL / m2.

[0012] The present invention also provides a saliva sampling stick for low-absorption THC detection, comprising a sampling head or a sampling sheet, wherein the sampling head or the sampling sheet adopts the medical sponge of the present invention.

[0013] Further, preferably: it includes a liquid-absorbing sponge head and an indicator tube, a test strip is arranged in the indicator tube, the liquid-absorbing sponge head is connected to the indicator 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 indicator tube is connected to a handle, a convex ring is arranged on the periphery of the connection between the handle and the indicator tube, and the handle is an air bag.

[0014] Beneficial effects of the present invention: The present invention uses SiO2 loading treatment on the sponge to effectively reduce the adsorption of THC by the PVA sponge, so that more THC enters the saliva and is collected; at the same time, the sponge is loaded with a composite enzyme and clove oil, the composite enzyme can effectively decompose organic matter in the oral cavity and saliva, and is helpful for releasing THC adsorbed by the organic matter into water, and the clove oil effectively improves the permeability of the oral mucosa, so that THC retained in the oral mucosa is 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. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0016] Figure 1 It is a schematic structural diagram of the saliva collection stick of the present invention. DETAILED DESCRIPTION

[0017] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only 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.

[0018] A saliva sampling sponge for low-absorption THC detection, wherein the sampling sponge is prepared by the following steps: (1) Sponge cleaning and impurity removal: Soak the medical sponge in ethanol and ultrasonically clean it twice, each time for 10 min, with an ultrasonic temperature of 25-30°C and a power of 150W. Then rinse it with deionized water until there is no alcohol smell, and finally dry it at 60°C. (2) SiO2 modification treatment: Add nano-SiO2 (A200) into water, perform ultrasound for 30 min, ultrasound temperature: 25-30°C, power: 150W, to form a silica dispersion, then put the cleaned medical sponge into the above solution, slowly squeeze and dip it twice, then take it out and dry it at 60°C. The concentration of the silica dispersion is 4 mg / mL; (3) Composite enzyme loading treatment: The modified medical sponge is sprayed with a composite enzyme solution, wherein the composite enzyme is a complex of protease and saccharifying enzyme. Specifically, the protease (papain, 1.5-2 million u / g) and saccharifying enzyme (50,000 u / g) are dissolved in deionized water to form a composite enzyme solution, and then the composite enzyme solution is evenly sprayed on the surface of the sponge after SiO2 modification and dried. The content of protease in the composite enzyme solution is 30 mg / L, and the content of saccharifying enzyme is 3 mg / L. The composite enzyme solution is heated at 30 mL / m 2 The amount of spray treatment; (4) Drying the medical sponge treated with complex enzyme loading at 40°C; (5) Clove oil loading treatment: The medical sponge after the complex enzyme loading treatment is sprayed with a 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 0.8%; the clove oil solution is heated at 10 mL / m 2 The amount was sprayed.

[0019] A saliva sampling sponge for low-absorption THC detection, wherein the sampling sponge is prepared by the following steps: (1) Sponge cleaning and impurity removal: Soak the medical sponge in ethanol and ultrasonically clean it twice, each time for 10 min, with an ultrasonic temperature of 25-30°C and a power of 150W. Then rinse it with deionized water until there is no alcohol smell, and finally dry it at 60°C. (2) SiO2 modification treatment: Add nano-SiO2 (A200) into water, perform ultrasound for 30 min, ultrasound temperature: 25-30°C, power: 150W, to form a silica dispersion, then put the cleaned medical sponge into the above solution, slowly squeeze and dip it twice, then take it out and dry it at 60°C. The concentration of the silica dispersion is 3 mg / mL; (3) Composite enzyme loading treatment: The modified medical sponge is sprayed with a composite enzyme solution, wherein the composite enzyme is a complex of protease and saccharifying enzyme. Specifically, protease (papain, 1.5-2 million u / g) and saccharifying enzyme (50,000 u / g) are dissolved in deionized water to form a composite enzyme solution, and then the composite enzyme solution is evenly sprayed on the surface of the sponge after SiO2 modification and dried. The content of protease in the composite enzyme solution is 30 mg / L, and the content of saccharifying enzyme is 4 mg / L. The composite enzyme solution is sprayed at a volume of 30 mL / m2. (4) Drying the medical sponge treated with complex enzyme loading at 40°C; (5) Clove oil loading treatment: The medical sponge after the complex enzyme loading treatment is sprayed with a 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 volume of 10 mL / m2.

[0020] A saliva sampling sponge for low-absorption THC detection, wherein the sampling sponge is prepared by the following steps: (1) Sponge cleaning and impurity removal: Soak the medical sponge in ethanol and ultrasonically clean it twice, each time for 10 min, with an ultrasonic temperature of 25-30°C and a power of 150W. Then rinse it with deionized water until there is no alcohol smell, and finally dry it at 60°C. (2) SiO2 modification treatment: Add nano-SiO2 (A200) into water, perform ultrasound for 30 min, ultrasound temperature: 25-30°C, power: 150W, to form a silica dispersion, then put the cleaned medical sponge into the above solution, slowly squeeze and dip it twice, then take it out and dry it at 60°C. The concentration of the silica dispersion is 5 mg / mL; (3) Composite enzyme loading treatment: The modified medical sponge is sprayed with a composite enzyme solution. The composite enzyme is a complex of protease and saccharifying enzyme. Specifically, the protease (papain, 1.5-2 million u / g) and saccharifying enzyme (50,000 u / g) are dissolved in deionized water to form a composite enzyme solution. The composite enzyme solution is then evenly sprayed on the surface of the sponge after SiO2 modification and dried. The content of protease in the composite enzyme solution is 40 mg / L, and the content of saccharifying enzyme is 4 mg / L. The composite enzyme solution is heated at 30 mL / m 2 The amount of spray treatment; (4) Drying the medical sponge treated with complex enzyme loading at 40°C; (5) Clove oil loading treatment: The medical sponge after the complex enzyme loading treatment is sprayed with a 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 0.6%; the clove oil solution is heated at 10 mL / m 2 The amount was sprayed.

[0021] A saliva sampling sponge for low-absorption THC detection, wherein the sampling sponge is prepared by the following steps: (1) Sponge cleaning and impurity removal: Soak the medical sponge in ethanol and ultrasonically clean it twice, each time for 10 min, with an ultrasonic temperature of 25-30°C and a power of 150W. Then rinse it with deionized water until there is no alcohol smell, and finally dry it at 60°C. (2) SiO2 modification treatment: Add nano-SiO2 (A200) into water, perform ultrasound for 30 min, ultrasound temperature: 25-30°C, power: 150W, to form a silica dispersion, then put the cleaned medical sponge into the above solution, slowly squeeze and dip it twice, then take it out and dry it at 60°C. The concentration of the silica dispersion is 5 mg / mL; (3) Composite enzyme loading treatment: The modified medical sponge is sprayed with a composite enzyme solution, wherein the composite enzyme is a complex of protease and saccharifying enzyme. Specifically, the protease (papain, 1.5-2 million u / g) and saccharifying enzyme (50,000 u / g) are dissolved in deionized water to form a composite enzyme solution, and then the composite enzyme solution is evenly sprayed on the surface of the sponge after SiO2 modification and dried. The content of protease in the composite enzyme solution is 50 mg / L, and the content of saccharifying enzyme is 5 mg / L. The composite enzyme solution is heated at 30 mL / m 2 The amount of spray treatment; (4) Drying the medical sponge treated with complex enzyme loading at 40°C; (5) Clove oil loading treatment: The medical sponge after the composite enzyme loading treatment is sprayed with a 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 1%; the clove oil solution is heated at 10 mL / m 2 The amount was sprayed.

[0022] like Figure 1 As shown, a saliva sampling stick for low absorption THC detection includes a sampling head or a sampling sheet, and the sampling head or the sampling sheet adopts the medical sponge of the present invention. The specific structure of the saliva collection stick can be designed according to actual needs. One structure provided by the present invention is: A saliva sampling stick for low-absorption THC detection comprises a liquid-absorbing sponge head 1 and an indicator tube 2, wherein a test strip 3 is arranged in the indicator tube 2, the liquid-absorbing sponge head 1 is connected to the indicator tube 2, a notch is arranged at the rear end of the liquid-absorbing sponge head 1, and the front end of the test strip 3 can be inserted into the notch; a handle 5 is connected to the rear end of the indicator tube 2, a convex ring 4 is arranged at the periphery of the connection between the handle 5 and the indicator tube 2, and the handle is an air bag.

[0023] The saliva sampler of the present invention adopts the modified sponge as the washing liquid sponge head, which can effectively improve the detection sensitivity and is conducive to the promotion and use of saliva THC detection.

[0024] In addition, the method of adding a test strip to the rear end of the liquid-absorbing sponge head of the present invention facilitates the observation of the amount of saliva collected during the sampling process, thereby achieving quantitative sampling and improving the efficiency of disease testing.

[0025] Comparative Examples 1-6 The method is basically the same as Example 1, except that: different loads are used to load the medical PVA sponge to investigate the effects of different loads on the THC detection sensitivity. The specific loads are shown in Table 1.

[0026] Table 1 Loading ratios of different comparison ratios SiO2 / (mg / mL) Protease / (50mg / L) Glucoamylase / (5mg / 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 Water absorption: At room temperature, soak the pre-weighed dry sponge samples (12 mm in diameter, 10 mm in height) prepared by different strengths in distilled water. After the sample was allowed to swell for 15 min, the excess liquid on the surface was removed with filter paper and weighed. The formula for calculating the water absorption rate of the sponge is as follows: Water absorption rate (%) = (sponge weight after water absorption - sponge weight before water absorption) / sponge weight before water absorption × 100.

[0027] The specific results are shown in Table 2.

[0028] Table 2 Absorption rate measurement results of sponges prepared in different embodiments Example 1 Example 2 Example 3 Example 4 Water absorption / % 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 / % 368 325 352 348 365 316 321 As shown in Table 1, the nanoparticle silica is mixed with the sponge to form a composite material, which makes the pores smaller or more irregular, improves the hydrophobicity of the sponge, and reduces the adsorption of oil molecules. However, the overall reduction is not high. Compared with Comparative Example 1, the water absorption rate of the modified sponge is reduced by 10.1%-19.0%, which has little effect on the water absorption rate of the overall sponge.

[0029] THC Detection Sensitivity Use artificial saliva (Saliva juice, pH 6.8) to prepare 20-100ng THC solution, then add 50mg porcine small intestinal mucosa powder per 100mL, mix evenly, and set aside. Preparation method of porcine small intestinal mucosa powder: After washing the porcine small intestine with distilled water, scrape the inner wall mucosa, absorb the water with absorbent paper, and vacuum freeze-dry it (temperature: -35~-50℃, time: 24h) to form small intestinal mucosa powder with a moisture content of less than 5%.

[0030] The sampling stick of the present invention (sponge head with a diameter of 12.5 mm and a height of 10 mm) was immersed in 10 mL of the above solution for 5 min, then taken out, the liquid in the sponge head was collected, and the THC colloidal gold immunochromatographic test strip with a detection sensitivity of 20 ng was used for determination. 200 μL of the solution was added to the micropores of the THC colloidal gold immunochromatographic test strip, and observed after 5 to 10 min of reaction. When the color of the T line was lighter than the G2 line or no T line appeared, it was positive. See Table 3 for details.

[0031] Blank control: Prepare 20-100 ng / mL THC standard working solution using 0.01 mol / L PBS solution (pH = 7.4) containing 10% TX-100.

[0032] Take 200 μL of standard working solution and add it to the micropores of THC colloidal gold immunochromatographic test strips, and observe after reacting for 5 to 10 minutes.

[0033] Table 3 Test strip test results of sponges prepared in different embodiments 20 ng / mL 30 ng / mL 40 ng / mL 50ng / mL 60ng / mL 70 ng / mL 80ng / mL 90ng / mL 100ng / 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 — — — + + + + + + In the embodiment of the present invention, pig small intestine mucosa powder is used to simulate oral mucosa, and artificial saliva (Salivajuice, pH 6.8) and pig small intestine mucosa powder are used to simulate the environment of THC in the oral cavity. As shown in Table 3, the present invention uses a composite enzyme to enzymolyze organic matter in the oral cavity, effectively reducing its adhesion so that it is more collected by sponges, and clove oil can effectively improve the permeability of the oral mucosa, so that THC retained in the oral mucosa is more dissolved with saliva and collected; silicon dioxide treatment can effectively reduce the lipophilicity of the sponge, so that the THC collected by the sponge is collected in the saliva with saliva, and through the above treatment, the collection of THC residual in the oral cavity can be effectively improved, and then the sensitivity of saliva THC detection is improved. Compared with the comparative example 1 without modification, the detection sensitivity of the present invention is increased from 80ng / mL to 30ng / mL, which greatly improves the sensitivity of saliva THC detection, which is conducive to the promotion and utilization of saliva THC detection.

[0034] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should 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 is prepared by the following steps: (1) Cleaning and removing impurities from the sponge: Clean and remove impurities from the medical sponge; (2) SiO2 modification: The cleaned medical sponge is modified with silicon dioxide; (3) Composite enzyme loading treatment: spraying the modified medical sponge with a composite enzyme solution, wherein the composite enzyme is a complex of protease and saccharifying enzyme; (4) Low-temperature drying of the medical sponge treated with complex enzyme loading; (5) Clove oil loading treatment: spray the medical sponge loaded with the complex enzyme with a clove oil solution, seal it and store it at low temperature.

2. A saliva sampling sponge for low absorption THC detection according to claim 1, characterized in that: The sponge cleaning is specifically as follows: the medical sponge is soaked in ethanol and ultrasonically washed 1-2 times, each time for 10 minutes, then rinsed with deionized water until there is no alcohol smell, and dried for later use.

3. A saliva sampling sponge for low absorption THC detection according to claim 1, characterized in that: The SiO2 modification treatment is specifically to add nano-SiO2 into water, ultrasonicate for 30 minutes to form a silicon dioxide dispersion, then put the cleaned medical sponge into the above solution, slowly squeeze and dip it 1-3 times, then take it out and dry it.

4. A saliva sampling sponge for low absorption THC detection according to claim 3, characterized in that: The concentration of the silicon dioxide dispersion is 3-5 mg / mL.

5. The saliva sampling sponge for low absorption THC detection according to claim 1, characterized in that: The composite enzyme loading treatment is specifically as follows: dissolving protease and saccharifying enzyme in deionized water to form a composite enzyme solution, and then evenly spraying the composite enzyme solution on the surface of the sponge soaked after SiO2 modification treatment, and drying.

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

7. A saliva sampling sponge for low absorption THC detection according to claim 6, characterized in that: The complex enzyme solution was prepared at 30 mL / min. 2 The amount was sprayed.

8. The 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 prepared by mixing at 10 mL / m 2 The amount was sprayed.

9. A saliva sampling stick for low absorption THC detection, characterized in that: It comprises a sampling head or a sampling sheet, wherein the sampling head or the sampling sheet adopts the medical sponge described in any one of claims 1-7.

10. The saliva sampling stick according to claim 9, characterized in that: It comprises a liquid-absorbing sponge head and an indicator tube, wherein a test strip is arranged in the indicator tube, the liquid-absorbing sponge head is connected to the indicator 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; a handle is connected to the rear end of the indicator tube, a convex ring is arranged at the periphery of the connection between the handle and the indicator tube, and the handle is an air bag.

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