Test kit and testing method

Abstract

This invention provides a test kit capable of selectively determining whether or not a sample contains Δ9-THC. [Solution] The test kit comprises a first reagent containing 2-hydroxybenzaldehyde and a water-soluble organic solvent capable of dissolving 2-hydroxybenzaldehyde, a second reagent which is hydrochloric acid, and a third reagent which contains an organic solvent that has a higher specific gravity than water and separates from water.

Description

[Technical Field]

[0001] This disclosure relates to a test kit and a test method. [Background technology]

[0002] Cannabis is one of the most widely abused drugs in the world. In Japan, cannabis is classified as a narcotic under the "Narcotics and Psychotropic Substances Control Act," its cultivation is regulated under the "Act on the Regulation of Cannabis Cultivation," and its possession is regulated under the same act.

[0003] Cannabis contains unique compounds collectively known as cannabinoids. The three main cannabinoids in cannabis are Δ9-tetrahydrocannabinol (Δ9-THC), cannabidiol (CBD), and cannabinol (CBN), with other known compounds including cannabigerol (CBG). Δ9-THC is biosynthesized in the plant as its acid form, Δ9-tetrahydrocannabinolic acid (Δ9-THCA), and is gradually decarboxylated to Δ9-THC. Similarly, CBD is biosynthesized in the plant as its acid form, cannabidiolic acid (CBDA), and is gradually decarboxylated to CBD.

[0004] Of the cannabinoids in cannabis, the main psychoactive component is Δ9-THC, which is classified as a narcotic under the "Narcotics and Psychotropic Substances Control Law." On the other hand, CBD, CBN, and CBG are not classified as narcotics. CBD, CBN, and CBG are said to have relaxing and analgesic effects, and e-cigarette liquids, foods, oils, and non-cannabis plant fragments containing them are sold on online shopping sites and in stores.

[0005] As described in Non-Patent Document 1, cannabis includes drug species with a high Δ9-THC content after decarboxylation and fiber species with a low Δ9-THC content and a high CBD content. Legal regulations regarding cannabis vary from country to country. For example, in the United States, cannabis with a total amount of Δ9-THC and Δ9-THCA (converted to Δ9-THC after decarboxylation) of 0.3% or less is excluded from the list of controlled substances. In contrast, in Japan, cannabis is treated as a narcotic regardless of its Δ9-THC content.

[0006] At crime scenes, plant fragments suspected of being cannabis undergo preliminary testing using a crime scene kit with a colorimetric reagent. Currently in Japan, the colorimetric reagent used for crime scene testing of cannabis is the Duquenois-Levine reagent, while in some countries overseas, the 4-aminophenol reagent is also used in addition to it.

[0007] As described in Non-Patent Document 1, the Duquenois-Levine reagent consists of three solutions: the first reagent: an ethanol solution of acetaldehyde and vanillin, the second reagent: concentrated hydrochloric acid, and the third reagent: chloroform. Also, as described in Non-Patent Document 1, if a small amount of sample is extracted with the first reagent, the second reagent is added to the extract and mixed, and then the third reagent is added, and the lower layer after extraction turns purple, the sample is presumed to be cannabis. [Prior art documents] [Non-patent literature]

[0008] [Non-Patent Document 1] "Recommended Methods for the IDENTIFICATION AND ANALYSIS OF CANNABIS AND CANNABIS PRODUCTS", United Nations Office on Drugs and Crime, Vienna, 2022. [Non-Patent Document 2] Journal of Forensic Sciences, vol. 23, 1978, p. 304-310 [Non-Patent Document 3] Journal of Forensic Sciences, vol. 66, 2021, p. 285-294 [Non-Patent Document 4] Forensic Toxicology, vol. 35, 2017, p. 185-189 [Overview of the project] [Problems that the invention aims to solve]

[0009] Non-patent document 2 reports that when Δ9-THC, CBD, CBG, and CBN were tested using the Duquenois-Levine reagent, the color of the lower layer after extraction of the third reagent was purple for Δ9-THC, CBD, and CBG, and blue for CBN. In other words, the Duquenois-Levine reagent cannot distinguish between Δ9-THC and CBD and CBG, and distinguishing between Δ9-THC and CBN is difficult because their color tones are similar.

[0010] As described in Non-Patent Document 3, the 4-aminophenol reagent consists of two solutions: a first reagent, a solution prepared by dissolving 4-aminophenol in ethanol and then adding 2M hydrochloric acid; and a second reagent, an aqueous ethanol solution of sodium hydroxide. Non-Patent Document 3 also reports that Δ9-THC, Δ9-THCA, CBN, and oregano (a type of herb) turned blue against the 4-aminophenol reagent, while CBD and CBG turned pink. In other words, the 4-aminophenol reagent can distinguish between Δ9-THC and CBD and CBG, but it cannot distinguish between Δ9-THC and CBN. Therefore, even if the Duquenois-Levine reagent and the 4-aminophenol reagent are used in combination, it is difficult to distinguish between Δ9-THC and CBN.

[0011] In Japan, the presence or absence of Δ9-THC is not a legal requirement for classifying a suspected sample as cannabis. On the other hand, plant fragments that are not cannabis but have CBD, CBN, and CBG added are circulating. From the perspective of preventing wrongful arrest, the test reagents used in investigation kits must be reagents that selectively produce a color change for Δ9-THC. As stated above, at present, there are no test reagents that can selectively determine whether or not a sample contains Δ9-THC.

[0012] This disclosure has been made in view of the problems of the prior art. The purpose of this disclosure is to provide a test kit and a test method that can selectively determine whether or not a sample contains Δ9-THC. [Means for solving the problem]

[0013] A test kit according to a first aspect of this disclosure comprises a first reagent containing 2-hydroxybenzaldehyde and a water-soluble organic solvent capable of dissolving 2-hydroxybenzaldehyde. The test kit comprises a second reagent which is hydrochloric acid. The test kit comprises a third reagent which contains an organic solvent having a specific gravity greater than water and separating from water.

[0014] A testing method according to a second aspect of this disclosure includes the step of preparing a first mixture by mixing a sample with a first reagent containing a water-soluble organic solvent in which 2-hydroxybenzaldehyde is dissolved. The testing method includes the step of preparing a second mixture by mixing the liquid contained in the first mixture with a second reagent which is hydrochloric acid. The testing method includes the step of mixing the second mixture with a third reagent which contains an organic solvent that has a specific gravity greater than water and separates from water. [Effects of the Invention]

[0015] This disclosure provides a test kit and a test method that can selectively determine whether or not a sample contains Δ9-THC. [Brief explanation of the drawing]

[0016] [Figure 1] It is the chemical structure of the coloring substance presumed to be generated by the reaction of the Duquenois-Levine reagent and Δ9-THC. [Figure 2] It is a photograph showing the color development after the addition of the second reagent and the color development after the addition of the third reagent when 0.25 mg each of Δ9-THC, CBD, CBN, Δ8-tetrahydrocannabinol (Δ8-THC), and CBG were examined in Experimental Example 1. [Figure 3] It is a photograph showing the color development after the addition of the second reagent and the color development after the addition of the third reagent when 0.25 mg each of Δ9-THC, CBD, CBN, Δ8-THC, and CBG were examined in Experimental Example 2. [Figure 4] It is a photograph showing the color development after the addition of the second reagent and the color development after the addition of the third reagent when 0.25 mg each of Δ9-THC, CBD, CBN, Δ8-THC, and CBG were examined in Experimental Example 3. [Figure 5] It is a photograph showing the color development after the addition of the second reagent and the color development after the addition of the third reagent when 1 mg each of Δ9-THC, CBD, CBN, Δ8-THC, and CBG and 0.1 mg of Δ9-THCA were examined in Experimental Example 4. [Figure 6] It is a photograph showing the color development after the addition of the third reagent when 0.25 mg of Δ9-THC was examined using an acetonitrile solution containing 2-hydroxybenzaldehyde as the first reagent at concentrations of 2, 5, 10, 20, 50, and 100 mg / mL in Experimental Example 5. [Figure 7] It is a photograph showing the color development after the addition of the third reagent when 0.25 mg of Δ9-THC was examined using concentrated hydrochloric acid, 4 mol / L phosphoric acid, and 6 mol / L sulfuric acid as the second reagent in Experimental Example 6. [Figure 8] It is a photograph showing the color development after the addition of the third reagent when 0.25 mg of Δ9-THC was examined using concentrated hydrochloric acid, 10 mol / L hydrochloric acid, 9 mol / L hydrochloric acid, 8 mol / L hydrochloric acid, and 7 mol / L hydrochloric acid as the second reagent in Experimental Example 7. [Figure 9]This is a photograph showing the color change after adding the third reagent when 0.25 mg of Δ9-THC was tested using chloroform, dichloromethane, 1,2-dichloroethane, ethyl acetate, and n-hexane as the third reagent in Experimental Example 8. [Figure 10] This is a photograph showing the color of the extract obtained when freeze-dried and pulverized cannabis was extracted with acetonitrile, ethanol, and methanol in Experimental Example 9. [Figure 11] This photograph shows the color change after adding the second reagent when testing with 1 mg each of Δ9-THC, CBD, CBN, Δ8-THC, and CBG in Experimental Example 10. [Figure 12] This photograph shows the color change after adding the third reagent when testing with 1 mg each of Δ9-THC, CBD, CBN, Δ8-THC, and CBG in Experimental Example 11. [Figure 13] This photograph shows the color changes after adding the third reagent when Δ9-THC (1, 0.25, 0.1, and 0.025 mg) and CBD (2.5, 1, 0.25, and 0.1 mg) were tested in Experimental Example 12. [Figure 14] This is a photograph showing the color change after adding the third reagent when freeze-dried and crushed cannabis samples 1-28 were tested in Experimental Example 13. [Figure 15] This is a photograph showing the color change after adding the third reagent when freeze-dried and pulverized cannabis samples 29-52 were tested in Experimental Example 13. [Figure 16] This photograph shows the color changes after adding the third reagent when two plant fragments containing CBN as the main component (CBN herb), two plant fragments containing CBD as the main component (CBD herb), six edible spices, two teas, and one tobacco sample were tested in Experiment Example 14. [Figure 17] This photograph shows the color change after adding the third reagent when testing two freeze-dried cannabis samples and one CBD herb sample with varying amounts of extract added, as in Experimental Example 15. [Figure 18] This photograph shows the color change after adding the third reagent to a single freeze-dried cannabis sample in Experimental Example 16, where the amount of the third reagent added was varied during the test. [Figure 19] This photograph shows the color change after adding the third reagent when two freeze-dried cannabis samples and one CBD herb sample were tested with varying ratios of extract, second reagent, and third reagent added, as in Experimental Example 17. [Figure 20] In Experimental Example 18, when the volume of the first reagent sealed in the first container was set to 0.8 mL, 0.9 mL, and 1.0 mL, the photograph shows the color change after adding the third reagent when two freeze-dried cannabis samples were tested using the test kit. [Figure 21] In Experimental Example 19, the amount of freeze-dried and pulverized cannabis added to the first container was 5, 10, 20, and 40 mg, and the test was performed using the test kit. The photograph shows the color change after the addition of the third reagent. [Figure 22] This photograph shows the color change after adding the third reagent when 28 freeze-dried and pulverized cannabis samples were placed in the first container and tested using a test kit in Experiment Example 20. [Figure 23] This photograph shows the color change after adding the third reagent when 16 freeze-dried and pulverized cannabis samples, 2 CBN herbs, 2 CBD herbs, 6 edible spices, 2 teas, and 1 tobacco sample were placed in the first container and tested using the test kit in Experiment Example 20. [Figure 24] This figure shows the first container, second container, and filter-equipped cap of a test kit according to an embodiment of the present invention. [Figure 25] This figure shows the third container, fourth container, fifth container, first spacer, second spacer, and cap of the test kit according to an embodiment of the present invention. [Figure 26] This figure shows a filter-equipped cap according to an embodiment of the present invention, where (b) is a cross-section of XXVIB-XXVIB in (a), and (c) is a view along the XXVIC arrow in (a). [Figure 27] This figure shows a paper slip and a sample used in a test kit according to an embodiment of the present invention. [Figure 28]This figure shows a test kit according to an embodiment of the present invention, in which the sample and the first container are placed in the second container, and a filter-equipped cap is installed at the opening of the second container. [Figure 29] This figure shows the operation of destroying the first container in the state shown in Figure 28. [Figure 30] This diagram shows the process of pouring the first mixed liquid from the second container, etc., as shown in Figure 29, into the fifth container, which contains the third container, the fourth container, the first spacer, and the second spacer. [Figure 31] This diagram shows the process of attaching a cap to the fifth container, as shown in Figure 30, and then destroying the third container. [Figure 32] This diagram shows the action of destroying the fourth container using the fifth container shown in Figure 31. [Modes for carrying out the invention]

[0017] The inspection kit and inspection method according to this embodiment will be described in detail below with reference to the drawings. Note that the dimensional ratios in the drawings are exaggerated for illustrative purposes and may differ from the actual ratios.

[0018] As mentioned above, the conventionally used Duquenois-Levine reagent consists of three solutions: the first reagent: an ethanol solution of acetaldehyde and vanillin, the second reagent: concentrated hydrochloric acid, and the third reagent: chloroform.

[0019] Non-patent document 4 proposes a color-producing mechanism for Δ9-THC using the Duquenois-Levine reagent, involving the conversion of Δ9-THC to Δ8-THC under strongly acidic conditions with concentrated hydrochloric acid, followed by the formation of a color-producing substance through the condensation of acetaldehyde and vanillin (4-hydroxy-3-methoxybenzaldehyde) into Δ8-THC. Figure 1 shows the chemical structure of the color-producing substance presumed to be produced by this reaction. In this case, CBD is also converted to Δ8-THC under strongly acidic conditions, similar to Δ9-THC. Therefore, theoretically, it is difficult to distinguish between Δ9-THC and CBD using the Duquenois-Levine reagent. Furthermore, the Duquenois-Levine reagent also produces a color-producing substance in CBN through the condensation of acetaldehyde and vanillin. Therefore, CBN is also thought to exhibit color, although its hue differs from that of Δ9-THC and CBD.

[0020] If the coloration mechanism proposed in Non-Patent Document 4 is correct, it is possible to change the color tone of the colorant produced by the reaction with the Duquenois-Levine reagent by altering the conjugated system of the colorant. It is also possible to increase the color difference between Δ9-THC and CBN. Possible methods for altering the conjugated system include replacing vanillin in the first reagent with an aldehyde whose hydroxyl group substitution position on the benzene ring is different, or omitting the addition of acetaldehyde.

[0021] Therefore, in this disclosure, we have found that by using 2-hydroxybenzaldehyde instead of vanillin, it is possible to selectively determine whether or not a sample contains Δ9-THC. The test kit and test method according to the first and second embodiments will be described in detail below.

[0022] [First Embodiment] (Test kit) First, the test kit according to the first embodiment will be described. The test kit according to this embodiment is a test kit for determining whether a sample contains Δ9-THC. According to the test kit and test method according to this embodiment, it is possible to selectively determine whether or not a sample contains Δ9-THC by checking the color change. The sample may contain chemical substances themselves such as Δ9-THC, CBD, CBN, Δ8-THC, and CBG, or it may contain plant fragments suspected to be cannabis. The test kit according to this embodiment comprises a first reagent, a second reagent, and a third reagent.

[0023] The first reagent contains 2-hydroxybenzaldehyde and a water-soluble organic solvent capable of dissolving 2-hydroxybenzaldehyde. The first reagent may contain 2-hydroxybenzaldehyde and the water-soluble organic solvent in separate containers. Alternatively, the first reagent may contain 2-hydroxybenzaldehyde dissolved in the water-soluble organic solvent in a container.

[0024] The water-soluble organic solvent may contain at least one of an alkyl cyanate and an alcohol. The alkyl cyanate may include, for example, acetonitrile. The alcohol may contain at least one of methanol and ethanol. The concentration of 2-hydroxybenzaldehyde in the first reagent is preferably 1 mg / mL or more, more preferably 5 mg / mL or more, and even more preferably 20 mg / mL or more. The concentration of 2-hydroxybenzaldehyde in the first reagent is not particularly limited, but may be, for example, 200 mg / mL or less, or 100 mg / mL or less.

[0025] If the sample contains plant fragments, the organic solvent is preferably an alkyl cyanide, and more preferably an acetonitrile. These organic solvents do not easily transfer chlorophyll, thus suppressing interference with color confirmation caused by chlorophyll.

[0026] The second reagent is hydrochloric acid. The hydrogen chloride concentration of the hydrochloric acid is preferably 7 mol / L or higher. A hydrogen chloride concentration of 7 mol / L or higher makes it easier to confirm the color change. There is no particular upper limit to the hydrogen chloride concentration, but it may be, for example, 15 mol / L or less. The hydrogen chloride concentration may be 8 mol / L or higher, 9 mol / L or higher, 10 mol / L or higher, or 11 mol / L or higher. The hydrogen chloride concentration may be 14 mol / L or less, 13 mol / L or less, or 12 mol / L or less.

[0027] The third reagent contains an organic solvent that has a higher specific gravity than water and separates from water. The third reagent may also be an organic solvent that separates from water. The density of the organic solvent at 20°C is 1.1 g / cm³. 3 More than 1.2g / cm 3 More than 1.3g / cm 3 Above, or 1.4 g / cm³ 3 The above is also acceptable. The density of the organic solvent at 20°C is 5 g / cm³. 3 Below, 2g / cm 3 Below 1.4g / cm 3 The following is also acceptable.

[0028] The third reagent may contain at least one chloroalkane selected from the group consisting of chloroform, dichloromethane, and 1,2-dichloroethane. Among these, the third reagent is preferably chloroform.

[0029] [Testing Method] Next, a testing method using the testing kit according to the first embodiment will be described. The testing method according to this embodiment includes a first reagent mixing step, a second reagent mixing step, and a third reagent mixing step. The testing method may also include a preparation step and a color confirmation step.

[0030] (preparation process) In the preparation step, the specimen to be tested using the testing method according to this embodiment is prepared. The specimen may be crushed manually or using an instrument. Since plant fragments examined on-site can be easily crushed, they can be easily crushed manually, for example, by hand while wearing disposable gloves.

[0031] The amount of sample used for the test is not particularly limited, but may be, for example, 10 mg to 100 mg. The sample amount may be 20 mg or more. Alternatively, the sample amount may be 70 mg or less, or 40 mg or less. The amount of sample to be used for the test can be estimated with some accuracy by spreading the sample within the frame indicated on the measuring platform. For example, if a circular frame with a diameter of 1.25 cm is indicated on the measuring platform, the sample amount can be kept within the range of approximately 20 mg to 40 mg by spreading the sample within the frame.

[0032] (First reagent mixing step) In the first reagent mixing step, the first reagent and the sample are mixed to prepare the first mixture. The first reagent contains a water-soluble organic solvent in which 2-hydroxybenzaldehyde is dissolved. The first reagent is the same as that described in the above-mentioned test kit, except that 2-hydroxybenzaldehyde is dissolved in a water-soluble organic solvent, so the explanation is omitted.

[0033] In the first reagent mixing step, the sample may be dissolved in the first reagent to prepare a dissolving solution, or components in the sample may be extracted to prepare an extract. That is, the first mixture may be a solution prepared by dissolving the sample in the first reagent, or it may be an extract containing components extracted from the sample by the first reagent.

[0034] For example, if the sample contains chemical substances such as Δ9-THC, CBD, CBN, Δ8-THC, and CBG, and the sample can be dissolved in the first reagent, the sample may be dissolved in the first reagent to prepare a dissolution.

[0035] Furthermore, the sample may contain plant fragments suspected to be cannabis, or other materials that do not completely dissolve in the first reagent. If the sample is a plant fragment, components such as Δ9-THC, CBD, CBN, Δ8-THC, and CBG may be extracted from the sample to prepare an extract. Methods for extracting components from the sample with the first reagent include grinding in a hand-operated simple grinding container and shaking within the container. The shaking time is not particularly limited and may be 5 seconds or more, 20 seconds or more, or 30 seconds or more. The shaking time may also be 10 minutes or less, 5 minutes or less, or 1 minute or less.

[0036] (Second reagent mixing step) In the second reagent mixing step, the liquid contained in the first mixture is mixed with the second reagent to prepare the second mixture. The liquid contained in the first mixture may be a dissolving solution obtained by dissolving the sample in the first reagent, as described above, or it may be an extract obtained by extracting components from the sample. To obtain the liquid contained in the first mixture, the first mixture may be filtered by methods such as filter filtration or cotton plug filtration. Alternatively, the second mixture may be prepared by mixing the first mixture and the second reagent, and the second mixture may contain the solid components contained in the first mixture. The second reagent is hydrochloric acid. The second reagent is the same as that used in the test kit described above, so its description is omitted.

[0037] The volume ratio of the liquid in the first mixture to the second reagent may be 0.1 or greater, 0.5 or greater, 1 or greater, 1.5 or greater, or 2 or greater. Alternatively, the volume ratio of the liquid in the first mixture to the second reagent may be 5 or less, 3 or less, 2.5 or less, 2 or less, 1.5 or less, or 1 or less.

[0038] After mixing the liquid in the first mixture with the second reagent, it may be allowed to stand for a certain period of time. The standing time after adding the second reagent may be 10 seconds or more, 30 seconds or more, 60 seconds or more, or 120 seconds or more. The longer the standing time, the stronger the color tends to become, but the intensity of the color may remain constant even if the standing time exceeds the specified time. There is no particular upper limit to the standing time, but it may be 60 minutes or less, 30 minutes or less, or 10 minutes or less.

[0039] (Third reagent mixing step) In the third reagent mixing step, the second mixture is mixed with the third reagent. The third reagent contains an organic solvent that has a higher specific gravity than water and separates from water. The third reagent is the same as that of the test kit described above, so its explanation is omitted. By mixing the second mixture with the third reagent, the color-developing component derived from Δ9-THC is extracted into the organic solvent of the third reagent. Since the third reagent separates from water, the color of the separated organic component can be easily confirmed.

[0040] The volume ratio of the third reagent to the second reagent may be 0.1 or greater, 0.5 or greater, 1 or greater, 1.5 or greater, or 2 or greater. Alternatively, the volume ratio of the third reagent to the second reagent may be 5 or less, 3 or less, 2.5 or less, 2 or less, 1.5 or less, or 1 or less.

[0041] (Color tone confirmation process) In the color confirmation step, after mixing the second mixture and the third reagent, the color of the liquid separated into two layers, an aqueous layer and an organic layer, is checked. In the color confirmation step, the color of the lower layer after separation into two layers (the upper layer being aqueous and the lower layer being organic) may also be checked. For example, if the lower layer after separation is blue or blue-green, the sample can be determined to contain Δ9-THC. Conversely, if the lower layer after separation is not blue or blue-green, the sample can be determined not to contain Δ9-THC.

[0042] After adding the third reagent, the mixture may be allowed to stand until the color change can be confirmed. The standing time may be 10 seconds or more, 30 seconds or more, 1 minute or more, 3 minutes or more, or 5 minutes or more. Better results may be obtained with a longer standing time of at least 5 minutes, but the color change can also be confirmed immediately after phase separation. There is no particular upper limit to the standing time, but it may be 60 minutes or less, 30 minutes or less, or 10 minutes or less.

[0043] Methods for checking the color tone of a phase-separated liquid include visually comparing it to a pre-created color sample, and taking a photograph with a smartphone or digital camera and then quantifying it using an RGB color system.

[0044] As described above, the test kit according to this embodiment comprises a first reagent containing 2-hydroxybenzaldehyde and a water-soluble organic solvent capable of dissolving 2-hydroxybenzaldehyde. The test kit comprises a second reagent which is hydrochloric acid. The test kit comprises a third reagent which contains an organic solvent having a specific gravity greater than water and separating from water.

[0045] Furthermore, the testing method according to this embodiment includes the step of preparing a first mixture by mixing a sample with a first reagent containing a water-soluble organic solvent in which 2-hydroxybenzaldehyde is dissolved. The testing method includes the step of preparing a second mixture by mixing the liquid contained in the first mixture with a second reagent which is hydrochloric acid. The testing method includes the step of mixing the second mixture with a third reagent which contains an organic solvent that has a higher specific gravity than water and separates from water.

[0046] According to the test kit and test method of this embodiment, it is possible to selectively determine whether or not a sample contains Δ9-THC.

[0047] [Second Embodiment] (Test kit) Next, the test kit according to the second embodiment will be described. Note that the same details as those of the first embodiment will be omitted from the description. The test kit 51 according to this embodiment comprises a first container 53, a second container 55, a third container 57, a fourth container 59, a fifth container 61, and a filter-equipped cap 89 (see Figures 24 to 26).

[0048] The first reagent 65 is sealed inside the first container 53. The first container 53 can be easily broken by stress generated by an externally applied force, allowing the first reagent 65 filled inside the first container 53 to leak out. The volume of the first reagent 65 inside the first container 53 may be in the range of 0.8 to 1.0 mL.

[0049] The second container 55 is a tubular body having an opening 67 at its top. The second container 55 is configured to accommodate the sample 69 and the first container 53. The second container 55 can break the first container 53 due to stress generated by an external force, but the resulting glass fragments of the first container 53 will not pierce the second container 55. A filter-equipped cap 89 can be fitted to the opening 67 of the second container 55.

[0050] The filter-equipped cap 89 is connectable to the opening 67 of the second container 55 and the opening 75 of the fifth container 61, and is configured to filter liquid from the second container 55 to the fifth container 61. The bottom surface of the filter-equipped cap 89 can be attached to the opening 67 of the second container 55 without causing liquid leakage. A filter 63 is attached to the tip of the filter-equipped cap 89, and the tip of the filter-equipped cap 89 can be inserted into the opening 75 of the fifth container 61.

[0051] The second reagent 71 is sealed inside the third container 57. The third container 57 can be easily broken by stress generated by an externally applied force, allowing the second reagent 71 filled inside the third container 57 to leak out. Considering the amount of sample extract obtained with the first reagent 65, the test can be performed with a volume of 0.177 mL of the second reagent 71 in the third container 57, but it is not limited to this amount.

[0052] The third reagent 73 is sealed inside the fourth container 59. The fourth container 59 can be easily broken by stress generated by an externally applied force, allowing the third reagent 73 filled inside the fourth container 59 to leak out. Considering the amount of sample extract obtained with the first reagent 65, the test can be performed with a volume of 0.177 mL of the third reagent 73 in the fourth container 59, but it is not limited to this amount.

[0053] The fifth container 61 has an opening 75 and is configured to accommodate the third container 57 and the fourth container 59. The fifth container 61 is a transparent or translucent tubular body made of resin. Inside the fifth container 61, the third container 57, the second spacer 125, the fourth container 59, and the first spacer 123 are stored in that order from top to bottom. To prevent the contents from falling out, a removable cap 127 is attached to the top of the fifth container 61. The stress generated by an externally applied force can cause the third container 57 and the fourth container 59 to break separately, and the resulting fragments will not pierce the fifth container 61. Since the fifth container 61 is sealed by the cap 127, the risk of direct contact with the contents is low. In addition, the color of the lower layer after the two layers have separated can be easily seen.

[0054] The test kit may include a measuring platform for estimating the amount of sample to be used for testing. The measuring platform is not particularly limited and may be a piece of paper with a frame printed on it (see Figure 27). The size and shape of the frame are not particularly limited. For example, a circular frame with a diameter of 1.25 cm may be printed on the measuring platform, and by spreading the sample within the frame, the amount of plant material sample can be kept within a range of approximately 20 mg to 40 mg.

[0055] The test kit may include instructions with color samples attached. The instructions may include several pieces of paper in shades of blue and blue-green, corresponding to the actual color of the test kit for cannabis containing Δ9-THC, as color samples.

[0056] (Testing method) Next, a testing method using the testing kit 51 according to the second embodiment will be described. Note that the same details as those in the first embodiment will be omitted from the explanation. The testing method according to this embodiment includes a first reagent mixing step, a second reagent mixing step, and a third reagent mixing step. The testing method may also include a first preparation step, a second preparation step, a filtration step, and a color confirmation step.

[0057] (1st preparation step) In the first preparation step, the sample 69 to be tested is prepared using the testing method, similar to the first embodiment. The sample 69 and the first container 53 are then placed in the second container 55 (see Figure 28). The first container 53 is filled with the first reagent 65.

[0058] (First reagent mixing step) In the first reagent mixing step, the first reagent 65 and the sample 69 are mixed to prepare the first mixed solution 77, similar to the first embodiment. However, in the second embodiment, force is applied to the second container 55 to generate stress, causing the first reagent 65 to flow out of the first container 53, and the first reagent 65 and the sample 69 are mixed in the second container 55 (see Figure 29). In the first reagent mixing step, force may be applied to the second container 55 to generate stress and break the first container 53. By breaking the first container 53, the first reagent 65 flows out of the first container 53, and the first reagent 65 and the sample 69 are mixed in the second container 55. From the viewpoint of suppressing leakage of the first reagent 65, it is preferable that the first container 53 breaks with the filter-equipped cap 89 attached to the opening 67 of the second container 55, but the first container 53 may also break with the opening 67 of the second container 55 open.

[0059] In the first reagent mixing step, the second container 55 may be shaken with the filter-equipped cap 89 attached to the second container 55 to mix the first reagent 65 and the sample 69. In the first reagent mixing step, the sample 69 may be dissolved in the first reagent 65 by shaking to prepare a dissolving solution, or components in the sample 69 may be extracted to prepare an extract.

[0060] (Second preparation process) In the second preparation step, a fifth container 61 is prepared in which a third glass container 57 filled with the second reagent 71 is inserted on top, and a fourth glass container 59 filled with the third reagent 73 is inserted on the bottom (see Figure 30). Inside the fifth container 61, the third container 57, the second spacer 125, the fourth container 59, and the first spacer 123 are arranged in that order from top to bottom. The second reagent 71 is sealed inside the third container 57. The third reagent 73 is sealed inside the fourth container 59.

[0061] (Filtration process) In the filtration process, the first mixed solution 77 is filtered by a filter-equipped cap 89 attached to the opening 67 of the second container 55, and the filtrate is transferred to the fifth container 61 (see Figure 30). This process removes plant fragments and other debris contained in the first mixed solution 77 and transfers the liquid contained in the first mixed solution 77 to the fifth container 61. After breaking the first container 53 with the filter-equipped cap 89 attached to the second container 55, the tip of the filter-equipped cap 89 is pointed downwards, and the center of the filter-equipped cap 89 is pressed firmly, causing the first reagent 65 inside the filter-equipped cap 89 to be filtered by the filter 63 and dispensed. Since the filtrate is transferred to the fifth container 61 through the filter-equipped cap 89, the risk of the tester directly contacting the filtrate is reduced. In this state, fragments originating from the first container 53 may fall into the filter-equipped cap 89, but even in this case, the fragments will not pierce the filter-equipped cap 89.

[0062] (Second reagent mixing step) In the second reagent mixing step, the liquid contained in the first mixture 77 and the second reagent 71 are mixed to prepare the second mixture 138, similar to the first embodiment. However, in the second embodiment, force is applied to the fifth container 61 to generate stress, causing the second reagent 71 to flow out of the third container 57, which is housed inside the fifth container 61 and filled with the second reagent 71, and mixing the liquid contained in the first mixture 77 with the second reagent 71 (see Figure 31). In the second reagent mixing step, force may be applied to the fifth container 61 to generate stress and break the third container 57. By breaking the third container 57, the second reagent 71 flows out of the third container 57, and the liquid contained in the first mixture 77 and the second reagent 71 are mixed inside the fifth container 61. From the viewpoint of suppressing leakage of the liquid contained in the first mixture 77 and the second reagent 71, it is preferable that the third container 57 breaks with the cap attached to the opening 75 of the fifth container 61; however, the third container 57 may also break with the opening 75 of the fifth container 61 open.

[0063] In the second reagent mixing step, the fifth container 61 may be shaken with the cap attached to the opening 75 of the fifth container 61 to mix the liquid contained in the first mixture 77 with the second reagent 71. The second reagent mixing step may also include a step of shaking the fifth container 61 up and down and then letting it stand for a certain period of time.

[0064] (Third reagent mixing step) In the third reagent mixing step, the second mixture 138 and the third reagent 73 are mixed, similar to the first embodiment. However, in the second embodiment, force is applied to the fifth container 61 to generate stress, causing the third reagent 73 to flow out of the fourth container 59, which is housed inside the fifth container 61 and filled with the third reagent 73, and the second mixture 138 and the third reagent 73 are mixed (see Figure 32). In the third reagent mixing step, force may be applied to the fifth container 61 to generate stress and break the fourth container 59. By breaking the fourth container 59, the third reagent 73 flows out of the fourth container 59 and the second mixture 138 and the third reagent 73 are mixed inside the fifth container 61. From the viewpoint of suppressing leakage of the second mixture 138 and the third reagent 73, it is preferable that the fourth container 59 breaks with the cap attached to the opening 75 of the fifth container 61; however, the fourth container 59 may also break with the opening 75 of the fifth container 61 open.

[0065] In the third reagent mixing step, the fifth container 61 may be shaken to mix the second mixture 138 and the third reagent 73. In the third reagent mixing step, it is preferable to shake the fifth container 61 with the cap attached to the opening 75, but it may also be shaken with the opening 75 open. The third reagent mixing step may include a step of shaking the fifth container 61 up and down and then letting it stand for a certain period of time.

[0066] (Color tone confirmation process) In the color confirmation step, similar to the first embodiment, the second mixture 138 and the third reagent 73 are mixed, and the color of the liquid, which has separated into two layers, an aqueous layer and an organic layer, is confirmed.

[0067] As described above, the test kit 51 according to this embodiment comprises a first container 53, a second container 55, a third container 57, a fourth container 59, a fifth container 61, and a filter-equipped cap 89. The first container 53 seals the first reagent 65. The second container 55 has an opening 67 and can accommodate the sample 69 and the first container 53. The third container 57 seals the second reagent 71. The fourth container 59 seals the third reagent 73. The fifth container 61 has an opening 75 and can accommodate the third container 57 and the fourth container 59. The filter-equipped cap 89 is connectable to the opening 67 of the second container 55 and the opening 75 of the fifth container 61, and can filter liquid from the second container 55 to the fifth container 61.

[0068] Furthermore, in the testing method according to this embodiment, in the step of mixing the first reagent 65, force is applied to the first container 53 filled with the first reagent 65 and the second container 55 containing the sample 69 to generate stress, causing the first reagent 65 to flow out of the first container 53, and the first reagent 65 and the sample 69 to be mixed in the second container 55. In the step of mixing the second reagent 71, force is applied to the fifth container 61 to generate stress, causing the second reagent 71 to flow out of the third container 57, which is housed inside the fifth container 61 and filled with the second reagent 71, and the liquid contained in the first mixture 77 and the second reagent 71 to be mixed. In the step of mixing the third reagent 73, force is applied to the fifth container 61 to generate stress, causing the third reagent 73 to flow out of the fourth container 59, which is housed inside the fifth container 61 and filled with the third reagent 73, and the second mixture 138 and the third reagent 73 to be mixed.

[0069] In the Duquenois-Levine reagent kit currently used in Japan for crime scene investigations (product name: Cannabis Reagent), the first and second reagents are contained in screw-capped glass vials, and the third reagent is in a glass ampoule. When performing the test with this kit, the sample is extracted in the glass vial of the first reagent, the extracted solution is transferred to the glass vial of the second reagent and mixed, the glass ampoule containing the third reagent is crushed, and the third reagent is transferred to the glass ampoule of the second reagent for extraction.

[0070] However, because the inner diameter of the glass vials in this kit is small (approximately 6 mm), it is difficult to place the sample into the vial containing the first reagent. Furthermore, since both the first and second reagents are contained in glass vials, it is difficult to transfer the extract from the first reagent into the glass vial of the second reagent. In addition, there is a risk of injury when crushing the glass ampoule of the third reagent, and there is a risk of the reagent leaking and contaminating the hands when transferring the third reagent into the glass vial of the second reagent. Thus, the structure of the current on-site cannabis testing kit raises concerns regarding usability and safety. Moreover, due to the reaction principle of the reagents, it is not possible to distinguish between Δ9-THC and CBD, and it is difficult to distinguish between Δ9-THC and CBN.

[0071] On the other hand, the test kit and test method according to this embodiment offer excellent operability, and can selectively determine whether or not a sample contains Δ9-THC in a simple and safe manner. Therefore, the test kit according to this embodiment can be used, for example, at a crime scene, and is suitable as a test kit for crime scenes. [Examples]

[0072] The following describes the test kit and test method in more detail with reference to experimental examples, but this disclosure is not limited to these experimental examples.

[0073] [Experimental Example 1] 0.25 mg each of Δ9-THC, CBD, CBN, Δ8-THC, and CBG compounds were placed in glass test tubes and allowed to stand for at least 5 minutes. Specifically, after placing the solution of each compound into the test tube, the solvent was removed under a nitrogen stream, and after stopping the nitrogen stream, the test tubes were allowed to stand for at least 5 minutes.

[0074] Next, 20 mL of an ethanol solution of 4-hydroxybenzaldehyde (20 mg / mL) was mixed with 0.5 mL of acetaldehyde to form the first reagent, which was then added to the compound to dissolve it in the first reagent. To this solution, 0.5 mL of concentrated hydrochloric acid (hydrogen chloride concentration of 35-37% by weight, equivalent to 12 mol / L) was added as the second reagent and immediately mixed. After mixing, the solution was allowed to stand for 30 seconds. The color change after standing is shown in Figure 2. Next, 1 mL of chloroform was added to the solution after standing as the third reagent, and the liquid in the glass tube separated into two layers. The color change immediately after the two-layer separation is shown in Figure 2.

[0075] As shown in Figure 2, the color of Δ9-THC after the addition of the second reagent was a dark purple, which was slightly different from the color of CBD (reddish-purple), but almost the same as the color of Δ8-THC (dark purple). Furthermore, the color of the lower layer after two-layer separation (after the addition of the third reagent) was dark purple for all three: Δ9-THC, CBD, and Δ8-THC. On the other hand, the color of CBN and CBG was clearly different from that of Δ9-THC, both after the addition of the second reagent and after two-layer separation (after the addition of the third reagent). From these results, it was considered that with the reagent composition tested in this experiment, it is possible to distinguish between Δ9-THC and CBN or CBG, but it is difficult to distinguish between Δ9-THC and Δ8-THC or CBD.

[0076] [Experimental Example 2] Except for using a solution of 20 mL of 2-hydroxybenzaldehyde in ethanol (20 mg / mL) with 0.5 mL of acetaldehyde added as the first reagent, the color changes after adding the second and third reagents were confirmed in the same manner as in Experimental Example 1. The results are shown in Figure 3.

[0077] As shown in Figure 3, after the addition of the second reagent, the colors of Δ9-THC, CBD, and Δ8-THC were all pink, but the color of Δ9-THC was the darkest. Furthermore, after the addition of the third reagent, the lower layer showed a dark brown color for Δ9-THC, CBD, and Δ8-THC, but Δ9-THC showed the strongest color. In addition, CBN turned white after the addition of the second reagent and no color was visible after the addition of the third reagent, and CBG turned a light pink color both after the addition of the second and third reagents, and all were clearly different from Δ9-THC. From these results, it is considered that when the first reagent contains 2-hydroxybenzaldehyde, the selectivity for Δ9-THC is improved compared to when the first reagent contains 4-hydroxybenzaldehyde.

[0078] [Experimental Example 3] Except for using 0.5 mL of an ethanol solution of 2-hydroxybenzaldehyde (20 mg / mL) as the first reagent, the color changes after adding the second and third reagents were confirmed in the same manner as in Experimental Example 1. The results are shown in Figure 4.

[0079] As shown in Figure 4, Δ9-THC exhibited a blue color both after the addition of the second reagent and the third reagent. On the other hand, among the other cannabinoids, CBD, Δ8-THC, and CBG exhibited an orange color after the addition of the second reagent, but showed no color change after the addition of the third reagent. CBN showed no color change either after the addition of the second reagent or the third reagent. From these results, it is considered that the reagent composition in this experiment exhibits selectivity for Δ9-THC.

[0080] [Experimental Example 4] One mg each of Δ9-THC, CBD, CBN, Δ8-THC, and CBG compounds, along with 0.1 mg of Δ9-THCA compound, were placed in glass test tubes and allowed to stand for at least 5 minutes. Additionally, 0.5 mL of 2-hydroxybenzaldehyde acetonitrile solution (20 mg / mL) was used as the first reagent. Except for the above, the color changes after adding the second and third reagents were observed in the same manner as in Experimental Example 1. The results are shown in Figure 5.

[0081] As shown in Figure 5, Δ9-THC exhibited a reddish-purple color after the addition of the second reagent, but turned blue in the lower layer after the addition of the third reagent. On the other hand, among the other cannabinoids, CBD, Δ8-THC, and CBG exhibited an orange color in the lower layer both after the addition of the second reagent and after the addition of the third reagent. CBN and Δ9-THCA showed no color change after the addition of either the second or third reagent. From these results, it is considered that the selectivity for Δ9-THC is maintained even when the solvent of the first reagent is replaced from ethanol to acetonitrile.

[0082] [Experimental Example 5] Δ9-THC 0.25 mg was placed in a glass test tube and allowed to stand for at least 5 minutes. Additionally, 0.5 mL of 2-hydroxybenzaldehyde acetonitrile solution (2, 5, 10, 20, 50, 100 mg / mL) was used as the first reagent. Except for the above, the color change after adding the third reagent was confirmed in the same manner as in Experimental Example 1. The results are shown in Figure 6.

[0083] As shown in Figure 6, the lower layer after the addition of the third reagent turned blue at all concentrations, but the blue coloration tended to be stronger as the concentration of 2-hydroxybenzaldehyde increased.

[0084] [Experimental Example 6] Δ9-THC 0.25 mg was placed in a glass test tube and allowed to stand for at least 5 minutes. 0.5 mL of 2-hydroxybenzaldehyde acetonitrile solution (20 mg / mL) was used as the first reagent. In addition, 0.5 mL each of concentrated hydrochloric acid, 4 mol / L phosphoric acid, and 6 mol / L sulfuric acid were used as the second reagent. Except for the above, the color change after adding the third reagent was confirmed in the same manner as in Experimental Example 1. The results are shown in Figure 7.

[0085] As shown in Figure 7, the lower layer after the addition of the third reagent turned blue only when concentrated hydrochloric acid was added as the second reagent; no color change was observed when any other acid was added.

[0086] [Experimental Example 7] Concentrated hydrochloric acid (hydrogen chloride concentration is 35 - 37% by weight percentage, corresponding to 12 mol / L), 10 mol / L hydrochloric acid, 9 mol / L hydrochloric acid, 8 mol / L hydrochloric acid, and 7 mol / L hydrochloric acid, each 0.5 mL were used as the second reagent. Except for the above, the color development after adding the third reagent was confirmed in the same manner as in Experimental Example 6. The results are shown in Figure 8.

[0087] As shown in Figure 8, the lower layer after adding the third reagent became bluish for all concentrations, but the higher the hydrogen chloride concentration, the stronger the tendency of the bluish color development.

[0088] [Experimental Example 8] 0.25 mg of Δ9-THC was placed in a glass test tube and allowed to stand for 5 minutes or more. 0.5 mL of a 2-hydroxybenzaldehyde acetonitrile solution (20 mg / mL) was used as the first reagent. Also, chloroform (1.48 g / cm 3 (20 °C)), dichloromethane (1.33 g / cm 3 (20 °C)), 1,2-dichloroethane (1.25 g / cm 3 (20 °C)), ethyl acetate (0.90 g / cm 3 (20 °C)), and n-hexane (0.66 g / cm 3 (20 °C)), each 1 mL were used as the third reagent. Except for the above, the color development after adding the third reagent was confirmed in the same manner as in Experimental Example 1. The results are shown in Figure 9.

[0089] As shown in Figure 9, when chloroform, dichloromethane, or 1,2-dichloroethane was added as the third reagent, the lower layer after adding the third reagent became bluish. Among these organic solvents, when chloroform was used as the third reagent, the visibility of layer separation was the best. On the other hand, when ethyl acetate was added as the third reagent, no two-layer separation occurred, and when n-hexane was added, the lower layer, which is the aqueous layer, showed a bluish color, while the upper layer, which is the organic layer, did not show color development.

[0090] [Experimental Example 9] Approximately 10 mg of freeze-dried cannabis powder was placed in a glass test tube, and 0.5 mL each of acetonitrile, ethanol, and methanol were added to the powder. Ultrasonographic extraction was then performed for 5 minutes. Afterward, centrifugation (3000 rpm, 1 minute) was performed, and the supernatant was transferred to another glass test tube to check the color of the extract. Figure 10 shows the colors of the cannabis extracts using acetonitrile, ethanol, and methanol.

[0091] As shown in Figure 10, all extracts exhibited a green tint derived from chlorophyll, but the extract with acetonitrile had the lightest color.

[0092] [Experimental Example 10] One mg each of Δ9-THC, CBD, CBN, Δ8-THC, and CBG compounds were placed in a glass test tube and allowed to stand for at least 5 minutes. Next, 0.5 mL of 2-hydroxybenzaldehyde acetonitrile solution (20 mg / mL) was added to the compounds as the first reagent to dissolve them in the first reagent. To this solution of the compounds dissolved in the first reagent, 0.5 mL of concentrated hydrochloric acid was added as the second reagent and immediately mixed. After mixing, the mixture was allowed to stand for 0 seconds, 10 seconds, 30 seconds, 60 seconds, and 120 seconds to observe the change in color over time. The change in color over time after the addition of the second reagent is shown in Figure 11.

[0093] As shown in Figure 11, Δ9-THC showed color development at all standing times, and the color intensified over time, but the intensity of the color remained generally constant after a standing time of 30 seconds or more.

[0094] [Experimental Example 11] One mg each of Δ9-THC, CBD, CBN, Δ8-THC, and CBG compounds were placed in glass test tubes and allowed to stand for at least 5 minutes. Furthermore, after adding the third reagent, the test tubes were allowed to stand for 0 seconds, 10 seconds, 30 seconds, 1 minute, and 5 minutes to observe the change in color over time. Except for the above, the change in color over time after adding the third reagent was observed in the same manner as in Experimental Example 4. Figure 12 shows the change in color over time after adding the third reagent.

[0095] As shown in Figure 12, the color of Δ9-THC was an orange-blue immediately after the addition of the third reagent, but it became a clear blue over time. On the other hand, the orange color of CBD, Δ8-THC, and CBG faded over time. CBN did not show any color change.

[0096] [Experimental Example 12] Δ9-THC 1 mg, 0.25 mg, 0.1 mg, and 0.025 mg, and CBD 2.5 mg, 1 mg, 0.25 mg, and 0.1 mg were placed in glass test tubes and allowed to stand for at least 5 minutes. In addition, the mixtures were allowed to stand for 0 minutes, 1 minute, and 5 minutes after adding the third reagent, and the change in color over time was observed. Except for the above, the change in color over time after adding the third reagent was observed in the same manner as in Experimental Example 4. The change in color over time after adding the third reagent is shown in Figure 13.

[0097] As shown in Figure 13, with Δ9-THC, a blue coloration was observed regardless of the elapsed time, depending on the amount of Δ9-THC added. On the other hand, with CBD, an orange coloration was observed at doses of 0.25 mg or more immediately after extraction, while it was observed at doses of 1 mg or more after 5 minutes. This result means that, with time elapsed after the addition of the third reagent, the detection sensitivity of Δ9-THC does not decrease, but the detection sensitivity of CBD does decrease.

[0098] [Experimental Example 13] Approximately 10 mg each of freeze-dried cannabis plant pulverized material (cannabis 1-52) was placed in a hand-operated simple grinding container (product name: Finger Masher, with filter cap; the same applies hereafter). Then, 0.5 mL of 2-hydroxybenzaldehyde acetonitrile solution (20 mg / mL) was added to the pulverized material as the first reagent, and the mixture was ground and extracted for 30 seconds. The resulting extract was filtered by discharging it into a glass test tube through a filter-equipped cap. 0.5 mL of concentrated hydrochloric acid was added to this filtrate as the second reagent and immediately mixed, and then allowed to stand for 30 seconds. After standing, 1 mL of chloroform was added to the solution as the third reagent, and the liquid in the glass tube separated into two layers. The color changes immediately after the two-layer separation, along with the cannabinoid concentrations of each cannabis plant, are shown in Figures 14 and 15. Note that, for space reasons, the "Δ9-" designation for Δ9-THC and Δ9-THCA is omitted in Figures 14 and 15.

[0099] As shown in Figures 14 and 15, in cannabis samples containing Δ9-THC but not CBD, or containing CBD at a lower concentration than Δ9-THC (cannabis 1-48), the lower layer after the addition of the third reagent showed a blue or blue-green color. On the other hand, in cannabis samples containing CBD but not Δ9-THC (cannabis 49-52), the color was generally light orange and did not show a blue or blue-green color.

[0100] [Experimental Example 14] Instead of freeze-dried cannabis, we used CBN Herb 1 and 2, CBD Herb 1 and 2, obtained via the internet, and approximately 10 mg each of six types of edible spices, green tea, black tea, and tobacco obtained from a supermarket as samples. Except for the above, the color change after adding the third reagent was confirmed in the same manner as in Experimental Example 13. The results are shown in Figure 16.

[0101] As shown in Figure 16, in CBD herb 1 and 2, the lower layer after the addition of the third reagent showed an orange coloration. However, no samples showed a blue or blue-green coloration.

[0102] [Experimental Example 15] Approximately 20 mg each of freeze-dried cannabis 1 and cannabis 25 used in Experimental Example 13, and CBD herb 2 used in Experimental Example 14, were placed in two hand-operated simple grinding containers. Then, 1 mL of 2-hydroxybenzaldehyde acetonitrile solution (20 mg / mL) was added as the first reagent, and the mixture was ground and extracted for 30 seconds. The resulting extract was filtered by discharging it into a glass test tube through a filter-equipped cap. After combining the filtered extracts from each sample, 0.08 mL, 0.16 mL, 0.24 mL, 0.32 mL, and 0.40 mL of the filtered extract were dispensed into small glass test tubes. 0.16 mL of concentrated hydrochloric acid was added to the dispensed filtrate as the second reagent and immediately mixed, then allowed to stand for 30 seconds. After standing, 0.32 mL of chloroform was added to the dissolved solution as the third reagent, and the liquid in the glass tube separated into two layers. Figure 17 shows the effect of the amount of extract added on the color development after the addition of the third reagent.

[0103] As shown in Figure 17, when cannabis 1 and cannabis 25 were used as samples, a blue or blue-green color was observed when the amount of extract added was in the range of 0.08 to 0.4 mL, with the strongest color observed at 0.32 mL. On the other hand, when CBD herb 2 was used as a sample, an orange color was observed when the amount of extract added was in the range of 0.08 to 0.4 mL, but the color did not become stronger even when the amount of the first reagent added increased. From these results, it was determined that when the volume ratio of the second reagent to the third reagent is 1:2, the test is possible if the volume ratio of the amount of extract added to the second reagent is in the range of at least 0.5:1 to 2.5:1.

[0104] [Experimental Example 16] Approximately 20 mg each of the freeze-dried cannabis 1 used in Experimental Example 13 was placed into two hand-operated simple grinding containers. Then, 1 mL of 2-hydroxybenzaldehyde acetonitrile solution (20 mg / mL) was added to the samples as the first reagent, and the mixture was ground and extracted for 30 seconds. The resulting extract was filtered by discharging it into a glass test tube through a filter-equipped cap. After combining the filtered extracts from each sample, 0.32 mL of the filtered extract was dispensed into a small glass test tube. 0.16 mL of concentrated hydrochloric acid was added to the dispensed filtrate as the second reagent and immediately mixed, then allowed to stand for 30 seconds. When 0.08 mL, 0.16 mL, and 0.32 mL of chloroform were added to the dissolved solution as the third reagent, the liquid in the glass tube separated into two layers. Figure 18 shows the effect of the amount of the third reagent added on the color development after the addition of the third reagent.

[0105] A bluish coloration was observed at all amounts of the third reagent added. Specifically, when 0.08 mL of the third reagent was added, the two-layer separation was unclear, but a bluish coloration was observed. Furthermore, when 0.16 mL and 0.32 mL of the third reagent were added, clear two-layer separation was observed in both cases. The bluish coloration of the lower layer was stronger when 0.16 mL of the third reagent was added. From these results, it was determined that when the volume ratio of the extract to the second reagent is 2:1, the test is possible with a volume ratio of the second reagent to the third reagent in the range of at least 2:1 to 1:2.

[0106] [Experimental Example 17] From Experimental Examples 15 and 16, we attempted to optimize the volume ratio of extract:second reagent:third reagent, and determined that 2:1:1 was optimal. Therefore, we evaluated the difference in the volume ratio of extract:second reagent:third reagent using the freeze-dried and pulverized cannabis 1 and 25 used in Experimental Example 13 and the CBD herb used in Experimental Example 14.

[0107] Freeze-dried and pulverized cannabis 1 and 25, along with approximately 10 mg each of CBD herb, were placed in a hand-operated grinding container. 0.5 mL of 2-hydroxybenzaldehyde acetonitrile solution (20 mg / mL) was added to the samples as the first reagent, and the mixture was ground and extracted for 30 seconds. The resulting extract was filtered by discharging it into a glass test tube through a filter-equipped cap. This filtrate was dispensed into small glass test tubes, and concentrated hydrochloric acid was added to the dispensed filtrate as the second reagent. The mixture was immediately mixed and allowed to stand for 30 seconds. Chloroform was added to the dissolved solution as the third reagent, causing the liquid in the glass tube to separate into two layers. The volumes of extract:second reagent:third reagent were 0.16 mL:0.16 mL:0.32 mL under condition 1, and 0.32 mL:0.16 mL:0.16 mL under condition 2. Figure 19 shows the color changes immediately after two-layer separation under conditions 1 and 2.

[0108] As shown in Figure 19, under Condition 2, compared to Condition 1, cannabis 1, which had a higher Δ9-THC concentration, showed a stronger blue hue. Also, cannabis 25, which had a relatively lower Δ9-THC concentration, showed a stronger blue-green hue. On the other hand, CBD herb 2 showed a weaker orange hue. From these results, it was determined that optimizing the volume ratio of extract:second reagent:third reagent would make it easier to determine that cannabis with a lower Δ9-THC concentration is Δ9-THC positive, while also mitigating the effects of CBD.

[0109] [Experimental Example 18] First container 53, second container 55, third container 57, fourth container 59, fifth container 61, filter-equipped cap 89, and first and second spacers 123 and 125 were prepared, each with the materials and dimensions shown in Table 1 (see Figures 24 to 26).

[0110] [Table 1]

[0111] 20 mg each of freeze-dried and pulverized cannabis 2 and 25 used in Experimental Example 13 were placed in the second container 55, which already contained the first container 53 (see Figure 28). 0.8 mL, 0.9 mL, and 1.0 mL of 2-hydroxybenzaldehyde acetonitrile solution (20 mg / mL) were sealed into the first container 53 as the first reagent 65. After attaching a filter-equipped cap 89 to the opening 67 of the second container 55, force was applied to the second container 55 to generate stress and break the first container 53, causing the first reagent 65 to flow out of the first container 53 (see Figure 29). The second container 55 was then shaken up and down for 30 seconds to extract the components in the sample 69 with the first reagent 65. Next, the extract was dropped through a filter-equipped cap 89 into a resin container 61, which had a third glass container 57 filled with 0.177 mL of concentrated hydrochloric acid as the second reagent 71 on top and a fourth glass container 59 filled with 0.177 mL of chloroform as the third reagent 73 on the bottom (see Figures 30(a) and (b)). After attaching the cap to the opening 75 of the fifth container 61, force was applied to the top of the fifth container 61 to generate stress, causing the third container 57 to break and the second reagent 71 to flow out of the third container 57 (see Figure 31). Then, the fifth container 61 was shaken up and down and left to stand for 30 seconds. Next, force was applied to the bottom of the fifth container 61 to generate stress, causing the fourth container 59 to break and the third reagent 73 to flow out of the fourth container 59 (see Figure 32). The fifth container 61 was shaken up and down, and the color change of the lower layer was immediately confirmed after the two layers separated. Figure 20 shows the color development of the lower layer after two-layer separation when the amount of the first reagent 65 in the first container 53 is 0.8 mL, 0.9 mL, and 1.0 mL.

[0112] As shown in Figure 20, the color change of the lower layer was observed with all insertion volumes, but the visibility was best when the insertion volume was 0.9 mL.

[0113] Table 2 shows the amount of extract solution dropped from the second container 55 to the fifth container 61 relative to the amount of the first reagent 65 sealed in the first container 53. The amount dropped was calculated using the following formula. Droplet volume (mL) = Weight of dropped substance (g) / Density of acetonitrile (0.79 g / mL)

[0114] [Table 2]

[0115] As shown in Table 2, when the amount of the first reagent 65 in the first container 53 was 0.9 mL, the average amount of drops dispensed was closest to twice the amount of the second reagent 71 in the third container 57 (0.177 mL). These results indicate that 0.9 mL is the optimal amount of the first reagent 65 in the first container 53, but testing is possible in the range of at least 0.8 to 1.0 mL.

[0116] [Experimental Example 19] 5 mg, 10 mg, 20 mg, and 40 mg of freeze-dried and pulverized cannabis 2 and 31, respectively, were placed into the second container 55, which already contained the first container 53. Additionally, 0.9 mL of 2-hydroxybenzaldehyde acetonitrile solution (20 mg / mL) was used as the first reagent 65. The color change after adding the third reagent 73 was confirmed in the same manner as in Experimental Example 18. These results are shown in Figure 21.

[0117] As shown in Figure 21, the larger the amount of sample used for testing, the clearer the blue or blue-green coloration of the lower layer after separation into two layers. Based on these results, it was determined that the optimal amount of sample to use for testing is 20 mg to 40 mg, but testing is possible with at least 10 mg.

[0118] [Experimental Example 20] The freeze-dried cannabis used in Experimental Example 13 (the cannabis used in Experimental Example 19 was not subjected to testing, and some of the other materials were also not subjected to testing due to remaining quantities), CBN Herb 1 and 2, CBD Herb 1 and 2, 20 mg each of six types of edible spices, green tea, black tea, and tobacco were placed in the second container 55, which already contained the first container 53. The color change after adding the third reagent 73 was confirmed in the same manner as in Experimental Example 19, except for the above. These results are shown in Figures 22 and 23. Note that in Figures 22 and 23, the "Δ9-" notation for Δ9-THC and Δ9-THCA is omitted due to space limitations.

[0119] As shown in Figures 22 and 23, in the freeze-dried cannabis material, the lower layer after two-layer separation exhibited a bluish to bluish-green color. Furthermore, none of the CBN herbs 1 and 2, CBD herbs 1 and 2, six types of edible spices, green tea, black tea, and tobacco exhibited a bluish to bluish-green color in the lower layer after two-layer separation. These results demonstrate that the test kit is highly specific for Δ9-THC, and does not produce false positives for samples that do not contain Δ9-THC.

[0120] [Experimental Example 21] Wearing rubber gloves, two pieces of dried cannabis (one leaf, the other flower head) that had been pre-crushed by hand were scattered within a 1.25 cm diameter circle pre-printed on a piece of paper (see Figure 27). This procedure was repeated four times for the cannabis leaves and three times for the flower heads, and the weight of each measurement was measured using an electronic balance. The results are shown in Table 3. The measured values ​​were 20.6-40.5 mg for cannabis leaves and 17.7-36.6 mg for flower heads, and it was determined that approximately 20-40 mg of cannabis could be extracted using this method without the use of an electronic balance.

[0121] [Table 3]

[0122] Here, the test kit 51 will be described further. As shown in Figures 24 and 25, the test kit 51 comprises a first container 53, a second container 55, a third container 57, a fourth container 59, a fifth container 61, and a filter-equipped cap 89.

[0123] The first container 53 is sealed with the first reagent 65. The second container 55 has an opening 67. The second container 55 is designed to hold the sample 69 and the first container 53. The third container 57 is sealed with the second reagent 71. The fourth container 59 is sealed with the third reagent 73.

[0124] The fifth container 61 has an opening 75. The fifth container 61 is designed to house the third container 57 and the fourth container 59. The filter cap 89 is for filtering the first mixture 77, which is prepared in the second container 55 by mixing the first reagent 65 and the sample 69, and for depositing the liquid components of the first mixture 77 into the fifth container 61.

[0125] Furthermore, given that it is frequently used at police and other investigative sites, test kit 51 may also be called a field test kit.

[0126] In the first container 53, a closed first space (closed space of the first container) 81 is formed on the inside by a thin first wall 79. The first reagent 65 is contained within the first space 81. The first wall 79 is made of a first material with a low fracture toughness value. The fracture toughness value of the first material is about the same as or lower than that of glass.

[0127] The first container 53 is formed, for example, by closing the opening of a bottomed cylindrical body with a disc-shaped lid. The first container 53 formed in this manner may also be called the first tubular body. In this specification, the shape of the bottom surface of the bottomed cylindrical container is not particularly limited and may be, for example, hemispherical or flat.

[0128] As the first material, for example, glass or a hard synthetic resin with low toughness may be listed. The first wall 79 of the first container 53 is transparent, but the first wall 79 may be semi-transparent, colored transparent, or opaque.

[0129] The enclosed first space 81 of the first container 53 and the space outside the first container 53 are completely separated. Furthermore, unless a part of the first wall 79 of the first container 53 is destroyed, the first reagent 65 inside the first container 53 will not escape from the first container 53.

[0130] In the second container 55, a second space 85 with an opening 67 is formed on the inside by a thin-walled second wall 83. The sample 69 and the first container 53 can be placed into the second space 85 through the opening 67.

[0131] The second wall 83 is made of a second material that has a higher fracture toughness value (larger deformation when force is applied) than the first container 53. The second container 55 is formed, for example, in the shape of a bottomed cylinder. The second container 55 formed in this manner may also be called a second tubular body.

[0132] As a second material, for example, a synthetic resin with high toughness and a certain degree of rigidity can be mentioned. The second wall 83 of the second container 55 is transparent, similar to the first wall 79 of the first container 53, but the second wall 83 may be semi-transparent, colored transparent, or opaque, similar to the first wall 79.

[0133] The diameter of the second space 85 is slightly larger than the outer diameter of the first container 53, and the height (depth) of the second space 85 is slightly larger than the height of the first container 53.

[0134] The second space 85 of the second container 55 is connected to the space outside the second container 55 through the opening 67. In the first container 53, which is placed into the second space 85 through the opening 67 of the second container 55, the first wall 79 is not damaged, and the first reagent 65 is inside the first space 81 and has not leaked out of the first container 53 at all.

[0135] With the first container 53 inside the second container 55, a force of a predetermined magnitude (for example, a bending moment of a human hand; an arm or fingers) is applied to the second container 55 (see arrow in Figure 29). This causes both the second container 55 and the first container 53 to deform. As shown in Figure 29, the first container 53 is destroyed inside the second container 55, and the first reagent 65 in the first space 81 of the first container 53 leaks into the second space 85 of the second container 55. As a result, the sample 69 is immersed in the first reagent 65, and predetermined components contained in the sample 69 are extracted by the first reagent 65 to prepare the first mixture 77.

[0136] Furthermore, even if a force of the predetermined magnitude is applied to the second container 55, the second container 55 will not break, but will only undergo elastic deformation, for example. When the predetermined magnitude of force is removed, the second container 55 will return to its shape as it was when no force was applied.

[0137] As shown in Figure 26, the filter 63, together with the cap body 87, forms a filter-equipped cap 89. The cap body 87 is made of a material (for example, a synthetic resin) that has a higher fracture toughness value (a larger amount of deformation when force is applied) than the first container 53. The cap body 87 is also installed in the opening 67 of the second container 55 to seal the opening 67 of the second container 55 (see Figure 28, etc.). The filter 63 is integrally provided with the cap body 87.

[0138] The filter 63 is provided in the cap body 87 within a small-diameter through-hole 91 provided in the cap body 87. When the cap body 87 of the filter-equipped cap 89 is used to seal the opening 67 of the second container 55, the second space 85 of the second container 55 is connected to the space outside the second container 55 only through the filter 63.

[0139] In other words, the filter-equipped cap 89 is installed at the opening 67 of the second container 55, and when both liquid and solid are present in the second space 85, only the liquid passes through the filter 63 of the filter-equipped cap 89 and exits the second container 55. The solid cannot pass through the filter 63 and remains inside the second container 55.

[0140] As shown in Figure 26, the cap body 87 is composed of a first part 93, a second part 95, a third part 97, and a fourth part 99. The first part 93, the second part 95, the third part 97, and the fourth part 99 are arranged in this order.

[0141] The first part 93 is formed in a cylindrical shape, but the outer side surface of the first part 93 is a tapered surface 101. This tapered surface 101 forms a fitting portion 105 that fits into the fitting portion 103 of the second container 55 when the filter-equipped cap 89 is installed in the second container 55. As shown in Figure 24, the fitting portion 103 of the second container 55 is formed by a tapered surface 107 that is located on the inner surface of the second container 55 at the opening 67 of the second container 55.

[0142] The second portion 95 of the cap body 87 is formed in a cylindrical shape. The outer diameter of the second portion 95 is slightly smaller than the outer diameter of the first portion 93. The inner diameter of the first portion 93 is equal to the inner diameter of the second portion 95.

[0143] The third portion 97 of the cap body 87 is formed in a frustoconical shape with a predetermined wall thickness. The inner diameter of the third portion 97 is equal to the inner diameter of the second portion 95 at the end on the second portion 95 side. The inner diameter of the third portion 97 gradually decreases as it moves away from the second portion 95. The outer diameter of the third portion 97 is equal to the outer diameter of the second portion 95 at the end on the second portion 95 side. The outer diameter of the third portion 97 also gradually decreases as it moves away from the second portion 95.

[0144] The fourth part 99 is formed in a cylindrical shape with a small diameter. The inner diameter of the fourth part 99 is equal to the inner diameter of the third part 97 at the end opposite to the second part 95. The outer diameter of the fourth part 99 is equal to the outer diameter of the third part 97 at the end opposite to the second part 95.

[0145] The space inside the cylinder of the fourth part 99 is the through-hole 91 of the cap body 87 described above, and the filter 63 is fitted into the through-hole 91. The central axes of the cylinder of the first part 93, the second part 95, the third part 97, and the fourth part 99 coincide with each other.

[0146] As shown in Figure 28, when the filter-equipped cap 89 is installed on the second container 55, the fitting portion 105 of the cap body 87 of the filter-equipped cap 89 is fitted into the fitting portion 103 of the second container 55 (engaged and tightly sealed in an tight fit state).

[0147] Furthermore, as shown in Figure 26, the cap body 87 is provided with a protrusion 109. This protrusion 109 is provided to prevent fragments of the broken first container 53 from falling into the filter-equipped cap 89 when filtering the first mixed liquid 77. The presence of the protrusion 109 causes the fragments of the broken first container 53 to strike the protrusion 109, suppressing their movement into the filter-equipped cap 89. This prevents fragments of the first container 53 from getting stuck in the filter-equipped cap 89 and preventing the liquid from flowing out.

[0148] The projection 109 is formed in an elongated, flat shape with a predetermined thickness. The longitudinal direction of the projection 109 coincides with the extension direction of the central axis of the cap body 87. The projection 109 protrudes to a predetermined height toward the center of the cylinders of the first part 93, second part 95, and third part 97 from a portion of the inner surface of the first part 93, a portion of the inner surface of the second part 95, and a portion of the inner surface of the third part 97. The tip of the projection 109 in the direction of projection does not reach, for example, the fourth part 99, but stops midway (see Figure 26(c)). The thickness direction of the projection 109 coincides with the circumferential direction of the cylinder of the cap body 87.

[0149] As shown in Figure 25, etc., in the third container 57, similar to the first container 53, a third space 113 is formed on the inside, enclosed by a thin third wall 111 made of a third material with a low fracture toughness value. The second reagent 71 is contained within this third space 113. The fracture toughness value of the third material is about the same as that of the first material, and is about the same as or lower than that of glass.

[0150] The third container 57, like the first container 53, is formed in such a way that, for example, the opening of the main body, which is formed in the shape of a bottomed cylindrical body, is closed with a disc-shaped lid. The third container 57 formed in this way may also be called the third tubular body.

[0151] In the fourth container 59, similar to the first container 53, a fourth space 117 is formed on the inside, enclosed by a thin fourth wall 115 made of a fourth material with a low fracture toughness value. The third reagent 73 is contained within this fourth space 117. The fracture toughness value of the fourth material is similar to that of the first material, and is similar to or lower than that of glass.

[0152] The fourth container 59, like the first container 53, is formed in such a way that, for example, the opening of the main body, which is formed in the shape of a bottomed cylindrical body, is closed with a disc-shaped lid. The fourth container 59 formed in this way may also be called the fourth tubular body.

[0153] In the fifth container 61, a fifth space 121 with an opening 75 is formed on the inside by a thin fifth wall 119 made of a fifth material (for example, a synthetic resin). The third container 57, the fourth container 59, and the first mixed liquid 77 can be placed into the fifth space 121 through the opening 75. The fifth material has a higher fracture toughness value (larger deformation when force is applied) than the first container 53, the third container 57, and the fourth container 59.

[0154] The fifth container 61 is formed, for example, in the shape of a bottomed cylinder. The fifth container 61 formed in this manner may also be called the fifth tubular body.

[0155] The diameter of the fifth space 121 is slightly larger than the outer diameter of the third container 57, and the height (depth) of the fifth space 121 is greater than the height of the third container. Also, the diameter of the fifth space 121 is slightly larger than the outer diameter of the fourth container 59, and the height (depth) of the fifth space 121 is greater than the height of the fourth container 59. The height (depth) of the fifth space 121 is greater than the sum of the heights of the third and fourth containers. For example, the outer diameters of the third container 57 and the fourth container 59 are equal.

[0156] Furthermore, as shown in Figure 25, the inspection kit 51 includes a first spacer 123, a second spacer 125, and a cap 127. The first spacer 123 is made of a material (for example, a synthetic resin) that has a higher fracture toughness value (larger deformation when force is applied) than the first container 53, the third container 57, and the fourth container 59. The second spacer 125 is also made of a material (for example, a synthetic resin) that has a higher fracture toughness value (larger deformation when force is applied) than the first container 53, the third container 57, and the fourth container 59. The first spacer 123 and the second spacer 125 can be placed into the fifth space 121 of the fifth container 61 through the opening 75.

[0157] Figure 30 shows the state in which the third container 57, the fourth container 59, the first spacer 123, and the second spacer 125 are located inside the fifth space 121. In this state, the first spacer 123, the fourth container 59, the second spacer 125, and the third container 57 are arranged in this order from the bottom of the fifth container 61 toward the opening 75.

[0158] The first spacer 123 and the second spacer 125 function as cushions. The first spacer 123 is formed in a cylindrical shape, for example, and the second spacer 125 is also formed in a cylindrical shape, for example.

[0159] The diameter of the fifth space 121 is slightly larger than the outer diameter of the first spacer 123, and the height dimension (depth dimension) of the fifth space 121 is larger than the height dimension of the first spacer 123. Furthermore, the diameter of the fifth space 121 is slightly larger than the outer diameter of the second spacer 125, and the height dimension (depth dimension) of the fifth space 121 is larger than the height dimension of the second spacer 125.

[0160] As is already understood, the first spacer 123, the fourth container 59, the second spacer 125, and the third container 57 are placed in this order into the fifth space 121 of the fifth container 61.

[0161] The cap 127 (see Figure 25) is made of a material (for example, a synthetic resin) that has a higher fracture toughness value (a larger amount of deformation when force is applied) than the first container 53, the third container 57, and the fourth container 59, and is installed at the opening 75 of the fifth container 61 to seal the opening 75 of the fifth container 61.

[0162] Next, we will explain how to use the test kit 51. The testing method described above is designed to be performed, for example, using the test kit 51.

[0163] In the initial state, as shown in Figures 24 and 25, the first container 53, second container 55, third container 57, fourth container 59, fifth container 61, filter-equipped cap 89, first spacer 123, second spacer 125, and cap 127 may be separate. Alternatively, the first container 53, second container 55, third container 57, fourth container 59, fifth container 61, filter-equipped cap 89, first spacer 123, second spacer 125, and cap 127 may be sealed separately in a storage container (not shown). This storage container may be formed, for example, in the shape of a bag, and can be easily broken by hand. By breaking it, the contents of the storage container can be removed. In addition, in the initial state, the kit may consist of the first container 53 inserted into the second space 85 of the second container 55. To prevent the first container 53 from falling out and from moving significantly within the second space 85 during transport, a filter-equipped cap 89 may be attached upside down to the opening 67. Alternatively, to prevent the first container 53 from falling out during transport, the opening 67 of the second container may be secured with tape or the like instead of the filter-equipped cap 89. In the initial state, the first spacer 123, fourth container 59, second spacer 125, and third container 57 may be placed in the fifth space 121 of the fifth container 61 in order from the bottom toward the opening 75. To prevent the first spacer 123, fourth container 59, second spacer 125, and third container 57 from falling out, the opening 75 of the fifth container 61 may be sealed with the cap 127.

[0164] Furthermore, the first container 53, the second container 55, the third container 57, the fourth container 59, the fifth container 61, the filter-equipped cap 89, the first spacer 123, the second spacer 125, and the cap 127 may be sealed in a single storage container (not shown). Also, the first container 53, the second container 55, and the filter-equipped cap 89 shown in Figure 24 may be sealed in a single storage container (not shown). The third container 57, the fourth container 59, the fifth container 61, the first spacer 123, the second spacer 125, and the cap 127 shown in Figure 25 may be sealed in another single storage container (not shown).

[0165] First, as shown in Figure 27, the sample 69 is placed on a circular frame 129, for example, and weighed. The sample 69 may be weighed directly using the frame 129 inside the paper piece 131, or a transparent or translucent paper piece may be placed on the paper piece 131, and after confirming the frame 129 inside the paper piece 131 by looking through the transparent or translucent paper piece, the sample 69 may be weighed using the frame 129 inside the paper piece 131. The weighed sample 69 may be manually crushed inside the paper piece 131 or the transparent or translucent paper piece. Next, as shown in Figure 28, with the first container 53 inserted into the second space 85 of the second container 55, the weighed sample 69 is placed into the second space 85 of the second container 55 and the filter-equipped cap 89 is placed on the second container 55.

[0166] Next, the force indicated by the arrow in Figure 29 is applied to the second container 55 and the first container 53 to break the first container 53. The broken line indicated by reference numeral 133 in Figure 29 shows the point of breakage of the first container 53. Due to the breakage of the first container 53, the first reagent 65 from the first container 53 comes out into the second container 55 and mixes with the sample 69. Then, by shaking the second container 55 appropriately, the first mixed solution 77 is prepared.

[0167] Next, as shown in Figure 30(a), with the first spacer 123, the fourth container 59, the second spacer 125, and the third container 57 placed inside the fifth container 61, the first mixed liquid 77 from the second container 55 is poured into the fifth container 61. When the first mixed liquid 77 from the second container 55 is poured into the fifth container 61, the filter cap 89 and the fifth container 61 are separated. However, as shown in Figure 30(b), when the first mixed liquid 77 from the second container 55 is poured into the fifth container 61, a portion of the filter cap 89 may be inside the fifth container 61. Furthermore, the filter cap 89 may be connectable to the opening 67 of the second container 55 and the opening 75 of the fifth container 61. Alternatively, with the filter-equipped cap 89 connected to the opening 67 of the second container 55 and the opening 75 of the fifth container 61, the first mixed liquid 77 in the second container 55 may be poured into the fifth container 61.

[0168] Next, as shown in Figure 31, a cap 127 is placed on the fifth container 61, closing the opening 75 of the fifth container 61. Then, in the same manner as shown in Figure 29, force is applied to the fifth container 61 and the third container 57 to break the third container 57. The broken line indicated by reference numeral 135 in Figure 31 indicates the point of destruction of the third container 57. Upon destruction of the third container 57, the second reagent 71 from the third container 57 flows into the fifth container 61 and mixes with the first mixture 77 to prepare the second mixture 138. The fifth container 61 may be shaken for the above mixing.

[0169] Next, as shown in Figure 32, force is applied to the fifth container 61 and the fourth container 59, and the fourth container 59 is destroyed in the same manner as shown in Figure 29. The broken line indicated by reference numeral 137 in Figure 32 indicates the location of the destruction of the fourth container 59. Upon destruction of the fourth container 59, the third reagent 73 from the fourth container 59 is released into the fifth container 61, and the third reagent 73 is mixed with the second mixture 138, which is a mixture of the first mixture 77 and the second reagent 71, to prepare the third mixture 139. The fifth container 61 may be shaken for the above mixing. The third mixture 139 may then be allowed to stand, and after separation into two layers, an upper layer which is aqueous and a lower layer which is organic, it may be determined whether or not the sample contains Δ9-THC based on the color of the lower layer.

[0170] Furthermore, since the color of the third mixture 139 is visible through the fifth wall 119 of the fifth container 61, it is desirable that the fifth wall 119 of the fifth container 61 be colorless and transparent, however, the fifth wall 119 of the fifth container 61 may be a white semi-transparent material with a high degree of light transmittance.

[0171] In the test kit 51, the first container 53 to the fifth container 61 are formed in a cylindrical shape. The first container 53, the third container 57, and the fourth container 59 are made of glass that is prone to brittle fracture. The second container 55 and the fifth container 61 are made of synthetic resin that is resistant to brittle fracture and has appropriate elasticity. In addition, the outer diameter of the cylindrical first container 53 is slightly smaller than the inner diameter of the cylindrical second container 55, and the outer diameters of the cylindrical third container 57 and the fourth container 59 are slightly smaller than the inner diameter of the cylindrical fifth container 61.

[0172] As a result, by applying a bending moment to the first container 53 that has entered the second container 55, the first container 53 can be easily destroyed. Similarly, by applying a bending moment to the third container 57 and the fourth container 59 that have entered the fifth container 61, the third container 57 and the fourth container 59 can be easily destroyed.

[0173] Furthermore, in the test kit 51, with the third container 57, the fourth container 59, the first spacer 123, and the second spacer 125 inside the fifth space 121, the first spacer 123, the fourth container 59, the second spacer 125, and the third container 57 are arranged in this order from the bottom of the fifth container 61 toward the opening 75. In other words, the second spacer 125 is located between the fragile third container 57 and the fourth container 59, preventing them from colliding with each other. Also, the first spacer 123 is located between the fifth container 61 and the fragile fourth container 59, preventing them from colliding with each other. As a result, there is no risk of the third container 57 and the fourth container 59 being damaged when they are placed inside the fifth container 61. Furthermore, the first spacer prevents container fragments from the third container 57 and the fourth container 59 from falling towards the bottom of the fifth container 61. This allows the color of the lower layer, which separates into an upper layer (aqueous layer) and an organic layer (above) after the third mixed liquid 139 has been allowed to stand, to be observed without interference from the container fragments.

[0174] Although this embodiment has been described above, this embodiment is not limited to these, and various modifications are possible within the scope of the gist of this embodiment. [Explanation of Symbols]

[0175] 51 Test Kits 53 1st container 55 Second container 57 Third container 59 4th container 61 5th container 63 Filters 65 Reagent 1 67 Opening 69 samples 71 Reagent 2 73 Reagent No. 3 75 Opening 77 First mixed liquid 79 1st wall 81 1st space 83 Second wall 85 Second space 87 Cap body 89 Filtered cap 111 Third wall 113 Third space 115 4th wall 117 4th space 119 5th wall 121 5th space 123 First Spacer 125 Second Spacer

Claims

1. A first reagent comprising 2-hydroxybenzaldehyde and a water-soluble organic solvent capable of dissolving the 2-hydroxybenzaldehyde, The second reagent is hydrochloric acid, A third reagent containing an organic solvent that has a higher specific gravity than water and separates from water, A test kit equipped with the following features.

2. The test kit according to claim 1, wherein the third reagent comprises an organic solvent that separates into two layers with water.

3. The test kit according to claim 1 or 2, wherein the third reagent comprises at least one selected from the group consisting of chloroform, dichloromethane, and 1,2-dichloroethane.

4. The test kit according to claim 1 or 2, wherein the test kit is a test kit for determining whether a sample contains Δ9-THC.

5. A first container in which the first reagent is sealed, A second container having an opening, which contains the specimen and the first container, The third container in which the second reagent is sealed, The third reagent is sealed in a fourth container, A fifth container having an opening, which houses the third container and the fourth container, A filter for filtering the first mixture prepared in the second container by mixing the first reagent and the sample and placing it into the fifth container, A test kit comprising the features of claim 1 or 2.

6. In the first container, a first space is formed, which is closed by a first wall made of a first material, and the first reagent is contained within this first space. In the second container, a second space with the opening is formed by a second wall made of a second material having a greater fracture toughness value than the first container, and the sample and the first container can be placed into the second space through the opening. The filter is provided on the cap body and together with the cap body forms a filter-equipped cap, the cap body is made of a material with a greater fracture toughness value than the first container, and is installed at the opening of the second container to seal the opening of the second container. In the third container, a third space is formed, which is closed by a third wall made of a third material, and the second reagent is contained within this third space. In the fourth container, a fourth space is formed, which is closed by a fourth wall made of a fourth material, and the third reagent is contained within this fourth space. The inspection kit according to claim 5, wherein the fifth container has a fifth space with an opening formed by a fifth wall made of a fifth material having a greater fracture toughness value than the third and fourth containers, and the third container, the fourth container and the first mixed liquid can be placed into the fifth space through the opening.

7. A first spacer is made of a material with a greater fracture toughness value than the third and fourth containers, A second spacer is made of a material with a greater fracture toughness value than the third and fourth containers, The first spacer and the second spacer can be inserted into the fifth space through the opening of the fifth container. The inspection kit according to claim 6, wherein, when the third container, the fourth container, the first spacer, and the second spacer are placed in the fifth space, the first spacer, the fourth container, the second spacer, and the third container are arranged in this order from the bottom of the fifth container toward the opening.

8. A step of preparing a first mixture by mixing a first reagent containing a water-soluble organic solvent in which 2-hydroxybenzaldehyde is dissolved with the sample, The process involves mixing the liquid contained in the first mixture with a second reagent, which is hydrochloric acid, to prepare a second mixture. The process involves mixing the second mixture with a third reagent containing an organic solvent that has a higher specific gravity than water and separates from water. Testing methods, including those mentioned above.

9. In the step of mixing the first reagent, force is applied to the first container filled with the first reagent and the second container containing the sample to generate stress, causing the first reagent to flow out of the first container, and the first reagent and the sample are mixed in the second container. In the step of mixing the second reagent, by applying force to the fifth container to generate stress, the second reagent is discharged from the third container, which is housed inside the fifth container and filled with the second reagent, and the liquid contained in the first mixture is mixed with the second reagent. The inspection method according to claim 8, wherein in the step of mixing the third reagent, force is applied to the fifth container to generate stress, thereby causing the third reagent to flow out from the fourth container, which is housed in the fifth container and filled with the third reagent, and the second mixture and the third reagent are mixed.

10. The inspection method according to claim 9, further comprising the steps of filtering the first mixture with a filter-equipped cap attached to the opening of the second container, and transferring the filtrate, which is the liquid contained in the first mixture, to the fifth container.

11. The testing method according to any one of claims 8 to 10, wherein the first mixed solution is prepared by dissolving the sample in the first reagent.

12. The aforementioned specimen includes plant fragments, The testing method according to any one of claims 8 to 10, wherein the liquid contained in the first mixed solution contains an extracted component extracted from the sample by the first reagent.

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