Rapid detection method for gamma-butyrolactone in beverage and application

Abstract

The invention discloses a method for enabling people to predict whether a beverage contains GBL or not in advance. Based on the principle that a purple complex is immediately formed by mixing and reacting GBL and an alkaline hydroxylamine solution and then adding acidic ferric chloride, reaction conditions are optimized, the relationship between the concentration of GBL in a beverage and the degree of chromogenic reaction is researched, and an optimal use method of portable qualitative test paper capable of being used on site is designed. The detection shows that the obvious change of colors of various drinks can be detected, and on the basis of not influencing continuous drinking, whether a sample contains GBL with the dosage exceeding the safe dosage or not can be detected within one minute, and the detection limit is 1 mu l / ml and is within the edible safety limit. The method is simple to operate, quick in reaction, low in cost and hopeful for large-scale production.

Description

Technical Field

[0001] The invention belongs to the field of food safety detection, and specifically relates to a rapid detection method for gamma-butyrolactone in various beverages. Background Art

[0002] Gamma-butyrolactone (GBL) is a precursor of the synthetic drug gamma-hydroxybutyric acid (GHB), commonly known as "obedient water". It is colorless and tasteless and easily soluble in drinks without obvious changes. Studies have shown that GBL has the same effect as GHB, and after being taken by the human body, it will relax the nerves and even cause deep sleep, making it impossible to make strong resistance behaviors. Gamma-hydroxybutyric acid (GHB) is an endogenous component in brain tissue and has an inhibitory effect on the central nervous system (CNS). It is reported that a dose of 0.5g of GHB produces a relaxed and stress-free state, a dose of 1g produces a euphoric effect, and a dose of 2-3g produces a deep sleep. In other words, in a 200ml drink, putting 1g of GHB to a concentration of about 5mg / ml GHB and drinking it all will produce a euphoric effect. GHB is colorless and tasteless, and can be easily added to drinks without the victim noticing, and it is quickly eliminated from the body and can only be detected in urine samples within 6-12 hours after ingestion. As a result, criminals have taken advantage of its effects to commit drug-facilitated sexual assault (DFSA), which refers to sexual assault without the victim's consent, and the victim is unable to take appropriate measures because they cannot distinguish whether GHB or GBL is in their drink. DFSA has become a global concern in recent years, with the number of cases increasing significantly.

[0003] In 2005, my country listed "γ-hydroxybutyric acid" as a Class II psychotropic drug for control, and changed it to Class I in 2007. It was still not lowered in 2013. γ-Butyrolactone (GBL) and 1,4-butanediol are precursors of γ-hydroxybutyric acid (GHB). Under alkaline conditions, they are converted into GHB by hydrolysis; under acidic conditions, GHB can be reversely converted into GBL by intramolecular esterification. On September 20, 2021, γ-Butyrolactone was listed as a drug that has been included in the "Regulations on the Management of Precursor Chemicals", but it is easier for criminals to obtain than GHB. The harm and anesthetic effects of GHB on the human body have been confirmed by many people, but whether GBL has a strong anesthetic effect has rarely been confirmed. Foreign studies have shown that in baboons and other organisms, the effectiveness of these three substances is from strong to weak: GBL, 1,4BD, GHB, and they differ in the onset and duration of the effect. The induction of behavioral damage is from fast to slow in the order of GBL, 1,4BD, and GHB. It can be seen that the anesthetic effect of GBL on the human body cannot be underestimated.

[0004] The quantitative and qualitative research steps of GHB and GBL by domestic and foreign researchers are complicated. The instrumental analysis methods include LC-MS, GC-MS, infrared spectroscopy, nuclear magnetic resonance (NMR), etc. Rapid detection in China has been at least seven years ago and has not been used in practical applications. Several potential miniaturized methods based on optical sensing and the use of chromogenic fluorescent probes have been reported abroad recently, but these methods still show certain shortcomings for the detection of GHB and GBL, such as the use of organic solvents, the application of screening technology, high detection limits in aqueous solutions, and difficulty in portable pretreatment. There is no detection method for GBL. In short, the simple and rapid on-site detection method of GBL in beverages required to prevent drug-assisted sexual assault through rapid detection methods has yet to be developed. Summary of the invention

[0005] In view of the defects of the prior art and the characteristics of the materials, the present invention provides a method for detecting γ-butyrolactone in beverages, which has the advantages of simple operation, good detection effect, excellent stability and low cost.

[0006] In order to achieve the above object, the present invention is implemented by the following technical solutions:

[0007] The method for detecting GHB is decomposed and optimized, the process of converting GHB into GBL is removed, hydroxylamine hydrochloride is replaced with hydroxylamine sulfate, and the liquid addition step is simplified. The specific preparation and operation method of the detection agent includes the following steps:

[0008] (1) 0.1-0.3 mol / L hydroxylamine sulfate solution and 1-3 mol / L sodium hydroxide solution were mixed in a ratio of 1:1, and ultrasonically treated at 70-90 kHz for 15 min to accelerate dissolution, thereby preparing reagent A.

[0009] (2) Mix 0.3-1.0 mol / L ferric chloride solution and 0.5-2 mol / L sulfuric acid solution in a ratio of 1:1, and perform ultrasonic treatment for 15-20 min to accelerate dissolution, thereby preparing reagent B.

[0010] (3) Take 1.0-5.0 ml of the GBL solution to be tested into a test tube, add 200 μl-1000 μl of reagent A, and gently shake the test tube to allow it to react fully.

[0011] (4) After waiting for 2-3 seconds, add 200μl-1000μl of reagent B and gently shake the test tube to allow for full reaction.

[0012] (5) Observe the experimental phenomena. If the color of the liquid is close to that of the ferric chloride solution or is still colorless, then the solution does not contain GBL. If the color of the liquid turns obviously purple-red, then it contains GBL.

[0013] (6) The purple-red reaction product was diluted five times and immediately scanned in the wavelength range of 300-800 nm using a UV-visible spectrophotometer. A characteristic peak was found at 496 nm, and the absorption peak was proportional to the concentration of GBL.

[0014] The above method is suitable for rapid qualitative and semi-quantitative detection in the laboratory. For daily safety detection, the present invention designs a portable detection scheme as follows.

[0015] Considering that the reagents used in the test plan are toxic to humans after ingestion, the drinks after testing cannot be drunk, so the inspection process requires the drinks to be taken out for testing. At the same time, considering the portability of the test agent, the design of the attached figure Figure 2 The portable solution of test strips + reaction tubes shown in the figure. The specific preparation and operation method of the detection agent includes the following steps:

[0016] (1) Use the single factor experiment method to determine the dosage of reagent B, and then determine the optimal pH of the reagent B solution to obtain the optimal ratio of the reagent B solution. Cut the base material in Table 1 into a square with a diameter of 10 cm × 10 cm, place it flat in the reagent B solution and soak it for 3h-24h, dry it in a constant temperature fume hood at 25℃ for 24h, use a rubber-tipped dropper to absorb the excess liquid, and then place it in a 40℃-50℃ oven to dry for 2h-4h, cut it into 5mm × 40mm, seal it and store it for use. Although the dried test strips are basically harmless to human skin, in order to ensure absolute safety, the test strips are wrapped in a plastic shell.

[0017] (2) Reagent A is stored in the reaction tube. Since it is an alkaline solution, a plastic container should be selected and stored in a refrigerator at 0℃-10℃, and the seal should be ensured to be good. The scale value is marked on the reaction tube to facilitate the control of the amount of drink required. The top of the reaction tube is designed as a dropper style, which can be directly dropped into the plastic shell of the test strip to observe the color change.

[0018] Table 1 Test paper parameters

[0019] BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is the ultraviolet spectroscopic spectrum of the product obtained by detecting GBL with a gradient concentration of 0-5 mg / ml using pure water as the solvent using the method of the present invention.

[0021] Figure 2 The absorbance (A) of the reaction product of the present invention for detecting GBL (using pure water as solvent) at 496 nm is the ordinate, and the concentration (C, mg·ml -1 ) is the horizontal axis for linear regression fitting results.

[0022] Figure 3 Schematic diagram of the operation of testing GBL in beverages using the portable solution of test strips + reaction tubes.

[0023] Figure 4 The UV spectra of the products obtained by liquid detection of GBL solutions in different solvents of the present invention are (A) Harbin Beer (B) Nongfu Oriental Leaf Jasmine Tea (C) Bairuka White Rum (D) Shabaohuang Vodka. DETAILED DESCRIPTION

[0024] The following examples are intended to further illustrate the above contents of the present invention in detail, but the protection scope of the present invention is not limited to the following specific examples.

[0025] Example 1

[0026] The method for rapid qualitative and semi-quantitative detection of γ-butyrolactone in beer-like beverages comprises the following steps:

[0027] (1) 0.2 mol / L hydroxylamine sulfate and 2 mol / L sodium hydroxide solution were mixed in a ratio of 1:1, and ultrasonically treated at 70-90 kHz for 15 min to accelerate dissolution, thereby preparing reagent A.

[0028] (2) Mix 0.7 mol / L ferric chloride solution and 1 mol / L sulfuric acid solution in a ratio of 1:1 and perform ultrasonic treatment for 15 min to accelerate dissolution to prepare reagent B.

[0029] (3) Prepare GBL beer samples containing the following gradient concentrations: 0 μl / hl, 0.2 μl / ml, 0.4 μl / ml, 0.6 μl / ml, 0.8 μl / ml, and 1.0 μl / ml.

[0030] (4) Take 1 ml of GBL of each concentration and place it in a test tube. Add 200 μl of reagent A and gently shake the test tube to allow for full reaction.

[0031] (5) After waiting for 2-3 seconds, add 200 μl of reagent B and gently shake the test tube to allow for full reaction.

[0032] (6) Observe the experimental phenomenon. As the concentration of GBL increases, the purple-red color gradually deepens. After the reaction product is diluted five times, the solution is immediately scanned in the wavelength range of 300-800 nm using a UV-visible spectrophotometer. It can be found that there is a characteristic peak at 496 nm. The absorption peak is proportional to the concentration of GBL. The fitting equation is recorded for subsequent quantitative detection.

[0033] Example 2

[0034] The portable method for rapid qualitative detection of γ-butyrolactone in beer-like beverages comprises the following steps:

[0035] (1) 0.3 mol / L hydroxylamine sulfate and 2 mol / L sodium hydroxide solution were mixed in a ratio of 1:1, and ultrasonically treated at 70-90 kHz for 15 min to accelerate dissolution, thereby preparing reagent A.

[0036] (2) Take 200 μl of reagent A and place it in a reaction tube for later use.

[0037] (3) Mix 0.8 mol / L ferric chloride solution and 1 mol / L sulfuric acid solution in a ratio of 1:1 and perform ultrasonic treatment for 15 min to accelerate dissolution to prepare reagent B.

[0038] (4) Cut the substrate material into a square with a diameter of 10 cm × 10 cm, place it flat in the reagent B solution and soak it for 8 h, dry it in a constant temperature fume hood at 25 °C for 24 h, absorb the excess liquid with a rubber-tipped dropper, and then place it in a 40 °C oven to dry for 2 h. Cut it into 5 mm × 40 mm squares and seal it in a plastic shell for storage.

[0039] (5) The above preparation process can be prepared in advance before the test, which is convenient for users to carry.

[0040] (6) Add Harbin beer containing GBL to a reaction tube. Control the amount of the sample to the 1 ml mark on the reaction tube and shake gently to ensure a complete reaction.

[0041] (7) Open the small opening at the top of the reaction tube and drop 3-4 drops of sample onto the test paper. It can be observed that the test paper changes from light yellow to purple-red immediately, which indicates that the Harbin beer sample contains GBL.

[0042] Example 3

[0043] The method for rapid qualitative and semi-quantitative detection of γ-butyrolactone in common foreign wines, and its specific preparation and operation method comprises the following steps:

[0044] (1) 0.2 mol / L hydroxylamine sulfate and 3 mol / L sodium hydroxide solution were mixed in a ratio of 1:1, and ultrasonically treated at 70-90 kHz for 15 min to accelerate dissolution, thereby preparing reagent A.

[0045] (2) Mix 0.7 mol / L ferric chloride solution and 2 mol / L sulfuric acid solution in a ratio of 1:1 and perform ultrasonic treatment for 15 min to accelerate dissolution to prepare reagent B.

[0046] (3) Prepare GBL liquor samples containing the following gradient concentrations: 0 μl / ml, 0.2 μl / ml, 0.4 μl / ml, 0.6 μl / ml, 0.8 μl / ml, 1.0 μl / ml.

[0047] (4) Take 1 ml of GBL of each concentration and place it in a test tube. Add 200 μl of reagent A and gently shake the test tube to allow for full reaction.

[0048] (5) After waiting for 2-3 seconds, add 200 μl of reagent B and gently shake the test tube to allow for full reaction.

[0049] (6) Observe the experimental phenomenon. As the concentration of GBL increases, the purple-red color gradually deepens. After the reaction product is diluted five times, the solution is immediately scanned in the wavelength range of 300-800 nm using a UV-visible spectrophotometer. It can be found that there is a characteristic peak at 496 nm. The absorption peak is proportional to the concentration of GBL. The fitting equation is recorded for subsequent quantitative detection.

[0050] Example 4

[0051] The portable method for rapid qualitative detection of γ-butyrolactone in foreign wines comprises the following steps:

[0052] (1) 0.2 mol / L hydroxylamine sulfate and 2 mol / L sodium hydroxide solution were mixed in a ratio of 1:1, and ultrasonically treated at 70-90 kHz for 15 min to accelerate dissolution, thereby preparing reagent A.

[0053] (2) Take 200 μl of reagent A and place it in a reaction tube for later use.

[0054] (3) Mix 0.7 mol / L ferric chloride solution and 1 mol / L sulfuric acid solution in a ratio of 1:1 and perform ultrasonic treatment for 15 min to accelerate dissolution to obtain reagent B.

[0055] (4) Cut the substrate material into a square with a diameter of 10 cm × 10 cm, place it flat in the reagent B solution and soak it for 3 h, dry it in a constant temperature fume hood at 25 °C for 24 h, absorb the excess liquid with a rubber-tipped dropper, and then place it in a 40 °C oven to dry for 2 h. Cut it into 5 mm × 40 mm squares and seal it in a plastic shell for storage.

[0056] (5) The above preparation process can be prepared in advance before the test, which is convenient for users to carry.

[0057] (6) Add the foreign liquor containing GBL to a reaction tube. Keep the sample volume at the 1 ml mark on the reaction tube and shake gently to ensure a complete reaction.

[0058] (7) Open the small opening at the top of the reaction tube and drop 3-4 drops of sample onto the test paper. If you can immediately observe that the test paper changes from light yellow to purple-red, it can be inferred that the foreign liquor contains GBL.

[0059] Example 5

[0060] The portable method for rapid qualitative and semi-quantitative detection of γ-butyrolactone in pure water comprises the following steps:

[0061] (1) 0.2 mol / L hydroxylamine sulfate and 2 mol / L sodium hydroxide solution were mixed in a ratio of 1:1, and ultrasonically treated at 70-90 kHz for 15 min to accelerate dissolution, thereby preparing reagent A.

[0062] (2) Take 200 μl of reagent A and place it in a reaction tube for later use.

[0063] (3) Mix 0.7 mol / L ferric chloride solution and 1 mol / L sulfuric acid solution in a ratio of 1:1 and perform ultrasonic treatment for 15 min to accelerate dissolution to obtain reagent B.

[0064] (4) Cut the substrate material into a square with a diameter of 10 cm × 10 cm, place it flat in the reagent B solution and soak it for 3 h, dry it in a constant temperature fume hood at 25 °C for 24 h, absorb the excess liquid with a rubber-tipped dropper, and then place it in a 40 °C oven to dry for 2 h. Cut it into 5 mm × 40 mm squares and seal it in a plastic shell for storage.

[0065] (5) The above preparation process can be prepared in advance before the test, which is convenient for users to carry.

[0066] (6) Using pure water as the solvent, prepare GBL samples containing the following gradient concentrations: 0 μl / ml, 0.2 μl / ml, 0.4 μl / ml, 0.6 μl / ml, 0.8 μl / ml, 1.0 μl / ml.

[0067] (7) Take GBL aqueous solutions of different concentrations and put them into reaction tubes. Keep the sample volume at the 1 ml mark in the reaction tube and shake gently to ensure complete reaction.

[0068] (8) Open the small opening at the top of the reaction tube and drop 3-4 drops of sample onto the test paper. You can immediately observe that the sample with GBL changes the test paper from light yellow to purple-red. As the concentration of GBL increases, the purple-red color gradually deepens. Take a photo to record the color change and make a colorimetric card that can be used for quantitative portable testing of pure water.

[0069] Obviously, the above embodiments are examples for clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived from them are still within the protection scope of the invention.

Claims

1. A method for detecting γ-butyrolactone in beverages and its application, characterized in that Based on the principle that a purple-red complex will be formed immediately after GBL is mixed with alkaline hydroxylamine solution and then acidic ferric chloride is added, an optimal method for using a portable qualitative test paper that can be used on site was designed. The specific preparation method comprises the following steps: (1) 0.1-0.3 mol / L hydroxylamine sulfate solution and 1-3 mol / L sodium hydroxide solution were mixed in a ratio of 1:1, and ultrasonically treated at 70-90 kHz for 15 min to accelerate dissolution, thereby preparing reagent A. (2) Mix 0.3-1.0 mol / L ferric chloride solution and 0.5-2 mol / L sulfuric acid solution in a ratio of 1:1, and perform ultrasonic treatment for 15-20 min to accelerate dissolution, thereby preparing reagent B. (3) Take 1.0-5.0 ml of the GBL solution to be tested into a test tube, add 200 μl-1000 μl of reagent A, and gently shake the test tube to allow it to react fully. (4) After waiting for 2-3 seconds, add 200μl-1000μl of reagent B and gently shake the test tube to allow for full reaction. (5) Observe the experimental phenomena. If the color of the liquid is close to that of the ferric chloride solution or is still colorless, then the solution does not contain GBL. If the color of the liquid turns obviously purple-red, then it contains GBL. (6) The purple-red reaction product was diluted five times and immediately scanned in the wavelength range of 300-800 nm using a UV-visible spectrophotometer. A characteristic peak was found at 496 nm, and the absorption peak was proportional to the concentration of GBL.

2. The method according to claim 1, characterized in that: In the steps (1) and (2), the solution concentration and ratio are fixed and effective. The amount of reagent A added should be 5%-30%, the amount of reagent B added should be 5%-30%, and the amount of the sample to be tested should be 50%-80%.

3. The method according to claim 1, characterized in that: In the steps (3) and (4), the order of adding the detection agents is: first adding reagent A, then adding reagent B.

4. The principle as claimed in claim 1, characterized in that: It can be used to design a portable test device of test strips + reaction tubes. The specific production process is as follows: cut the base material into a square with a diameter of 10cm×10cm, place it flat in the reagent B solution and soak it for 3h-24h, dry it in a constant temperature fume hood at 25℃ for 24h, use a rubber-tipped dropper to absorb the excess liquid, and then place it in a 40℃-50℃ oven to dry for 2h-4h, cut it into 5mm×40mm, seal it and store it for use, and wrap the test strip in a plastic shell to ensure absolute safety. Reagent A is stored in the reaction tube. Because it is an alkaline solution, a plastic container is selected and stored in a refrigerator at 0℃-10℃, and the seal is ensured to be good. The scale value is marked on the reaction tube to facilitate the control of the amount of drink required. The top of the reaction tube is designed as a dropper style, which can be directly dropped into the plastic shell of the test strip to observe the color change.

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