A method for the determination of cooling agents in a mouthpiece

Abstract

The application belongs to the technical field of oral products, and particularly relates to a method for determining a cooling agent in an oral product. The method comprises the following steps: mixing a base and a dispersing agent in the oral product to obtain a mixture; performing dynamic headspace extraction on the mixture to obtain an extract, and delivering the extract to a GC-MS to obtain a cooling agent peak area A1; obtaining multiple cooling agent peak areas after the above steps are repeated according to a target extraction number; and obtaining a cooling agent content according to the multiple cooling agent peak areas. The method combines dynamic headspace extraction and GC-MS, and utilizes multiple dynamic headspace extractions and concentration, so that the recovery rate of menthol is increased, and the accuracy of the cooling agent measurement is ensured.

Description

Technical Field

[0001] This application belongs to the field of detection technology, specifically relating to a method for determining cooling agents in oral products. Background Technology

[0002] To enhance the taste of lozenges, cooling agents such as menthol are often added. Determining the content of these cooling agents is crucial for product quality control, batch stability, formula development, and compliance assessment.

[0003] Oral supplements consist of a matrix including purified water, active ingredients, microcrystalline cellulose, pH adjusters, and cooling agents. The cooling agent is encapsulated within the matrix, making complete separation difficult with conventional solvent extraction. This results in incomplete extraction, low recovery rates, and ultimately affects the accuracy of cooling agent measurement. Furthermore, due to the high water content in oral supplements, conventional GC-MS cannot be used to directly determine the cooling agent content, while GC-MS alone fails to meet the detection limit requirements. Summary of the Invention

[0004] This application aims to provide a method for determining cooling agents in oral products, in order to solve the problem of low accuracy in the measurement of cooling agents in the prior art.

[0005] To address the aforementioned technical problems, this application provides a method for determining cooling agents in oral products. The method includes: mixing a matrix and a dispersant in the oral product to obtain a mixture; performing dynamic headspace extraction on the mixture to obtain an extract, and sending the extract to GC-MS to obtain the peak area A1 of the cooling agent; repeating the above steps according to a target number of extractions to obtain multiple peak areas of the cooling agent, and obtaining the cooling agent content based on the multiple peak areas of the cooling agent.

[0006] This method uses dynamic headspace extraction coupled with GC-MS, and by performing multiple dynamic headspace extractions and concentrations, it increases the recovery rate of cooling agents and improves the accuracy of cooling agent measurement.

[0007] In one embodiment, the matrix and dispersant in the oral product are mixed to obtain a mixture, including: mixing 0.3~0.7g of matrix and 5~15mL of dispersant to obtain a mixture. This dosage setting ensures that the sample is fully wetted and dispersed, taking into account both the sensitivity of detection and the capacity of the headspace vial, thereby ensuring extraction efficiency and method stability.

[0008] In one embodiment, the dispersant includes at least one of the following: water, ethanol, an aqueous ethanol solution, a saturated aqueous sodium chloride solution, or a low-co-solubility dispersant. These dispersants can improve the dispersibility and surface area of ​​the oral product being tested during dynamic headspace extraction, thereby promoting the release and escape of the cooling agent from the matrix.

[0009] Specifically, the mass of the matrix is ​​0.3g, 0.5g, 0.7g, or any value within the above range; the volume of the dispersant is 5mL, 10mL, 15mL, or any value within the above range.

[0010] In one embodiment, the parameters of the dynamic headspace extraction include: column oven temperature of 80-140°C, equilibration time of 15-60 min, quantitative loop temperature of 140-180°C, nitrogen purging time of 3-15 min, and nitrogen purging flow rate of 20-50 mL / min; and transfer line temperature of 130-160°C. The moderate column oven temperature promotes the full volatilization of the cooling agent without decomposition, the longer equilibration time ensures solid-gas distribution balance, and the higher quantitative loop and transfer line temperatures prevent the cooling agent from condensing and adsorbing. Combined with the appropriate nitrogen purging time and flow rate, efficient, stable, and repeatable extraction and transfer are achieved, thereby obtaining a stable and highly responsive cooling agent peak area, supporting the accuracy of multi-step cumulative extraction.

[0011] Specifically, the column oven temperature is 80℃, 100℃, 120℃, 140℃ or any value within the above range; the equilibration time is 15min, 30min, 45min, 60min or any value within the above range; the metering loop temperature is 140℃, 160℃, 180℃ or any value within the above range; the nitrogen purging time is 3min, 5min, 10min, 15min or any value within the above range; the nitrogen purging flow rate is 20mL / min, 30mL / min, 40mL / min, 50mL / min or any value within the above range; and the transfer line temperature is 130℃, 140℃, 150℃, 160℃ or any value within the above range.

[0012] In one embodiment, determining the target number of extractions includes: performing an nth dynamic headspace extraction on the mixture to obtain an extract, and then sending the extract to GC-MS to obtain the peak area A of the cooling agent. n Determine the peak area A of the cooling agent. n The ratio of the peak area A1 of the cooling agent to the peak area of ​​the cooling agent is used. If the ratio is less than 0.01, the cycle ends, and n times is taken as the target number of extractions for the determination of cooling agent in oral products.

[0013] In one embodiment, the method further includes: obtaining a cooling agent attenuation coefficient; calculating a matrix effect factor Q based on the target number of extractions and the cooling agent attenuation coefficient; wherein the matrix effect factor Q is used to characterize the adsorption effect of the matrix on the cooling agent. The larger the Q, the greater the adsorption effect of the matrix on the cooling agent, and the more difficult it is to measure in oral products.

[0014] In one embodiment, the formula for calculating the attenuation coefficient of the cooling agent is: Where β is the attenuation coefficient; A nA represents the peak area of ​​the cooling agent in the nth extraction; n-1 The peak area of ​​the cooling agent in the (n-1)th extraction is denoted by ; n is the number of extractions.

[0015] In one embodiment, the matrix effect factor Q is calculated using the following formula: Where Q is the matrix effect factor; β is the attenuation coefficient; and n is the number of extractions.

[0016] In one embodiment, the cooling agent includes at least one of the following: menthol, menthone, and menthol.

[0017] In one embodiment, the ethanol-water solution contains 5-15% ethanol by mass. This ratio promotes the release of the cooling agent from the sample while maintaining appropriate polarity to match the distribution behavior of the cooling agent, thereby improving the recovery and reproducibility of multiple dynamic headspace extractions.

[0018] Specifically, in the ethanol-water solution, the mass fraction of ethanol is 5%, 10%, 15%, or any value within the above range.

[0019] In one embodiment, the eutectic dispersant is a mixture of saturated sodium chloride aqueous solution and decanoic acid. The saturated brine reduces the solubility of the cooling agent in water through the salting-out effect, while decanoic acid enhances the affinity extraction of the cooling agent. The two work synergistically to improve the release efficiency of the cooling agent into the headspace without interfering with GC-MS detection, thereby improving recovery rate and accuracy.

[0020] In one embodiment, the mass ratio of saturated sodium chloride aqueous solution to decanoic acid in the eutectic dispersant is 4:1 to 19:1. Within this ratio range, the eutectic dispersant maintains a homogeneous and stable liquid system, enabling continuous and uniform wetting and dispersion of the oral product matrix, ensuring a stable release of the cooling agent during multiple dynamic headspace extraction processes.

[0021] Specifically, in the eutectic dispersant, the mass ratio of saturated sodium chloride aqueous solution to decanoic acid is 4:1, 9:1, 14:1, 19:1, or any value within the above range.

[0022] In one embodiment, the matrix further includes at least one of the following: an active substance, a pH adjuster, a flavoring, or a sweetener.

[0023] Specifically, the active substance includes at least one of nicotine and nicotine derivatives. For example, nicotine includes natural nicotine and / or synthetic nicotine, and nicotine derivatives include one or more of nicotine salts, nicotine in a matrix such as a glycobase or an organometallic complex, nicotine-resin combinations, nicotine inclusion complexes, and non-covalently bound nicotine.

[0024] Non-covalently bonded nicotine includes nicotine lactate, nicotine malate, nicotine salicylate, nicotine cyclodextrin encapsulated complex, nicotine hydrochloride, nicotine dihydrochloride, nicotine tartrate, nicotine tartrate dihydrate, nicotine sulfate, nicotine zinc chloride, and nicotine benzoate. Nicotine derivatives also include nicotine with substituents, such as hexamethylnicotine, hexamethylnicotine lactate, hexamethylnicotine malate, hexamethylnicotine salicylate, hexamethylnicotine cyclodextrin encapsulated complex, hexamethylnicotine hydrochloride, hexamethylnicotine dihydrochloride, hexamethylnicotine tartrate, hexamethylnicotine tartrate dihydrate, hexamethylnicotine sulfate, hexamethylnicotine zinc chloride, and hexamethylnicotine benzoate, among one or more mixtures thereof.

[0025] Specifically, the pH adjuster includes at least one of the following: citric acid, malic acid, tartaric acid, fumaric acid, phosphoric acid, and sodium bicarbonate.

[0026] Specifically, the flavorings include at least one of the following: bergamot flavoring, eucalyptus flavoring, citrus flavoring, lemon flavoring, peppermint flavoring, peppermint flavoring, menthol, licorice flavoring, wintergreen flavoring, tobacco flavoring, coffee flavoring, vanilla flavoring, lime flavoring, apple flavoring, peach flavoring, mango flavoring, cherry flavoring, blueberry flavoring, strawberry flavoring, cola flavoring, cinnamon flavoring, pandan flavoring, and watermelon flavoring.

[0027] Specifically, the sweetener includes at least one of the following: xylitol, sorbitol, mannitol, iodine, lactitol, maltitol, isomaltitol, hydrogenated starch hydrolysate, erythritol, maltodextrin, aspartame, acesulfame potassium, sodium saccharin, sucralose, neotame, cyclamate, alitane, steviol glycosides, arabinitol, and monk fruit sweetener.

[0028] Specifically, the filler includes at least one of the following: isomaltitol, sorbitol, mannitol, xylitol, erythritol, maltitol, lactose, microcrystalline cellulose, pregelatinized starch, dextrin, polydextrose, resistant dextrin, inulin, and oat fiber.

[0029] In one embodiment, the oral product includes a dry-prepared oral product and a wet-prepared oral product; wherein the content of the cooling agent in the dry-prepared oral product is greater than 0.084 mg / kg, and the content of the cooling agent in the wet-prepared oral product is greater than 0.108 mg / kg.

[0030] In other words, the detection limit of this cooling agent determination method is 0.084 mg / kg in oral products prepared by the dry method, and 0.108 mg / kg in oral products prepared by the wet method, which is much higher than the existing technology.

[0031] Additional aspects and advantages of this application will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of this application. Attached Figure Description

[0032] The above and / or additional aspects and advantages of this application will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which: Figure 1 This is a flowchart of the method for determining the cooling agent in oral products. Detailed Implementation

[0033] The embodiments of this application will be described in detail below. These embodiments are exemplary and are only used to explain this application, and should not be construed as limiting this application. Based on the embodiments in this application, all other embodiments obtained by those skilled in the art without inventive effort are within the scope of protection of this application.

[0034] The terms "first" and "second" in the specification and claims of this application may explicitly or implicitly include one or more of the features. In the description of this application, unless otherwise stated, "multiple" means two or more. Furthermore, "and / or" in the specification and claims indicates at least one of the connected objects, and the character " / " generally indicates that the preceding and following objects are in an "or" relationship.

[0035] In the description of this application, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", "axial", "radial", "circumferential", etc., indicating orientation or positional relationships are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of this application.

[0036] In the description of this application, it should be noted that, unless otherwise expressly specified and limited, the terms "installation," "connection," and "linking" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal connection between two components. Those skilled in the art can understand the specific meaning of the above terms in this application based on the specific circumstances.

[0037] This application provides a method for determining the cooling agent in oral products, such as... Figure 1As shown, the method includes: S1. Mix the matrix and dispersant in the oral product to obtain a mixture.

[0038] To better describe the scheme, the cooling agent is menthol, and the eutectic dispersant is a saturated sodium chloride aqueous solution and decanoic acid in a mass ratio of 9:1.

[0039] Step S1 is as follows: Pour 0.5g of the matrix from the oral product into a headspace vial, add 10mL of a low-cosolubility dispersant containing 10mg / L 2-methylquinoline internal standard solution into the headspace vial, and then seal the headspace vial.

[0040] It should also be noted that the dry and wet methods for preparing oral products are different. Therefore, this scheme selects oral products prepared by the dry method (dry oral product) and oral products prepared by the wet method (wet oral product). The content of menthol in the matrix of the dry oral product is 2000 mg / kg (mass of the matrix of the dry oral product is 0.4439 g), and the content of menthol in the matrix of the wet oral product is 2000 mg / kg (mass of the matrix of the wet oral product is 0.5192 g).

[0041] S2. Dynamic headspace extraction is performed on the mixture to obtain the extract, and the extract is sent to GC-MS to obtain the peak area A1 of the cooling agent.

[0042] Dynamic headspace extraction (MHE), often referred to as purge-trap headspace extraction, is a solvent-free sample pretreatment technique for volatile / semi-volatile substances.

[0043] In this scheme, the condition parameters for dynamic headspace extraction are shown in Table 1, and the condition parameters for GC-MS are shown in Table 2.

[0044] Table 1. Conditional parameters for dynamic headspace extraction

[0045] Table 2. Conditional Parameters for GC-MS

[0046] S3. Repeat the above steps to obtain multiple cooling agent peak areas, and obtain the cooling agent content based on the multiple cooling agent peak areas.

[0047] Step S3 specifically includes: S31. Establish the standard curve for menthol.

[0048] Menthol (CAS No.: 1490-04-6) was diluted with 10 mL of a 10 mg / L 2-methylquinoline internal standard solution as a eutectic dispersant to prepare menthol concentrations of 1, 5, 10, 20, 50, 100, and 200 mg / L. These dilutions were then poured into headspace vials, and tests were performed according to steps S2-S3 above. The menthol standard curve was obtained by plotting the ratio of the menthol peak area to the 2-methylquinoline peak area as the ordinate (Y) and the ratio of the menthol concentration to the 2-methylquinoline concentration as the abscissa (X): Y = 1.0009X - 0.0505.

[0049] S32. For dry-process oral products, the peak areas of the cooling agent were measured after three MHE extractions as follows: A1=150036, A2=13501, A3=1249. The ratio of A3 to A1 was 0.008. Since 0.008 is less than 0.01, the cycle can be terminated, indicating that the target number of extractions for dry-process oral products is 3.

[0050] To verify the feasibility of the extraction times in this scheme, the total concentration of menthol detected in the solution was calculated to be 88.5 mg / L based on the peak areas of the cooling agents in the three extractions and the menthol standard curve. Since the weight of the dry-process oral product is 0.4439 g and the dilution volume is 10 mL, the menthol content in the dry-process oral product is 88.5 * 10 / 0.4439 = 1993.7 mg / kg, which is close to the theoretical menthol content of 2000 mg / kg in the dry-process oral product, achieving a recovery rate of 99.68%. Therefore, for the dry-process oral product, a target extraction number of 3 times can achieve the recovery requirement.

[0051] In addition, the adsorption effect of the matrix on menthol in dry oral products can be calculated and represented by the matrix effect factor Q. The larger the Q is, the greater the adsorption effect of the matrix on menthol, and the more difficult the sample is to measure.

[0052] The matrix effect factor Q is calculated as follows: β1=13501 / 150036=0.090, β2=1249 / 13501=0.0925; Q=[(1-0.09)+(1-0.0925)] / 2=0.9087.

[0053] For wet-process oral products, the peak areas of the cooling agent were measured after 5 MHE extractions as follows: A1=120255, A2=59123, A3=9469, A4=1855, A5=569. The ratio of A5 to A1 was 0.005. Since 0.005 is less than 0.01, the cycle can be terminated, indicating that the target number of extractions for wet-process oral products is 5.

[0054] To verify the feasibility of the extraction number in this scheme, based on the peak area of ​​the cooling agent in five extractions and the menthol standard curve, the total concentration of menthol detected in the solution was calculated to be 102.7 mg / L. Since the weight of the wet-process oral product is 0.5192 g and the dilution volume is 10 mL, the menthol content in the wet-process oral product is 102.7 * 10 / 0.5192 = 1978 mg / kg, which is close to the theoretical menthol content of 2000 mg / kg in the wet-process oral product, achieving a recovery rate of 98.9%. Therefore, for the wet-process oral product, a target extraction number of 5 times can achieve the recovery requirement.

[0055] In addition, the adsorption effect of the matrix on menthol in wet-process oral products can be calculated and represented by the matrix effect factor Q. The larger the Q is, the greater the adsorption effect of the matrix on menthol, and the more difficult the sample is to measure.

[0056] The matrix effect factor Q is calculated as follows: β1=59123 / 120255=0.4916, β2=9469 / 59123=0.1602, β3=1855 / 9469=0.1959, β4=569 / 1855=0.3067; Q=[(1-0.4916)+(1-0.1602)+(1-0.1959)+(1-0.3067)] / 4=0.7113.

[0057] According to the matrix effect factor Q, the matrix effect factor Q of the wet process is larger. Therefore, the recovery rate of menthol in wet-process oral products is theoretically lower than that of menthol in wet-process oral products.

[0058] To illustrate the universality of this solution, examples and comparative examples are provided below.

[0059] Example 1 A dry-process oral product is provided, wherein the menthol content in the dry-process oral product is 1.8 mg / g.

[0060] The menthol content of the dry-process oral product was determined using the above steps S1-S3. The determined menthol content was 1.77 mg / g, and the recovery rate reached 98.3%.

[0061] Comparative Example 1 The difference from Example 1 is that: in step S1, the low co-soluble dispersant is replaced with 95% aqueous ethanol solution; in steps S2 and S3, dynamic headspace extraction is replaced with room temperature ultrasonication for 30 minutes, and the extraction is performed once.

[0062] The menthol content determined by this method was 1.36 mg / g, with a recovery rate of 75.8%.

[0063] As can be seen from Example 1 and Comparative Example 1, the scheme in Example 1 uses a low co-soluble dispersant that can effectively disperse the extracted particles. Combined with the multiple headspace extraction and concentration technology of MHE, the recovery rate of menthol is greatly improved.

[0064] Example 2 A wet-process oral product is provided, wherein the menthol content in the wet-process oral product is 1.0 mg / g.

[0065] The menthol content of the wet-process oral product was determined by using the above steps S1-S3. The determined menthol content was 0.953 mg / g, and the recovery rate reached 95.3%.

[0066] Comparative Example 2 The difference from Example 2 is that: in step S1, the low co-soluble dispersant is replaced with 95% aqueous ethanol solution; in steps S2 and S3, dynamic headspace extraction is replaced with room temperature ultrasonication for 30 minutes, and the extraction is performed once.

[0067] The menthol content determined by this method was 0.631 mg / g, with a recovery rate of 63.1%.

[0068] As can be seen from Example 2 and Comparative Example 2, the scheme in Example 2 uses a low co-soluble dispersant that can effectively disperse the extracted particles. Combined with the multiple headspace extraction and concentration technology of MHE, the recovery rate of menthol is greatly improved.

[0069] Example 3 To verify the detection limit of this method in dry-process oral products, a dry-process oral product is provided, wherein the menthol content in the dry-process oral product is 0.10 mg / kg.

[0070] The menthol content of the dry-process oral product was determined using the above steps S1-S3. The determined menthol content was 0.0971 mg / g, and the recovery rate reached 97.1%.

[0071] Example 4 To verify the detection limit of this method in wet-process oral products, a wet-process oral product is provided, wherein the menthol content in the wet-process oral product is 0.10 mg / kg.

[0072] The menthol content of the dry-process oral product was determined by using the above steps S1-S3. The determined menthol content was 0.0963 mg / g, and the recovery rate reached 96.3%.

[0073] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "illustrative embodiment," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of this application. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0074] Although embodiments of this application have been shown and described, those skilled in the art will understand that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of this application, the scope of which is defined by the claims and their equivalents.

Claims

1. A method for determining the cooling agent in a lozenge, characterized in that, The method includes: The matrix and dispersant in a lozenge are mixed to obtain a mixture; The mixture was subjected to dynamic headspace extraction to obtain an extract, and the extract was sent to GC-MS to obtain the peak area A1 of the cooling agent; After repeating the above steps according to the target number of extractions, multiple cooling agent peak areas are obtained, and the cooling agent content is obtained based on the multiple cooling agent peak areas.

2. The method according to claim 1, characterized in that, The step of mixing the matrix and dispersant in the oral product to obtain a mixture includes: mixing 0.3~0.7g of the matrix and 5~15mL of the dispersant to obtain a mixture; The dispersant includes at least one of the following: water, ethanol, aqueous ethanol solution, saturated sodium chloride aqueous solution, and low co-solubility dispersant.

3. The method according to claim 1, characterized in that, The parameters for the dynamic headspace extraction include: column oven temperature of 80~140℃, equilibration time of 15~60min, quantitative loop temperature of 140~180℃, nitrogen purging time of 3~15min, nitrogen purging flow rate of 20~50mL / min, and transfer line temperature of 130~160℃.

4. The method according to claim 1, characterized in that, Determining the target number of extractions includes: performing an nth dynamic headspace extraction on the mixture to obtain an extract, and delivering the extract to the GC-MS to obtain a cooling agent peak area A n ; the ratio of the peak area of the cooling agent A1 to the peak area of the cooling agent A, and if the ratio is less than 0.01, the cycle is ended and the number n is taken as the target extraction number for the determination of the cooling agent in the oral product. n the ratio of the peak area of the cooling agent A1 to the peak area of the cooling agent A, and if the ratio is less than 0.01, the cycle is ended and the number n is taken as the target extraction number for the determination 5. The method according to claim 4, characterized in that, The method further includes: Obtain the attenuation coefficient of the cooling agent; Calculate the matrix effect factor Q based on the target number of extractions and the attenuation coefficient of the cooling agent; The matrix effect factor Q is used to characterize the adsorption effect of the matrix on the cooling agent.

6. The method according to claim 5, characterized in that, The formula for calculating the attenuation coefficient of the cooling agent is as follows: ； Where β is the attenuation coefficient; A n A represents the peak area of ​​the cooling agent in the nth extraction; n-1 The peak area of ​​the cooling agent in the (n-1)th extraction is denoted by ; n is the number of extractions.

7. The method according to claim 6, characterized in that, The formula for calculating the matrix effect factor Q is as follows: ； Where Q is the matrix effect factor; β is the attenuation coefficient; and n is the number of extractions.

8. The method according to claim 2, characterized in that, The ethanol aqueous solution contains 5-15% ethanol by mass.

9. The method according to claim 2, characterized in that, The eutectic dispersant is a mixture of saturated sodium chloride aqueous solution and decanoic acid.

10. The method according to claim 9, characterized in that, In the aforementioned eutectic dispersant, the mass ratio of saturated sodium chloride aqueous solution to decanoic acid is 4:1 to 19:1.

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