Method and equipment for evaluating dissolution behavior of active ingredients in oral product

Abstract

The invention provides a method and equipment for evaluating the dissolution behavior of active ingredients in an oral product. The method comprises the following steps: placing the oral product in a sample chamber; conveying the artificial saliva into the sample chamber, so that the artificial saliva is in contact with the oral product, and active ingredients of the oral product are dissolved in the artificial saliva to form a dissolution solution; detecting the contents of the active components in the dissolution liquid at different time through liquid chromatography to obtain dissolution data of a plurality of groups of active components; and evaluating the dissolution behavior of the oral product according to the multiple groups of dissolution data and a preset similarity factor calculation formula. The active components in the oral product are detected and separated through liquid chromatography and are not easily interfered by other impurity components in the oral product, and the detection efficiency is high. And the evaluation equipment is assembled by only using conventional experimental apparatuses without using flow cell equipment, so that the operation is simple, and the detection time is shortened. Finally, the evaluation method has good discrimination and reproducibility in the detection process, and can stably distinguish the active ingredient dissolution behaviors of the oral products from different manufacturers.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of chemical analysis, in particular to a method and device for evaluating the dissolution behavior of active ingredients in oral products. BACKGROUND

[0002] Oral products are a new type of tobacco product, which are consumed by oral cavity without burning process. The active ingredients of oral products include nicotine and its derivatives (such as nicotine salt). In addition to active ingredients, oral products also contain other ingredients, such as fillers (such as microcrystalline cellulose), sweeteners (such as xylitol or maltitol), other flavors and preservatives, etc.

[0003] Oral products are usually packaged in small synthetic fiber bags. During use, these fiber bags are generally placed between the gums and the upper lip. The active ingredients are dissolved from the bag into the saliva and then absorbed into the body through the oral cavity.

[0004] However, nicotine bags produced by different manufacturers may have different dissolution behaviors. Therefore, a method is needed to evaluate the dissolution behavior of nicotine in nicotine bags.

[0005] The utility model patent of Chinese patent CN208872662U designs a set of in vitro release device for simulating the release of bagged oral tobacco in the oral cavity, which includes a drug dissolution tester with an optical fiber probe. The lower end of the rotating shaft of the dissolution tester is connected with a narrow release unit of the drip type. The rotating shaft is controlled by a stirring controller. In combination with optical fiber transmission technology, ultraviolet spectrum analysis technology and computer data processing technology, the device performs in vitro simulation test on the release of nicotine in bagged oral tobacco. Although this method can detect in real time, if there are other ultraviolet absorbing substances in the bagged oral tobacco, it is easy to interfere with the detection results.

[0006] The invention patent of Chinese patent CN106932229B provides a simulation oral cavity device for collecting oral tobacco dissolution and its use method, which belongs to the technical field of simulation oral cavity device. In the simulation oral cavity state, the device dissolves and collects the substances in the oral tobacco. The device includes two layers of film, the edges of which are sealed to form a film body clamping sleeve. A clamping layer for placing the oral tobacco sample is arranged between the two layers of film. The device uses film permeation or capillary flow method to guide the infiltration liquid into the clamping layer to infiltrate the oral tobacco sample. However, the operation of the device is relatively complex, which may result in poor reproducibility of the experiment.

[0007] Fadi Aldeek et al. published an article in the journal Chromatography, titled "Dissolution Testing of Nicotine Release from OTDN Pouches: Product Characterization and Product-to-Product Comparison", which describes the use of a flow cell device to determine the dissolution curve of 7 flavors and 5 specifications of nicotine pouches, to quantitatively evaluate the dissolution behavior of nicotine in nicotine pouches. The results show that the dissolution behavior of nicotine pouches of different flavors is similar, and the dissolution of nicotine is related to the product characteristics. The nicotine pouch is placed in the sample cell, and the constant flow pump pumps the artificial saliva into the sample cell at a certain flow rate to contact the sample to be tested. Due to the action of the glass beads at the bottom of the sample cell, the artificial saliva flows through the nicotine pouch in a relatively mild laminar flow state, and the nicotine slowly dissolves from the nicotine pouch into the artificial saliva. The sample solution is filtered at the top of the flow cell and then flows out, and the sample solution is collected for analysis and detection. However, this method takes 60 minutes, which is time-consuming, and the dissolution conditions are quite different from the secretion conditions of human oral saliva, which has certain shortcomings. SUMMARY

[0008] The embodiments of the present application provide a method and device for evaluating the dissolution behavior of active ingredients in oral products, which is fast, simple, specific and reproducible.

[0009] In a first aspect, the present application provides a method for evaluating the dissolution behavior of active ingredients in oral products, comprising: placing an oral product in a sample chamber; delivering artificial saliva into the sample chamber so that the artificial saliva contacts the oral product, and the active ingredients of the oral product dissolve in the artificial saliva to form a dissolution liquid; detecting the content of active ingredients in the dissolution liquid at different times by liquid chromatography to obtain a plurality of dissolution data of the active ingredients; evaluating the dissolution behavior of the oral product according to the plurality of dissolution data and a preset similarity factor calculation formula.

[0010] Optionally, the evaluation of the dissolution behavior of the oral product according to the plurality of dissolution data and the preset similarity factor calculation formula comprises: determining the target dissolution data of the active ingredients of the target oral product, the sampling time of the target dissolution data being consistent with the sampling time of the dissolution data; calculating a similarity factor according to the target dissolution data, the dissolution data, and the similarity factor calculation formula; determining whether the similarity factor is less than a similarity threshold value; If yes, it is evaluated that the dissolution behavior of the oral product is not similar to that of the target oral product. If no, it is evaluated that the dissolution behavior of the oral product is similar to that of the target oral product.

[0011] Optionally, the similarity factor calculation formula is: .

[0012] Optionally, the mobile phase of the liquid chromatography is a mixed solution of ammonium formate and methanol.

[0013] Optionally, the pH of the mobile phase is 3.8 to 5.0.

[0014] Optionally, the mobile phase is further added with at least one of triethylamine, diethylamine or ammonia water.

[0015] Optionally, the active ingredient includes nicotine and its derivatives.

[0016] Optionally, the artificial saliva is heated to 36.3 to 37.2℃ before being contacted with the oral product, and the flow rate of the artificial saliva delivered to the sample cell is 0.4 mL / min.

[0017] Optionally, the dissolution time of the oral product is not more than 15 minutes.

[0018] In a second aspect, the present application further provides an evaluation device for the dissolution behavior of an active ingredient in an oral product, which is used to perform the evaluation method according to any one of the first aspect, comprising: a sample chamber for placing the oral product, a liquid inlet is arranged at the top end of the sample chamber, the liquid inlet is used to deliver the artificial saliva, and a closable sampling port is arranged at the bottom end of the sample chamber, the sampling port is used to collect the dissolution liquid; an artificial saliva heating device for storing and heating the artificial saliva; a pump connected between the liquid inlet of the sample chamber and the artificial saliva heating device, for delivering the artificial saliva in the artificial saliva heating device to the sample chamber.

[0019] The embodiment of the present application can achieve the following technical effects: the active ingredient in the oral product is detected and separated by liquid chromatography, the active ingredient is not easily interfered by other impurity components in the oral product, and the detection efficiency is high. Secondly, the evaluation method provided by the embodiment of the present application only uses conventional experimental equipment to assemble an evaluation device, without using a special flow cell device, and the operation is simple and the detection time is shortened. Finally, the evaluation method provided by the embodiment of the present application has good distinguishability and reproducibility in the detection process, and can stably distinguish the dissolution behavior of the active ingredient of oral products of different manufacturers. BRIEF DESCRIPTION OF DRAWINGS

[0020] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings needed to be used in the description of the embodiments of the present application will be briefly introduced. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without creative labor.

[0021] Figure 1 The process schematic diagram of the active ingredient dissolution behavior evaluation method of the oral product provided by an embodiment of the present application is shown in the figure. Figure 2 The standard sample liquid chromatogram provided by an embodiment of the present application is shown in the figure. Figure 3 The standard sample dissolution curve diagram provided by an embodiment of the present application is shown in the figure. Figure 4 The dissolution curve diagram of the peach flavored nicotine bag of manufacturer A provided by an embodiment of the present application is shown in the figure. Figure 5 The different time dissolution curve diagram of the peach flavored nicotine bag of manufacturer A provided by an embodiment of the present application is shown in the figure. Figure 6 The dissolution curve diagram of the peach flavored nicotine bag and the watermelon flavored nicotine bag of manufacturer A provided by an embodiment of the present application is shown in the figure. Figure 7 The dissolution curve diagram of the peach flavored nicotine bag of manufacturer A and the ice flavored nicotine bag of manufacturer B provided by an embodiment of the present application is shown in the figure. Figure 8 The dissolution curve diagram of the peach flavored nicotine bag of manufacturer A and the watermelon flavored nicotine bag of manufacturer C provided by an embodiment of the present application is shown in the figure. Figure 9 The dissolution curve diagram of the different dissolution methods of the peach flavored nicotine bag of manufacturer A provided by an embodiment of the present application is shown in the figure. DETAILED DESCRIPTION

[0022] In order to make the purposes, technical solutions and advantages of the present application clearer, further detailed description will be made to the present application in combination with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present application and not used to limit the present application. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the protection scope of the present application.

[0023] It should be noted that, if there is no conflict, each feature in the embodiments of the present application can be combined with each other, and all fall within the protection scope of the present application. In addition, although the functional modules are divided in the device schematic diagram, and the logical order is shown in the flowchart, in some cases, the steps shown or described can be executed in a different order from the module division in the device or the order in the flowchart. Furthermore, the "first", "second", "third" and the like used in the present application do not limit the data and execution order, but only distinguish the same items or similar items with basically the same function and effect.

[0024] As analyzed in the technical background of the present application, in addition to the active ingredient, the oral preparation also contains other impurity ingredients, if directly dissolved and then used ultraviolet spectrum, interference is easy to produce, so that the detection result is not accurate. And if using other flow cell or self-made dissolution equipment, it may be due to the complex operation that the reproducibility is poor, or the detection time is too long and so on.

[0025] Please refer to Figure 1 , Figure 1 The flowchart of the method for evaluating the dissolution behavior of the active ingredient in the oral preparation provided by an embodiment of the present application is shown in the figure. In order to solve the problem, the embodiment of the present application provides a method for evaluating the dissolution behavior of the active ingredient in the oral preparation, comprising: S1, placing the oral preparation in the sample chamber.

[0026] S2, conveying artificial saliva into the sample chamber, so that the artificial saliva contacts with the oral preparation, the active ingredient of the oral preparation is dissolved in the artificial saliva and forms a dissolution liquid; S3, detecting the content of the active ingredient in the dissolution liquid at different times by liquid chromatography to obtain a plurality of dissolution data of the active ingredient; S4, evaluating the dissolution behavior of the oral preparation according to the plurality of dissolution data and a preset similarity factor calculation formula.

[0027] In step S1, the artificial saliva is first placed in a temperature maintaining device, and heated to a temperature between 36.3°C and 37.2°C by means of constant temperature water bath to simulate the temperature of the saliva in the oral cavity. The oral product is placed in a sample chamber. The sample chamber is overall inverted conical, the oral product is horizontally placed in the sample chamber, and the edge of the oral product is clamped to the inclined side wall in the sample chamber by friction. The top of the sample chamber is provided with a liquid inlet connected to the temperature maintaining device by a liquid delivery pipeline for receiving the artificial saliva. The bottom of the sample chamber is provided with a closable sampling port, when the sampling port is closed, the artificial saliva accumulates in the bottom space of the sample chamber; when the sampling port is opened, the artificial saliva flows out through the sampling port to facilitate the experimenter to collect the artificial saliva for detection and analysis.

[0028] In step S2, the temperature maintaining device is connected to a peristaltic pump, the peristaltic pump inputs the artificial saliva in the temperature maintaining device into the sample chamber at a certain flow rate. The liquid delivery pipelines connected among the temperature maintaining device, the peristaltic pump and the sample chamber are provided with a temperature maintaining layer to reduce the temperature loss of the artificial saliva in the transmission process. The artificial saliva vertically drops to the surface of the oral product through the liquid inlet at the top of the sample chamber, and gradually spreads and covers the entire surface of the oral product to simulate the dissolution state of the oral product contained in the user's oral cavity (when the oral product is used, it is clamped between the inner wall of the user's oral cavity and the gums, saliva is secreted from the inner wall of the oral cavity and gradually spreads and covers the surface of the oral product, at this time the active ingredient in the oral product gradually dissolves and is absorbed into the oral cavity through saliva). After the active ingredient is dissolved into the artificial saliva, a dissolution liquid is formed, which accumulates in the bottom conical space of the sample chamber. It should be noted that in the embodiments of the present application, the active ingredient includes nicotine or nicotine derivatives. In specific embodiments, the sampling time is set to 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes and the 15th minute after the dissolution starts, a total of 9 samples are collected.

[0029] In step S3, the sampling port of the sample chamber is opened according to the set time to receive the dissolution liquid. The volume of the dissolution liquid is measured, and the dissolution liquid is quantitatively detected using a liquid chromatograph to obtain the dissolution degree corresponding to each sampling time. In specific embodiments, the detection conditions are as follows: The chromatographic column selects octadecylsilane bonded silica gel as the filler or a chromatographic column with equivalent performance, for example, Agilent 5 HC-C18, with a specification of 250mm x 4.6mm, 5μm. The mobile phase selects a mixed solution of 0.02mol / L ammonium formate and methanol to obtain a buffer solution with a pH of about 4.3, wherein the mass ratio of ammonium formate to methanol is 9:1, and 0.05% triethylamine is added. The flow rate of the mobile phase is set to 0.8mL / min. The temperature of the chromatographic column is set to 35°C. The detector is a diode array detector, the detection wavelength is 260nm, and the running time is set to 10min.

[0030] In step S4, the similarity factor evaluation formula is: where t is the dissolution time, Rt is the average dissolution amount of the reference sample within t time, Tt is the average dissolution amount of the target sample within t time, and n is the number of sampling time points. The target sample is an oral preparation whose dissolution behavior is to be evaluated, and the reference sample is an oral preparation whose dissolution behavior is known. The dissolution behaviors of the target sample and the reference sample both refer to the dissolution behavior of the same active ingredient.

[0031] In some embodiments, after the sampling is completed, the loss of the active ingredient also needs to be evaluated. When the loss of the active ingredient is small, it can be considered that the active ingredient is completely dissolved; otherwise, if the loss of the active ingredient is large, it is obvious that the active ingredient is not completely dissolved, and thus the evaluation result is abnormal.

[0032] In specific embodiments, first, the residual oral preparation in the sample chamber is taken out, crushed, and placed in a 50 mL centrifuge tube, and then 10 mL of solvent is added to accelerate the dissolution of the remaining active ingredient in the oral preparation. The solvent in this embodiment is a mixed solution of 0.02 mol / L ammonium dihydrogen phosphate solution (pH 4.3, and 0.05% triethylamine by mass is added) and methanol solution, wherein the volume ratio of the ammonium dihydrogen phosphate solution to the methanol solution is 9:1. After the solvent is added, the centrifuge tube is shaken for 15 minutes to completely dissolve the active ingredient. After the active ingredient is completely dissolved, the solution is filtered, and the filtrate is also subjected to liquid chromatography detection to determine the residual active ingredient content in the filtrate. The residual active ingredient content and the dissolution amount at the last dissolution time are added, and the difference between the original content and the added content is compared to determine the loss value of the active ingredient. For example, the dissolution amount of active ingredient A at 15 min is 8.5 mg, the residual active ingredient content obtained by filtrate detection is 0.3 mg, and the original content is 9 mg, so the loss value of the active ingredient is 9-8.5-0.3=0.2 mg, and the loss ratio is 0.2 / 9=2.2%.

[0033] By the evaluation method provided in the embodiments of the present application, the experimental environment is closer to the oral environment of the human body, and the dissolution result obtained is more accurate. At the same time, when detecting the content of the active ingredient, the operation is relatively simple, the experimental time is short, and only 15 minutes is needed for a single experiment, which greatly shortens the experimental time compared with 60 minutes in the reference. At the same time, the evaluation method used in the embodiments of the present application uses liquid chromatography detection, which has good separation of the active ingredient. Compared with ultraviolet absorption detection, it can avoid the interference of other substances on ultraviolet absorption, and thus the detection result is more accurate and reliable.

[0034] The effectiveness and accuracy of the liquid chromatography detection method provided by the embodiments of the present application for detecting active substances are described below.

[0035] The nicotine content detection method is described in the 11.0 version of the "Guidelines for Sample Handling Techniques for Smokeless Tobacco and Non-Extended Tobacco Products" published by the International Tobacco Science Research Cooperation Center. This method is an acid-base titration method, which uses perchloric acid titrant to titrate the solution containing nicotine to the end point, and calculates the nicotine content by calculating the consumption of titrant. However, in the embodiments of the present application, the dissolution liquid contains artificial saliva, and the artificial saliva is configured with ammonium formate. The ammonium formate solution, as a pH buffer solution, has certain resistance to pH change and can seriously interfere with the results detected by the acid-base titration method. Therefore, the detection method adopted in the embodiments of the present application is not suitable for evaluation by the acid-base titration method.

[0036] In the United States Pharmacopoeia 2024 edition, a nicotine content detection method is also described, which uses high performance liquid chromatography, uses ammonium acetate buffer solution with pH 10.0 and acetonitrile as the mobile phase, performs gradient elution, and the running time is 40 minutes. The method can be used to evaluate the effectiveness and accuracy of the detection method in the embodiments of the present application.

[0037] Specifically, according to the properties of nicotine molecules, a liquid chromatography column with high carbon load is selected, for example, Agilent 5 HC-C18, 250mm x 4.6mm, 5μm or other chromatography columns with equivalent performance. The mobile phase is a mixture of ammonium formate and methanol with pH 3.8-pH 5.0 (preferably pH 4.3) as the mobile phase, and an appropriate amount of tailing agent (such as triethylamine, diethylamine or ammonia water, preferably 0.05% triethylamine) is added to improve the detection peak shape. The flow rate of the mobile phase is set to 0.8mL / min, the column temperature is 35℃, the detection wavelength is 260nm, and the running time is 10 minutes.

[0038] Please refer to Figure 2 , Figure 2 The standard sample liquid chromatogram provided by an embodiment of the present application is shown in the figure. As can be seen from the figure, nicotine produces a peak shape at 5.462 minutes, and the peak shape is good, which proves that the detection method can effectively detect nicotine. It is further necessary to prove that the method can quantitatively detect the nicotine content.

[0039] Please refer to Figure 3 , Figure 3 The standard sample dissolution curve provided by an embodiment of the present application is shown in the figure. Specifically, by detecting the dissolution peak area of the standard sample with different nicotine concentrations, the relationship between the nicotine concentration and the corresponding detection peak area is obtained. The corresponding relationship between the nicotine concentration of the standard sample and the dissolution peak area is shown in Table 1: Table 1 Nicotine concentration and dissolution peak area of different standard samples From Figure 3 The relationship between nicotine concentration and dissolution peak area can be calculated from Table 2 as y = 13.7366x + 0.6235, where y is the dissolution peak area and x is the nicotine concentration, and the correlation coefficient R 2 is 1.0000, i.e., there is a strong linear relationship between nicotine concentration and dissolution peak area.

[0040] Further, the accuracy can be verified by the standard addition recovery rate, and the precision can be verified by the standard deviation of different standard addition recovery results. The detection can be blank standard addition recovery or sample standard addition recovery, which is not limited, and in the embodiments of the present application, it is blank standard addition recovery.

[0041] Specifically, first, a blank sample containing no nicotine is provided, a fixed amount of nicotine is added, and then the nicotine content in the sample is detected, and the actual detection value is divided by the theoretical addition value to obtain the recovery rate. To verify the detection accuracy under different concentration conditions, the experiment provides nicotine with three different addition amounts, so that the samples after addition are sorted from low to high in nicotine concentration and are named as low concentration sample, medium concentration sample, and high concentration sample. At the same time, in order to reduce experimental error, each group of samples is additionally provided with 2 groups of parallel samples, i.e., a total of 3 groups of samples, and finally the average value of the recovery rates of the 3 groups of samples is taken as the recovery rate under the concentration condition. The standard addition recovery rate and relative standard deviation of each group of samples are shown in Table 2.

[0042] Table 2 Standard addition recovery rate and relative standard deviation of each group of samples As can be seen from Table 2, under different concentration conditions, the standard addition recovery rate of the sample is 98.8% to 102.1%, so the detection method has good accuracy. At the same time, after calculation, the relative standard deviation (RSD) of the standard addition recovery rate is ≤2.0%, which proves that the detection method has good precision.

[0043] The application of the evaluation method provided in the embodiments of the present application will be described below with respect to Examples 1 to 4.

[0044] Example 1 In Example 1, the flow rate of the artificial saliva was set to 0.4 mL / min, the temperature was set to 37°C, the oral product used was a peach-flavored nicotine pouch provided by Manufacturer A, with a nicotine content of 9 mg, and the quantitative detection method for the dissolution sample was high-performance liquid chromatography, with an octadecylsilane-bonded silica gel packed column, such as an Agilent 5HC-C18, 250 mm x 4.6 mm, 5 μm. The mobile phase was a mixture of ammonium formate and methanol, with ammonium formate at 0.02 mol / L, pH 4.3, and the addition of 0.05% triethylamine, at a ratio of 9:1. The flow rate of the mobile phase was set to 0.8 mL / min, the temperature of the column was set to 35°C, and the run time was set to 10 minutes.

[0045] Before the evaluation began, the artificial saliva was first placed in the incubation device, and then the temperature of the incubation device was set to 37°C, and the water bath heating was turned on. Then the nicotine pouch was placed in the sample chamber, the flow rate of the peristaltic pump was set, and the artificial saliva in the incubation device was delivered to the sample chamber, so that the artificial saliva was vertically dripped onto the surface of the nicotine pouch through the liquid inlet at the top of the sample chamber. At the 3rd minute, 4th minute, 5th minute, 6th minute, 7th minute, 8th minute, 9th minute, 10th minute, and 15th minute after the start of liquid feeding, samples were collected from the sampling port of the sample chamber, and the sample dissolution liquid was quantitatively detected by high-performance liquid chromatography, and the nicotine dissolution amount of the sample was calculated according to the detection results, and the nicotine dissolution curve was drawn. The dissolution results of the peach-flavored nicotine pouch of Manufacturer A are shown in Table 3.

[0046] Table 3 Dissolution results of the peach-flavored nicotine pouch of Manufacturer A Figure 4 The dissolution curve of the peach-flavored nicotine pouch of Manufacturer A provided in an embodiment of the present application is shown in FIG. 2. According to the data in Table 3 and FIG. 2, the dissolution speed of the peach-flavored nicotine pouch of Manufacturer A was relatively fast, with 90% of the nicotine dissolved at the 8th minute and nearly complete dissolution at the 10th minute. Figure 4 According to the data in Table 3 and FIG. 2, the dissolution speed of the peach-flavored nicotine pouch of Manufacturer A was relatively fast, with 90% of the nicotine dissolved at the 8th minute and nearly complete dissolution at the 10th minute.

[0047] After 15 minutes of dissolution, the remaining nicotine pouch was removed, cut open, and placed in a 50 mL centrifuge tube. 10 mL of dissolution medium was added to continue dissolution. The dissolution medium was a mixture of ammonium formate and methanol, with an ammonium formate concentration of 0.02 mol / L, pH 4.3, and containing 0.05% triethylamine. The volume ratio of ammonium formate to methanol was 9:1. The centrifuge tube with the added dissolution medium was shaken for 15 minutes, and the dissolved liquid in the centrifuge tube was quantitatively measured to calculate the cumulative amount of nicotine dissolved. Specifically, in this example, the peach-flavored nicotine pouch from Manufacturer A contained a total of 9 mg of nicotine. After 15 minutes, a total of 8.94 mg was dissolved, and the remaining sample dissolved 0.23 mg after 15 minutes of shaking. Therefore, the total amount of nicotine dissolved and the remaining amount were 9.17 mg, which deviates from the nominal 9 mg of nicotine by 0.17 mg, meeting the error standard.

[0048] Example 2 In Example 2, the sample was still another nicotine packet with peach flavor from Manufacturer A, and the nicotine content was still 9 mg. All other testing conditions and experimental methods were the same, the only difference being the experimental date and the personnel involved. This was to verify whether the evaluation method provided in this application's examples is reproducible. The specific dissolution results of the peach flavor nicotine packet from Manufacturer A are shown in Table 4.

[0049] Table 4 Comparison of two leaching results for nicotine pouches from Manufacturer A (peach flavor) Figure 5 This is a dissolution curve at different times for a peach-flavored nicotine pouch from Manufacturer A provided in an embodiment of this application. (See Table 4 or...) Figure 5 The data from the first and second cumulative dissolution measurements were substituted into the similarity factor calculation formula, yielding a similarity factor f2 of 83. Since f2 > 50, the two samples were determined to have similar dissolution behaviors. This verifies that the evaluation method, when testing the dissolution behavior of the same sample, can identify samples with identical dissolution behaviors, demonstrating good reproducibility.

[0050] Similar to Example 1, the residual sample was dissolved, and it was found that a total of 8.69 mg was dissolved in 15 minutes. After shaking for 15 minutes, 0.32 mg of the residual sample was dissolved. Therefore, the amount of nicotine dissolved and the amount of residue were 9.01 mg, which is 0.01 mg different from the nominal 9 mg of nicotine, and meets the error standard.

[0051] Example 3 The only difference between Example 3 and Example 1 is that the oral product used was a 9mg nicotine pouch with watermelon flavor from Manufacturer A; all other conditions were the same as in Example 1. The specific dissolution results of the nicotine pouch with watermelon flavor from Manufacturer A are shown in Table 5.

[0052] Table 5 Comparison of nicotine leaching results between watermelon and peach flavored nicotine packets from Manufacturer A Figure 6 Dissolution curves of nicotine pouches with peach and watermelon flavors provided by Manufacturer A in one embodiment of this application. (See Table 5 or...) Figure 6 Substituting the data—specifically, the cumulative dissolution amounts of the peach flavor and the watermelon flavor—into the similarity factor calculation formula, we can calculate a similarity factor f2 of 84. Since f2 > 50, we determine that the two have similar dissolution behaviors; that is, manufacturer A's peach-flavored nicotine pouch and watermelon-flavored nicotine pouch exhibit similar nicotine dissolution behaviors.

[0053] Similar to Example 1, the residual sample was dissolved. It was found that 8.52 mg of nicotine was dissolved from the watermelon-flavored nicotine bag in 15 minutes, and 0.40 mg of nicotine was dissolved from the residual sample after shaking for 15 minutes. Therefore, the amount of nicotine dissolved and the amount of residue were 8.91 mg, which is 0.09 mg different from the nominal 9 mg of nicotine, and meets the error standard.

[0054] Example 4 The only difference between Example 4 and Example 1 is that the oral product used was a 15mg nicotine pouch with the "Ice Flavor" from Manufacturer B; all other conditions were the same as in Example 1. The specific dissolution results of the nicotine pouch with the "Ice Flavor" from Manufacturer B are shown in Table 6.

[0055] Table 6 Comparison of nicotine leaching results between Peach Flavor (Manufacturer A) and Icy Flavor (Manufacturer B) Figure 7 Dissolution curves of nicotine packets with peach flavor from manufacturer A and ice flavor from manufacturer B, provided for an embodiment of this application. (See Table 6 or...) Figure 7 Substituting the data—namely, the cumulative dissolution amounts of peach flavor from manufacturer A and ice flavor from manufacturer B—into the similarity factor calculation formula, we can calculate a similarity factor f2 of 83. Since f2 > 50, we determine that their dissolution behaviors are similar; that is, the nicotine pouches of peach flavor from manufacturer A and ice flavor from manufacturer B have similar nicotine dissolution behaviors.

[0056] Similar to Example 1, the residual sample was dissolved. It was found that 14.80 mg of nicotine was dissolved from the nicotine bag of Manufacturer B's Ice Flavor in 15 minutes. After shaking for 15 minutes, 0.21 mg of the residual sample was dissolved. Therefore, the amount of nicotine dissolved and the amount of residue were 15.01 mg, which is 0.01 mg different from the nominal 15 mg of nicotine and meets the error standard.

[0057] Example 5 Example 5 differs from Example 1 only in that the oral product used is a 6 mg nicotine pouch of brand C watermelon flavor, and the rest of the conditions are the same as in Example 1. The dissolution results of the specific brand C watermelon flavor nicotine pouch are shown in Table 7.

[0058] Table 7 Comparison of dissolution results of brand A peach flavor and brand C watermelon flavor nicotine pouches Substituting the data in Table 7, i.e. the cumulative dissolution amount of brand A peach flavor and the cumulative dissolution amount of brand C watermelon flavor, into the similarity factor calculation formula, the similarity factor f2 can be calculated to be 18. The similarity factor f2 < 50, so it is judged that the dissolution behaviors are not similar, i.e. the nicotine dissolution behaviors of the brand A peach flavor nicotine pouch and the brand C watermelon flavor nicotine pouch are not similar.

[0059] Please refer to Table 7 and Figure 8 , Figure 8 The dissolution curve graph of the brand A peach flavor and the brand C watermelon flavor nicotine pouches provided in an embodiment of the present application. From the Figure 8 , it can also be determined that the dissolution curves of the brand A peach flavor and the brand C watermelon flavor nicotine pouches are completely dissimilar, wherein the brand A peach flavor nicotine pouch has looser content particles and faster nicotine dissolution rate, and is basically completely dissolved at 10 minutes; the brand C watermelon flavor nicotine pouch has tighter content particles and slower nicotine dissolution rate, and dissolves about 60% at 10 minutes, and the content particle states of the two are greatly different.

[0060] As in Example 1, the residual sample was subjected to dissolution, and it was detected that the brand C watermelon flavor nicotine pouch dissolved 4.74 mg in 15 minutes, and the residual sample dissolved 1.18 mg after 15 minutes of shaking, so the dissolution amount and the residual amount of nicotine were 5.92 mg, which had a 0.08 mg deviation from the nominal 6 mg of nicotine, and met the error standard.

[0061] Comparative Example 1 The comparative example 1 uses the soaking method to dissolve nicotine. Specifically, the oral product of the comparative example 1 is selected as a factory A peach flavor 9 mg nicotine bag. First, 10 mL of 0.02 mol / L ammonium dihydrogen phosphate buffer (pH 7.4, the pH is adjusted by potassium hydroxide) is taken in a 15 mL centrifuge tube, the centrifuge tube is placed in a water bath and heated to 37℃, after the temperature is stable for 2 min, the nicotine bag sample is soaked, when the soaking time is over, the nicotine bag is taken out for measurement. The soaking time of the sample is set to 1 min, 2 min, 5 min, 10 min, 30 min and 60 min, and the sample is taken for detection. For the nicotine bag corresponding to each soaking time, it needs to be cut and placed in a new centrifuge tube, and then water solution is added to extract nicotine, and then liquid chromatograph is used for detection. The dissolution results of the nicotine bag of different dissolution methods are shown in Table 8.

[0062] Table 8 Dissolution results of the nicotine bag of different dissolution methods Figure 9 The dissolution curve of the factory A peach flavor nicotine bag of different dissolution methods provided in an embodiment of the present application is provided. By comparing two different dissolution methods, although the dissolution rate of the two methods reaches 90% at about 8 minutes, the dissolution rate rises rapidly in the early stage (within 3 minutes) of the soaking method, it is difficult to analyze the release behavior, and the dissolution reagent used is more and the time is longer, the experimental cost is higher and the efficiency is lower.

[0063] In summary, the embodiment of the present application uses high performance liquid chromatograph to quantitatively detect nicotine in oral products, which is not easily interfered by substances with ultraviolet absorption in the formula compared with the titration detection method for detecting nicotine; the linearity is good, the precision is high, the accuracy is good, and the quantitative requirements of nicotine can be met. At the same time, the embodiment of the present application also designs an efficient dissolution device, and designs a dissolution method in matching, compared with the evaluation method provided in the United States Pharmacopoeia method, the single solution detection time is shortened by 75%, which greatly improves the detection efficiency. Further, in the 15-minute dissolution process, the dissolution curve of nicotine is more gentle, which is more conducive to analyzing the dissolution behavior of nicotine, and realizes more scientific and reliable evaluation of the dissolution behavior of nicotine in a shorter detection time.

[0064] The above only discloses the preferred embodiments of the present application, and of course cannot limit the scope of the rights of the present application, therefore the equivalent changes made according to the claims of the present application still belong to the scope covered by the present application.

Claims

1. A method for evaluating the dissolution behavior of active ingredients in oral products, characterized in that, include: Place oral products in the sample chamber; Artificial saliva is delivered into the sample chamber, so that the artificial saliva comes into contact with the oral product, and the active ingredient of the oral product dissolves in the artificial saliva to form a dissolution solution; The content of active ingredients in the dissolution solution at different times was detected by liquid chromatography, and multiple sets of dissolution data of the active ingredients were obtained. The dissolution behavior of the oral product is evaluated based on multiple sets of dissolution data and a preset similarity factor calculation formula.

2. The method according to claim 1, characterized in that, The evaluation of the dissolution behavior of the oral product based on multiple sets of dissolution data and a preset similarity factor calculation formula includes: Determine the target dissolution data of the active ingredient in the target oral product, wherein the sampling time of the target dissolution data is consistent with the sampling time of the dissolution data; Calculate the similarity factor based on the target dissolution data, the dissolution data, and the similarity factor calculation formula; Determine whether the similarity factor is less than the similarity threshold; If so, the dissolution behavior of the oral product is assessed to be dissimilar to that of the target oral product; If not, then assess that the dissolution behavior of the oral product is similar to that of the target oral product.

3. The method according to claim 1 or 2, characterized in that, The formula for calculating the similarity factor is: 。 4. The method according to claim 1, characterized in that, The mobile phase of the liquid chromatography is a mixed solution of ammonium formate and methanol.

5. The method according to claim 4, characterized in that, The pH of the mobile phase is 3.8 to 5.

0.

6. The method according to claim 4, characterized in that, The mobile phase also contains at least one of triethylamine, diethylamine, or ammonia.

7. The method according to claim 1, characterized in that, The active ingredients include nicotine and its derivatives.

8. The method according to claim 1, characterized in that, The artificial saliva is heated to a constant temperature of 36.3°C to 37.2°C before contacting the oral product, and the artificial saliva is delivered to the sample cell at a flow rate of 0.4 mL / min.

9. The method according to claim 1, characterized in that, The dissolution time of the oral product shall not exceed 15 minutes.

10. An evaluation device for the dissolution behavior of an active ingredient in an oral product, said evaluation device being used to perform the evaluation method as described in any one of claims 1 to 9, characterized in that, include: A sample chamber for holding the oral product, a liquid inlet is provided at the top of the sample chamber for delivering the artificial saliva, and an openable and closable sampling port is provided at the bottom of the sample chamber for collecting the dissolution solution. An artificial saliva warming device for storing and heating the artificial saliva; A pump is connected between the inlet of the sample chamber and the artificial saliva warming device to deliver artificial saliva from the artificial saliva warming device to the sample chamber.

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