CN109030398B - Method for detecting nicotine release behavior of buccal cigarettes and special test instrument thereof - Google Patents
Method for detecting nicotine release behavior of buccal cigarettes and special test instrument thereof Download PDFInfo
- Publication number
- CN109030398B CN109030398B CN201810987568.5A CN201810987568A CN109030398B CN 109030398 B CN109030398 B CN 109030398B CN 201810987568 A CN201810987568 A CN 201810987568A CN 109030398 B CN109030398 B CN 109030398B
- Authority
- CN
- China
- Prior art keywords
- nicotine
- release
- rotating shaft
- buccal
- instrument
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- SNICXCGAKADSCV-JTQLQIEISA-N (-)-Nicotine Chemical compound CN1CCC[C@H]1C1=CC=CN=C1 SNICXCGAKADSCV-JTQLQIEISA-N 0.000 title claims abstract description 119
- SNICXCGAKADSCV-UHFFFAOYSA-N nicotine Natural products CN1CCCC1C1=CC=CN=C1 SNICXCGAKADSCV-UHFFFAOYSA-N 0.000 title claims abstract description 119
- 229960002715 nicotine Drugs 0.000 title claims abstract description 119
- 235000019504 cigarettes Nutrition 0.000 title claims abstract description 65
- 238000000034 method Methods 0.000 title claims abstract description 34
- 238000012360 testing method Methods 0.000 title claims abstract description 26
- 238000001514 detection method Methods 0.000 claims abstract description 48
- 239000003814 drug Substances 0.000 claims abstract description 36
- 238000004090 dissolution Methods 0.000 claims abstract description 31
- 238000000338 in vitro Methods 0.000 claims abstract description 21
- 238000005516 engineering process Methods 0.000 claims abstract description 20
- 239000013307 optical fiber Substances 0.000 claims abstract description 19
- 241000208125 Nicotiana Species 0.000 claims abstract description 17
- 235000002637 Nicotiana tabacum Nutrition 0.000 claims abstract description 17
- 238000004458 analytical method Methods 0.000 claims abstract description 13
- 238000012545 processing Methods 0.000 claims abstract description 6
- 230000005540 biological transmission Effects 0.000 claims abstract description 5
- 238000004088 simulation Methods 0.000 claims abstract description 5
- 238000002211 ultraviolet spectrum Methods 0.000 claims abstract description 5
- 239000000523 sample Substances 0.000 claims description 50
- 238000002835 absorbance Methods 0.000 claims description 34
- 230000006399 behavior Effects 0.000 claims description 31
- 238000003756 stirring Methods 0.000 claims description 27
- 239000007788 liquid Substances 0.000 claims description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 22
- 229940079593 drug Drugs 0.000 claims description 17
- 239000000120 Artificial Saliva Substances 0.000 claims description 15
- 239000012535 impurity Substances 0.000 claims description 15
- 230000008569 process Effects 0.000 claims description 10
- 230000001186 cumulative effect Effects 0.000 claims description 8
- 239000000463 material Substances 0.000 claims description 8
- 230000000149 penetrating effect Effects 0.000 claims description 4
- 238000011156 evaluation Methods 0.000 claims description 3
- 239000011521 glass Substances 0.000 claims description 3
- 238000013178 mathematical model Methods 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 239000007769 metal material Substances 0.000 claims 1
- 239000002861 polymer material Substances 0.000 claims 1
- 235000019505 tobacco product Nutrition 0.000 abstract description 6
- 238000003908 quality control method Methods 0.000 abstract description 4
- 238000013461 design Methods 0.000 abstract description 2
- 230000006872 improvement Effects 0.000 abstract description 2
- 238000010948 quality risk assessment Methods 0.000 abstract description 2
- 238000010223 real-time analysis Methods 0.000 abstract description 2
- 238000002791 soaking Methods 0.000 description 21
- 238000002474 experimental method Methods 0.000 description 19
- 239000000243 solution Substances 0.000 description 11
- 239000000779 smoke Substances 0.000 description 8
- 210000000214 mouth Anatomy 0.000 description 7
- 239000012086 standard solution Substances 0.000 description 6
- 238000010521 absorption reaction Methods 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 238000000502 dialysis Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000003287 bathing Methods 0.000 description 2
- 238000011088 calibration curve Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 238000004806 packaging method and process Methods 0.000 description 2
- 230000002572 peristaltic effect Effects 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000008213 purified water Substances 0.000 description 2
- 210000003296 saliva Anatomy 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- AQCRXZYYMOXFAN-UHFFFAOYSA-N 2-(1-methyl-2-pyrrolidinyl)-pyridine Chemical compound CN1CCCC1C1=CC=CC=N1 AQCRXZYYMOXFAN-UHFFFAOYSA-N 0.000 description 1
- 238000012356 Product development Methods 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 150000003797 alkaloid derivatives Chemical class 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 230000001055 chewing effect Effects 0.000 description 1
- 238000001723 curing Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 210000002200 mouth mucosa Anatomy 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000012502 risk assessment Methods 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 230000000391 smoking effect Effects 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
Landscapes
- Physics & Mathematics (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention belongs to the technical field of smokeless tobacco product testing, in particular to a buccal tobacco nicotine release behavior detection method and a special testing instrument, which are characterized in that: according to the consumption characteristics of the bagged cigarette, the structure improvement suitable for the buccal cigarette test is carried out on the existing medicine dissolution instrument, an in-vitro detection method capable of simulating the buccal cigarette nicotine release behavior is provided, and in-vitro simulation test is carried out on the bagged buccal cigarette nicotine release behavior by combining an optical fiber transmission technology, an ultraviolet spectrum analysis technology and a computer data processing technology. The method has the following maximum advantages: the method realizes in-vitro continuous analysis, real-time analysis and online analysis of the release behavior of the nicotine of the buccal cigarette, has good reproducibility and high degree of automation, and provides important technical support and scientific basis for product design, quality control, risk assessment and policy establishment of buccal cigarette products.
Description
Technical Field
The invention relates to the technical field of in-vitro detection of smoke-free tobacco product component release behaviors, in particular to a buccal smoke nicotine release behavior detection method and a special test instrument bag thereof, which can be used for in-vitro continuous on-line real-time qualitative and quantitative detection of nicotine component release in buccal smoke-free tobacco products, namely, the real-time on-line detection of the release degree (%) and release speed (mug.s -1) of nicotine component in bagged buccal smoke-free tobacco products.
Background
The bagged buccal cigarette is prepared by packaging the processed tobacco wet powder in a moisture-permeable pouch, and placing the pouch between the upper lip and gum of the oral cavity during consumption without chewing, and the tobacco component is dissolved by saliva and then released into the oral cavity and absorbed by oral mucosa. In recent years, with the continuous strictness of worldwide smoke control policies, smoking places are increasingly reduced, and oral consumption of smoke-free air bags filled with cigarettes as an important supplement form for tobacco consumption has occupied an important market share in North America and North Europe, and the domestic and foreign countries have the same type of products on the market, so that related researches such as chemical composition analysis, formula analysis and process molding technology of the cigarettes are becoming hot spots for tobacco research in recent years.
Nicotine is the major alkaloid in tobacco, and the rate of release (μg·s -1) and degree of release (or cumulative release,%) of nicotine are one of the important indicators of the quality of buccal tobacco products. The nicotine release behavior in vitro simulation experiment can screen reasonable raw material formula and production process on one hand, and can measure the risk assessment of the buccal cigarette on the other hand, and strengthen the product quality control means.
In order to achieve effective control of nicotine release from buccal cigarettes, several laboratory studies have been conducted in various countries on the release of nicotine. In 1998, nasr et al studied the release of buccal nicotine using a self-contained dialysis device. The method places the buccal cigarette in a dialysis bag, then places the buccal cigarette in artificial saliva, and researches the release characteristics of nicotine at different time points through sampling at intervals. The method is simpler, but the release rate of nicotine measured by the device is limited by a dialysis bag, so that the release condition of nicotine in a sample is difficult to reflect. In 1999, luque-Perez et al placed the buccal cigarettes in a supported liquid film device, and combined with an ultraviolet spectrophotometer to detect the release of nicotine. Although the method can achieve the purpose of detecting the release amount of nicotine, the supported liquid film is unstable, and the fluctuation of the measurement result is large. In 2011, zhang Jie and the like developed an in-vitro release device capable of simulating the release of buccal nicotine in the oral cavity, and analyzed the release of the nicotine and free nicotine in the buccal smoke by adopting the device, the device is a domestic first-set in-vitro release experimental device for buccal nicotine, can simulate release behaviors and distinguish the release difference of nicotine between different samples, is a laboratory zero-loading device, is complex in operation and is not easy to popularize and apply.
The drug dissolution rate meter is a device for detecting dissolution rate data of a preparation such as a tablet, which is a device commonly used in the pharmaceutical field, but is not found to be used in other fields, particularly in the tobacco field.
Disclosure of Invention
The invention aims at overcoming the defects of the existing method for detecting the nicotine release behavior in the bagged buccal smokeless tobacco products (buccal cigarettes for short), and provides a detection method capable of continuously measuring the nicotine content in the buccal tobacco in-vitro release liquid on line and a special test instrument suitable for the method. The invention aims to provide technical support and data reference for the nicotine release detection and regulation of the bagged buccal cigarettes, and simultaneously provides an accurate detection method for the buccal cigarette nicotine release behavior and a technical platform convenient for popularization and application for relevant personnel working in the tobacco industry such as buccal cigarette product development, quality control and the like.
The invention mechanism of the method is as follows: according to the consumption characteristics of the bagged buccal cigarettes, the structure improvement suitable for buccal cigarettes test is carried out on the existing medicine dissolution instrument, an in-vitro detection method capable of simulating release of the bagged buccal cigarettes in the oral cavity is provided, and the release behavior of the bagged buccal cigarettes is depicted by combining an optical fiber transmission technology, an ultraviolet spectrum analysis technology and a computer data processing technology. The theoretical principle on which the data processing software is concerned is as follows:
since the buccal cigarette releasing liquid is a multi-component mixed system, the principle of adding absorbance can be known:
1 (1)
In formula 1, ε is the absorbance coefficient, C is the concentration, and l is the optical path.
And subtracting the spectrogram of the nicotine standard from the spectrogram of the soaking solution to obtain the spectrogram of the impurity. Namely:
Δa=a Soaking in water -ANic _2
In formula 2, Δa is absorbance of impurities in the soaking solution, a Soaking in water is absorbance of the soaking solution, and a Nic is absorbance of the nicotine standard.
Let k= (Δa 1-ΔA3)/( ΔA2-ΔA3) 3
In the formula 3, deltaA 1 is the absorbance of impurities at the wavelength lambda 1, and the wavelength selection principle is the maximum absorption wavelength of nicotine, namely 260nm; Δa 2 is the absorbance of the impurity at the reference wavelength λ 2, the wavelength selection principle is that nicotine is not absorbed, and the impurity has stronger absorption, namely 290nm; Δa 3 is the absorbance of the impurity at reference wavelength λ 3, and the wavelength selection principle is that nicotine is not absorbed, and the impurity absorption is weak or not absorbed, i.e. 550nm.
Substituting formula 1 into formula 3, namely:
k=(ΔA1-ΔA3)/( ΔA2-ΔA3)=[（ε1-ε3）/（ε2-ε3）]*C*l, Since C, l is a fixed value, k is a constant and can be calculated from the value of Δa 1、ΔA2、ΔA3.
From formula 2, A 1 Nic=A1 Soaking in water -ΔA1 is formula 4
Wherein, A 1 Nic is the absorbance of the nicotine standard at the wavelength lambda 1, A 1 Soaking in water is the absorbance of the sample soaking solution at the wavelength lambda 1, and delta A 1 is the absorbance of the impurity in the sample soaking solution at the wavelength lambda 1.
From formula 3, ΔA 1=k（ΔA2-ΔA3）+ΔA3=kΔA2-(k-1)ΔA3 is shown in formula 5
Substituting formula 5 into formula 4, namely:
A 1 Nic=A1 Soaking in water - kΔA2-(k-1)ΔA3 type 6
In the formula 6, since delta A 2 is the absorbance of impurities at the reference wavelength lambda 2, the wavelength selection principle is that nicotine is not absorbed, and impurities are strongly absorbed; Δa 3 is the absorbance of the impurity at reference wavelength λ 3, and the wavelength selection principle is that nicotine is not absorbed, and the impurity absorption is weak or not absorbed. It can be considered that: Δa 2= A2 Soaking in water ,ΔA3= A3 Soaking in water . Thus, equation 6 can be expressed as:
A 1 Nic=A1 Soaking in water - k A2 Soaking in water -(k-1) A3 Soaking in water type 7
In summary, the instant nicotine concentration can be calculated according to the absorbance values at the positions of lambda 1、λ2 and lambda 3, and based on the powerful calculation function of the computer, the nicotine concentration in the system can be obtained immediately on the data processing software, and a time-varying curve of the nicotine concentration, namely a pouch buccal tobacco nicotine release behavior curve, can be drawn.
The aim of the invention is realized by the following technical scheme:
The utility model provides a buccal cigarette nicotine release behavior detection method, which is to carry out structural transformation suitable for buccal cigarette test on the existing medicine dissolution instrument according to the consumption characteristics of the buccal cigarette in bags, and provides an in vitro detection method capable of simulating buccal cigarette nicotine release behavior in the oral cavity, and carrying out in vitro simulation test on the buccal cigarette nicotine release behavior in bags by combining an optical fiber transmission technology, an ultraviolet spectrum analysis technology and a computer data processing technology, and the specific method is as follows:
1) The existing medicine dissolution instrument is modified to obtain a special instrument suitable for detecting the release behavior of the nicotine of the buccal cigarettes;
2) The 1-small-pack bagged cigarette sample is placed in a draining type narrow release unit (positioned on the liquid surface) of a modified special instrument, the artificial saliva flows through the sample and then brings out the component to be tested, and then the component to be tested is dripped into a detection cup, and an optical fiber probe is placed below the liquid surface of the detection cup to detect the absorbance value of a release liquid system. According to the relation between the nicotine concentration and the nicotine absorbance, the method accords with the Lambert-Beer law, the influence of impurities is deducted by using a drug dissolution data processing software (a drug dissolution instrument is provided with the drug dissolution instrument), the instant nicotine concentration in a system is obtained, the nicotine concentration of the system per second is recorded along with time, and then a buccal cigarette nicotine release behavior curve is drawn;
3) Carrying out mathematical evaluation on the accumulated release rate of nicotine by utilizing a Weibull model; the Weibull model is a mathematical model for evaluating the release of drugs, which is commonly used in the pharmaceutical field, and is used for describing the cumulative release rate of drugs at the time t, and the form is as follows: y=1-exp (- α×t) μmu, where Y is the cumulative release rate, α is the time scale of the release process determined by the scale parameter, μmu is the shape characteristic of the curve determined by the shape parameter, μ1 is an exponential shape, μ1 is an S-shape, and μ1 is an exponential shape with a steeper starting slope; in the method, [ mu ] =1, the shape of the fitting curve mainly depends on alpha, namely the accumulated release rate of nicotine along with time is determined by alpha, and the magnitude of alpha can reflect the release speed of nicotine, so that alpha is defined as a nicotine release rate coefficient (min -1), and the release speed of nicotine of different samples can be judged by comparing the values of alpha; finally, the whole process of in-vitro detection of the nicotine release behavior of the buccal cigarette is realized.
The utility model provides a be suitable for special test instrument of above-mentioned detection method, includes the medicine dissolution appearance that has fiber probe, and the pivot lower extreme of medicine dissolution appearance is connected with the narrow and small release unit of waterlogging caused by excessive rainfall formula that is used for placing the sample, and the pivot is controlled by stirring controller, wherein: the rotating shaft of the medicine dissolution instrument is transformed into a hollow rotating shaft, the lower end of the hollow rotating shaft is combined with the release unit through a clamp type connecting piece, and the release unit rotates along with the hollow rotating shaft. An artificial saliva instilling tube controlled by a flow controller is arranged in the hollow rotating shaft in a penetrating way, the lower end of the instilling tube extends out from the lower end of the hollow rotating shaft to the releasing unit, a detecting cup is arranged below the releasing unit, and the detecting cup is arranged in the constant-temperature water bath box. (see FIG. 1 for details)
The capacity of the draining type narrow release unit is 5-15cm 3, preferably 10cm 3,, and draining holes are uniformly distributed at the bottom; the releasing unit may be cylindrical, cuboid or square, and may be made of metal, polymer, glass, etc.
The bottom of the release unit is provided with a stirring part which extends downwards and is used for stirring and uniformly mixing the liquid in the detection cup. The stirring part can be a stirring rod, and the bottom of the release unit is directly screwed by a screw thread mode (see fig. 1); or a clamping ring multi-claw stirring piece which is directly clamped on the outer circumferential wall surface of the release unit in a clamping mode. (see FIG. 2)
The detection cup has a capacity of 100-300cm 3, preferably 200cm 3,, and can be cylindrical, cuboid or cube, and the detection cup can be made of metal, polymer, glass and other materials.
The slit on the optical fiber probe is 0.1-5mm, preferably 1mm, and the optical fiber probe is positioned on one side of the detection cup and fixed in position.
The special test instrument is used for simulating the consumption characteristic that nicotine in tobacco is dissolved by saliva and then released into the oral cavity when the buccal cigarette is used, taking 1 small bag of buccal cigarette sample to be placed in a water draining type narrow release unit, taking out components to be tested after artificial saliva flows through the sample at a constant speed, dripping the components to be tested into a detection cup, placing an optical fiber probe under the liquid surface of the detection cup, detecting the absorbance of the release liquid, conforming to Lambert-Beer law according to the relationship between the concentration of nicotine and the absorbance of the nicotine, namely, the relationship between the concentration of the nicotine and the absorbance of the nicotine is linear, obtaining the concentration of the nicotine in the system after data processing is carried out by utilizing data processing software (a drug dissolution instrument is self-contained), and drawing a time-varying curve of the nicotine, namely, a buccal cigarette nicotine release behavior curve. The detailed experimental procedure is as follows:
a. Starting up, preheating, water-bathing to 37 ℃, taking a proper amount of purified water, adding into a detection cup, weighing 1 bag of sample, placing under the liquid surface of the detection cup, stirring and soaking for one hour under the condition of constant-temperature water bath, taking out the sample, obtaining a soaking solution, and detecting the absorbance value at the lambda 1、λ2、λ3.
B. and taking the soaking solution into a chromatographic bottle, and detecting by high performance liquid chromatography to obtain the nicotine concentration.
C. Preparing a nicotine standard solution with the same concentration, measuring an absorbance value at a lambda 1、λ2、λ3, calculating a k value and inputting the k value into software.
D. Starting up, preheating, water-bathing to 37 ℃, and taking a proper amount of purified water into the detection cup, wherein the optical fiber probe is immersed under the water surface. Weighing 1 bag of sample, and horizontally placing at the bottom of a narrow releasing unit which is positioned above the water surface. The peristaltic pump is turned on to allow the artificial saliva to drop at a uniform rate in the middle of the sample at a suitable flow rate.
E. at the same time stirring was turned on and timing was started and the instrument started to detect and record.
F. After a period of time, the experiment is stopped and the file is saved.
G. and calculating the value of the accumulated release rate% and the release rate coefficient alpha of the nicotine according to experimental detection data.
The invention has the advantages that:
1. The experimental method disclosed by the invention has stronger novelty, integrates an optical fiber technology, a spectrum technology, an analysis technology and a computer technology, accurately calculates the sampling time and the detection time at second level intervals, and innovatively realizes in-vitro continuous analysis, real-time analysis and online analysis of the release behavior of the nicotine containing the cigarette.
2. The determination method provided by the invention has the characteristics of good reproducibility, high automation degree and convenience in popularization, can effectively simulate the release behavior of the buccal cigarette nicotine in vitro, has strong practicability, and provides important technical support and scientific basis for product design, quality control, risk assessment and policy establishment of buccal cigarette products.
Drawings
FIG. 1 is a schematic diagram of a special test instrument for nicotine release behavior of a buccal cigarette according to the present invention
FIG. 2 is a schematic view showing the structure of the draining type narrow release unit of FIG. 1 connected with other parts
In the figure, 1, a medicine dissolution instrument, 2, an optical fiber probe, 3, a hollow rotating shaft, 4, a narrow release unit, 5, a clamp type connector, 6, a instillation tube, 7, a stirring controller, 8, artificial saliva, 9, a flow controller, 10, an artificial saliva drop inlet, 11, a stirring component, 11-1, a clamping ring multi-claw stirring piece, 12, a sample, 13, a constant-temperature water bath box and 14, and a detection cup.
Fig. 3 is a full-wave scan of the soak solution for domestic single-material smoke G1# sample.
FIG. 4 is a full-wave scan of 49.9 μg/ml nicotine standard.
Figure 5 in vitro release profile of nicotine from domestic single-dose smoke G1# samples.
Detailed Description
The detection method is further described with reference to the accompanying drawings:
The invention provides a method for detecting the release behavior of buccal cigarette nicotine, which is to carry out structural transformation suitable for buccal cigarette test on the existing medicine dissolution instrument according to the consumption characteristics of the buccal cigarette in bags, and provides an in-vitro detection method capable of simulating the release behavior of buccal cigarette nicotine in the oral cavity, and carry out in-vitro simulation test on the release behavior of the buccal cigarette in bags by combining an optical fiber transmission technology, an ultraviolet spectrum analysis technology and a computer data processing technology, and the specific method is as follows:
1) Modifying the existing medicine dissolution instrument to obtain a special instrument suitable for detecting the release behavior of the nicotine of the buccal cigarette (see figure 1);
2) The 1-small-pack packaged buccal cigarette sample 12 is placed in a draining type narrow release unit 4 (positioned on the liquid level) of a modified special instrument, the artificial saliva flows through the sample and then brings out the components to be tested to drop into a detection cup, and the optical fiber probe 2 is placed below the liquid level of the detection cup 14 to detect the absorbance value of a release liquid system. According to the relation between the nicotine concentration and the nicotine absorbance, the method accords with the Lambert-Beer law, the influence of impurities is deducted by using a drug dissolution data processing software (a drug dissolution instrument is provided with the drug dissolution instrument), the instant nicotine concentration in a system is obtained, the nicotine concentration of the system per second is recorded along with time, and then a buccal cigarette nicotine release behavior curve is drawn;
3) Carrying out mathematical evaluation on the accumulated release rate of nicotine by utilizing a Weibull model; the Weibull model is a mathematical model for evaluating the release of drugs, which is commonly used in the pharmaceutical field, and is used for describing the cumulative release rate of drugs at the time t, and the form is as follows: y=1-exp (- α×t) μmu, where Y is the cumulative release rate, α is the time scale of the release process determined by the scale parameter, μmu is the shape characteristic of the curve determined by the shape parameter, μ1 is an exponential shape, μ1 is an S-shape, and μ1 is an exponential shape with a steeper starting slope; in the method, [ mu ] =1, the shape of the fitting curve mainly depends on alpha, namely the accumulated release rate of nicotine along with time is determined by alpha, and the magnitude of alpha can reflect the release speed of nicotine, so that alpha is defined as a nicotine release rate coefficient (min -1), and the release speed of nicotine of different samples can be judged by comparing the values of alpha; finally, the whole process of in-vitro detection of the nicotine release behavior of the buccal cigarette is realized.
The special test instrument of the invention is described below with reference to the accompanying drawings:
As shown in fig. 1 and 2: the special test instrument suitable for the detection method comprises a medicine dissolution instrument 1 with an optical fiber probe, wherein the lower end of a rotating shaft of the medicine dissolution instrument is connected with a draining type narrow release unit 4 for placing a sample, and the rotating shaft is controlled by a stirring controller 7, wherein: the rotating shaft of the medicine dissolution instrument is transformed into a hollow rotating shaft 3, the lower end of the hollow rotating shaft 3 is combined with the release unit 4 through a clamp type connecting piece 5, and the release unit rotates along with the hollow rotating shaft. An artificial saliva instilling tube 6 controlled by a flow controller 9 is arranged in the hollow rotating shaft 3 in a penetrating way, the lower end of the instilling tube extends out from the lower end of the hollow rotating shaft to the releasing unit 4, a detecting cup 14 is arranged below the releasing unit, and the detecting cup is arranged in the constant-temperature water bath 13.
A stirring part 11 extending downwards is arranged at the bottom of the release unit and is used for stirring and uniformly mixing the liquid in the detection cup. The stirring part 11 may be a stirring rod, and the bottom of the releasing unit is directly screwed by a screw thread manner (see fig. 1); or a clamping ring multi-claw stirring piece 11-1 which is directly clamped on the outer circumferential wall surface of the release unit 4 in a clamping manner. (see FIG. 2)
Specific test and implementation effect
The following examples are only illustrative of the present application and are not intended to limit the scope of the application. Further, it is understood that various changes and modifications may be made by those skilled in the art after reading the description of the application, and such equivalents are intended to fall within the scope of the application as defined in the appended claims.
1. Experimental procedure for domestic single-material smoke
1. Sample preparation: 9 domestic flue-cured tobacco, burley tobacco and sun-cured tobacco samples produced in Guangxi, hubei, sichuan, yunnan and other areas. After being processed by the processes of crushing, sieving, curing, packaging and the like, the experimental sample of the buccal cigarette is self-made, and all samples are stored in a sealed and frozen manner (‒ ℃ C.) before the experiment.
2. Taking a domestic single-material tobacco sample with the number of G1, soaking for 1 hour at the constant temperature of 37 ℃, and taking out the sample to obtain a soaking solution. The absorbance values were measured by full wavelength scanning (as shown in FIG. 3), and the absorbance values at 260nm, 290nm and 550m were read out.
3. And detecting the nicotine concentration of the G1# sample soaking solution by using a high performance liquid chromatography to obtain the nicotine concentration of 49.9 mug/ml.
4. A nicotine standard solution was prepared with artificial saliva at a concentration of approximately 49.9 μg/ml. The absorbance values were measured by full wavelength scanning (as shown in FIG. 4), and the absorbance values at 260nm, 290nm and 550m were read out.
5. K=1.24 was calculated from absorbance values of the soak solution and the standard solution at wavelengths of 260nm, 290nm, 550 nm.
6. Repeating the steps 2-5 for 3 times, and taking the average value 1.24 of the 3 parallel experiments as the k value. (this step may be omitted)
7. And taking 1 bag of sample, horizontally arranging at the bottom of a release unit, wherein the release unit is positioned on the liquid level, and the optical fiber probe is positioned below the liquid level. The stirring part at the lower part of the release unit is positioned below the liquid surface.
8. After the cover of the detection cup is covered, the peristaltic pump is opened to drive the artificial saliva to drop in the middle of the sample at a constant speed of 0.2ml/min, and the artificial saliva is dropped into the detection cup after the smoke-containing release component is taken out. Simultaneously, stirring is started, and the whole experiment process keeps the closed state of the cover of the detection cup so as to reduce the volatilization of water in the experiment process.
9. After the experiment started, the instrument started to detect and time.
After 10.45 min, the absorbance value of the released liquid system can be seen to be gentle through an experiment view window, and at the moment, the experiment is stopped and the file is saved.
11. From the experimental data, the data were processed using excel or origin software and the nicotine release profile was plotted as shown in fig. 5.
12. According to experimental data, the nicotine release rate alpha of the domestic single-material smoke G1# sample is calculated to be 0.04. (see Table 2 for details)
13. The nicotine cumulative release profile for the remaining 8 samples can be obtained by repeating experimental steps 2-10.
14. The nicotine release rate a for the remaining 8 samples can be obtained by repeating experiment step 12. (see Table 2 for details)
2. Experimental procedure for the commercially available finished cigarette at home and abroad
Taking foreign commercial buccal cigarette samples with the number of 1-10# and repeating the step 2-12 of the step 2 of the experiment of the first and the second single cigarettes to obtain a nicotine accumulated release curve graph and a nicotine release rate alpha of 10 samples. (see Table 2 for details)
3. Effect of the invention
1. Absorbance stability, linearity and range of the experimental method of the invention
And (3) taking the prepared serial concentration nicotine standard solution, and detecting by using the special testing instrument to obtain a calibration curve, a linear equation and a correlation coefficient. Experimental results show that in the range of 10-80ug/ml, the absorbance stabilization time of the nicotine standard solution at 260nm is greater than 1 hour. In the concentration range of 10-80ug/ml, the linear relation between the absorbance and the concentration of the nicotine standard solution at 260nm is good, and the regression equation of the calibration curve is: y=270.88x+0.3522, r=0.9993.
2. Repeatability of the experimental method according to the invention
Thawing a sample to be tested at room temperature for 2 hours, taking a 100 small bag to calculate the average mass of the sample, sorting the sample according to +/-1% of the average mass, taking a qualified sample, carrying out a nicotine release degree experiment according to an experiment flow of 1.2.3, detecting for 6 times in parallel, and calculating the Relative Standard Deviation (RSD) of the accumulated release rate of nicotine in each time period, wherein the result is shown in Table 1.
Table 1 method reproducibility (n=6, rsd%)
The relative standard deviation of the sample release amount in each time period is between 3.4% and 7.4%, which shows that the method has better repeatability.
3. Analysis of results of the experimental methods of the invention
The results of the Weibull model fitting are shown in Table 2, and the goodness of fit of each sample is greater than 0.96, so that the Weibull model is suitable for the nicotine release method, and the Weibull model can be used for evaluating the nicotine release rate of the buccal cigarette and predicting the nicotine release amount in each time.
TABLE 2 Nicotine Release Rate coefficient for different buccal cigarettes
| Sample number | Moisture content (%) | pH | Nicotine (mg/g) on a wet basis | Degree of Release (%) | Coefficient of release rate lambda (min -1) |
| Domestic single-material cigarette 1 | 22.2 | 4.78 | 27.5 | 89 | 0.04 |
| 2 | 24.7 | 5.07 | 18.7 | 96 | 0.07 |
| 3 | 26.9 | 4.91 | 31.9 | 79 | 0.03 |
| 4 | 26.5 | 4.64 | 23.3 | 96 | 0.07 |
| 5 | 25.1 | 4.76 | 33.1 | 85 | 0.03 |
| 6 | 25.4 | 7.35 | 16.3 | 97 | 0.06 |
| 7 | 28.4 | 7.00 | 27.7 | 80 | 0.02 |
| 8 | 24.9 | 7.06 | 19.0 | 70 | 0.02 |
| 9 | 23.6 | 6.98 | 26.2 | 94 | 0.05 |
| Foreign finished cigarette 1 | 41.0 | 6.68 | 8.0 | 94 | 0.06 |
| 2 | 45.4 | 6.33 | 8.0 | 85 | 0.04 |
| 3 | 38.1 | 7.35 | 11.1 | 51 | 0.02 |
| 4 | 31.0 | 6.02 | 10.4 | 97 | 0.06 |
| 5 | 37.1 | 6.67 | 15.0 | 83 | 0.04 |
| 6 | 18.2 | 6.22 | 14.3 | 96 | 0.07 |
| 7 | 26.8 | 6.33 | 16.5 | 62 | 0.02 |
| 8 | 24.8 | 7.48 | 7.7 | 99 | 0.11 |
| 9 | 44.6 | 6.84 | 14.2 | 96 | 0.08 |
| 10 | 47.7 | 6.57 | 20.0 | 88 | 0.04 |
Claims (5)
1. A method for detecting nicotine release behavior of a buccal cigarette, which is characterized by comprising the following steps: according to the consumption characteristics of the bagged buccal cigarettes, the traditional medicine dissolution instrument is structurally modified for buccal cigarettes, a detection method capable of simulating buccal cigarette nicotine release behaviors is provided, and the bagged buccal cigarette nicotine release behaviors are subjected to in vitro simulation tests by combining an optical fiber transmission technology, an ultraviolet spectrum analysis technology and a computer data processing technology, wherein the specific method is as follows:
The existing medicine dissolution instrument is modified to obtain a special instrument suitable for detecting the release behavior of the nicotine of the buccal cigarettes; the special instrument comprises a medicine dissolution instrument with an optical fiber probe, wherein the lower end of a rotating shaft of the medicine dissolution instrument is connected with a draining type narrow release unit for placing a sample, and the rotating shaft is controlled by a stirring controller, and is characterized in that: the rotating shaft of the medicine dissolution instrument is a hollow rotating shaft, the lower end of the hollow rotating shaft is combined with the release unit through a clamp type connecting piece, and the release unit rotates along with the hollow rotating shaft; an artificial saliva instilling tube controlled by a flow controller is arranged in the hollow rotating shaft in a penetrating way, the lower end of the instilling tube extends out from the lower end of the hollow rotating shaft to a release unit, a detection cup is arranged below the release unit, and the detection cup is arranged in a constant-temperature water bath box;
Placing a 1-small-package bagged buccal tobacco sample into a draining type narrow release unit of a modified special instrument, taking out components to be detected after artificial saliva flows through the sample, dripping the components to be detected into a detection cup, placing an optical fiber probe below the liquid level of the detection cup, detecting the absorbance value of a release liquid system, conforming to the Lambert-Beer law according to the relationship between the nicotine concentration and the nicotine absorbance, deducting the influence of impurities by utilizing drug dissolution data processing software to obtain the instant nicotine concentration in the system, recording the nicotine concentration of the system every second with time, and further drawing a buccal tobacco nicotine release behavior curve;
Carrying out mathematical evaluation on the accumulated release rate of nicotine by utilizing a Weibull model; the Weibull model is a mathematical model for evaluating the release of drugs, which is commonly used in the pharmaceutical field, and is used for describing the cumulative release rate of drugs at the time t, and the form is as follows: y=1-exp (- α×t) μmu, where Y is the cumulative release rate, α is the time scale of the release process determined by the scale parameter, μmu is the shape characteristic of the curve determined by the shape parameter, μmu=1 curve is exponential, μ1 is S-shaped, and μ1 is exponential curve with steep starting slope; in the method, [ mu ] =1, the shape of the fitting curve mainly depends on alpha, namely the accumulated release rate of nicotine along with time is determined by alpha, and the magnitude of alpha can reflect the release speed of nicotine, so that alpha is defined as a nicotine release rate coefficient min -1, and the release speed of nicotine of different samples is judged by comparing the values of alpha; finally, the whole process of in-vitro detection of the nicotine release behavior of the buccal cigarette is realized.
2. The special test instrument suitable for the detection method of claim 1, comprising a drug dissolution instrument with an optical fiber probe, wherein the lower end of a rotating shaft of the drug dissolution instrument is connected with a draining type narrow release unit for placing a sample, and the rotating shaft is controlled by a stirring controller, and the special test instrument is characterized in that: the rotating shaft of the medicine dissolution instrument is a hollow rotating shaft, the lower end of the hollow rotating shaft is combined with the release unit through a clamp type connecting piece, and the release unit rotates along with the hollow rotating shaft; an artificial saliva instilling tube controlled by a flow controller is arranged in the hollow rotating shaft in a penetrating way, the lower end of the instilling tube extends out from the lower end of the hollow rotating shaft to a release unit, a detection cup is arranged below the release unit, the detection cup is arranged in a constant-temperature water bath box, the draining type narrow release unit is positioned above the liquid level of the system and is not contacted with the liquid level, an optical fiber probe is positioned below the liquid level and is soaked in the liquid system, and the volume of the draining type narrow release unit is 5-10 cubic centimeters; the shape is barrel-shaped, cuboid-shaped or cube-shaped, and the material is metal material, polymer material or glass material.
3. The special test instrument according to claim 2, wherein: the bottom of the draining type narrow release unit is provided with a stirring part for stirring and uniformly mixing the liquid in the detection cup.
4. A special test instrument according to claim 3, characterized in that: the stirring part is a stirring rod and is directly screwed at the bottom of the release unit in a threaded mode.
5. A special test instrument according to claim 3, characterized in that: the stirring part is a clamping ring multi-claw stirring part and is directly clamped on the outer circumferential wall surface of the bottom of the release unit in a clamping mode.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810987568.5A CN109030398B (en) | 2018-08-28 | 2018-08-28 | Method for detecting nicotine release behavior of buccal cigarettes and special test instrument thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810987568.5A CN109030398B (en) | 2018-08-28 | 2018-08-28 | Method for detecting nicotine release behavior of buccal cigarettes and special test instrument thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109030398A CN109030398A (en) | 2018-12-18 |
| CN109030398B true CN109030398B (en) | 2024-05-17 |
Family
ID=64625725
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810987568.5A Active CN109030398B (en) | 2018-08-28 | 2018-08-28 | Method for detecting nicotine release behavior of buccal cigarettes and special test instrument thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109030398B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111855933A (en) * | 2019-04-28 | 2020-10-30 | 中国石油化工股份有限公司 | Solid corrosion inhibitor release rate evaluation device and method |
| CN112444466B (en) * | 2019-09-05 | 2023-02-28 | 上海新型烟草制品研究院有限公司 | Method for evaluating in-vitro absorption of nicotine buccal cigarette |
| CN114646718A (en) * | 2022-04-07 | 2022-06-21 | 国家烟草质量监督检验中心 | Method for evaluating nicotine metabolism dynamics absorbed by oral route and digestive tract route when people use buccal tobacco products |
Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4681858A (en) * | 1986-06-19 | 1987-07-21 | Warner-Lambert Company | Dissolution cell and method for determining the in-vitro release of a drug |
| JPH05249097A (en) * | 1992-03-04 | 1993-09-28 | Yamanouchi Pharmaceut Co Ltd | Elution test method for estimating in vivo elution of tablet |
| CA2371524A1 (en) * | 2002-02-07 | 2003-08-07 | William E. Gemmell | Craving reduction method and toothpick delivery system for active e.g. nicotine |
| WO2003098199A1 (en) * | 2002-05-17 | 2003-11-27 | Delphian Technology, Inc. | In situ methods for measuring the release of a substance from a dosage form |
| CN1558213A (en) * | 2004-01-23 | 2004-12-29 | 新疆富科思生物技术发展有限公司 | Optical fiber in situ tester for dissolution and release of medicine |
| CN102507790A (en) * | 2011-11-17 | 2012-06-20 | 中国烟草总公司郑州烟草研究院 | In-vitro simulation detection method for bioavailability of nicotine in tobacco gum base product |
| CN103323393A (en) * | 2013-05-24 | 2013-09-25 | 新疆富科思生物技术发展有限公司 | A real-time optical-fiber medicine intrinsic dissolution determinator |
| CN103543225A (en) * | 2013-11-06 | 2014-01-29 | 云南烟草科学研究院 | Determining method and device of nicotine in oral smokeless tobacco products |
| CN104833641A (en) * | 2015-05-14 | 2015-08-12 | 云南中烟工业有限责任公司 | In-vitro simulated dissolution and continuous on-line detection device and method for nicotine in bagged oral tobacco products |
| CN106770246A (en) * | 2016-12-26 | 2017-05-31 | 重庆中烟工业有限责任公司 | Nicotine detecting system in one tobacco articles dissolution fluid |
| CN208872662U (en) * | 2018-08-28 | 2019-05-17 | 中国烟草总公司郑州烟草研究院 | A kind of buccal cigarette nicotine release behavior detection device |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7470545B2 (en) * | 2001-11-05 | 2008-12-30 | Rohm And Haas Company | Buccal dissolution of active substances |
| FI20050385A0 (en) * | 2005-04-14 | 2005-04-14 | Jukka Rantanen | Method and apparatus for dissolving a solid in a liquid |
| CA2646942C (en) * | 2006-03-16 | 2014-07-29 | Niconovum Ab | Improved snuff composition |
-
2018
- 2018-08-28 CN CN201810987568.5A patent/CN109030398B/en active Active
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4681858A (en) * | 1986-06-19 | 1987-07-21 | Warner-Lambert Company | Dissolution cell and method for determining the in-vitro release of a drug |
| JPH05249097A (en) * | 1992-03-04 | 1993-09-28 | Yamanouchi Pharmaceut Co Ltd | Elution test method for estimating in vivo elution of tablet |
| CA2371524A1 (en) * | 2002-02-07 | 2003-08-07 | William E. Gemmell | Craving reduction method and toothpick delivery system for active e.g. nicotine |
| WO2003098199A1 (en) * | 2002-05-17 | 2003-11-27 | Delphian Technology, Inc. | In situ methods for measuring the release of a substance from a dosage form |
| CN1558213A (en) * | 2004-01-23 | 2004-12-29 | 新疆富科思生物技术发展有限公司 | Optical fiber in situ tester for dissolution and release of medicine |
| CN102507790A (en) * | 2011-11-17 | 2012-06-20 | 中国烟草总公司郑州烟草研究院 | In-vitro simulation detection method for bioavailability of nicotine in tobacco gum base product |
| CN103323393A (en) * | 2013-05-24 | 2013-09-25 | 新疆富科思生物技术发展有限公司 | A real-time optical-fiber medicine intrinsic dissolution determinator |
| CN103543225A (en) * | 2013-11-06 | 2014-01-29 | 云南烟草科学研究院 | Determining method and device of nicotine in oral smokeless tobacco products |
| CN104833641A (en) * | 2015-05-14 | 2015-08-12 | 云南中烟工业有限责任公司 | In-vitro simulated dissolution and continuous on-line detection device and method for nicotine in bagged oral tobacco products |
| CN106770246A (en) * | 2016-12-26 | 2017-05-31 | 重庆中烟工业有限责任公司 | Nicotine detecting system in one tobacco articles dissolution fluid |
| CN208872662U (en) * | 2018-08-28 | 2019-05-17 | 中国烟草总公司郑州烟草研究院 | A kind of buccal cigarette nicotine release behavior detection device |
Non-Patent Citations (3)
| Title |
|---|
| 紫外-可见分光光度法测定袋装口含烟溶入人工唾液中的烟碱含量;袁大林;杨继;汤建国;杨柳;吴俊;段沅杏;巩效伟;陈永宽;;理化检验(化学分册)(第04期);全文 * |
| 茶碱缓释片体外释放度过程分析;李建光, 李新霞, 熊海江, 陈坚;新疆医科大学学报(第06期);全文 * |
| 袋装口含型无烟气烟草制品烟碱体外释放分析;张杰;李鹏;彭书海;宗永立;孙世豪;钱发成;谢剑平;;烟草科技(第05期);全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109030398A (en) | 2018-12-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109030398B (en) | Method for detecting nicotine release behavior of buccal cigarettes and special test instrument thereof | |
| CN102156178B (en) | Method for detecting release situation of nicotine in buccal tobacco products | |
| CN101393099B (en) | Continuous measurement method for tobacco moisture percentage and apparatus thereof | |
| CN101393114B (en) | Rapid measuring method for physical property of flavouring essences for tobacco | |
| US7310995B2 (en) | Method and device for determination of moisture content and solid state phase of solids using moisture sorption gravimetry and near infrared or Raman spectroscopy | |
| WO2012065330A1 (en) | Test method of moisturizing properties of tobacco | |
| CN107367483A (en) | A kind of online throwing of cigarette mixes the method for determination and evaluation of silk uniformity | |
| CN102183613B (en) | Method for detecting release condition of tobacco specific nitrosamines in buccal tobacco product | |
| CN102323178B (en) | Method and device for measuring physical property indexes of soil body | |
| CN105606793B (en) | Fine aggregate soil-water characteristic curve estimation method based on methylene blue value | |
| Parrish et al. | Measurement of formaldehyde in a single puff of cigarette smoke using tunable diode laser infrared spectroscopy | |
| CN107941925A (en) | A kind of liquid phase adsorption assay method in real time in situ | |
| Luo et al. | Near infrared spectroscopy combination with PLS to monitor the parameters of naproxen tablet preparation process | |
| CN208872662U (en) | A kind of buccal cigarette nicotine release behavior detection device | |
| Li et al. | Real-Time Monitoring of Nicotine Release Behavior from Smokeless Tobacco (Snus) Based on Fiber Optic Sensing Technology | |
| CN107247014B (en) | Device and method for measuring particle size distribution of vacuum intermediate infrared interference smoke screen | |
| CN110174371A (en) | It is a kind of based near infrared technology pipe tobacco processing in mass change characterizing method | |
| CN101285767A (en) | Method and its device for enhancing tobacco powder near infrared spectra collection accuracy | |
| CN109738556A (en) | A kind of preparation method of nicotine standard substance and its nicotine standard substance of preparation | |
| CN106770987B (en) | The prediction technique of NNK release amount in a kind of pipe tobacco flue gas | |
| CN105547902A (en) | Method for determination of moisture-proof curve of tobacco capsules | |
| CN117191732A (en) | Method for rapidly and accurately detecting water content of cut tobacco in heated cigarettes | |
| CN116609233B (en) | Evaluation method for permeation condition of reconstituted tobacco coating liquid | |
| WO2024192719A1 (en) | Method for rapidly measuring content of berberine in phellodendri amurensis cortex | |
| CN108956848B (en) | Method for measuring nicotine in tobacco smoke by infrared mirror reverberatory furnace-continuous flow method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |