CN115015453A - Pretreatment method for detecting sweetener in oral smokeless tobacco product - Google Patents
Pretreatment method for detecting sweetener in oral smokeless tobacco product Download PDFInfo
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- CN115015453A CN115015453A CN202210685298.9A CN202210685298A CN115015453A CN 115015453 A CN115015453 A CN 115015453A CN 202210685298 A CN202210685298 A CN 202210685298A CN 115015453 A CN115015453 A CN 115015453A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
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Abstract
The invention discloses a pretreatment method for detecting a sweetening agent in an oral smokeless tobacco product, which is characterized in that the pretreatment method comprises the steps of cutting or grinding the oral smokeless tobacco product into powder, extracting the powder in a constant-temperature ultrasonic water bath for a certain time by using ultrapure water, and detecting the sweetening agent in the oral smokeless tobacco product by using a high performance liquid chromatography-diode array detector, a high performance liquid chromatography-evaporative light scattering detector or a high performance liquid chromatography-tandem mass spectrometry machine. The application has the advantages of simple and convenient sample pretreatment process and higher efficiency, and is suitable for the pretreatment of the sweetener in the oral smokeless tobacco products.
Description
Technical Field
The invention relates to the technical field of tobacco chemical component determination, in particular to a pretreatment method for detecting a sweetener in an oral smokeless tobacco product.
Background
Oral smokeless tobacco products, which include mainly gum-based, pouched mouth-containing and tobacco-containing products, are typically consumed by being placed between the lips and the gums, or chewed in the mouth. The gum base cigarette is a smokeless tobacco product in the form of tablet or granule prepared by adding certain amount of tobacco powder, gum base, sugar, additive, essence and other edible additives into gum base, and is chewed in oral cavity. The bagged buccal cigarette is a wet tobacco powder or tobacco shred tobacco product which is prepared by crushing, heat treating and packaging air-cured tobacco or open fire cured tobacco, and is contained between lips and gums when in use. The buccal tobacco is a smokeless tobacco product which is pressed by tobacco powder added with components such as mint, eucalyptus and the like, can be dissolved in the oral cavity and is usually put into the oral cavity for buccal use.
Some sweeteners are usually added into oral smokeless tobacco products to achieve the purposes of improving flavor and increasing sweetness, and the oral smokeless tobacco products can be divided into natural sweeteners and artificial sweeteners according to sources. The common natural sweetening agents comprise stevioside, momordica grosvenori glucoside V and the like, the common artificial sweetening agents mainly comprise acesulfame potassium, saccharin sodium, alitame, sodium cyclamate, sucralose and the like, and the sweetness of the sweetening agents is generally 30-700 times that of sucrose. GB2760-2014 national food safety standard food additive use standard specifies 21 food additives allowed to be added, and specifies the maximum addition amount of the food additives in different food varieties. In the oral smokeless tobacco product, the type and the addition amount of the sweetener have great influence on the stability of the sensory quality of the oral smokeless tobacco product, and the supervision and the measurement of the sweetener in the oral smokeless tobacco product are realized.
Therefore, a need exists for an efficient pretreatment method for detecting sweeteners in oral smokeless tobacco products.
Disclosure of Invention
The invention aims to solve the technical problem of providing a pretreatment method for detecting a sweetening agent in an oral smokeless tobacco product, which lays a foundation for detecting and controlling the content of related substances in the tobacco product.
The technical problem to be solved by the invention is realized by the following technical scheme:
a pretreatment method for detecting a sweetening agent in an oral smokeless tobacco product comprises the steps of cutting or grinding the oral smokeless tobacco product into powder, extracting the powder in a constant-temperature ultrasonic water bath for a certain time by using ultrapure water, and detecting the powder on a high-performance liquid chromatography-diode array detector, a high-performance liquid chromatography-evaporative light scattering detector or a high-performance liquid chromatography-tandem mass spectrometry machine to complete the determination of the sweetening agent in the oral smokeless tobacco product.
Preferably, in the technical scheme, the water bath temperature is 40 ℃, and the extraction time is 20 min.
Preferably, in the above technical solution, the oral smokeless tobacco product is gum-based tobacco, bagged buccal tobacco or buccal tobacco.
Preferably, in the above technical solution, when the oral smokeless tobacco product is gum-based tobacco, the pretreatment method includes the following steps:
(1) cutting a gum base cigarette sample, weighing the sample, placing the sample in a triangular flask with a plug, and adding an extraction liquid;
(2) extracting in 40 deg.C constant temperature ultrasonic water bath for 20min to obtain extractive solution;
(3) and (3) sucking a proper amount of extract, filtering the extract by a 0.22 mu m aqueous phase filter membrane into a chromatographic bottle, and performing high performance liquid chromatography analysis by taking the extract as a sample to be detected.
Preferably, in the above technical scheme, when the oral smokeless tobacco product is a bagged mouth cigarette, the pretreatment method comprises the following steps:
(1) cutting a non-woven fabric layer in the bagged mouth-holding cigarette, grinding the non-woven fabric layer into powder, weighing a sample, placing the sample in a triangular bottle with a plug, and adding an extraction liquid;
(2) extracting in 40 deg.C constant temperature ultrasonic water bath for 20min to obtain extractive solution;
(3) and (3) sucking a proper amount of extract, filtering the extract by a 0.22 mu m aqueous phase filter membrane into a chromatographic bottle, and performing high performance liquid chromatography analysis by taking the extract as a sample to be detected.
Preferably, in the above technical solution, when the oral smokeless tobacco product is a tobacco product containing chemical substances, the pretreatment method includes the steps of:
(1) grinding a tobacco sample containing chemical tobacco into powder, weighing the sample, placing the sample in a triangular flask with a plug, and adding an extraction liquid;
(2) extracting in 40 deg.C constant temperature ultrasonic water bath for 20 min;
(3) and (3) sucking a proper amount of extract, filtering the extract by a 0.22 mu m aqueous phase filter membrane into a chromatographic bottle, and performing high performance liquid chromatography analysis by taking the extract as a sample to be detected.
Preferably, in the above technical solution, in the step (1), a material ratio of the sample to the extraction solution is 1: 10.
preferably, in the above technical solution, in the step (1), the extraction liquid is ultrapure water.
The technical scheme of the invention has the following beneficial effects:
(1) the method is used for the efficient pretreatment method for measuring the sweetener in the oral smokeless tobacco product, the sample to be analyzed can be obtained without sample transfer in the whole sample pretreatment process, and the experimental operation is simplified.
(2) The invention optimizes the extraction mode, extraction time, volume of the extracting agent, extraction temperature and the like, and improves the extraction efficiency.
(3) The extraction solvent used in the invention is ultrapure water, has no pollution to the environment, and accords with the green development concept.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description, serve to explain the principles of the invention.
FIG. 1 shows the effect of different extraction methods on the extraction efficiency.
FIG. 2 shows the effect of different extractant volumes on the extraction.
FIG. 3 shows the effect of different extraction times on the extraction results.
FIG. 4 shows the effect of different extraction temperatures on the extraction performance.
FIG. 5 is a flow chart of the measurement method of the present invention.
Detailed Description
Various exemplary embodiments of the present invention will now be described in detail with reference to the accompanying drawings. It should be noted that: the relative arrangement of the components and steps, the numerical expressions and numerical values set forth in these embodiments do not limit the scope of the present invention unless specifically stated otherwise.
Example 1
A pretreatment method for detecting a sweetener in an oral smokeless tobacco product comprises the following steps:
(1) gum base tobacco: the gum-based tobacco sample was chopped, 1g of the sample was weighed into a 25mL stoppered flask, 10mL of extract was added: ultrapure water;
(2) extracting in 40 deg.C constant temperature ultrasonic water bath for 20 min;
(3) and (3) sucking a proper amount of extract, filtering the extract by a 0.22 mu m aqueous phase filter membrane into a chromatographic bottle, and performing high performance liquid chromatography analysis by taking the extract as a sample to be detected.
Example 2
A pretreatment method for detecting a sweetener in an oral smokeless tobacco product comprises the following steps:
(1) bagging buccal cigarettes: cutting a non-woven fabric layer in the bagged buccal cigarette, grinding the non-woven fabric layer into powder, weighing 1g of sample, placing the sample in a 25mL triangular flask with a plug, and adding 10mL of extraction liquid: ultrapure water;
(2) extracting in 40 deg.C constant temperature ultrasonic water bath for 20 min;
(3) and (3) sucking a proper amount of extract, filtering the extract by a 0.22 mu m aqueous phase filter membrane into a chromatographic bottle, and performing high performance liquid chromatography analysis by taking the extract as a sample to be detected.
Example 3
A pretreatment method for detecting a sweetener in an oral smokeless tobacco product comprises the following steps:
(1) smoke is dissolved: grinding the tobacco sample into powder, weighing 1g of the sample, placing the sample in a 25mL triangular flask with a plug, adding 10mL of extraction liquid: ultrapure water;
(2) extracting in 40 deg.C constant temperature ultrasonic water bath for 20 min;
(3) and (3) sucking a proper amount of extract, filtering the extract by a 0.22 mu m aqueous phase filter membrane into a chromatographic bottle, and performing high performance liquid chromatography analysis by taking the extract as a sample to be detected.
In the research, the oral smokeless tobacco product is cut or ground into powder, and then 10mL of ultrapure water is used for 20min in a constant-temperature ultrasonic water bath at 40 ℃ to complete extraction. The method is simple to operate, and provides a technical means for detecting the sweetener in the oral smokeless tobacco product.
Although the present invention has been described with reference to the above embodiments, it should be understood that the present invention is not limited thereto, and various changes and modifications may be made by those skilled in the art without departing from the spirit and scope of the present invention.
Claims (8)
1. A pretreatment method for detecting a sweetening agent in an oral smokeless tobacco product is characterized in that the pretreatment method comprises the steps of cutting or grinding the oral smokeless tobacco product into powder, extracting the powder in a constant-temperature ultrasonic water bath for a certain time by using ultrapure water, and detecting the powder on a computer by using a high performance liquid chromatography-diode array detector, a high performance liquid chromatography-evaporative light scattering detector or a high performance liquid chromatography tandem mass spectrometry to complete the determination of the sweetening agent in the oral smokeless tobacco product.
2. The method of claim 1, wherein the water bath temperature is 40 ℃ and the extraction time is 20 min.
3. The method of claim 2, wherein the oral smokeless tobacco product is gum-based, pouched, or chemically-containing.
4. The method of claim 3, wherein when the oral smokeless tobacco product is gum-based, the method comprises the steps of:
(1) cutting a gum base cigarette sample, weighing the sample, placing the sample in a triangular flask with a plug, and adding an extraction liquid;
(2) extracting in 40 deg.C constant temperature ultrasonic water bath for 20min to obtain extractive solution;
(3) and (3) sucking a proper amount of extract, filtering the extract by a 0.22 mu m aqueous phase filter membrane into a chromatographic bottle, and performing high performance liquid chromatography analysis by taking the extract as a sample to be detected.
5. The method of claim 3, wherein when the oral smokeless tobacco product is a pouched mouth cigarette, the method comprises the steps of:
(1) cutting a non-woven fabric layer in the bagged mouth-holding cigarette, grinding the non-woven fabric layer into powder, weighing a sample, placing the sample in a triangular bottle with a plug, and adding an extraction liquid;
(2) extracting in 40 deg.C constant temperature ultrasonic water bath for 20min to obtain extractive solution;
(3) and (3) sucking a proper amount of extract, filtering the extract by a 0.22 mu m aqueous phase filter membrane into a chromatographic bottle, and performing high performance liquid chromatography analysis by taking the extract as a sample to be detected.
6. The method of claim 3, wherein when the oral smokeless tobacco product is a tobacco product containing chemical, the method comprises the steps of:
(1) grinding a tobacco sample containing chemical tobacco into powder, weighing the sample, placing the sample in a triangular flask with a plug, and adding an extraction liquid;
(2) extracting in 40 deg.C constant temperature ultrasonic water bath for 20 min;
(3) and (3) sucking a proper amount of extract, filtering the extract by a 0.22 mu m aqueous phase filter membrane into a chromatographic bottle, and performing high performance liquid chromatography analysis by taking the extract as a sample to be detected.
7. The method for pretreating a sweetener in an oral smokeless tobacco product according to claim 4, 5 or 6, wherein in the step (1), the material ratio of the sample to the extract is 1: 10.
8. the method for pretreating a sweetener in an oral smokeless tobacco product according to claim 4, 5 or 6, wherein the extraction liquid in step (1) is ultrapure water.
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| CN202210685298.9A CN115015453A (en) | 2022-06-15 | 2022-06-15 | Pretreatment method for detecting sweetener in oral smokeless tobacco product |
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| CN202210685298.9A CN115015453A (en) | 2022-06-15 | 2022-06-15 | Pretreatment method for detecting sweetener in oral smokeless tobacco product |
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