Solid phase extraction agent, method of making and use thereof
By preparing solid-phase extractants from cigar stems, shavings, and grape stems, the problems of complex pretreatment and high time cost in detecting benzo[a]pyrene in cigarette smoke were solved, and rapid, stable, and efficient adsorption and detection of benzo[a]pyrene were achieved.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- HUBEI CHINA TOBACCO INDUSTRY CO LTD
- Filing Date
- 2024-01-11
- Publication Date
- 2026-06-26
AI Technical Summary
Existing methods for detecting benzo[a]pyrene in cigarette smoke involve complex pretreatment and are time-consuming.
Solid-phase extractants were prepared using cigar stems, shavings, and grape stems as raw materials. The solid-phase extractants were prepared through steps such as acidic water washing, microwave heating and soaking, grinding, organic solvent heat treatment, and freeze drying, and were used for the detection of benzo[a]pyrene in cigarette smoke.
It achieves rapid, stable adsorption and efficient enrichment of benzo[a]pyrene in cigarette smoke, reducing detection costs and improving detection efficiency and accuracy.
Smart Images
Figure BDA0004659675730000071
Abstract
Description
Technical Field
[0001] This application relates to the field of rapid pretreatment technology for cigarette smoke, and in particular to a solid-phase extractant, its preparation method, and its application. Background Technology
[0002] Benzo[a]pyrene (BaP) is the most potent carcinogen among polycyclic aromatic hydrocarbons (PAHs), and accurate determination of BaP content in cigarette smoke is crucial for assessing the harmfulness of cigarettes. Currently, reported methods for BaP content analysis include fluorescence spectrophotometry, gas chromatography, gas chromatography / mass spectrometry (GC / MS), paper chromatography, and high-performance liquid chromatography (HPLC). However, commonly used methods such as GC / MS and HPLC suffer from drawbacks such as complex pretreatment procedures and high time costs. Summary of the Invention
[0003] This invention provides a solid-phase extractant, its preparation method, and its application to solve the technical problems of complex pretreatment and high time cost in existing methods for detecting benzo[a]pyrene content in cigarette smoke.
[0004] To achieve the above objectives, the technical solution provided by the present invention is as follows:
[0005] In a first aspect, the present invention provides a solid-phase extractant, the raw materials of which include cigar stems and shavings, wherein the mass ratio of the cigar stems to the shavings is 1:1 to 2.
[0006] Furthermore, the raw materials for the solid phase extractant also include grape stems, and the mass ratio of the grape stems to the shavings is 1:1.
[0007] In a second aspect, the present invention provides a method for preparing the above-mentioned solid-phase extractant, comprising the following steps: washing the raw material of the solid-phase extractant with acidic water, heating and soaking it, grinding it into powder, then heat-treating it with an organic solvent, evaporating it, and freeze-drying it to obtain the solid-phase extractant.
[0008] Furthermore, in the acidic water washing step, the weight of the acidic water is 4 to 15 times the total weight of the raw materials of the solid phase extractant; the pH value of the acidic water is 1 to 3.
[0009] Furthermore, the heating step uses microwave heating with a microwave power of 500W to 1500W and a heating time of 10min to 30min.
[0010] Further, the organic solvent includes ethyl acetate and butyl acetate, wherein the volume ratio of ethyl acetate to butyl acetate is 0.5 to 2:1; and the mass ratio of the organic solvent to the powder is 4 to 15:1.
[0011] Furthermore, in the heat treatment step, the heating temperature is 70℃~80℃, and the heating time is 0.5h~1.5h.
[0012] A third aspect of the present invention provides the application of the above-mentioned solid-phase extractant in the detection of benzo[a]pyrene in cigarette smoke.
[0013] Furthermore, the method for detecting benzo[a]pyrene in cigarette smoke includes the following steps:
[0014] S1. The solid-phase extractant is placed in an empty column tube as a packing material to obtain a solid-phase extraction column;
[0015] S2. Smoke cigarettes and use acetic acid to adsorb benzo[a]pyrene in the smoke, and use the adsorbed solution as the sample loading solution;
[0016] S3. The sample solution obtained in step S2 is added to and flows through the solid phase extraction column of step S1, and then ethanol and acetic acid are added to the solid phase extraction column for elution, and the eluent is collected.
[0017] S4. The eluent is concentrated by nitrogen blowing and then analyzed.
[0018] Furthermore, between step S2 and step S3, the following step is also included: passing dichloromethane, acetic acid, and water sequentially through the solid-phase extraction column, wherein the volume ratio of dichloromethane, acetic acid, and water is 3-5:3-5:1, and the flow rate is less than 0.5 mL / min to 1 mL / min.
[0019] Further, in step S2, after smoking the cigarette, benzo[a]pyrene in the smoke is continuously adsorbed by glass fiber filter and acetic acid. The adsorbed glass fiber filter is extracted with acetic acid to obtain an extract. The adsorbed acetic acid and the extract are combined as a loading solution.
[0020] Furthermore, in step S3, the volume ratio of ethanol to acetic acid in the eluent is 3:1 to 9.
[0021] The solid-phase extractant provided by this invention uses cigar stems and shavings as raw materials, both of which are natural plants. In addition to being rich in vascular systems and cellulose, the xylem and phloem form a bundled structure, and the various capillary pore structures make the pores abundant. It also has abundant polar hydroxyl and hydrophobic aromatic functional groups. After being made into a solid-phase extractant, it can form π-π bonds and hydrogen bonds with benzo[a]pyrene, and has a strong adsorption capacity and high adsorption stability for benzo[a]pyrene. Due to its rich pores and good permeability, benzo[a]pyrene in the adsorbed solution passes through the extraction column faster. It has high adsorption, enrichment and purification efficiency for benzo[a]pyrene in the adsorbed solution, and is low in cost and environmentally friendly. Detailed Implementation
[0022] To enable those skilled in the art to better understand the technical solutions in this application, the technical solutions in the embodiments of this application will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of this application, and not all of the embodiments. Based on the embodiments in this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.
[0023] In a first aspect of this application, a solid-phase extractant is provided. The raw materials of the solid-phase extractant include cigar stems and shavings, and the mass ratio of cigar stems to shavings is 1:1 to 2.
[0024] Removing the stems from cigar tobacco leaves is a crucial step in handmade cigar production. Overly large cigar stems (the coarse, hard veins in the center of the tobacco leaf) negatively impact the cigar's appearance, affect its combustibility (causing uneven burning), and also affect its flavor, making it overly strong and causing a higher burning temperature, thus hindering the smoothness of the draw. Therefore, cigar stems, which affect both the cigar's appearance and the smoking experience, are discarded during handmade cigar production. Cigar stems comprise approximately 25% to 37% of the tobacco leaf's weight. This waste contains toxic compounds such as nicotine and cannot be disposed of with ordinary waste. Improper disposal of cigar stem waste causes environmental pollution and represents a significant waste of resources.
[0025] The waste cigar stems used in this application refer to the coarse stems and fragments of tobacco leaves discarded in handmade cigar production, while the wood shavings are commonly used waste wood shavings from commercial processing, thus being reused as waste resources. Cigar stems are generally larger in diameter than flue-cured tobacco stems and contain a richer vascular system. The processed porous structure has a higher adsorption efficiency for benzo[a]pyrene, while also providing efficient elution, making the detection of benzo[a]pyrene concentration in the smoke more accurate and faster.
[0026] The solid-phase extractant provided in this application includes cigar stems and shavings as raw materials, both of which are natural plants. In addition to being rich in vascular systems and cellulose, the xylem and phloem form a bundled structure, and the various capillary pore structures make the pores abundant. It also has abundant polar hydroxyl and hydrophobic aromatic functional groups, which can form π-π bonds and hydrogen bonds with benzo[a]pyrene. It has a strong adsorption capacity for benzo[a]pyrene and high adsorption stability. Due to its abundant pores and good permeability, benzo[a]pyrene in the adsorbed solution passes through the extraction column faster. It has high adsorption, enrichment and purification efficiency for benzo[a]pyrene in the adsorbed solution, and is low in cost and environmentally friendly.
[0027] In some embodiments, the raw materials for the solid-phase extractant also include grape stems, with a grape stem to shaving mass ratio of 1:1. Grape destemming is an important step in wine fermentation, and the resulting waste grape stems are a byproduct of grape processing, receiving little attention or development. In this application, grape stems are added, referring to the grape stems discarded before wine fermentation. Similar to cigar stems, they are rich in vascular systems and cellulose, and their xylem and phloem form a bundled structure with various capillary pores, abundant porosity, and rich polar hydroxyl and hydrophobic aromatic functional groups. These groups form π-π bonds and hydrogen bonds with benzo[a]pyrene, exhibiting strong adsorption capacity for benzo[a]pyrene.
[0028] A second aspect of the embodiments of this application provides a method for preparing the above-mentioned solid phase extractant, comprising the following steps: washing the raw material of the solid phase extractant with acidic water, heating and soaking it, grinding it into powder, then heat-treating it with an organic solvent, evaporating it, and freeze-drying it to obtain the solid phase extractant.
[0029] Cigar stems, grape stems, and shavings all contain impurities such as natural pigments, proteins, tannins, bitter resins, and tannic acid. To improve utilization, the embodiments of this application first subject them to acidic water washing and microwave heating soaking to quickly remove natural pigments, proteins, tannins, and other flavor substances from their surfaces. Then, the filter residue is ground into powder using a pulverizer and then heat-treated with an organic solvent to further remove natural pigments, proteins, and flavor components from the surface. Most of the solvent can be removed by rotary evaporation, and then vacuum freeze-drying is performed to obtain a solid-phase extractant that can rapidly pre-treat cigar smoke.
[0030] The solid-phase extractant preparation method of this application embodiment is convenient, fast and safe, requires few experimental equipment and procedures, is environmentally friendly and low in cost, uses waste natural raw materials with stable performance, improves the utilization value of waste resources such as waste tobacco stems and grape stems, and has high adsorption efficiency for benzo[a]pyrene in mainstream cigarette smoke.
[0031] A third aspect of this application provides the application of the aforementioned solid-phase extractant in the detection of benzo[a]pyrene in cigarette smoke. Solid-phase extraction is a widely used sample pretreatment method that can be coupled with techniques such as chromatography, both offline and online, to achieve the separation and analysis of substances in complex samples. The solid-phase extractant of this application can rapidly pretreat benzo[a]pyrene in cigarette smoke, thereby improving the detection efficiency of benzo[a]pyrene.
[0032] Furthermore, the method for detecting benzo[a]pyrene in cigarette smoke includes the following steps:
[0033] S1. A solid-phase extraction column is obtained by placing a solid-phase extraction agent as a packing material in an empty column tube.
[0034] S2. Smoke cigarettes and use acetic acid to adsorb benzo[a]pyrene in the smoke, and use the adsorbed solution as the sample loading solution;
[0035] S3. The sample solution obtained in step S2 is added to and flows through the solid phase extraction column of step S1. Ethanol and acetic acid are then added to the solid phase extraction column for elution, and the eluent is collected.
[0036] S4. The eluent is concentrated by nitrogen blowing and then analyzed.
[0037] In this embodiment, after the extraction column is packed in step S1, it is ultrasonically filled to ensure uniform packing, and then the top is covered with a sieve plate and pressed tightly. In step S2, acetic acid can be used to adsorb benzo[a]pyrene in the flue gas, or a glass fiber filter and acetic acid can be used for continuous solid-liquid adsorption of benzo[a]pyrene in the flue gas. The adsorbed glass fiber filter is extracted with acetic acid to obtain an extract, and the adsorbed acetic acid and the extract are combined as the loading solution. Glass fiber filter is a commonly used adsorption method, and will not be described in detail here. If the flue gas still contains some benzo[a]pyrene after adsorption by the glass fiber filter, it will be adsorbed by the subsequent acetic acid. The eluent contains impurities such as ethanol and acetic acid, which are collected and concentrated by nitrogen blowing for subsequent detection.
[0038] In this embodiment, the instrument used for detection and analysis in step S4 can be a commonly used instrument such as a high-performance liquid chromatograph, gas chromatograph, or fluorescence spectrophotometer, all of which can be used offline or online in conjunction with the solid-phase extraction in this embodiment.
[0039] All reagents used in the following examples are commercially available.
[0040] Example 1
[0041] A solid-phase extractant, the raw materials of which include cigar stems, grape stems and wood shavings, wherein the mass ratio of cigar stems, grape stems and wood shavings is 1:1:2.
[0042] The preparation method of the above solid-phase extractant includes the following steps:
[0043] 1. Mix cigar stems, grape stems, and wood shavings according to the above mass ratio, and wash with acidic water. The acidic water is purified water with a pH of 3 adjusted by hydrochloric acid, and its mass is 10 times the total mass of the cigar stems, grape stems, and wood shavings. While soaking the raw materials in the acidic water, microwave them for 18 minutes at a power of 1000W. Then, vacuum filter, air dry, and grind the filter residue into coarse powder using a grinder. The coarse powder should be 10-50 mesh.
[0044] 2. Add the above crude powder to an organic solvent, including ethyl acetate and butyl acetate, both of chromatographic purity, in a volume ratio of 1:1; the mass ratio of organic solvent to crude powder is 10:1. Heat to 75°C for 1 hour, reflux, then concentrate at least a small amount of solvent by rotary evaporation, and freeze-dry under vacuum to obtain a solid-phase extractant.
[0045] The application of the above-mentioned solid-phase extractant in the detection of benzo[a]pyrene in cigarette smoke. The method for detecting benzo[a]pyrene in cigarette smoke includes the following steps:
[0046] 1. Weigh 0.5g of solid phase extraction agent as packing material and load it into a 6mL solid phase extraction empty column tube with a sieve plate. Use an ultrasonic instrument to sonicate the packing material evenly. The ultrasonic frequency is 25kHz and the time is 5min. Cover the top with the sieve plate and press it tightly to obtain a solid phase extraction column.
[0047] 2. A deep-suction method was used to inhale cigarette smoke through a smoking machine. Benzo[a]pyrene in the smoke was adsorbed using a glass fiber filter and acetic acid in a continuous solid-liquid process. The adsorbed glass fiber filter was then extracted with acetic acid to obtain an extract. The adsorbed acetic acid and the extract were combined as the loading solution. The acetic acid was of chromatographic grade.
[0048] 3. Under vacuum, 8 mL of dichloromethane, 8 mL of acetic acid, and 2 mL of water are sequentially passed through the solid-phase extraction column prepared in step 1 at a flow rate of less than 4 mL / min. Then, 1 mL of the sample loading solution from step 2 is added and passed through the solid-phase extraction column. After completion, 2 mL of water is added as a washing solution and passed through the extraction column to remove residual matrix adsorbed on the extraction column. The water is then dried under vacuum. Finally, 2 mL of ethanol and 2 mL of acetic acid are added as eluents, respectively, and the mixture is passed through the extraction column under gravity and dried under vacuum. The eluent is collected in a centrifuge tube and combined, then concentrated to dryness under nitrogen blowing at 40°C.
[0049] 4. Dissolve in 200 μL of methanol, and add 10 μL to a high-performance liquid chromatography-ultraviolet detector for detection and analysis. The chromatographic column is a C18 column (250 mm * 4.6 mm id, 5 μm); mobile phase A: water, mobile phase B: methanol, mobile phase ratio A / B (5 / 95, v / v), flow rate is 1.0 mL / min; column temperature is 40℃; detector wavelength is 295 nm.
[0050] Example 2
[0051] This embodiment is basically the same as Embodiment 1, except that the raw materials of the solid phase extractant include cigar stems and shavings, and the mass ratio of cigar stems to shavings is 1:2.
[0052] Comparative Example 1
[0053] The content of benzo[α]pyrene was detected using a C18 extraction column in accordance with GB / T 21130 method for collecting total particulate matter in cigarette smoke and HJ956-2018 method for determining benzo[α]pyrene content by HPLC.
[0054] Comparative Example 2
[0055] Using a commercially available ALN neutral alumina column, the total particulate matter in cigarette smoke was collected according to GB / T 21130 and the benzo[α]pyrene content was determined by HPLC according to HJ956-2018.
[0056] Comparative Example 3
[0057] Using commercially available phenyl columns, the total particulate matter in cigarette smoke was collected according to GB / T 21130 and the benzo[α]pyrene content was determined by HPLC according to HJ956-2018.
[0058] The detection results of benzo[α]pyrene content in cigarette smoke in Examples 1 and 2, and Comparative Examples 1 to 3 are shown in Table 1. The results in Table 1 indicate that, in the examples of this application, solid-phase extraction columns prepared using cigar stems, grape stems, and shavings as raw materials resulted in higher extraction efficiency, shorter throughput, and greater adsorption of benzo[α]pyrene in cigarette smoke when the sample solution passed through the aforementioned solid-phase extraction column. Elution was also smoother, with elution efficiency tens of times higher, and the detection results of benzo[α]pyrene content were more accurate. The average detection value in the examples was 8.59 ng / cigarette, and the average value in the comparative examples was 7.69 ng / cigarette, representing an average increase of 11.7% and a maximum increase of 12.28%.
[0059] Table 1. Detection results of benzo[α]pyrene content in cigarette smoke.
[0060]
[0061] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims
1. The application of a solid-phase extractant in the detection of benzo[a]pyrene in cigarette smoke, wherein, The raw materials of the solid phase extractant include cigar stems and shavings, with the mass ratio of cigar stems to shavings being 1:1 to 2; the raw materials of the solid phase extractant also include grape stems, with the mass ratio of grape stems to shavings being 1:
1. The preparation method of the solid phase extractant includes the following steps: washing the raw material of the solid phase extractant with acidic water, heating and soaking it, grinding it into powder, then heat-treating it with an organic solvent, evaporating it, and freeze-drying it to obtain the solid phase extractant.
2. The application according to claim 1, characterized in that, In the acidic water washing step, the weight of the acidic water is 4 to 15 times the total weight of the raw materials of the solid phase extractant; the pH value of the acidic water is 1 to 3.
3. The application according to claim 1, characterized in that, The heating step uses microwave heating with a power of 500W to 1500W and a heating time of 10min to 30min.
4. The application according to claim 1, characterized in that, The organic solvent includes ethyl acetate and butyl acetate, wherein the volume ratio of ethyl acetate to butyl acetate is 0.5 to 2:1; and the mass ratio of the organic solvent to the powder is 4 to 15:
1.
5. The application according to claim 1, characterized in that, In the heat treatment step, the heating temperature is 70℃~80℃ and the heating time is 0.5h~1.5h.
6. The application according to claim 1, characterized in that, The method for detecting benzo[a]pyrene in cigarette smoke includes the following steps: S1. The solid-phase extractant is placed in an empty column tube as a packing material to obtain a solid-phase extraction column; S2. Smoke cigarettes and use acetic acid to adsorb benzo[a]pyrene in the smoke, and use the adsorbed solution as the sample loading solution; S3. The sample solution obtained in step S2 is added to and flows through the solid phase extraction column of step S1, and then ethanol and acetic acid are added to the solid phase extraction column for elution, and the eluent is collected. S4. The eluent is concentrated by nitrogen blowing and then analyzed.
7. The application according to claim 6, characterized in that, Between step S2 and step S3, the following step is also included: passing dichloromethane, acetic acid, and water sequentially through the solid-phase extraction column, wherein the volume ratio of dichloromethane, acetic acid, and water is 3-5:3-5:1, and the flow rate is less than 0.5 mL / min to 1 mL / min.
8. The application according to claim 6, characterized in that, In step S2, after smoking the cigarette, benzo[a]pyrene in the smoke is continuously adsorbed by glass fiber filter and acetic acid. The adsorbed glass fiber filter is then extracted with acetic acid to obtain an extract. The adsorbed acetic acid and the extract are combined as a loading solution.
9. The application according to claim 6, characterized in that, The volume ratio of ethanol to acetic acid in the eluent of step S3 is 3:1 to 9.