CN109738567B - Method for evaluating uniformity of cut tobacco flavoring mode - Google Patents
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Abstract
The invention relates to a method for evaluating the uniformity of a cut tobacco flavoring mode, which comprises the following steps: SPME-GC-MS analysis is carried out on the added essence and spice, SPME-GC-MS analysis is carried out on the on-line raw material tobacco shred, the characteristic marker of the essence and spice is determined, solid phase micro-extraction analysis is carried out on the flavored tobacco shred obtained by different flavoring modes, the elliptical area of PCA (principal component analysis) images of different flavoring modes is calculated, and the uniformity evaluation of the different flavoring modes is carried out. The method adopts solid-phase microextraction, can effectively measure volatile and semi-volatile components in the flavors and fragrances and the tobacco shreds, selects specific components in the flavors and fragrances as characteristic markers by removing common components of the flavors and the tobacco shreds, and selects peaks as common mode peaks for main component analysis, so that the result is more accurate.
Description
Technical Field
The invention relates to a uniformity evaluation method, in particular to a uniformity evaluation method of a cut tobacco perfuming mode.
Background
Cigarette formula is always regarded as the core technology of cigarette enterprises, however, the addition of flavors and fragrances in cigarettes is more regarded as a key technology. The technology of fingerprint spectrum is widely applied to the fields of Chinese medicine quality judgment and analysis, agricultural and sideline product traceability and the like. The chemical fingerprint spectrum has the characteristics of fingerprint characteristic analysis, macroscopic inference analysis and the like. In recent years, the application research of the fingerprint in the tobacco field has been carried out with a great deal of progress, and reports on the aspect are more and more, and the fingerprint has many applications in the aspects of quality monitoring, quality evaluation, quality consistency and the like of tobacco at present.
The quantitative characterization aspect of the perfuming uniformity of the tobacco shred perfuming equipment is always a difficult problem for evaluating the perfuming uniformity of the tobacco shred perfuming equipment, and the industry develops a large amount of work for evaluating the perfuming uniformity of the tobacco shred perfuming equipment, wherein the perfuming uniformity is deduced by theoretical calculation and a formula through previous research reports, and a tobacco essence marker is additionally added to evaluate the perfuming uniformity. However, the chemical composition of the tobacco essence perfume is complex, the varieties are various, and the essence added on the tobacco shreds of the cigarettes is the same in composition, only has different contents, and the content of the added essence is less. When one or more characteristic compounds are used as markers for evaluating the perfuming uniformity, the markers are difficult to find, and the markers are difficult to add in large-scale production, so that a reasonable comprehensive evaluation result cannot be obtained.
SPME has the effect of selectively concentrating and extracting trace components, so that specific trace compounds in cigarettes have good analysis accuracy.
Disclosure of Invention
In order to solve the above-mentioned problems, an object of the present invention is to provide a method for evaluating the uniformity of a mode of adding flavor to tobacco shreds. The purpose of the invention is realized by the following technical scheme:
a method for evaluating the uniformity of a cut tobacco flavoring mode is characterized by comprising the following steps: the method comprises the following steps:
step (1), SPME-GC-MS analysis is carried out on the added essence and flavor
Performing solid phase microextraction analysis on the used essence and flavor to obtain SPME-GC-MS spectrum of the added essence and flavor;
step (2), SPME-GC-MS analysis is carried out on the on-line raw material cut tobacco
Performing solid phase micro-extraction analysis on the on-line raw material cut tobacco to obtain an SPME-GC-MS spectrum of the on-line raw material cut tobacco;
step (3) determination of flavor and fragrance characteristic markers
Selecting specific components in the essence and spice as characteristic markers by comparing SPME-GC-MS spectrums of the on-line raw material tobacco shreds obtained in the step (2) and the essence and spice obtained in the step (1);
SPME-GC-MS analysis of flavored cut tobacco obtained in different flavoring modes in step (4)
Randomly selecting a plurality of flavored cut tobacco samples obtained in different online flavoring modes to perform SPME-GC-MS analysis to obtain SPME-GC-MS spectra in different flavoring modes;
step (5) calculating the elliptical areas of different perfuming modes in the PCA score chart
In the solid phase micro-extraction maps with different perfuming modes in the step (4), respectively taking a sample with the best internal standard recovery rate as a base number, and comparing the internal standard peak areas of other samples with the base number to obtain a ratio which is a weighting coefficient; using the formula: weighted peak area = original integrated peak area/weighting coefficient, peak area is quantified;
selecting peaks of the characteristic markers of the flavors and fragrances for calculation, quantifying the peak areas of the samples according to the method, determining to obtain common peaks, and establishing a chemical fingerprint spectrum common mode of the tobacco shred flavored samples;
performing dimensionality reduction treatment on the essence and spice characteristic marker peak data in the SPME-GC-MS spectrum obtained in the step (4) by adopting a principal component analysis method, and drawing a PCA score map; the elliptical area is then calculated as follows:
A=π*std(X)*std(Y)；
wherein std (X) and std (Y) respectively represent the major axis and the minor axis of the ellipse, and X, Y respectively represent the PC1 direction and the PC2 direction in the PCA score chart; the standard deviation of X, Y was calculated as follows, and the elliptical area was used to characterize the dispersion of each set of samples:
wherein, XiAnd YiIs the spatial location coordinate of each element,
and
is the arithmetic mean center;
step (6), uniformity evaluation of different perfuming modes
The differences are compared by the sizes of the elliptical areas, and the larger the elliptical area is, the poorer the perfuming uniformity of the perfuming mode is.
Further, in the step (1), randomly selecting online essence and spice samples, wherein the sampling frequency is not less than 5 times, and the sampling amount is not less than 10g each time; accurately weighing 0.7g of each sample, placing the sample in a 20mL brown headspace bottle, adding 0.25ug/ul of deuterated toluene as an internal standard, screwing the bottle cap tightly, sealing, and measuring by an SPME-GC-MS method to obtain the solid phase microextraction spectrum of the essence and spice.
Further, in the step (2), on-line raw material tobacco shred samples are randomly selected, the sampling frequency is not less than 5 times, and the sampling amount is not less than 10g each time; accurately weighing 0.7g of each sample, placing the sample in a 20mL brown headspace bottle, adding 0.25ug/ul of deuterated toluene as an internal standard, screwing the bottle cap tightly, sealing, and measuring by an SPME-GC-MS method to obtain the solid phase microextraction spectrum of the on-line raw material cut tobacco.
Further, in the step (4), the sampling frequency of each perfuming mode is not less than 10 times, and the sampling amount of each time is not less than 10 g; accurately weighing 0.7g of each sample, placing the sample in a 20mL brown headspace bottle, adding 0.25ug/ul of deuterated toluene as an internal standard, screwing the bottle cap tightly, sealing, and measuring according to an SPME-GC-MS method to obtain solid phase microextraction spectra in different perfuming modes.
Further, the solid phase microextraction conditions were as follows:
an extraction head: 85 μm Carboxen/PDMS; the extraction temperature is as follows: 80 ℃; extraction time: 60 min; desorption time: and (3) 30 min.
Further, the gas chromatography/mass spectrometry conditions were as follows:
sample inlet temperature: shunting and injecting samples at 250 ℃; carrier gas: 99.99% helium; constant flow rate: 1.0 mL/min; a chromatographic column: HP-INNOWAX; the split ratio is as follows: 5: 1; temperature rising procedure: maintaining the initial temperature at 50 deg.C for 2 min; the heating rate is as follows: 5 ℃/min to 220 ℃, then 10 ℃/min to 250 ℃, and keeping for 1 min;
an ionization mode: EI (El)+(ii) a Ion source temperature: 230 ℃; transmission line temperature: 250 ℃; scanning range: 40amu to 350 amu; solvent retardation: 3.5 min.
Further, in the step (5), the interpretation rate of the accumulated variance of the principal components obtained by PCA analysis is more than 85%.
Compared with the prior art, the invention has the following beneficial effects:
(1) the method adopts solid-phase microextraction, can effectively measure volatile and semi-volatile components in the flavors and fragrances and the tobacco shreds, selects specific components in the flavors and fragrances as characteristic markers by removing common components of the flavors and the tobacco shreds, and selects peaks as common mode peaks for main component analysis, so that the result is more accurate.
(2) The invention calculates the elliptical area of the main component score chart, quantifies the perfuming uniformity through the elliptical area, further evaluates the perfuming uniformity of different perfuming modes, has objective and intuitive result and can accurately evaluate the perfuming uniformity of tobacco shreds in different perfuming modes.
(3) The invention can evaluate the perfuming mode of online large-scale production without adding a characteristic marker.
(4) The method does not need specific analysis on one or a certain class of compounds, and is suitable for evaluating the perfuming uniformity of all types of tobacco shreds.
Drawings
FIG. 1 is a diagram of PCA for different modes of perfuming of the examples;
wherein, in the procedure of perfuming in a workshop, perfuming is carried out through a vertical material pipe, and in the procedure of perfuming in the workshop, perfuming is carried out through a roller material pipe.
Detailed Description
The invention is further described in detail below with reference to the drawings and examples, but the invention is not limited in any way, and any changes or modifications made based on the teaching of the invention fall within the scope of the invention. The examples do not specify particular techniques or conditions, and are performed according to the techniques or conditions described in the literature in the art or according to the product specifications. The materials or equipment used are not indicated by manufacturers, and all are conventional products available by purchase.
Example 1
The embodiment aims at two perfuming modes of a vertical material pipe and a roller material pipe in the perfuming process of the existing workshop, wherein the essence and the spice of the two perfuming modes are the same as those of the raw material cut tobacco.
The uniformity evaluation method of the two tobacco shred flavoring modes comprises the following steps:
step (1), performing SPME-GC-MS (solid phase microextraction-gas chromatography-mass spectrometry) analysis on the used flavors and fragrances
Randomly selecting an online essence and spice sample, and sampling for 8 times, wherein the sample amount is 20g each time. Accurately weighing 0.7g of sample (about 1 cigarette tobacco shred with conventional specification) per sample, placing the sample in a 20mL brown headspace bottle, adding 0.25ug/ul of deuterated toluene (n-hexane solution) as an internal standard, then screwing the bottle cap tightly, sealing, and measuring according to the SPME-GC-MS method to obtain the SPME-GC-MS spectrum of the essence and spice.
Wherein, the solid phase micro-extraction conditions are as follows:
an extraction head: 85 μm Carboxen/PDMS; the extraction temperature is as follows: 80 ℃; extraction time: 60 min; desorption time: and (3) 30 min.
The gas chromatography/mass spectrometry conditions were as follows:
sample inlet temperature: shunting and injecting samples at 250 ℃; carrier gas: 99.99% helium; constant flow rate: 1.0 mL/min; a chromatographic column: HP-INNOWAX (30 m.times.0.25 mm id. times.0.25. mu. mdf, Agilent, USA); the split ratio is as follows: 5: 1; temperature rising procedure: maintaining the initial temperature at 50 deg.C for 2 min; the heating rate is as follows: 5 ℃/min to 220 ℃, then 10 ℃/min to 250 ℃, for 1 min.
An ionization mode: EI (El)+(ii) a Ion source temperature: 230 ℃; transmission line temperature: 250 ℃; scanning range: 40amu to 350 amu; solvent retardation: 3.5 min.
Step (2), SPME-GC-MS analysis of on-line raw material cut tobacco
And randomly selecting an online raw material tobacco shred sample, wherein the sampling time is 8 times, and the sample amount is 20g each time. Accurately weighing 0.7g of sample (about 1 cigarette tobacco shred with conventional specification) per sample, placing the sample in a 20mL brown headspace bottle, adding 0.25ug/ul of deuterated toluene (n-hexane solution) as an internal standard, then screwing the bottle cap tightly, sealing, and measuring by an SPME-GC-MS method to obtain an SPME-GC-MS spectrum of the on-line raw material tobacco shred. The solid phase microextraction conditions and gas chromatography/mass spectrometry conditions were the same as in step (1).
Step (3), determination of specific component of essence and spice, namely marker
By comparing the solid phase micro-extraction maps of the on-line raw material cut tobacco and the essence perfume, the specific components of vanillin, longifolene, menthol, citronellol methyl ester, citronellol ethyl ester, geraniol, nerol, eugenol, phenethyl alcohol and acetic acid in the essence perfume are selected as characteristic markers.
SPME-GC-MS analysis of flavored cut tobacco obtained in different flavoring modes in step (4)
Respectively and randomly selecting 30 flavored tobacco shred samples obtained by two flavoring modes of a vertical material pipe and a roller material pipe, wherein the amount of each sample is not less than 10 g. Accurately weighing 0.7g of sample (about 1 cigarette tobacco shred with conventional specification) per sample, placing the sample in a 20mL brown headspace bottle, adding 0.25ug/ul of deuterated toluene (n-hexane solution) as an internal standard, then screwing the bottle cap, sealing, and measuring according to an SPME-GC-MS method to obtain two perfuming SPME-GC-MS spectra of a vertical material tube and a roller material tube. The solid phase microextraction conditions and gas chromatography/mass spectrometry conditions were the same as in step (1).
Step (5) calculating the elliptical areas of different perfuming modes in the PCA score chart
In the two perfuming solid phase micro-extraction spectra of the vertical material tube and the roller material tube obtained in the step (4), the sample with the best internal standard recovery rate is taken as a base number, the internal standard peak areas of other samples are compared with the base number, and the ratio is obtained as a weighting coefficient.
Using the formula: weighted peak area = raw integrated peak area/weighting coefficient, resulting in quantification of peak area. And (3) sequentially introducing two perfuming solid-phase microextraction spectrums of the vertical material tube and the roller material tube into Chemmind Chempattern 2017 in a CDF format, only selecting the peaks of specific components, namely markers, of vanillin, longifolene, menthol, citronellol methyl ester, citronellol ethyl ester, geraniol, nerol, eugenol, phenethyl alcohol and an acetic acid essence perfume, calculating, quantifying the peak area of each sample according to the method, determining a common peak, and establishing a chemical fingerprint spectrum common mode of the cut tobacco perfuming sample.
And (3) performing dimensionality reduction treatment on the flavor and fragrance characteristic marker peak data in the SPME-GC-MS spectrum in the step (4) by adopting a Principal Component Analysis (PCA) method, and drawing a PCA diagram, wherein the X axis is a PC1 score, and the Y axis is a PC2 score, as shown in figure 1.
The elliptical area is then calculated as follows:
A=π*std(X)*std(Y)；
wherein std (X) and std (Y) respectively represent the major axis and the minor axis of the ellipse, and X, Y respectively represent the PC1 direction and the PC2 direction in the PCA score chart; the standard deviation of X, Y was calculated as follows, and the elliptical area was used to characterize the dispersion of each set of samples:
wherein, XiAnd YiIs the spatial location coordinate of each element,
and
is the arithmetic mean center.
The elliptical areas for both modes of perfuming are shown in table 1:
TABLE 1 elliptical area for perfuming vertical tubes and for perfuming roller tubes
Step (6), uniformity evaluation of different perfuming modes
As can be seen from table 1: the elliptical area of the vertical material pipe is small, and the perfuming uniformity of the cut tobacco is good.
In order to verify the results, the two groups of tobacco shreds are prepared into cigarettes by adopting a cloud cigarette (purple) three-paper one-rod rolling process, the cigarettes are divided into 10 groups for smoking evaluation and scoring, and the results are shown in table 2 in the specification of GB 5606.4-2005:
TABLE 2 evaluation and analysis of cigarettes with different flavoring methods
As can be seen from the standard deviation calculation in Table 2, the standard deviation of the cut tobacco cigarette with the vertical pipe is 0.158, and the standard deviation of the cut tobacco with the roller pipe is 0.789, which shows that the uniformity of the cut tobacco with the vertical pipe is superior to that of the cut tobacco with the roller pipe, and the result is consistent with the result obtained by the calculation of the invention, which shows that the method has better accuracy in the uniformity evaluation of the cut tobacco flavoring mode.
Comparative example 1
In order to verify the superiority and effectiveness of the invention, the same method steps as the example are adopted for the sample of the above example 1, wherein only the detection method is changed into the HS-GC-MS method, and the specific test conditions are as follows:
head space conditions
Furnace temperature: 110 ℃, sampling needle temperature: 140 ℃, transmission line temperature: 190 ℃, carrier gas: helium (99.99%), 20Psi, incubation time: 40min, sample injection time: 0.2min, needle pulling time: 0.5min, pressurization time: 2min, GC analysis cycle time: and (5) 50 min. Sample introduction mode: time, operation mode: is constant.
Gas chromatography/mass spectrometry conditions:
sample inlet temperature: 250 ℃, no-split-flow sample injection, carrier gas: helium (99.99%), constant flow rate 1.0mL/min, column: DB-WAX (30 m × 0.25 mm id × 0.25 μm df, Agilent Corp., USA), split ratio: 20: 1, temperature rising program: the initial temperature is kept at 40 ℃ for 1min, the heating rate is 5 ℃/min to 220 ℃, and the temperature is kept for 3 min. An ionization mode: EI +, ion source temperature: 230 ℃, transmission line temperature: 240 ℃, scan range: 33amu-400amu, solvent retardation: 3.5 min.
The data processing method used in example 1 was used to calculate the elliptical area, and the results are shown in table 3.
TABLE 3 elliptical area for perfuming vertical tubes and for perfuming roller tubes
As can be seen from Table 3, the elliptical area of the roller tube is small and inconsistent with the result of the smoke panel test.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (4)
1. A method for evaluating the uniformity of a cut tobacco flavoring mode is characterized by comprising the following steps: the method comprises the following steps:
step (1), SPME-GC-MS analysis is carried out on the added essence and flavor
Performing solid phase microextraction analysis on the used essence and flavor to obtain SPME-GC-MS spectrum of the added essence and flavor;
randomly selecting online essence and spice samples, wherein the sampling frequency is not less than 5 times, and the sampling amount is not less than 10g each time; accurately weighing 0.7g of each sample, placing the sample in a 20mL brown headspace bottle, adding 0.25ug/ul of deuterated toluene as an internal standard, immediately screwing a bottle cap, sealing, and measuring according to an SPME-GC-MS method to obtain a solid phase microextraction spectrum of the essence and the spice;
step (2), SPME-GC-MS analysis is carried out on the on-line raw material cut tobacco
Performing solid phase micro-extraction analysis on the on-line raw material cut tobacco to obtain an SPME-GC-MS spectrum of the on-line raw material cut tobacco;
step (3) determination of flavor and fragrance characteristic markers
Selecting specific components in the essence and spice as characteristic markers by comparing SPME-GC-MS spectrums of the on-line raw material tobacco shreds obtained in the step (2) and the essence and spice obtained in the step (1);
SPME-GC-MS analysis of flavored cut tobacco obtained in different flavoring modes in step (4)
Randomly selecting a plurality of flavored cut tobacco samples obtained in different online flavoring modes to perform SPME-GC-MS analysis to obtain SPME-GC-MS spectra in different flavoring modes;
the solid phase microextraction conditions were as follows:
an extraction head: 85 μm Carboxen/PDMS; the extraction temperature is as follows: 80 ℃; extraction time: 60 min; desorption time: 30 min;
the gas chromatography/mass spectrometry conditions were as follows:
sample inlet temperature: shunting and injecting samples at 250 ℃; carrier gas: 99.99% helium; constant flow rate: 1.0 mL/min; a chromatographic column: HP-INNOWAX; the split ratio is as follows: 5: 1; temperature rising procedure: maintaining the initial temperature at 50 deg.C for 2 min; the heating rate is as follows: 5 ℃/min to 220 ℃, then 10 ℃/min to 250 ℃, and keeping for 1 min;
an ionization mode: EI (El)+(ii) a Ion source temperature: 230 ℃; transmission line temperature: 250 ℃; scanning range: 40amu to 350 amu; solvent retardation: 3.5 min;
step (5) calculating the elliptical areas of different perfuming modes in the PCA score chart
In the solid phase micro-extraction maps with different perfuming modes in the step (4), respectively taking a sample with the best internal standard recovery rate as a base number, and comparing the internal standard peak areas of other samples with the base number to obtain a ratio which is a weighting coefficient; using the formula: quantifying the peak area as the original integral peak area/weighting coefficient;
selecting peaks of the characteristic markers of the flavors and fragrances for calculation, quantifying the peak areas of the samples according to the method, determining to obtain common peaks, and establishing a chemical fingerprint spectrum common mode of the tobacco shred flavored samples;
performing dimensionality reduction treatment on the essence and spice characteristic marker peak data in the SPME-GC-MS spectrum obtained in the step (4) by adopting a principal component analysis method, and drawing a PCA score map; the elliptical area A is then calculated as:
A＝π*std(X)*std(Y)；
wherein std (X) and std (Y) respectively represent the major axis and the minor axis of the ellipse, and X, Y respectively represent the PC1 direction and the PC2 direction in the PCA score chart; the standard deviation of X, Y was calculated as follows, and the elliptical area was used to characterize the dispersion of each set of samples:
wherein, XiAnd YiIs the spatial location coordinate of each element,
and
is the arithmetic mean center;
step (6), uniformity evaluation of different perfuming modes
The differences are compared by the sizes of the elliptical areas, and the larger the elliptical area is, the poorer the perfuming uniformity of the perfuming mode is.
2. The uniformity evaluation method according to claim 1, characterized in that: in the step (2), randomly selecting an online raw material tobacco shred sample, wherein the sampling frequency is not less than 5 times, and the sampling amount is not less than 10g each time; accurately weighing 0.7g of each sample, placing the sample in a 20mL brown headspace bottle, adding 0.25ug/ul of deuterated toluene as an internal standard, screwing the bottle cap tightly, sealing, and measuring by an SPME-GC-MS method to obtain the solid phase microextraction spectrum of the on-line raw material cut tobacco.
3. The uniformity evaluation method according to claim 1, characterized in that: in the step (4), the sampling frequency of each perfuming mode is not less than 10 times, and the sampling amount of each time is not less than 10 g; accurately weighing 0.7g of each sample, placing the sample in a 20mL brown headspace bottle, adding 0.25ug/ul of deuterated toluene as an internal standard, screwing the bottle cap tightly, sealing, and measuring according to an SPME-GC-MS method to obtain solid phase microextraction spectra in different perfuming modes.
4. The uniformity evaluation method according to claim 1, characterized in that: in the step (5), the interpretation rate of the accumulated variance of the principal components obtained by PCA analysis is more than 85%.
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| CN111426714B (en) * | 2020-05-15 | 2023-03-31 | 云南中烟工业有限责任公司 | Cut tobacco perfuming uniformity detection method based on scanning electron microscope energy spectrum |
| CN114755318B (en) * | 2022-03-07 | 2024-05-03 | 浙江中烟工业有限责任公司 | Tracing analysis method for natural components in flavoring essence |
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