CN103235047B - Method for determining fluorescent whitening agent migration in cigarette paper - Google Patents
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Abstract
The invention relates to the technical field of cigarette material detection, and specifically discloses a method for determining fluorescent whitening agent migration in cigarette paper. The method comprises the following steps: S1, a fluorescent whitening agent migration test: clipping cigarette paper and filter paper, accurately weighing, soaking the cigarette paper with a migration solution, covering the cigarette paper with the filter paper, completely clamping the filter paper covering the cigarette paper, flattening, and standing; and S2, quantitative determination: taking the filter paper, placing into a solvent to extract, carrying out centrifugation on the extracted solution, filtering the supernatant with a filtration membrane, and adopting a high performance liquid chromatography method to determine a fluorescent whitening agent content. The invention provides a quantitative determination method for fluorescent whitening agent migration in cigarette paper, wherein the method has characteristics of accurate testing and convenient operation, and migration analysis on the fluorescent whitening agent migration in the cigarette paper is conveniently performed so as to reduce harm of the fluorescent whitening agent on consumer body.
Description
Technical field
The present invention relates to smoking material detection technique field, more specifically, relate to a kind of method of measuring fluorescer migration in Cigarette paper.
Background technology
It is yellow to canescence that paper pulp fiber is always, even through general bleaching, still can not eliminate this micro-yellow hue.In paper pulp, add after fluorescer, whitening agent absorbs ultraviolet light and sends blue-fluorescence, according to optics complementary principle, makes paper pulp become pure white, makes paper pulp produce brighter, more gorgeous effect simultaneously.Although fluorescer is not yet found the obvious harm of human body, but fluorescer has harm to a certain degree to environment, and very easily absorbed by the mouth and nose mucous membrane tissue of human body, be not easy to be decomposed very much once enter human body, and can make cell produce variation, become potential carcinogenic factor.For Cigarette paper, the most influential principal element of health is not lain in to fluorescent brightener levels, and be whether paper can contact with human body skin the animal migration of rear fluorescer.
The standard (EN648:1993) of " with the paper of Food Contact and the mensuration of cardboard-fluorescer fastness " has been formulated in Europe in 1993, is mainly used in packaging for foodstuff paper using.The method has been simulated the process of food wrapper and Food Contact by uviol lamp observational technique, be the method for measuring fluorescer and food wrapper binding strength.Japan has formulated the bioassay standard (JIS S3104-1992) of fluorescer animal migration in " face tissue " for 1992.Whether under uviol lamp, detect equally, observing migration gauze has fluorescence reaction to determine that the fluorescer in sample has or not animal migration.Practical measurement be the fastness that also fluorescer is combined with paper.In these two kinds of method of testings, test process more complicated, the test duration is long, more than generally needing 12h, and observes with under uviol lamp, and human factor is more, and the accurate evaluation of testing result is had a certain impact.And the current detection method that also there is no fluorescer migration in Cigarette paper.
Summary of the invention
Technical matters to be solved by this invention is, in order to overcome the deficiency of Cigarette paper fluorescer migration detection technique in prior art, to provide a kind of method of measuring fluorescer migration in Cigarette paper.
Above-mentioned technical matters of the present invention is achieved by the following technical programs:
A method of measuring fluorescer migration in Cigarette paper, comprises the steps:
S1. fluorescer migration test: clip Cigarette paper, filter paper, accurately weighs, migration immersion profit for Cigarette paper, then with filter paper, Cigarette paper is coated to (upper and lower surface of Cigarette paper is all coated with filter paper), then the filter paper that is coated with Cigarette paper is all clamped and flattened, leave standstill and place;
S2. quantitative measurement: take out filter paper, put into solvent and extract, then by the solution centrifugal after extraction, supernatant membrane filtration, with the content of high effective liquid chromatography for measuring fluorescer;
Wherein high-efficient liquid phase chromatogram condition is: chromatographic column is selected carbon 18 posts; Mobile phase A is methyl alcohol, and Mobile phase B is 4.5～5.5mmol/L n-octyl amine aqueous solution; The volume ratio of mobile phase A and B is 40～50:60～50; Flow velocity 0.6～1.5ml/min; Column temperature is 20～30 DEG C; Maximum excitation wavelength is 330～360nm; Emission wavelength is 400～450nm, sample size 1～15 μ l.
This method, tests fluorescer migration method for quantitatively determining in Cigarette paper accurate, easy and simple to handle, conveniently fluorescer in Cigarette paper is carried out to migration analysis.
As a kind of preferred version, the filter paper by being coated with Cigarette paper described in S1 is all clamped and flattens, and is the two blocks of glass of filter paper that are coated with Cigarette paper are all clamped and flattened.
The area of described Cigarette paper is 3～6cm × 3～6cm.
As a kind of preferred version, the area of described Cigarette paper is 5cm × 5cm.
The size of described filter paper is 3～6cm × 6～15cm.
As a kind of preferred version, the size of described filter paper is 5cm × 10cm.
The size of described filter paper can just be coated Cigarette paper.
As a kind of preferred version, described filter paper size can envelope Cigarette paper during for doubling.
As a kind of preferred version, two glass plates of filter paper that are coated with Cigarette paper are all clamped and flattened, leave standstill and place.
As a kind of preferred version, described filter paper is qualitative filter paper.
As a kind of preferred version, described migration liquid is pure water, saturated salt solution or organic solvent.
As the further preferred version of one, described saturated salt solution is saturated NaHCO
3solution, described organic solvent is 40～70(volume) ethanolic solution of %.
As most preferably scheme of one, described organic solvent is 50(volume) ethanolic solution of %.
As a kind of preferred version, the consumption of migration liquid is 1～4ml.
As the further preferred version of one, the consumption of migration liquid is 2～3ml.
As most preferably scheme of one, the consumption of migration liquid is 1.5ml.
As a kind of preferred version, described standing placement refers to leave standstill places 1～4h.
As the further preferred version of one, described standing placement refers to leave standstill under the condition of lucifuge places 1～4h.
As most preferably scheme of one, described standing placement refers in temperature at 0～40 DEG C, with leaving standstill placement 2～3h under the condition of tinfoil lucifuge.
The fluorescer of 15 μ g/ml (APC) standard solution is divided into three parts, is placed in respectively not lucifuge of normal temperature, normal temperature tinfoil lucifuge, and 2h under environment in 4 DEG C of refrigerators; After 2h, these three parts of APC standard solution and the APC standard solution of new preparation are together carried out to HPLC detection, observe the impact of each condition on fluorescer stability; Measure spectrogram result as shown in Fig. 1～4; From Fig. 1～4, illumination is very large on the impact of fluorescer, and the speed that under illumination, fluorescer decomposes is very fast, and temperature does not affect substantially on the stable of fluorescer; Consider the ease for operation of experiment, suggestion selects the condition of tinfoil lucifuge under room temperature as the experiment condition of pattern migration.
As a kind of preferred version, the extraction described in S2 is lucifuge ultrasonic extraction, described ultrasonic time 0.5～1h.
As most preferably scheme of one, described ultrasonic time 0.5h.
As a kind of preferred version, the solvent described in S2 is DMSO; The consumption of DMSO is 8～12ml; In S2, the speed of centrifugal use is 5000～10000r/min.
As most preferably scheme of one, the consumption of DMSO is 10ml; In S2, the speed of centrifugal use is 7000r/min.
As a kind of preferred version, the above-mentioned centrifugal time is 5～10min.
As a kind of preferred version, wherein high-efficient liquid phase chromatogram condition is: chromatographic column is selected carbon 18 posts; Described high-efficient liquid phase chromatogram condition is: chromatographic column is selected Hypersil BDS C18; Mobile phase A is methyl alcohol, and Mobile phase B is 5mmol/L n-octyl amine aqueous solution; The volume ratio of mobile phase A and B is 45:55; Flow velocity 0.6～1.0ml/min; Column temperature is 20～30 DEG C.
As the further preferred version of one, wherein high-efficient liquid phase chromatogram condition is: chromatographic column is selected carbon 18 posts; High-efficient liquid phase chromatogram condition described in mobile phase is: chromatographic column is selected Hypersil BDS C18; Mobile phase A is methyl alcohol, and Mobile phase B is 5mmol/L n-octyl amine aqueous solution; The volume ratio of mobile phase A and B is 45:55; Flow velocity 0.6～1.0ml/min; Column temperature is 20～30 DEG C; Maximum excitation wavelength is 330～360nm; Emission wavelength is 400～450nm, sample size 1～5 μ l.
As most preferably scheme of one, described high-efficient liquid phase chromatogram condition is: chromatographic column is selected Hypersil BDS C18; High-efficient liquid phase chromatogram condition described in mobile phase is: chromatographic column is selected Hypersil BDS C18; Mobile phase A is methyl alcohol, and Mobile phase B is 5mmol/L n-octyl amine aqueous solution; The volume ratio of mobile phase A and B is 45:55; Flow velocity 0.8ml/min; Column temperature is 25 DEG C; Maximum excitation wavelength is 350nm; Emission wavelength is 425nm, sample size 2 μ l.
Cigarette paper of the present invention refers to the body paper of cigarette tipping paper, internal lining paper, seal paper, bar and box packaging paper, frame paper or above-mentioned paper.
Compared with prior art, the present invention has following beneficial effect: (1) provides a kind of fluorescer migration method for quantitatively determining in Cigarette paper accurate, easy and simple to handle of testing, conveniently fluorescer in Cigarette paper is carried out to migration analysis, to reduce the fluorescer harm healthy to consumer; (2) method of migration experiment of the present invention, simulate the contact process of Cigarette paper and human body, improve the mobility of fluorescer, especially when using 50% ethanol as migration liquid, can obviously improve the mobility of fluorescer, be conducive to judge whether the fluorescer migration in Cigarette paper is qualified.
Brief description of the drawings
Fig. 1 is the not impact of lucifuge condition on fluorescer stability of normal temperature.
Fig. 2 is the impact of normal temperature tinfoil lucifuge condition on fluorescer stability.
Fig. 3 is the impacts of 4 DEG C of static placement conditions of refrigerator on fluorescer stability.
Fig. 4 is without the impact on fluorescer stability under any treatment conditions.
Embodiment
Further explain the present invention below in conjunction with specific embodiment, but embodiment does not limit in any form to the present invention.
Embodiment 1
S1. migration test:
Clip area is 5 of 5 of the patterns of 5cm × 5cm, quantitative filter papers that area is 5cm × 10cm, all accurately weighs and numbers; Outturn infiltrates with pure water 1.5ml, pattern reference numeral is coated with the quantitative filter paper of doubling; Every suit paper is all clamped and is flattened with two glass plates, leaves standstill and place 2h under tinfoil lucifuge condition;
S1. quantitative measurement
By the pattern taking-up being clipped in the middle, 5 quantitative filter papers are placed in respectively to 20ml tool plug test tube with the tweezers of wash clean, add respectively 10mlDMSO; Lucifuge ultrasonic extraction 0.5h, gets turn/min of solution high speed centrifugation 7000,5min, and clear liquid is used the membrane filtration of 0.45 μ m again, gets supernatant and carries out HPLC analysis; Replicate determination five times; Utilize typical curve equation y=84.066x+11.899, R2=0.9995, calculates the concentration of each pattern migration fluorescer;
Described chromatographic condition is: chromatographic column is Hypersil BDS C18, and mobile phase A is methyl alcohol, and Mobile phase B is 5mmol/L n-octyl amine aqueous solution; The volume ratio of mobile phase A and B is 45:55, flow velocity 0.8ml/min, room temperature, maximum excitation wavelength 350nm, emission wavelength 425nm, sample size 2 μ l.
Result is calculated:
Data analysis is calculated: the content of fluorescer
In formula: fluorescer mobility in X-sample, unit is %;
The concentration of the fluorescer that C-calculates from working curve, unit is every milliliter of microgram (μ g/ml);
The cumulative volume of V-extract and migration liquid, unit is milliliter (mL).
M-sample mass, unit is gram (g);
Final fluorescer detectable concentration is 1.024 μ g/ml, is 1.22% by calculating certain box packaging paper body paper fluorescer mobility in pure water.The relative standard deviation of 5 measurement results is 3.8%.
Embodiment 2
S1. migration test:
Clip area is 3 of 3 of the patterns of 5cm × 5cm, quantitative filter papers that area is 5cm × 10cm, all accurately weighs and numbers; Outturn is used respectively pure water 1.5ml water, the saturated NaHCO of 1.5ml
3the ethanolic solution of solution, 1.5ml50% infiltrates, and pattern reference numeral is coated with the quantitative filter paper of doubling.Every suit paper is all clamped and is flattened with two glass plates, leaves standstill and place 2h under normal temperature tinfoil lucifuge condition;
S2. the method for quantitative measurement is identical with S2 in embodiment 1.
The chromatographic peak area of target compound will be recorded, the typical curve that substitution is drawn, obtain fluorescer in sample concentration, computing method are with the computing method of embodiment 1, in different migration solution, the mobility result of calculation of paper fluorescer is listed in table 1.As can be seen from Table 1, for water and saturated NaHCO
3solution is as migration solution, although there is part fluorescer to move on filter paper, mobility ratio is lower, even at 50(volume) in the migration system of the ethanol of %, the mobility of fluorescer is 15.8%, apparently higher than the mobility of other migration liquid.Fluorescer is the extremely unsettled material of one, and it has more shown the character of easy migration in ethanolic solution, illustrating when using 50(volume) ethanol of % can significantly improve the mobility of fluorescer during as migration liquid, is conducive to judge whether the fluorescer in Cigarette paper moves.
The mobility of paper fluorescer in the different migration of table 1 solution
Embodiment 2
S1. migration test:
Clip area is 3 of 3 of the patterns of 5cm × 5cm, quantitative filter papers that area is 5cm × 10cm, all accurately weighs and numbers; Outturn 70(volume) % ethanol 1.5ml infiltrates, pattern reference numeral is coated with the quantitative filter paper of doubling; Every suit paper is all clamped and is flattened with two glass plates, leaves standstill and place 2h under 20 DEG C of tinfoil lucifuge conditions;
S1. quantitative measurement
By the pattern taking-up being clipped in the middle, 3 quantitative filter papers are placed in respectively to 20ml tool plug test tube with the tweezers of wash clean, add respectively 15mlDMSO; Lucifuge ultrasonic extraction 0.5h, gets turn/min of solution high speed centrifugation 10000,5min, and clear liquid is used the membrane filtration of 0.45 μ m again, gets supernatant and carries out HPLC analysis; Replicate determination three times; Utilize typical curve equation to calculate the concentration of each pattern migration fluorescer;
Described chromatographic condition is: chromatographic column: Hypersil BDS C18, and mobile phase A is methyl alcohol, Mobile phase B is 5mmol/L n-octyl amine aqueous solution; The volume ratio of mobile phase A and B is 45:55, flow velocity 1.2ml/min, 25 DEG C, maximum excitation wavelength 350nm, emission wavelength 425nm, sample size 4 μ l.
Final fluorescer detectable concentration is 12.87 μ g/ml, by calculating certain box packaging paper body paper at 70(volume) in % ethanol fluorescer mobility be that 12.94%(computing method are with embodiment 1).
Embodiment 4
S1. migration test:
Clip area is 3 of 3 of the patterns of 5cm × 5cm, quantitative filter papers that area is 5cm × 10cm, all accurately weighs and numbers; Outturn 40(volume) % ethanol 3ml infiltrates, pattern reference numeral is coated with the quantitative filter paper of doubling; Every suit paper is all clamped and is flattened with two glass plates, leaves standstill and place 2h under 20 DEG C of tinfoil lucifuge conditions;
S1. quantitative measurement
By the pattern taking-up being clipped in the middle, 3 quantitative filter papers are placed in respectively to 20ml tool plug test tube with the tweezers of wash clean, add respectively 5mlDMSO; Lucifuge ultrasonic extraction 0.5h, gets turn/min of solution high speed centrifugation 5000,5min, and clear liquid is used the membrane filtration of 0.45 μ m again, gets supernatant and carries out HPLC analysis; Replicate determination three times; Utilize typical curve equation to calculate the concentration of each pattern migration fluorescer;
Described chromatographic condition is: chromatographic column: Hypersil BDS C18, and mobile phase A is methyl alcohol, Mobile phase B is 5mmol/L n-octyl amine aqueous solution; The volume ratio of mobile phase A and B is 45:55, flow velocity 1.2ml/min, 25 DEG C, maximum excitation wavelength 350nm, emission wavelength 425nm, sample size 1 μ l.
Final fluorescer detectable concentration is 9.14 μ g/ml, by calculating certain box packaging paper body paper at 40(volume) in % ethanol fluorescer mobility be that 8.89%(computing method are with embodiment 1).
Claims (7)
1. a method of measuring fluorescer migration in Cigarette paper, is characterized in that, comprises the steps:
S1. fluorescer migration test: clip Cigarette paper, filter paper, accurately weighs, and migration immersion profit for Cigarette paper is then coated by Cigarette paper with filter paper, then the filter paper that is coated with Cigarette paper is all clamped and flattened, and leaves standstill and places;
S2. quantitative measurement: take out filter paper, put into solvent and extract, then by the solution centrifugal after extraction, supernatant membrane filtration, with the content of high effective liquid chromatography for measuring fluorescer; Wherein high-efficient liquid phase chromatogram condition is: chromatographic column is selected carbon 18 posts; Mobile phase A is methyl alcohol, and Mobile phase B is 4.5～5.5mmol/L n-octyl amine aqueous solution; The volume ratio of mobile phase A and B is 40～50:60～50, flow velocity 0.6～1.5ml/min; Column temperature is 20～30 DEG C, and maximum excitation wavelength is 330～360nm; Emission wavelength is 400～450nm, sample size 1～5 μ l,
The ethanolic solution that described migration liquid is 50%.
2. method according to claim 1, is characterized in that, described filter paper is qualitative filter paper.
3. method according to claim 1, is characterized in that, described standing placement refers to leave standstill under the condition of lucifuge places 1～4h.
4. method according to claim 1, is characterized in that, the solvent described in S2 is DMSO; The consumption of DMSO is 8～12ml; In S2, the speed of centrifugal use is 5000～10000r/min.
5. method according to claim 1, is characterized in that, chromatographic column is selected carbon 18 posts; Mobile phase A is methyl alcohol, and Mobile phase B is 5mmol/L n-octyl amine aqueous solution; The volume ratio of mobile phase A and B is 45:55; Flow velocity 0.6～1.0ml/min; Column temperature is 20～30 DEG C.
6. method according to claim 5, is characterized in that, described high-efficient liquid phase chromatogram condition is: chromatographic column is selected Hypersil BDS C18; Mobile phase A is methyl alcohol, and Mobile phase B is 5mmol/L n-octyl amine aqueous solution; The volume ratio of mobile phase A and B is 45:55; Flow velocity 0.8/min; Column temperature is 25 DEG C; Maximum excitation wavelength is 350nm; Emission wavelength is 425nm, sample size 2 μ l.
7. method according to claim 1, is characterized in that, described Cigarette paper refers to the body paper of cigarette tipping paper, internal lining paper, seal paper, bar and box packaging paper or above-mentioned paper.
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| CN107084936A (en) * | 2017-04-24 | 2017-08-22 | 沃奇中 | The assay method of transportable property fluorescent whitening agent in toilet paper |
| CN113418816B (en) * | 2021-05-21 | 2023-01-03 | 江苏中烟工业有限责任公司 | Method for measuring inter-paper mobility of saccharin sodium in tipping paper under storage state |
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