CN101071104B - Method for determining chloride content in tobacco - Google Patents
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Abstract
This invention discloses a method to measure chlorine content in the tobacco, providing a continuous flow analysis of the chlorine content in tobacco optimum conditions to obtain the most appropriate way to measure tobacco chlorine content. It is not only consistent with Mohr, Xinhua-titration, ion chromatography, and other classical methods of analysis' results with a high degree of consistency, but compared with the existing continuous flow analyzer test efficiency, samples testing cycle is shortened by nearly 40 percent. It is simple, fast, easy to operate, and has accurate test results and the determination of high credibility.
Description
Technical field
The present invention relates to a kind of analytical approach, especially relate to the Continuous Flow Analysis method of chlorinity in the tobacco.
Background technology
The quality of chlorinity and tobacco is closely related, is an important technology index during tobacco and goods overall quality thereof are estimated.Domestic in recent years many tobacco enterprises all adopt the Continuous Flow Analysis instrument to measure Chlorine in Tobacco.The ultimate principle of Continuous Flow Analysis method is to discharge thiocyanate radical by chlorine and mercuric thiocyanate reaction, and then develops the color with the ferric iron complexing, adopts photometer to measure again, and this is exactly a kind of spectrophotometry in fact.Concrete operation method is the method draw samples according to " GB/T5606.1-1996 " regulation, according to " YC/T31-1996 " preparation offal sample, does the water percentage experiment simultaneously.Take by weighing about 0.5g (being accurate to 0.1mg) offal, place 100mL5% (volume fraction) acetic acid, oscillation extraction 30min filters with qualitative filter paper then, and filtrate is measured with automatic analyzer.Automatic analyzer and detection method commonly used have: German AAII/AAIII, Dutch SKALAR, French ALLIANCE, U.S. API, YC/T162-2002 and GB13580.9-92, the principle of these instruments and method chlorine detection is basically identical all.Its concrete workflow can be with reference to figure 1---and flow analysis is surveyed the current methods of chlorinity in the tobacco in the Chinese tobacco industry standard.
The whole operation process comprise the extraction and the instrument spectrophotometric analysis all under the common air-conditioning environment of 20 ℃-30 ℃ of temperature, carry out.
But on the concrete scheme of instrument spectrophotometric analysis, but be subjected to all multifactor impacts, for the detection of chlorinity in the tobacco, Continuous Flow Analysis method and classical analysis the method for example testing result of Moire technique, potentiometric titration, the chromatography of ions etc. have than big-difference.
The applicant is by repeatedly testing and summing up, having drawn the Continuous Flow Analysis method, to adopt dialysis treatment, 37 ℃ of colour developings and 450-490nm to detect wavelength be the suitable condition that the Continuous Flow Analysis method is measured the tobacco chlorinity, write out relevant paper, according to further discovering, the result's of Continuous Flow Analysis method accuracy and agility still are subjected to other correlative factors and disturb.
The Continuous Flow Analysis technology has advantages such as easy, quick, easy to operate, if finding more, the assay method of system is applied in the Continuous Flow Analysis instrument, make the Continuous Flow Analysis technology can possess testing result advantage accurate, with a high credibility simultaneously, in tobacco business, will have widely and use, and certainly will become the method that the pertinent instruments producer follows.
Summary of the invention
The objective of the invention is deficiency, a kind of assay method of Chlorine in Tobacco is provided at the Continuous Flow Analysis technology.
Technical scheme of the present invention provides a kind of assay method of Chlorine in Tobacco, discharge thiocyanate radical by chlorine and mercuric thiocyanate reaction, and then develop the color with the ferric iron complexing, use spectrophotometry again, adopt dialysis treatment during spectrophotometry, 37 ℃ of colour developings, 450-490nm detects wavelength, select the 660-670 chromogenic reaction time of second, especially by adopting 485nm ± 4nm to detect wavelength, the adjustment of dialysis solvent, the flow velocity proportioning of developer and main flow solvent and diluted sample ratio, the combination of appropriate size colorimetric pool, thus acquisition is more accurate, measurement result fast.
Preferred detection wavelength of the present invention is 485nm.
Colorimetric pool of the present invention adopts not de-bubble of 10mm colorimetric pool, cooperate to adopt the 1M salpeter solution as dialysis solvent, water as main solvent, the flow velocity proportioning of developer and main flow solvent is 1.20 ml/min: 1.00 ml/min, it is 0.80 ml/min that dilution ratio adopts sample size and dialysis solvent flux proportioning: 0.32 ml/min.
The instrumental response value of not de-bubble of 10mm colorimetric pool and common 10mm colorimetric pool close, but be subjected to the influence of solute diffusional effect to want little many, thereby its cycle detection time is also the shortest, the present invention adopts not de-bubble of 10mm colorimetric pool.
But then, adopt the little 10mm colorimetric pool of optical path length, can cause the sensitivity of instrument to descend, influence the detection of the low tobacco sample of chloride ion content.Therefore, adopt the 1M salpeter solution as dialysis solvent, water as main solvent.And the flow velocity proportioning of regulating colour-developing solvent and main flow solvent is 1.20 ml/min: 1.00 ml/min, the throughput ratio of diluted sample ratio sample size and dialysis solvent is 0.32 ml/min: 0.80 ml/min.
The invention has the beneficial effects as follows and define the suitable condition that the Continuous Flow Analysis method is measured chlorinity in the tobacco, thereby draw the optimum method of measuring Chlorine in Tobacco, not only have consistance---RSD＜3%, the t＜t of height with the testing result of classical analysis methods such as Moire technique, potentiometric titration, the chromatography of ions
1-a=0.10/2(n-1), and with adjust before compare the sample detection cycle and shortened closely 40%, be a kind of easy, quick, easy to operate, assay method that testing result is accurate, with a high credibility.
Description of drawings
Fig. 1 China's current tobacco business standard flow assay chlorinity workflow reference diagram
The chromatic diagram of the standard solution after Fig. 2 embodiment of the invention 1 sample extraction liquid and the colour developing
The accompanying drawing of the absorbance of the different sample extraction liquid of Fig. 3 embodiment of the invention 1 table 1
The chromatic diagram of the sample extraction liquid of Fig. 4 embodiment of the invention 1 different disposal mode
The chromatic diagram of the sample extraction liquid after 2 dialysis of Fig. 5 embodiment of the invention are handled
The sample chromatic diagram of 3 different chromogenic reaction times of Fig. 6 embodiment of the invention
Absorbance spectrogram during the different light path of Fig. 7 embodiment of the invention 4
Embodiment
Fig. 8 assay method workflow of the present invention reference diagram
Further describe the present invention below in conjunction with concrete experimental data and embodiment.
The selection experiment that embodiment 1 dialysis is handled
1, instrument, sample and main agents Skalar SAN
++Type Continuous Flow Analysis instrument (Holland, SKALAR), Lambda850 type ultraviolet-visible spectrophotometer (U.S., PE), H110 type electronic balance (Germany, Sartorius, sensibility reciprocal: 0.1mg); Crosslinked aromatic polyamide composite semipermeable membrane (SKALAR-5283), polyethoxy bay ether (30%, Dutch Skalar), mercuric thiocyanate, methyl alcohol, ferric nitrate and nitric acid are analytical reagent.Standard tobacco sample GBW08514 and GBW08515 (Qingzhou Dohanykutato Intezet).
2, the processing of sample and analysis
According to the method draw samples of " GB/T5606.1-1996 " regulation,, do the water percentage experiment simultaneously according to " YC/T31-1996 " preparation offal sample.Take by weighing about 0.5g (being accurate to 0.1mg) offal, place 100mL5% (volume fraction) acetic acid, oscillation extraction 30min filters with qualitative filter paper then, and filtrate is measured with automatic analyzer.The whole operation process comprise the extraction and instrumental analysis all under 20 ℃-30 ℃ common air-conditioning environment, carry out.
3, experimental analysis:
Be to investigate dialysis and handle influence to testing result, experiment is carried out absorbance scanning to handling through dialysis with the tobacco sample extract of handling without dialysis, and the absorbance spectrogram after the chlorion colour developing relatively.
When 1. not adopting dialysis to handle, the background color of tobacco sample extract can cause serious interference to the chlorion testing result, and it is strong more to measure the short more undesired signal of wavelength, and chromatic diagram is seen Fig. 2.
Among Fig. 2, ordinate is an absorbance, and horizontal ordinate is a wavelength value; The chlorion standard solution curve of Standard2 after for colour developing, Sample1 is not for developing the color and without dialysis treatment sample extraction liquid.
2. under identical mensuration wavelength, the extract of different cultivars tobacco sample, it is strong and weak different that it produces undesired signal; And the dark more interference that produces of the color of extract is serious more, the results are shown in Table 1 and Fig. 3.
The absorbance of table 1 sample extraction liquid (480nm place)
Among Fig. 3, ordinate goes out peak heights numerical value for each sample, and horizontal ordinate is an appearance time, and unit is second.
3. the interference that can effectively eliminate tobacco sample extract background color is handled in dialysis, and chromatic diagram is seen Fig. 4.
Among Fig. 4, ordinate is an absorbance, and horizontal ordinate is a wavelength value; Sample1 is the sample extraction liquid curve of handling without dialysis, and Sample4 is the sample extraction liquid curve of handling through dialysis.
Experiment conclusion: the background color of tobacco sample extract can be disturbed the testing result of continuous flow method to chlorion, and this disturbing factor can be effectively eliminated in the dialysis processing, so, though the standard (YC/T162-2002 of domestic current, " the mensuration continuous flow method of tobacco and tobacco product chlorine " [S]) in recommend to use dialysis membrane, but adopted in the present invention.
The processing of instrument, sample and main agents and sample and analysis are with embodiment 1.
Experimental analysis: determining to need to adopt dialysis membrane to come the tobacco sample extract is carried out on the basis of pre-treatment, by further research to the absorbance spectrogram, reach the analysis of each being measured standard tobacco sample testing result under wavelength, with the suitable wavelength of determining that the tobacco sample chlorinity detects.
1. as seen from Figure 2, the maximum absorbance after the chlorion colour developing is near the 460nm wavelength.In the 450nm-490nm scope, absorbance is less with wavelength change, and measuring wavelength regulation 5nm influences Δ A/2＜3% to the sample absorbance; Outside this interval, occur measuring absorbance changes delta A/2＞3% of wavelength regulation 5nm than great fluctuation process with measuring the wavelength change absorbance.That is to say that the luminosity range of wavelengths of 450nm-490nm does not comprise endpoint value 450nm and 490nm, be the characteristic absorption wave band of chlorion colour developing.
2. the absorbance spectrogram of handling the tobacco sample extract through dialysis is amplified, see Fig. 5, can see that sample extraction liquid background color does not produce the interference absorbance substantially when mensuration wavelength 〉=489nm.Be reduced to the 489nm-480nm place when measuring wavelength, then begin to occur faint interference absorbance; Detect wavelength≤480nm, disturb the amplification of absorbance to begin to rise, and increasing; Again move towards relaxation near the 400nm, and reach the maximum interference absorbance.Judge thus, for the tobacco sample extract, even still have bigger interference luminosity to absorb when≤480nm the wavelength because it handles the back through dialysis.So the suitable luminosity range of wavelengths that detects for the tobacco sample chlorion should be 480nm-490nm, does not comprise two end points of 480nm and 490nm.
Among Fig. 5, ordinate is an absorbance, and horizontal ordinate is a wavelength value, and wavelength value is 488.99nm in the horizontal ordinate square frame.Curve S ample4 is through the dialysis processing but the sample extraction liquid curve that does not develop the color as yet,
3. the testing result by the standard tobacco sample sees Table 2, can see, when measuring wavelength and be 481nm, 485nm, 489nm and 490nm, each test result of samples is all in calibration range.All the other are measured under the wavelength, and the testing result of part round exceeds calibration value, the sample of especially high chlorinity, and it is obvious to depart from situation.
Different measured value (the units: %) that measure tobacco standard sample under the wavelength of table 2
Experiment conclusion: though 460nm is the maximum absorption wavelength of chlorion in the thiocyanate radical development process, also be for example GB13580.9-92 and HJ/T27-1999 of most standard, the detection wavelength that suggestion is adopted, but near this luminosity wavelength, exist than strong jamming for tobacco sample, even the original 480nm wavelength place that adopts of tobacco examination criteria (YC/T162-2002), still have certain interference and exist, thereby all be not suitable for adopting.Comprehensively 1. 2. 3. point analysis can be judged 481nm to 489nm, comprises 481nm and 489nm, is to carry out the suitable mensuration wavelength of tobacco sample chlorinity Continuous Flow Analysis method and selected 485nm is optimal selection of the present invention.
The experiment of 3 chromogenic reaction times of embodiment
The processing of instrument, sample and main agents and sample and analysis are with embodiment 1.
Experimental analysis: we regulate chromogenic reaction time of sample by the mode that increases spiral pipe, and the sample chromatic diagram is seen Fig. 6.
1. the reaction time variant can influence the absorbance that develops the color of sample, and with 664 seconds, i.e. absorbance maximum under the spiral pipe 10 circle conditions.
2. after the chromogenic reaction time was increased to 660 seconds, the variation that prolongs absorbance with the reaction time was very little, and absorbance remains unchanged substantially near detecting wavelength 485nm.
Curve among Fig. 6 from top to bottom is the chromatic diagram of Sample3-10 circle spiral pipe, Sample5-20 circle spiral pipe, Sample4-0 circle spiral pipe, Sample1-5 circle spiral pipe successively, and developing time was respectively 664 seconds, 701 seconds, 630 seconds, 648 seconds.
Experiment conclusion: guarantee that chromogenic reaction time 660-670 can guarantee the stability of testing result second, and obtain higher instrumental response value.The term harmonization of this and most of chlorinity color developing detection is not subjected to detecting the too big influence that wavelength is adjusted.
The selection experiment of embodiment 4 colorimetric pools
The processing of instrument, sample and main agents and sample and analysis are with embodiment 1
Experimental analysis: the size of the response signal by more different light path colorimetric pools, and the size of the examined cycle influence of testing result when relatively adopting dissimilar colorimetric pool, understand the effect of colorimetric pool.
Ordinate is the peak height value among Fig. 7, and horizontal ordinate is the time, and unit is second.Be followed successively by the absorption curve of 50mm, 30mm, 10mm colorimetric pool from top to down.
1. as seen from Figure 7, along with the colorimetric pool light path is the increasing of optical path length, instrumental response value is also corresponding can be strengthened, and the ratio of response approaches the ratio of optical path length.Simultaneously, the optical path length of colorimetric pool is big more, and instrument is subjected to the influence of solute diffusional effect also obvious more---and it is the most sharp-pointed that the response of 50mm colorimetric pool goes out the peak, lacks to walk flat process, thereby influence the testing result accuracy; And very long one section flat peak has appearred in the 10mm colorimetric pool, shows that instrumental response value is stable, thus powerful guarantee the accuracy and the stability of testing result; The 30mm colorimetric pool is then between between the above two, and the instrument detecting value is basicly stable.
2. can judge from the situation of table 3 that for the 30mm colorimetric pool, it should be that sample injection time and sampling interval time are 80s+80s that its suitable sample detection time cycle is provided with; For the 10mm colorimetric pool, suitable setting then is 70s+70s; And the instrumental response value of not de-bubble of 10mm colorimetric pool and common 10mm colorimetric pool is close, but is subjected to the influence of solute diffusional effect to want little many, thereby its cycle detection time is also the shortest, and it is set to 60s+30s.
The influence of table 3 colorimetric pool
Remarks: 1, expression signal peak top lacks plateau, and reaction does not reach static balancing.Zero expression reaction has reached static balancing.
2, ◆ the hangover of expression signal peak, back one test result of samples is subjected to the interference of previous sample.◇ represents not disturb.
Experiment conclusion:
1. adopt not de-bubble of 10mm colorimetric pool can effectively reduce the influence of solute diffusional effect, significantly improve the efficient of instrument detecting.
2. the instrumental response value of not de-bubble of 10mm colorimetric pool is very low, therefore must manage to improve the signal response of instrument, promptly selects the dialysis solvent, adjusts dilution ratio.
Embodiment 5 dialysis solvents are selected experiment
The processing of instrument, sample and main agents and sample and analysis are with embodiment 1
Experimental analysis: chlorinity is that the solvent of using has: the solvent of water---all reagent in detecting, the extract of 5% acetum---sample and the washing fluid of instrument, wherein a kind of solvent of 1M salpeter solution---developer, the another kind of solvent of methanol solution---developer.Can cause when using because of methanol solution has bubble to produce in the pipeline, be unfavorable for detecting and carry out, so do not consider in this experiment.In the experiment at first with water as main solvent, the check different solutions the dialysis result; Behind definite dialysis solvent, the situation when attempting using different solvents respectively again as main solvent.
The analysis of table 4 pair dialysis solution
Note: every group of test with the mean value of three testing results as making a report on data.
Highlight with boldface letter for the data that do not meet relevant requirements.
The analysis of table 5 pair main flow solution
Note: every group of test with the mean value of three testing results as making a report on data.
Highlight with boldface letter for the data that do not meet relevant requirements.
1. can judge by the data of table 4 that when selecting water as the dialysis solvent, instrumental response value is obviously on the low side; And when selecting 5% acetum, the undulatory property of testing result is bigger; Only with salpeter solution during as the dialysis solvent, the response of instrument goes out the peak value maximum, and testing result is also very stable.
2. can judge by the data of table 5 that when selecting 5% acetum as main solvent, instrumental response value is obviously on the low side; And the instrumental response value of water is the highest, and it also is the simplest solvent.
Experiment conclusion: owing to will be that purpose and guarantee have certain stability to obtain the highest sensitivity of instrument, so adopt the 1M salpeter solution as dialysis solvent, water during as main solvent with this understanding, the effect that dialysis is handled be the most desirable, and instrumental response value is the highest.
Embodiment 6 dilution ratio adjustment experiment
The processing of instrument, sample and main agents and sample and analysis are with embodiment 1
Experimental analysis: constant in order to guarantee in the whole testing process chromogenic reaction time, control solvent total flow (=colour-developing solvent flow+main solvent flow) is that 2.20 ml/min are constant in the test.Test is regulated the dilution of sample ratio by regulating sample size and dialysis solvent flux, then relatively detect the typical curve facies relationship numerical value that is obtained, wherein, concentration of standard solution coverage 0.142%-1.424%, effectively include the possible chloride ion content of tobacco sample, go out peak value with sample response, with chloride ion content be 0.641% standard solution as sample, and the spectrogram situation of analytical standard curve.Experimental result sees Table 6.
The analysis of table 6 pair flow design
Note: the numerical value of goal condition represented to meet in boldface letter.
Analyze according to table 6, can obtain to draw a conclusion:
1. be that the facies relationship numerical value of typical curve can reach more than 0.999 in certain diluted sample proportional range.
2. (employed colour-developing solvent has been a saturated solution in the test to improve the colour-developing solvent consumption, therefore have only by improving flow and realize this purpose), the response that can effectively improve sample goes out peak value (peak height＞500bit), and make mark Qu Buhui be parabolic shape than low dilution ratio when example, facies relationship numerical value thereby can reach more than 0.999.
Experiment conclusion: under the condition of colour-developing solvent flow 1.20 milliliters/second, main solvent flow 1.00CC/M, dialysis solvent flux 0.80CC/M, sample size 0.32CC/M, analyser can obtain higher response, and to the typical curve of effective covering tobacco sample chlorine content ranges, its facies relationship numerical value can reach more than 0.999.
Embodiment 7 the inventive method measurement result accuracy confirmatory experiments
Experimental analysis: by standard addition method survey time yield, promptly adopt the sample of known content, adopt standard offal sample GBW08514 and GBW08515 in the present embodiment, the standard solution of the chloride ion-containing by adding certain volume detects then.With detected value and calculated value relatively, thus obtain the recovery of standard addition of method.Detect according to assay method of the present invention, concrete workflow can obtain detected value with reference to figure 8, and calculate recovery rate the results are shown in Table 7.
Table 7 recovery test
Note: every group of test carried out 3 rounds and detected, and with in and with median as making a report on data.
More same sample, in commensurate not, result's accuracy is verified the accuracy of detection scheme when detecting with distinct methods.Experimental result sees Table 8,9.
Table 8 contrast test
Note: 1, the detection method of Qingzhou tobacco research institute employing is the chromatography of ions, spectrophotometric method and Moire technique (silver nitrate volumetric method).
2, new detection method is an automatic potentiometric titration in the analytical test of Guangzhou.
3, analyser detects mean value with three measurement results for making a report on data.
Table 9 back-to-back test
Note: 1, analyser detects mean value with three measurement results for making a report on data.
2, new detection method is an automatic potentiometric titration in the analytical test of Guangzhou.
1. the recovery of detection scheme of the present invention shows that the measurement result of this programme is more accurate in the 101.7%-104.6% scope.
2. by this programme sample is detected resultant data, authorize quality inspection organization very approaching, show that the detection data of this scheme are comparatively credible, the accuracy height with result that different detection method is obtained with country.
2. use back-to-back test hypothesis test (
, d
1=x
i-y
1) test data is done further to analyze, obtaining the t value is 1.6278, less than t
1-a/2(n-1)=1.7341 (a=0.10, n=19).Both on the level of significance of a=0.10, assay method of the present invention and classical automatic potentiometric titration, its testing result difference is less.
Experiment conclusion: the assay method of chlorinity and existing Continuous Flow Analysis method are relatively in the tobacco of the present invention, determined to guarantee that the Continuous Flow Analysis method measures in the tobacco chlorinity optimal conditions of accuracy as a result, having increased dialysis processing, 37 ℃ of colour developings, having measured wavelength is on the basis of 485nm ± 4nm, the clear and definite sample chromogenic reaction time is near 670 seconds, and not de-bubble of employing 10mm colorimetric pool, with be provided with suitable dialysis solvent and dilution ratio, thereby sense cycle was shortened to 90 seconds by original 160 seconds, improved efficient greatly; Other detection scheme testing results such as the inventive method and classical detection method, the chromatography of ions are more close, reached accurate, stable, quick, the with a high credibility requirement of data.
Claims (2)
1. the assay method of a Chlorine in Tobacco, discharge thiocyanate radical by chlorine and mercuric thiocyanate reaction, and then with developer in ferric iron complexing colour developing, use spectrophotometry again, adopt the detection wavelength of dialysis treatment, 37 ℃ of colour developings, OK range during spectrophotometry and select 660～670 seconds chromogenic reaction time, it is characterized in that:
Adopt 485nm ± 4nm to detect wavelength;
The dialysis solvent adopts the 1M salpeter solution, and water is as main solvent;
The flow velocity proportioning of developer and main solvent is 1.20 ml/min: 1.00 ml/min;
The diluted sample ratio is that sample size and dialysis solvent flow rate proportioning are 0.32 ml/min: 0.80 ml/min;
Colorimetric pool is not de-bubble of a 10mm colorimetric pool.
2. the assay method of Chlorine in Tobacco according to claim 1 is characterized in that described detection wavelength is 485nm.
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| CN101285831B (en) * | 2008-05-22 | 2012-09-05 | 重庆烟草工业有限责任公司 | Flowing analysis technique for determining tobacco cellulose |
| CN104034678B (en) * | 2014-06-09 | 2016-04-27 | 华能国际电力股份有限公司 | Reagent formula for measuring trace chloride ions in water |
| CN108508223A (en) * | 2018-02-12 | 2018-09-07 | 云南中烟工业有限责任公司 | A kind of high-throughput Continuous Flow Analysis method for chlorinity in tobacco gene editor's material |
| CN110646414A (en) * | 2019-09-27 | 2020-01-03 | 常州罗盘星检测科技有限公司 | Reagent for rapidly detecting concentration of chloride ions in water and use method |
| CN112525895A (en) * | 2020-11-20 | 2021-03-19 | 云南省烟草质量监督检测站 | Continuous flow method for measuring content of calcium carbonate in cigarette paper |
| CN112326576A (en) * | 2020-11-20 | 2021-02-05 | 云南省烟草质量监督检测站 | Continuous flow method for determining content of calcium carbonate in paper-making reconstituted tobacco |
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