CN105372371A - Ion chromatography method for simultaneous determination of oxalic ion, succinate ion, sulfate ion and phosphate ion in reconstituted tobacco and application thereof - Google Patents
Ion chromatography method for simultaneous determination of oxalic ion, succinate ion, sulfate ion and phosphate ion in reconstituted tobacco and application thereof Download PDFInfo
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- CN105372371A CN105372371A CN201510870988.1A CN201510870988A CN105372371A CN 105372371 A CN105372371 A CN 105372371A CN 201510870988 A CN201510870988 A CN 201510870988A CN 105372371 A CN105372371 A CN 105372371A
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Abstract
The invention discloses an ion chromatography method for simultaneous determination of oxalic ion, succinate ion, sulfate ion and phosphate ion in reconstituted tobacco The method established in the invention uses methanesulfonic acid as an extraction liquid to guarantee the extraction efficiency of each group to be tested in a sample reach above 99.7%; a mixed solution of sodium carbonate and sodium bicarbonate is used as a mobile phase, in order to effectively eliminate the effects of degradation of vitamin C on the quantification of oxalic acid; sample treatment and sampling at a time can simultaneously detect four components including oxalic ion, succinate ion, sulfate ion and phosphate ion in reconstituted tobacco; and the relative standard deviation of content of each group to be tested is 0.27%-0.99%, the adding standard recovery is 99.3%-106.2%. The method is accurate, simple and reliable, and is applicable to the determination of relative groups in tobacco samples and reconstituted tobacco samples.
Description
Technical field
The detection technique field of reconstituted tobacco of the present invention, more specifically, relates to chromatography of ions detection method and the application of a kind of Simultaneously test reconstituted tobacco mesoxalic acid, succinic acid, sulfuric acid and phosphate radical.
Background technology
Papermaking-method reconstituted tobaccos has balance tobacco structure because of it, stablizes cigarette quality, effectively reduces the advantage of tar and part objectionable constituent burst size and be subject to increasing utilization.In papermaking-method reconstituted tobaccos process, need to add various auxiliary agent, wherein succinic acid (succinic acid) and oxalic acid are added into as extraction auxiliary agent as process assistant, phosphoric acid.Based on reconstituted tobacco constant product quality and security consideration, the addition of each analog assistant will strictly control, and therefore needs to set up a set of accurate and effective method of testing and tests the auxiliary agent in reconstituted tobacco is residual.
It is good, highly sensitive that the chromatography of ions (IC) has selectivity, the simple feature of pre-treatment and being used widely.In tobacco and goods thereof, negative ion, kation and organic acid measure and adopt the chromatography of ions more, but there are no reporting the method for Simultaneously test reconstituted tobacco mesoxalic acid, succinic acid, sulfate radical and phosphate content.A lot of ion-chromatographic determination is had to comprise tobacco containing the plant of vitamin C (ascorbic acid) and food mesoxalic acid content method at present, but the impact that vitamin C measures oxalic acid all do not considered by above document, in biological fluid medical science detects, only consider the impact that vitamin C measures oxalic acid.The method of current mensuration tobacco mesoxalic acid all also exists two problems: one is extract the incomplete of oxalic acid using pure water as extract; Two is use the stronger NaOH of alkalescence to make leacheate, causes vitamin C in tobacco (ascorbic acid) be converted into oxalic acid and make measurement result higher.
Summary of the invention
The technical problem to be solved in the present invention is, for the deficiency of existing reconstituted tobacco mesoxalic acid, succinic acid, sulfuric acid and phosphoric acid detection technique, establish the ion chromatographic method of a kind of energy Simultaneously test reconstituted tobacco oxalate, amber acid radical, sulfate radical and phosphate content, be intended to provide technical basis to the control of chemical substance addition in reconstituted tobacco process.
Another technical matters that the present invention will solve is to provide the application of described detection method.
Goal of the invention of the present invention is achieved by the following technical programs:
The ion chromatographic method of a kind of Simultaneously test reconstituted tobacco oxalate, amber acid radical, sulfate radical and phosphate radical is provided, by reconstituted tobacco sample using pyrovinic acid after extract oscillation extraction through membrane filtration, then carry out ion chromatography;
Chromatographiccondition, analytical column: AS22-4mm; Column temperature: 30 DEG C; Leacheate: 4.5mmol/LNaCO
3+ 1.4mmol/LNaHCO
3; Flow velocity: 1.2mL/min; Sample size: 50 μ L; Rejector electric current: 31mA.Detecting device: self-loopa negative ion Suppressor conductivity detection.
Preferably, also comprise filtrate solid-phase extraction column purified treatment step before described ion chromatography.Due to reconstituted tobacco complicated component, sample extraction liquid removes the macromolecular substances that chromatographic column may be caused to pollute through purification, then carries out ion chromatography.
The present invention preferably adopts Solid-Phase Extraction RP post can impurity screening composition have the higher pre-treatment recovery effectively.
Preferably, the flow velocity of filtrate described in purified treatment is 0.5mL/min.Described RP post activates with the flow velocity of 1mL/min with 5mL methyl alcohol and 10ml pure water before using respectively.
Preferably, the concentration of described pyrovinic acid is 40mmol/L.
Preferably, the time of described oscillation extraction is 20 ~ 60 minutes, further preferred 30min.
Four kinds of objects to be measured are all water soluble anions, detect for chromatography of ions, and conventional Extraction solvent is pure water, acid solution and sodium hydroxide solution.But for pure water for extraction solvent, it is to reconstituted tobacco sample and once extract experimental summary, and the extraction efficiency of oxalic acid only has 37.7%, cannot single extraction complete.The present invention adopts 40mmol/LMSA well can extract four kinds of object ions, in extraction residue, do not detect oxalic acid, and extraction efficiency reaches 99.7% or more.
The present invention through carefully studying summary, the preferred analytical column of AS22 as this method, four kinds of target components have good separating effect on this post; And comparing the impact of flow rate of mobile phase and sample size further, flow velocity is 1.2mL/min, sample size is promoted to the sensitivity that 50 μ L can significantly improve composition to be measured, and sample introduction repeatability RSD reaches 0.13% ~ 0.7%.
The inventive method can be advantageously applied to the mensuration of the mensuration related component of tobacco sample and reconstituted tobacco sample mesoxalic acid, succinic acid, sulfuric acid and/or phosphoric acid components; And be applicable to the mensuration of all kinds of organic additive mesoxalic acid, succinic acid, sulfuric acid and/or phosphoric acid components in reconstituted tobacco.
Beneficial effect of the present invention:
The present invention has filled up the blank of prior art, provides the chromatography of ions detection method of a kind of Simultaneously test reconstituted tobacco mesoxalic acid, succinic acid, sulfuric acid and phosphate radical.Establish with sodium carbonate-bicarbonate mixed system for leacheate, 40mmol/L pyrovinic acid (MSA) is extract, and AS22 is the method that the IC of separating column measures reconstituted tobacco oxalate, amber acid radical, phosphate radical and sulfuric acid acid content.This method solve with water is extract and the irrationality making leacheate mensuration oxalic acid content with NaOH, make the oxalic acid in sample quantitatively more accurate, reliably, moreover, be applied to Simultaneously test reconstituted tobacco mesoxalic acid, succinic acid, when sulfuric acid and phosphoric acid, the relative standard deviation (RSD) of each component test content to be measured is 0.27% ~ 0.99%, recovery of standard addition is between 99.3% ~ 106.2%, accurately, simply, reliably, be applicable to the mensuration of related component in tobacco sample and reconstituted tobacco sample simultaneously and be applicable to the mensuration of all kinds of organic additive in reconstituted tobacco.
Accompanying drawing explanation
Under Fig. 1 variable concentrations NaOH solution wash-out, vitamin C and oxalic acid standard solution are separated spectrogram on AS18 post.
Wherein, spectrogram 1,3,5,7-vitamin C standard solution; Spectrogram 2,4,6,8-oxalic acid standard solution.
Fig. 2 vitamin C standard solution produces oxalic acid content comparative result (ug/mL) under different solvents matrix and variable concentrations leacheate condition.
The extraction efficiency comparative result of Fig. 3 variable concentrations pyrovinic acid extract.
Different extraction time test result comparative result (peak area) of Fig. 4.
Fig. 5 vitamin C is at 4.5mMNaCO
3+ 1.4mMNaHCO
3chromatogram under leacheate.
Wherein, spectrogram 1-vitamin C standard solution; Spectrogram 2-four kinds of desired anion mixed standard solutions.
Fig. 6 actual sample test color spectrogram.
Embodiment
The present invention is further illustrated below in conjunction with the drawings and specific embodiments.Following embodiment, only for exemplary illustration, can not be interpreted as limitation of the present invention.Unless stated otherwise, the reagent used in following embodiment is the reagent that routine is commercial or commercial sources obtains, unless stated otherwise, and the method and apparatus that the method and apparatus used in following embodiment uses for this area routine.
Materials and methods:
1.1 materials, reagent and instrument
The present invention is described for 26 of random selecting reconstituted tobacco trial targets and finished product; Aqueous phase filter membrane (0.45um, uncommon Podbielniak); Solid phase extraction column (U.S. wears peace for RP post, 1cc); Stratographic analysis post (AS22-4mm, the U.S. wears peace).
Pyrovinic acid (MSA) (99%, Fluka company of the U.S.); Natrium carbonicum calcinatum (benchmark product, Tianjin red rock chemical reagent work); Sodium carbonate (100%, purechemicalAnalysisCo.Ltd); Vitamin C (ascorbic acid) (99%, Sigma-Aldrich); Oxalic acid (99%, Acros company of the U.S.); Phosphate radical, sulfate radical standard solution (1000ug/ml, State center for standard matter); Six water succinic acids (99%, Sigma-Aldrich); Ultrapure water (resistivity >=18.2M Ω cm).
ICS5000 ion chromatograph (Dai An company of the U.S.); Milli-Q ultrapure water instrument (U.S. Millipore); CP224S electronic balance (sensibility reciprocal 0.0001g, German Sartorius company); HY-5 oscillator (domestic Jiangsu).
The impact experiment that embodiment 1 vitamin C measures oxalic acid
Much comprising tobacco at present in the chromatography of ions including ascorbic plant mesoxalic acid assay is all use NaOH solution to be separated oxalate and other negative ion as leacheate.This research finds that vitamin C can be converted into oxalic acid at a high ph.Take water as solvent preparation 100ug/ml vitamin C standard solution, AS18 anion analysis post is used respectively 10mmol/L, 20mmol/L, 30mmol/LNaOH solution isocratic elution and 11mmol/L ~ 50mmol/LNaOH solution gradient wash-out as mobile phase program, measure vitamin C standard solution, (retention time is qualitative, as shown in Figure 1) test color spectrogram all to have oxalate go out peak.Figure 2 shows that the 100ug/mL vitamin C standard solution prepared under adopting pure water and pyrovinic acid two kinds of solvent substrates produces the amount of oxalic acid when variable concentrations NaOH solution wash-out.Above result shows: nearly 50% vitamin C is converted into oxalic acid, therefore NaOH can not be used as extract and leacheate when measuring and containing ascorbic sample mesoxalic acid content, otherwise, test result oxalic acid content can be caused higher.
The research experiment of embodiment 2 sample-pretreating method
Four kinds of objects to be measured are all water soluble anions, detect for chromatography of ions, and conventional Extraction solvent is pure water, acid solution and sodium hydroxide solution.Reason due to 2.1 can not use strong alkali solution to extract.Test pure water is that extraction solvent carries out concussion extraction in 30 minutes to reconstituted tobacco sample and the residue after once extracting thereof, and measures four kinds of target components.Result show adopt pure water be extractant, the extraction efficiency of oxalic acid only has 37.7%, cannot single extraction complete.Because the analytical column that the method is used is anion column, conventional acid ion can bring negative ion to disturb, and therefore test have employed variable concentrations pyrovinic acid (MSA) is extraction solvent.Result shows (as shown in Figure 3): adopt 40mmol/LMSA well can extract four kinds of object ions, in extraction residue, do not detect oxalic acid, extraction efficiency reaches 99.7%.
Take 40mmol/LMSA as extraction solvent, have studied the impact of different duration of oscillation on test result, result shows that four kinds of target component test results difference in 20 ~ 60 minutes duration of oscillations is little, and comprehensive various factors, determines that Best Times is 30 minutes (as shown in Figure 4).
Due to reconstituted tobacco complicated component, sample extraction liquid needs to remove the macromolecular substances that chromatographic column may be caused to pollute through purification, then carries out ion chromatography.Experiment shows that Solid-Phase Extraction RP post can impurity screening composition have the higher pre-treatment recovery effectively.
The determination experiment of embodiment 3 chromatographic condition
To study for a long period of time experimental summary and previous reasons based on the present invention, NaOH solution can not be adopted as leacheate, Test Study use NaCO
3+ NaHCO
3as the possibility of mobile phase, result shows (see Fig. 5), adopts 4.5mmol/LNaCO
3+ 1.4mmol/LNaHCO
3make leacheate, vitamin C standard solution is not converted into oxalic acid.Test is the multiple chromatographic column of study tour simultaneously, such as AS14, AS22 isochromatic spectrum post, compare analysis from multiple factors such as degree of separation, peak shape symmetry, peak width and sensitivity, preferably determine the analytical column of AS22 as this method, four kinds of target components have good separation on this post; Compare the impact of flow rate of mobile phase and sample size, flow velocity is 1.2ml/min, sample size is promoted to the sensitivity that 50 μ L can significantly improve succinic acid, and sample introduction repeatability RSD reaches 0.13% ~ 0.7%.
The development test of embodiment 4 interfering ion
The present invention has investigated the impact of the organic acids such as Common Anions and formic acid, acetic acid, propionic acid, lactic acid, malic acid, malonic acid and the distinctive fumaric acid of reconstituted tobacco such as fluorine, chlorine, nitric acid, nitrite anions in tobacco on experiment simultaneously.Result shows that fluorine, chlorine, nitric acid, nitrite anions and formic acid, acetic acid, propionic acid, lactic acid go out peak before all concentrating on 10min, malic acid malonic acid and fumaric acid go out peak simultaneously in the middle of sulfate radical and succinic acid, have good being separated (as shown in Figure 6) with determinand.
Embodiment 5 determines working curve and detection limit
Use 40mmol/LMSA preparing standard solution respectively, the linear response under MSA matrix of the oxalate in four kinds of target components and succinate ion, but sulfate radical and phosphate radical are then quafric curve response under MSA matrix.
Use the phosphate radical of 1000ug/mL, sulfate radical, amber acid radical and oxalate standard reserving solution respectively, be mixed with the series standard working solution of table 1, carry out ion chromatography.With the peak area of object (Y), matching recurrence is carried out to the concentration (X, ug/mL) that it responds.Get the standard solution point of least concentration in series standard solution, continuous sample introduction 10 times, calculate standard deviation (SD), the detection limit being method with 3 times of SD.Regression equation, related coefficient and detection limit are shown in Table 2.
Table 1 hybrid standard working solution concentration (ug/ml)
Table 2 regression equation, related coefficient and detection limit
Embodiment 6 recovery and Precision Experiment
Accurately take reconstituted tobacco sample, measure according to the inventive method, each sample repeats 5 times, and result shows (shown in table 3), sample object component content relative standard deviation (RSD) is 0.27% ~ 0.99%, and the repeatability of illustration method is better.Get the sample solution of known content, the standard solution adding three concentration levels respectively carries out IC analysis, calculates the recovery, result display (shown in table 4), the recovery is between 99.3% ~ 106.2%, and the accuracy of illustration method is higher, can meet the requirement of quantitative test.
Table 3 method reperformance test result (n=5)
Table 4 method recovery of standard addition
Embodiment 7 sample pre-treatments and stratographic analysis
Sample and working sample moisture is prepared according to GB/T19616 and YC/T31-1996, accurately take 0.1g sample (being accurate to 0.0001g) in 150mL ground triangular flask, add 50.0mL40mmol/LMSA extract, cover stopper, be placed on oscillator, oscillation extraction 30min.Sample solution through membrane filtration, filtrate again with the flow velocity of 0.5mL/min through RP Solid-Phase Extraction column purification (RP post use before activate with the flow velocity of 1mL/min with 5ml methyl alcohol and 10ml pure water respectively), finally carry out ion chromatography.
Chromatographiccondition, analytical column: AS22-4mm; Column temperature: 30 DEG C; Leacheate: 4.5mmol/LNaCO
3+ 1.4mmol/LNaHCO
3; Flow velocity: 1.2mL/min; Sample size: 50 μ L; Rejector electric current: 31mA.Detecting device: self-loopa negative ion Suppressor conductivity detection.
Adopt this method to measure test agent in the reconstituted tobacco of 26 random selecting and tobacco finished product sample, the results are shown in Table shown in 5.MSA extract may contain a small amount of sulfate radical and phosphate anion, carries out blank test deduction if desired.The inventive method is applicable to the mensuration of tobacco simultaneously.
Table 5 actual sample measures (mg/g)
The present invention establishes with sodium carbonate-bicarbonate mixed system for leacheate, and 40mmol/L pyrovinic acid (MSA) is extract, and AS22 is the method that the IC of separating column measures reconstituted tobacco oxalate, amber acid radical, phosphate radical and sulfuric acid acid content.This method solving with water is extract and the irrationality making leacheate mensuration oxalic acid content with NaOH, makes the oxalic acid in sample quantitatively more accurate, reliable, is applicable to the mensuration of all kinds of organic additive in reconstituted tobacco.
Claims (10)
1. an ion chromatographic method for Simultaneously test reconstituted tobacco mesoxalic acid, succinic acid, sulfuric acid and phosphate radical, is characterized in that, be by reconstituted tobacco sample using pyrovinic acid after extract oscillation extraction filtrate through membrane filtration, then carry out ion chromatography;
Described chromatographiccondition, analytical column: AS22-4mm; Column temperature: 30 DEG C; Leacheate: 4.5mmol/LNaCO
3+ 1.4mmol/LNaHCO
3; Flow velocity: 1.2mL/min; Sample size: 50 μ L; Rejector electric current: 31mA; Detecting device: self-loopa negative ion Suppressor conductivity detection.
2. the ion chromatographic method of Simultaneously test reconstituted tobacco mesoxalic acid, succinic acid, sulfuric acid and phosphate radical according to claim 1, is characterized in that, also comprise and adopt solid-phase extraction column to carry out purified treatment step filtrate before described ion chromatography.
3. the ion chromatographic method of Simultaneously test reconstituted tobacco mesoxalic acid, succinic acid, sulfuric acid and phosphate radical according to claim 2, is characterized in that, described solid-phase extraction column purified treatment adopts Solid-Phase Extraction RP post.
4. the ion chromatographic method of Simultaneously test reconstituted tobacco mesoxalic acid, succinic acid, sulfuric acid and phosphate radical according to claim 2, is characterized in that, described RP post activates with the flow velocity of 1mL/min with 5mL methyl alcohol and 10ml pure water before using respectively.
5. the ion chromatographic method of Simultaneously test reconstituted tobacco mesoxalic acid, succinic acid, sulfuric acid and phosphate radical according to claim 2, it is characterized in that, the flow velocity of filtrate described in described purified treatment is 0.5mL/min.
6. the ion chromatographic method of Simultaneously test reconstituted tobacco mesoxalic acid, succinic acid, sulfuric acid and phosphate radical according to claim 1, it is characterized in that, the concentration of described pyrovinic acid is 40mmol/L.
7. the ion chromatographic method of Simultaneously test reconstituted tobacco mesoxalic acid, succinic acid, sulfuric acid and phosphate radical according to claim 1, it is characterized in that, the time of described oscillation extraction is 20 ~ 60 minutes.
8. the ion chromatographic method of Simultaneously test reconstituted tobacco mesoxalic acid, succinic acid, sulfuric acid and phosphate radical according to claim 1, it is characterized in that, the time of described oscillation extraction is 30min.
9. the application of method described in any one of claim 1 to 8, is characterized in that, is applied to the mensuration of tobacco sample and reconstituted tobacco sample mesoxalic acid, succinic acid, sulfuric acid and/or phosphoric acid components.
10. the application of method described in any one of claim 1 to 8, is characterized in that, is applied to the mensuration of organic additive mesoxalic acid, succinic acid, sulfuric acid and/or phosphoric acid components in reconstituted tobacco.
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