CN114200041A - Method for rapidly determining polysaccharide and sugar alcohol in food additive - Google Patents
Method for rapidly determining polysaccharide and sugar alcohol in food additive Download PDFInfo
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- 150000005846 sugar alcohols Chemical class 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 33
- 235000013373 food additive Nutrition 0.000 title claims abstract description 20
- 239000002778 food additive Substances 0.000 title claims abstract description 20
- 150000004676 glycans Chemical class 0.000 title claims description 13
- 229920001282 polysaccharide Polymers 0.000 title claims description 13
- 239000005017 polysaccharide Substances 0.000 title claims description 13
- 238000001514 detection method Methods 0.000 claims abstract description 45
- 235000003599 food sweetener Nutrition 0.000 claims abstract description 30
- 239000003765 sweetening agent Substances 0.000 claims abstract description 30
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 14
- 239000008103 glucose Substances 0.000 claims abstract description 14
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims abstract description 11
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims abstract description 11
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims abstract description 11
- 238000004255 ion exchange chromatography Methods 0.000 claims abstract description 11
- 239000000845 maltitol Substances 0.000 claims abstract description 11
- VQHSOMBJVWLPSR-WUJBLJFYSA-N maltitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H]([C@H](O)CO)O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O VQHSOMBJVWLPSR-WUJBLJFYSA-N 0.000 claims abstract description 11
- 235000010449 maltitol Nutrition 0.000 claims abstract description 11
- 229940035436 maltitol Drugs 0.000 claims abstract description 11
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims abstract description 10
- 229930195725 Mannitol Natural products 0.000 claims abstract description 10
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims abstract description 10
- 229930006000 Sucrose Natural products 0.000 claims abstract description 10
- 239000000594 mannitol Substances 0.000 claims abstract description 10
- 235000010355 mannitol Nutrition 0.000 claims abstract description 10
- 239000005720 sucrose Substances 0.000 claims abstract description 10
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 claims abstract description 9
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 claims abstract description 9
- 239000005715 Fructose Substances 0.000 claims abstract description 9
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 claims abstract description 9
- 229930091371 Fructose Natural products 0.000 claims abstract description 9
- 239000000600 sorbitol Substances 0.000 claims abstract description 9
- 235000010356 sorbitol Nutrition 0.000 claims abstract description 9
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims abstract description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 63
- 239000000243 solution Substances 0.000 claims description 34
- 239000012086 standard solution Substances 0.000 claims description 25
- 238000002386 leaching Methods 0.000 claims description 17
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 17
- 239000012498 ultrapure water Substances 0.000 claims description 17
- 239000000523 sample Substances 0.000 claims description 12
- 239000003085 diluting agent Substances 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 238000011088 calibration curve Methods 0.000 claims description 5
- 239000012528 membrane Substances 0.000 claims description 5
- 238000002156 mixing Methods 0.000 claims description 5
- 238000004587 chromatography analysis Methods 0.000 claims description 4
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 4
- 229910021607 Silver chloride Inorganic materials 0.000 claims description 3
- 238000004364 calculation method Methods 0.000 claims description 3
- 239000010931 gold Substances 0.000 claims description 3
- 229910052737 gold Inorganic materials 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 claims description 3
- 238000010812 external standard method Methods 0.000 claims description 2
- 239000012488 sample solution Substances 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims 1
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- 235000000346 sugar Nutrition 0.000 abstract description 26
- 238000004458 analytical method Methods 0.000 abstract description 20
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- 150000001720 carbohydrates Chemical class 0.000 description 8
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- 238000002474 experimental method Methods 0.000 description 3
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- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 150000001298 alcohols Chemical class 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
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- 230000003247 decreasing effect Effects 0.000 description 2
- 238000004817 gas chromatography Methods 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 125000001183 hydrocarbyl group Chemical group 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
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- 238000004519 manufacturing process Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000001488 sodium phosphate Substances 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241001089723 Metaphycus omega Species 0.000 description 1
- 208000008589 Obesity Diseases 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 125000003172 aldehyde group Chemical group 0.000 description 1
- LKDRXBCSQODPBY-ZXXMMSQZSA-N alpha-D-fructopyranose Chemical compound OC[C@]1(O)OC[C@@H](O)[C@@H](O)[C@@H]1O LKDRXBCSQODPBY-ZXXMMSQZSA-N 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
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- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
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- 229910052799 carbon Chemical group 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 125000000468 ketone group Chemical group 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 235000020824 obesity Nutrition 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 235000021309 simple sugar Nutrition 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/30—Control of physical parameters of the fluid carrier of temperature
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
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- G—PHYSICS
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- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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- G01N30/62—Detectors specially adapted therefor
- G01N30/64—Electrical detectors
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- G—PHYSICS
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- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
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- G01N30/28—Control of physical parameters of the fluid carrier
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- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
Abstract
The invention relates to a method for detecting sugar and sugar alcohol in a food additive, which belongs to the field of analytical chemistry, adopts an integral pulse ampere-ion chromatography, researches experimental factors influencing component separation by changing detection conditions, establishes a method for simultaneously detecting sugar and sugar alcohol (sorbitol, trehalose, maltitol, glucose, sucrose, mannitol and fructose) in a sweetening agent at one time, solves the technical problem of accurately analyzing multiple components in the food additive, is quick, accurate and sensitive, realizes the simultaneous analysis of multiple classes and components, and makes an important contribution to the detection of the components of the food additive.
Description
Technical Field
The invention relates to detection of sugar and sugar alcohol in a food additive, in particular to a method for rapidly determining polysaccharide and sugar alcohol in the food additive.
Background
With the rapid increase of the economy of China and the continuous improvement of the living standard of people, the food additive also puts forward higher requirements on the quality of food. The frequent food safety problem makes food safety become a focus of attention for people once. The food inspection and detection are important means for guaranteeing food safety and are also important ways for improving food quality. Among them, sugar compounds are the main source of energy required for people to sustain life activities. Sugar alcohols are substances formed by reducing the aldehyde group or ketone group of saccharides to hydroxyl group, are sweeteners mainly generated by reaction of corresponding sugars, do not cause elevation of blood sugar, and thus have an effect of preventing diabetes and obesity. Alcohols are compounds whose molecules contain a hydrocarbon group bound to a hydrocarbon group or a carbon on a side chain of a benzene ring, and many alcohols are beneficial to human health, but some have some toxicity. The detection of components of sugar and sugar alcohols is a difficult point in the field of analytical chemistry and a research hotspot in the field of analytical testing, and the sugar content of food must be labeled on a food label according to the regulations of national food safety standard 'nutrition label general rules for prepackaged food'.
The content of sugar and sugar alcohol in the sweetener directly affects the quality of the sweetener. Currently, the main detection methods for saccharides and sugar alcohols include gas chromatography, liquid chromatography, enzyme electrode method, ion chromatography, and the like. The gas chromatography has the advantages of rapidness and sensitivity, but the pretreatment steps are complex, and the quantification is not accurate enough. The liquid chromatography has the characteristics of accuracy and good selectivity, but has low sensitivity. The enzyme electrode method generally detects only one kind of saccharide, and has good specificity but low detection efficiency. The ion chromatography has the advantages of simplicity, convenience, rapidness, sensitivity, accuracy and the like, and is a novel ideal method for simultaneously detecting multiple components.
At present, although people have researches on methods for simultaneously detecting sugar and sugar alcohol in sweeteners, only a certain kind of components (such as simple sugar or alcohol) or a small number of components simultaneously detected can be searched in a current database, and the increasing detection requirements cannot be met, so that the research and establishment of a method for simultaneously detecting multiple kinds of sugar and sugar alcohol with strong applicability, good selectivity and high sensitivity has important practical significance and application value.
Disclosure of Invention
The invention provides a method for rapidly determining polysaccharide and sugar alcohol in a food additive, which adopts an integral pulse ampere-ion chromatography method, optimizes a pretreatment detection method, establishes a method for simultaneously detecting 7 saccharides and sugar alcohol in a sweetener at one time by researching influence factors of component separation, has the advantages of rapidness, simplicity, convenience, high sensitivity and good stability, realizes rapid analysis and green analysis, solves the technical problem of simultaneous analysis of multiple components of the saccharides and the sugar alcohol, and is suitable for determining the saccharides and the sugar alcohol in the sweetener.
In order to achieve the purpose, the invention adopts the following technical scheme:
a detection method of polysaccharide and sugar alcohol in food additive, wherein the food additive is sweetener, and the detection steps are as follows;
step one, preparation of a mixed standard solution:
preparing mixed standard stock solution from standard solution of sorbitol, trehalose, maltitol, glucose, sucrose, mannitol and fructose according to the requirement;
step two, pretreatment of a sample to be detected:
taking 0.1000g of a sweetener sample to be detected in a 100mL beaker, adding about 50mL of ultrapure water, uniformly mixing and standing for 10min, adjusting the pH value of the diluent to 7.0-8.5 by adopting 0.10mol/L sodium hydroxide solution, transferring all the diluent to a 500mL volumetric flask, fixing the volume to the scale by adopting the ultrapure water, shaking up, standing for 15min, taking 10.0mL of solution, firstly passing through a 0.22um filter membrane, collecting the later-stage clear liquid, and detecting.
Step three, ion chromatography conditions and multi-stage gradient leaching conditions:
and (3) analyzing the column: CarboPacPA20 (3X 150 mm);
protection of the column: CarboPacPA20 (3X 30 mm);
a working electrode: gold;
reference electrode: AgCl electrode
Mobile phase: a:200mmol/L NaOH solution, B: ultrapure water;
step four, drawing a calibration curve:
detecting the mixed standard solution prepared in the step one by using an ion chromatograph according to the chromatographic analysis condition and the multi-stage gradient leaching condition given in the step three, drawing a calibration curve by taking the peak area of the component to be detected as a vertical coordinate, and quantifying by using an external standard method;
step five, detecting the sample solution:
detecting the clear liquid obtained in the step two by using an ion chromatograph according to the chromatographic analysis conditions and the multi-stage gradient leaching conditions given in the step three, and calculating a result;
step six, calculating a result:
and correcting the calculation result measured in the fifth step according to the result measured by mixing the standard solution in the fourth step, thereby calculating the content of each component in the measured food additive which is the sweetener.
Preferably, the above solutions are each prepared using ultrapure water having a resistivity of 18.2 M.OMEGA.cm.
Preferably, in the second step, 0.1000g of sweetener is weighed in a 100mL beaker, 50mL of ultrapure water is added, the mixture is uniformly mixed and kept stand for 10min to obtain a diluent, the diluent is completely transferred to a 500mL volumetric flask, the volume is constant to a scale by adopting the ultrapure water, the mixture is shaken up and kept stand for 15min, 10.0mL of solution is taken and firstly passes through a 0.22um filter membrane, and clear liquid is collected and detected.
Further preferably, in the second step, the pH value of the diluent is adjusted to 7-8.5 by using 0.10mol/L sodium hydroxide solution.
Preferably, in the third step, the leaching is multi-step gradient leaching, and the multi-step gradient leaching procedure is as follows: the concentration of the leacheate NaOH is 200.0mmol/L when (-40.0) - (-30.05) min, and the concentration of the leacheate NaOH is 50.0mmol/L when (-30.05) - (8.00) min.
TABLE 1 ion chromatography elution conditions
| Time/min | A NaOH solution (200mmol/L) | B ultrapure water |
| -40.00 | 100％ | 0 |
| -30.05 | 100％ | 0 |
| -30.00 | 25％ | 75％ |
| 0.00 | 25％ | 75％ |
| 8.00 | 25％ | 75％ |
Preferably, in the third step, the flow rate: 0.50 mL/min.
Preferably, in the third step, the column temperature: at 30 ℃.
The invention has the following beneficial effects:
1. in actual production, chemical molecular formulas and properties of sugar and sugar alcohol in a plurality of sweeteners are similar, and because the detection method and detection contents are limited, the polysaccharide and the sugar alcohol are difficult to detect and process quickly and accurately.
2. In order to simultaneously separate and detect 7 sugar and sugar alcohol components of a sweetening agent and improve the overall separation degree of the components, through experimental analysis and comparison, the method provided by the invention selects the steps of dissolving a sample and adjusting the pH value of the solution to 7.0-8.5 during sample pretreatment, adopts a high-concentration leaching method within the first 10min, uses 200mmol/L sodium hydroxide solution to carry out powerful washing on a chromatographic column, removes interferents influencing the components to be detected, and can greatly improve the detection accuracy. The system was then flushed with 50mmol/L NaOH solution for 30min to ensure system stability.
3. According to the invention, through multi-factor multi-matrix investigation and analysis, the multi-level gradient elution condition, the initial elution concentration, the elution liquid flow rate and the chromatographic column temperature are researched and analyzed in sequence, and the accuracy and the applicability of the method for simultaneously measuring multiple components are improved to the greatest extent.
4. The invention researches the important influence of the temperature of the chromatographic column on the detection, and the small change of the temperature of the chromatographic column can also cause component overlapping. At the column temperature of 30 ℃, the qualitative and quantitative requirements of 7 components can be just met.
5. The detection method is simple, convenient and quick to operate and high in detection sensitivity, and can be used for on-machine detection through simple pretreatment operation.
6. The detection method does not use or generate solvents, reagents and byproducts which are harmful to human health and ecological environment, and accords with the concept of green analysis.
Drawings
FIG. 1 is an ion chromatogram of a 7-sugar alcohol mixed standard solution.
FIG. 2 is an ion chromatogram of a mixed standard solution at different initial leacheate concentrations.
FIG. 3 is an ion chromatogram of a mixed standard solution at different column temperatures.
Figure 4 is an ion chromatogram of 3 compound sweeteners.
The materials corresponding to the numbers in the attached drawings: 1. sorbitol; 2. maltitol; 3. trehalose; 4. mannitol; 5. glucose; 6. sucrose; 7. fructose.
FIG. 5 is an ion chromatogram of a standard solution of 7 sugar and sugar alcohol mixtures without pre-washing.
Detailed Description
The invention is further illustrated by the following examples, but the scope of the invention as claimed is not limited to the scope of the examples.
Example 1
A method for simultaneously detecting sugar and sugar alcohol in 7 in a sweetener comprises the following steps:
1. instruments and reagents
An ICS6000+ ion chromatography system equipped with an electrochemical detector and an autosampler (Thermo Scientific, usa); MIlli-Q ultra pure water apparatus (Millipore, USA); electronic balance (Mettler ML 802/02); water for experiment: ultrapure water (resistivity 18.2M Ω · cm); sugar and sugar alcohol standard substance or standard reagent (purity > 98%); sodium hydroxide solution (50% w/w, Fisher Chemical); hydrochloric acid (guaranteed purity);
2. ion chromatography conditions
CarboPacPA20 (3X 150mm) analytical column, CarboPacPA20 (3X 30mm) guard column; a working electrode: gold (au); reference electrode: an AgCl electrode; mobile phase: NaOH leacheate (nitrogen blanket); flow rate: 0.50 mL/min; column temperature: 30 ℃; sample introduction amount: 25 uL. Initial leacheate concentration: 50mmol/L NaOH solution.
3. Multi-stage gradient leaching conditions:
the multi-stage gradient elution procedure is as follows: the concentration of the leacheate NaOH is 200.0mmol/L when (-40.0) - (-30.05) min, and the concentration of the leacheate NaOH is 50.0mmol/L when (-30.05) - (8.00) min;
4. preparation of mixed standard solution:
preparing a mixed standard solution with a certain concentration from standard solutions or standard products of sorbitol, trehalose, maltitol, glucose, sucrose, mannitol and fructose according to the needs, and adjusting the pH value of the solution to prepare the mixed standard solution with the pH value of 7.0 in the process of preparing the mixed standard solution.
5. Pretreatment of a sample to be detected:
taking 0.1000g of a compound sweetener sample, adding about 50mL of ultrapure water into a 100mL beaker, uniformly mixing and standing for 10min, adjusting the pH value of the diluent to 7.0 by adopting 0.10mol/L sodium hydroxide solution, completely transferring the diluent into a 500mL volumetric flask, fixing the volume to the scale by adopting the ultrapure water, shaking up and standing for 15min, taking 10.0mL of solution, firstly passing through a 0.22um filter membrane, collecting the later-stage clear liquid, and detecting.
The results of 7 kinds of sugars and sugar alcohols in the mixed standard solution were examined under the conditions of the above experimental analysis, and are shown in FIG. 1.
Example 2: effect of flow velocity on detection
According to the conditions of the experimental analysis in example 1, the separation of the components of the mixed standard solution was successively tested under the conditions of the flow rates of 0.30mL/min, 0.40mL/min, 0.50mL/min and 0.60mL/min of the eluents, and the relationship between the flow rate of the eluents and the system pressure was as shown in Table 2.
TABLE 2 relationship of flow rate of rinsing liquid to system pressure
| Flow rate of flow | 0.30mL/min | 0.40mL/min | 0.50mL/min | 0.60mL/min |
| System pressure | 1678psi | 2248psi | 2739psi | 3125psi |
From Table 2, it can be seen that the system pressure was 1678psi at a 0.30mL/min eluent flow rate, because the flow rate was lower, the pressure was lower, the peak width increased, the peak sensitivity decreased, and the overall separation was not high. At a flow rate of 0.40mL/min, the system pressure reached 2248psi, and although the sensitivity of the components was improved, the degree of separation between individual components was not ideal. At a flow rate of 0.60mL/min, the system pressure reaches 3125psi, the system pressure is greater, exceeding the upper recommended pressure limit of the column of 3000psi, the overall analysis time of the components is shortened, but the overall separation of the components is reduced, and the pressure is greater, which may affect the useful life of the column to some extent. The initial pressure is 2739psi at the flow rate of 0.50mL/min, the pressure is moderate, and the sensitivity and the separation degree of the components can meet the detection requirements. Therefore, the present invention chooses to perform the experiment at a flow rate of 0.5mL/min, in view of system stability, resolution, and sensitivity.
Example 3: effect of Multi-stage gradient elution conditions on detection
According to the conditions of the experimental analysis in example 1, the separation of the components of the mixed standard solution with the initial leacheate concentration of 10, 20, 50, 100 and 200mmol/L is tested in sequence under the condition that other conditions are not changed, because the initial concentration has little influence on the separation degree of the three components of the components 1, 2 and 3, the influence of the initial concentration on the latter 4 components is mainly discussed, and the separation of the components of the mixed standard solution is shown in FIG. 2.
As can be seen from FIG. 2, when the initial leaching concentrations are 100 and 200mmol/L, respectively, trehalose of the component 2 is not separated from maltitol of the component 3, and mannitol of the component 4 is not separated from glucose of the component 5; the lower the initial elution concentration, the more desirable the overall separation and sensitivity of these 7 components, but taking into account the peak pattern and time cost. Therefore, the invention selects the initial leaching concentration of 50mmol/L for experimental detection.
Example 4: effect of column temperature on detection
According to the conditions of the experimental analysis in example 1, the separation effect of 7 kinds of saccharides and sugar alcohols under the conditions of 25 ℃, 27.5 ℃, 30 ℃, 32.5 ℃ and 35 ℃ column temperature was tested in sequence without changing other conditions, and the test results are shown in FIG. 3.
As can be seen from FIG. 3, the temperature of the chromatographic column is low at 25 ℃ and 27.5 ℃, the separation effect of most components is good, but the overall peak-off time is delayed, and the sucrose component 6 and the fructose component 7 also have slight tailing conditions, which is not favorable for the quantitative accuracy calculation. When the temperature of the chromatographic column is 32.5 ℃ and 35 ℃, the ion exchange rate is increased, the column efficiency is improved, and the analysis time is shortened, but the peak production time of the component 2 maltitol and the component 3 trehalose is very similar, the separation effect is not ideal, and the quantitative accuracy cannot be ensured. At the column temperature of 30 ℃, the analysis rate is moderate, the separation degree is good, and the overall separation effect of 7 components is good, so that the temperature has obvious influence on the simultaneous detection of the 7 components, and the experimental analysis is performed at the column temperature of 30 ℃ in the invention from the aspects of improving the separation degree, sensitivity and selectivity of the method.
Example 5: linear range and detection limit
Mixed standard solutions of different concentrations were prepared and a calibration curve fit was performed using the chromatographic conditions in example 1. The detection limit of the component to be measured (S/N ═ 3) was calculated from the chromatographic peak signal-to-noise ratio, and detection and linear fitting were performed for 8 concentration levels (0.10, 0.20, 0.50, 1.0, 2.0, 5.0, 10.0, and 20.0mg/L) of the mixed standard solutions in the concentration range of 0.10mg/L to 20.0mg/L, and the results are shown in table 3.
TABLE 3 Linear Range, Linear equation, correlation coefficient and detection Limit for the test Components (S/N ═ 3)
Y: peak area (nC · min), X: mass concentration (mg/L)
As can be seen from Table 3, the linear correlation coefficients for all 7 components are greater than 0.999, indicating that the components are simultaneously linear well under the experimental conditions. The detection limit of the 7 components is between 0.0059 and 0.2059 mg/L.
Example 6: recovery and precision
Respectively selecting 3 uniformly mixed samples, sequentially adding mixed standard solutions with concentration levels of 0.50mg/L, 1.00mg/L and 5.00mg/L into the 3 samples, carrying out 6 times of parallel determination experiments on each concentration level to verify the recovery rate and the precision of different sweetener matrix detection methods, and calculating the recovery rate according to the matrix concentration, the added concentration and the detection concentration after addition. Table 4 lists the three concentration levels, the average recovery of six tests and RSD data.
Table 4 recovery and precision (RSD) of assay components (n ═ 6)
As can be seen from Table 4, the recovery rates of the 7 sugars and sugar alcohols are between 90% and 110%, and all the recovery rate data meet the technical requirements of physicochemical inspection of foods (the detection content is less than 0.10mg/L, and the recovery rate should be between 60% and 120%), which fully indicates that the recovery rates of the 7 sugars and sugar alcohols are good. At three concentration levels, the RSD of 7 components is in the range of 1.03-4.85%. In general, the recovery rate data results of three matrixes and six parallel detections meet the physicochemical detection requirements of foods, and the detection method is high in accuracy and good in precision.
Example 7: detection of actual samples
3 compound sweeteners (formula A, formula B and formula C) were extracted, and detection and analysis of the contents of 7 sugars and sugar alcohols of actual samples were performed according to the ion chromatography conditions and pretreatment steps in example 1, and Table 5 shows the results of the detection component data, and FIG. 4 shows the ion chromatogram for the actual sample detection.
TABLE 5 detection results (mg/L) in sweetener
| Serial number | Components | Formulation A | Formulation | Formulation C | |
| 1 | Sorbitol | 8.9078 | 6.3787 | 4.9426 | |
| 2 | Trehalose | 8.5321 | 7.3322 | 5.6153 | |
| 3 | Maltitol | 7.7706 | 8.2063 | 8.8948 | |
| 4 | Glucose | 7.5887 | 9.7671 | 8.4372 | |
| 5 | Sucrose | 0.3834 | 9.9246 | 9.2247 | |
| 6 | Mannitol | 6.9526 | 7.2723 | 7.8559 | |
| 7 | Fructose | 8.0460 | 9.7619 | 7.2114 |
Comparing 3 sweetener spectrograms, wherein 7 components are detected in 7 sweeteners, main saccharides contained in the sweeteners are glucose, trehalose, sucrose and fructose, and sugar alcohols are mainly sorbitol, maltitol and mannitol; the formula A contains a large amount of phosphate and a small amount of sodium carbonate, and the solution is weakly alkaline; the formula B contains a large amount of sodium chloride, sodium bicarbonate and phosphate, and the solution is neutral; formula C contains a large amount of sodium citrate and phosphate, the solution is acidic, but the accuracy of the measurement result is not affected, and the method is completely satisfactory for detecting the medium sugar and the sugar alcohol in a complex matrix of the sweetener.
Example 8: influence of pre-washing on detection in pretreatment of sweetener to-be-detected sample
According to the experimental analysis conditions in example 1, the conditions of the multi-step gradient elution are the same under other conditions:
the multi-stage gradient elution procedure is as follows: when (-30.05) - (8.00) min, the concentration of the leacheate NaOH is 50.0 mmol/L; the map is shown in figure 5, and the separation effect is poor when the components of glucose, sucrose and mannitol are overlapped. The method of high-concentration leaching is adopted within the first 10min, and the strong washing of the chromatographic column is carried out by using 200mmol/L sodium hydroxide solution, so that the interferents influencing the component to be detected are removed, and the detection accuracy can be greatly improved.
Example 9: influence of pH on detection in pretreatment of sweetener to-be-detected sample
According to the experimental analysis conditions in example 1, mixed standard solutions with different pH values (3.0, 5.5, 7.0, 8.5 and 10.0) and the same theoretical concentration are prepared in sequence and tested under the condition that other conditions are not changed, and the experimental result shows that the content of 6 components, namely, the sorbitol component 1, the maltitol component 2, the trehalose component 3, the glucose component 5, the sucrose component 6 and the fructose component 7, is greatly influenced by the pH value of the solution, the mannitol is slightly influenced by the pH value, and the test influence of different pH values on each component is shown in Table 6.
As can be seen from Table 6, the measured glucose content was increased to some extent under both strong acid and strong alkali conditions, and the recovery rates of other sugars and sugar alcohols were decreased, because most of the sugars and sugar alcohols were easily decomposed into glucose under both strong acid and strong alkali conditions, and the measured glucose content was increased. Under the conditions that the pH value of sorbitol, maltitol, trehalose, glucose and fructose is more than 8.5 and the pH value is less than 7.0, the recovery rate is greatly reduced, and the quantitative analysis is influenced. As is clear from Table 6, under the weak alkaline condition of pH value of 7.0-8.5, the content change of the 7 components is small, the fluctuation is within 2.36%, which shows that the content of each component in the solution is stable at the moment.
TABLE 6 influence of pH value of solution on test results
According to the invention, through deep research on various experimental influence factors such as the concentration, the flow rate, the column temperature and the like of the initial leacheate, the chromatographic strip separation condition for simultaneously detecting sugar and sugar alcohol in the sweetener is explored, and the analytical method for simultaneously detecting 7 kinds of sugar and sugar alcohol by the integral pulse amperometric-ion chromatography is established. The method is simple, rapid, sensitive and accurate, and is completely suitable for rapid analysis of sugar and sugar alcohol in sweetener.
The above-described embodiments are merely preferred embodiments of the present invention, and should not be construed as limiting the present invention, and features in the embodiments and examples in the present application may be arbitrarily combined with each other without conflict. The protection scope of the present invention is defined by the claims, and includes equivalents of technical features of the claims. I.e., equivalent alterations and modifications within the scope hereof, are also intended to be within the scope of the invention.
Claims (7)
1. A method for rapidly determining polysaccharide and sugar alcohol in a food additive, wherein the food additive is a sweetener, and the method comprises the following steps:
step one, preparation of a mixed standard solution:
preparing mixed standard solutions of sorbitol, trehalose, maltitol, glucose, sucrose, mannitol and fructose according to the requirements;
step two, pretreatment of a sample to be detected:
weighing a sweetening agent in a beaker, adding ultrapure water, uniformly mixing and standing, adjusting the pH value of a diluent by using a sodium hydroxide solution, completely transferring the diluent to a volumetric flask, fixing the volume to a scale by using the ultrapure water, shaking up and standing, taking a solution, filtering the solution by using a membrane, collecting a clear solution, and detecting to be detected;
step three, ion chromatography conditions and leaching:
and (3) analyzing the column: CarboPacPA20 (3X 150 mm);
protection of the column: CarboPacPA20 (3X 30 mm);
a working electrode: gold;
reference electrode: an AgCl electrode;
mobile phase: a, 200mmol/L NaOH solution, B: ultrapure water;
sample introduction amount: 25 uL;
step four, drawing a calibration curve:
detecting the mixed standard solution prepared in the step one by using an ion chromatograph according to the chromatographic analysis condition and the multi-stage gradient leaching condition given in the step three, drawing a calibration curve by taking the mass concentration of the component to be detected as a horizontal coordinate and the peak area of the component to be detected as a vertical coordinate, and quantifying by using an external standard method;
step five, detecting the sample solution:
detecting the clear liquid obtained in the step two by using an ion chromatograph according to the chromatographic analysis conditions and the multi-stage gradient leaching conditions given in the step three, and calculating a result;
step six, calculating a result:
and correcting the calculation result obtained in the step five by using the result obtained by mixing the standard solution in the step four, so as to calculate the component contents of the polysaccharide and the sugar alcohol in the measured food additive.
2. The method of claim 1, wherein the solutions are prepared using ultrapure water having a resistivity of 18.2M Ω -cm.
3. The detection method for rapidly determining the polysaccharides and the sugar alcohols in the food additives as claimed in claim 1, wherein in the second step, 0.1000g of the sweetener is weighed into a 100mL beaker, 50mL of ultrapure water is added, the mixture is mixed and kept stand for 10min to obtain a diluted solution, the diluted solution is completely transferred into a 500mL volumetric flask, the volume is determined to the scale by adopting the ultrapure water, the diluted solution is shaken and kept stand for 15min, 10.0mL of the solution is taken and firstly passed through a 0.22um filter membrane, and a clear solution is collected and detected.
4. The method for rapidly detecting polysaccharides and sugar alcohols in food additives as claimed in claim 1, wherein in the second step, the pH value of the diluent is adjusted to 7.0-8.5 by 0.10mol/L sodium hydroxide solution.
5. The method for rapidly detecting polysaccharides and sugar alcohols in food additives according to claim 1, wherein: in the third step, the leaching is multi-stage gradient leaching, and the multi-stage gradient leaching procedure is as follows: the concentration of the leacheate NaOH is 200.0mmol/L when (-40.0) - (-30.05) min, and the concentration of the leacheate NaOH is 50.0mmol/L when (-30.05) - (8.00) min.
6. The method for rapidly detecting polysaccharides and sugar alcohols in food additives according to claim 1, wherein: in the third step, the flow rate is as follows: 0.50 mL/min.
7. The method for rapidly detecting polysaccharides and sugar alcohols in food additives according to claim 1, wherein: in the third step, the column temperature: at 30 ℃.
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