CN112305094A - Method for detecting maltose syrup doped in honey - Google Patents
Method for detecting maltose syrup doped in honey Download PDFInfo
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- CN112305094A CN112305094A CN202011002075.5A CN202011002075A CN112305094A CN 112305094 A CN112305094 A CN 112305094A CN 202011002075 A CN202011002075 A CN 202011002075A CN 112305094 A CN112305094 A CN 112305094A
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- 238000000034 method Methods 0.000 title claims abstract description 35
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- 238000004587 chromatography analysis Methods 0.000 claims abstract description 27
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- 238000001514 detection method Methods 0.000 claims abstract description 23
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- 235000010232 propyl p-hydroxybenzoate Nutrition 0.000 claims abstract description 12
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- BNABBHGYYMZMOA-AHIHXIOASA-N alpha-maltoheptaose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)O[C@H](O[C@@H]2[C@H](O[C@H](O[C@@H]3[C@H](O[C@H](O[C@@H]4[C@H](O[C@H](O[C@@H]5[C@H](O[C@H](O[C@@H]6[C@H](O[C@H](O)[C@H](O)[C@H]6O)CO)[C@H](O)[C@H]5O)CO)[C@H](O)[C@H]4O)CO)[C@H](O)[C@H]3O)CO)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O BNABBHGYYMZMOA-AHIHXIOASA-N 0.000 claims description 7
- FJCUPROCOFFUSR-UHFFFAOYSA-N malto-pentaose Natural products OC1C(O)C(OC(C(O)CO)C(O)C(O)C=O)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(OC3C(C(O)C(O)C(CO)O3)O)C(CO)O2)O)C(CO)O1 FJCUPROCOFFUSR-UHFFFAOYSA-N 0.000 claims description 7
- FJCUPROCOFFUSR-GMMZZHHDSA-N maltopentaose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O[C@H]([C@H](O)CO)[C@H](O)[C@@H](O)C=O)O[C@H](CO)[C@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O[C@@H]2[C@@H]([C@@H](O)[C@H](O[C@@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)[C@@H](CO)O2)O)[C@@H](CO)O1 FJCUPROCOFFUSR-GMMZZHHDSA-N 0.000 claims description 7
- DBTMGCOVALSLOR-UHFFFAOYSA-N 32-alpha-galactosyl-3-alpha-galactosyl-galactose Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(OC2C(C(CO)OC(O)C2O)O)OC(CO)C1O DBTMGCOVALSLOR-UHFFFAOYSA-N 0.000 claims description 6
- RXVWSYJTUUKTEA-UHFFFAOYSA-N D-maltotriose Natural products OC1C(O)C(OC(C(O)CO)C(O)C(O)C=O)OC(CO)C1OC1C(O)C(O)C(O)C(CO)O1 RXVWSYJTUUKTEA-UHFFFAOYSA-N 0.000 claims description 6
- LUEWUZLMQUOBSB-UHFFFAOYSA-N UNPD55895 Natural products OC1C(O)C(O)C(CO)OC1OC1C(CO)OC(OC2C(OC(OC3C(OC(O)C(O)C3O)CO)C(O)C2O)CO)C(O)C1O LUEWUZLMQUOBSB-UHFFFAOYSA-N 0.000 claims description 6
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 claims description 6
- UYQJCPNSAVWAFU-UHFFFAOYSA-N malto-tetraose Natural products OC1C(O)C(OC(C(O)CO)C(O)C(O)C=O)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(O)C(CO)O2)O)C(CO)O1 UYQJCPNSAVWAFU-UHFFFAOYSA-N 0.000 claims description 6
- LUEWUZLMQUOBSB-OUBHKODOSA-N maltotetraose Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@H](CO)O[C@@H](O[C@@H]2[C@@H](O[C@@H](O[C@@H]3[C@@H](O[C@@H](O)[C@H](O)[C@H]3O)CO)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O LUEWUZLMQUOBSB-OUBHKODOSA-N 0.000 claims description 6
- FYGDTMLNYKFZSV-UHFFFAOYSA-N mannotriose Natural products OC1C(O)C(O)C(CO)OC1OC1C(CO)OC(OC2C(OC(O)C(O)C2O)CO)C(O)C1O FYGDTMLNYKFZSV-UHFFFAOYSA-N 0.000 claims description 6
- FYGDTMLNYKFZSV-BYLHFPJWSA-N β-1,4-galactotrioside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@H](CO)O[C@@H](O[C@@H]2[C@@H](O[C@@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-BYLHFPJWSA-N 0.000 claims description 6
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 claims description 5
- 229960003415 propylparaben Drugs 0.000 claims description 2
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Images
Classifications
-
- 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
-
- 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
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
-
- 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
- G01N30/96—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 using ion-exchange
Abstract
The invention discloses a method for detecting maltose syrup doped in honey, which comprises the following steps: s1, adding a mixture of vitamin E and ethanol into a honey sample, extracting and oscillating at-10-4 ℃, and filtering to obtain a filtrate; s2, adding propyl p-hydroxybenzoate and sodium sulfite into the filtrate, diluting with a saturated sodium chloride aqueous solution, and filtering with a water-phase filter membrane to obtain a detection solution; and S3, obtaining an ion chromatogram of the detection solution by using ion chromatographic analysis, qualitatively analyzing whether the honey sample contains pentasaccharide and above malto polysaccharide according to retention time, mixing the honey sample with maltose syrup if the honey sample contains pentasaccharide and above malto polysaccharide, and judging whether the honey is mixed with maltose syrup if the honey sample does not contain pentasaccharide and above malto polysaccharide.
Description
Technical Field
The invention relates to the technical field of honey detection, in particular to a method for detecting maltose syrup mixed in honey.
Background
The honey is a natural sweet substance which is obtained by sufficiently brewing the nectar collected from flowers of flowering plants in a honeycomb by bees, has fragrant and strong smell and pure and sweet taste, and the honeybees convert polysaccharides in the nectar into monosaccharides glucose and fructose which can be directly absorbed by human bodies. Honey is supersaturated sugar solution, and can generate crystallization at low temperature, wherein the generated crystallization is glucose, and the part without crystallization is mainly fructose.
Part of the false honey is prepared by decocting malt syrup, etc., and contains no honey or little honey. The quality is characterized in that the honey-sugar syrup does not have natural honey flower odor, has a syrup boiling odor, has no moist taste when tasted and has the taste of white sugar water. In order to identify whether honey is counterfeited by adding syrup such as maltose syrup, GB/T21533-. The method is based on the following principle: the honey does not contain oligosaccharides with more than pentasaccharide, various maltose syrups contain oligosaccharides with more than pentasaccharide, fructose and glucose in a sample are removed by using a gel size exclusion method, the oligosaccharides are enriched and then are directly detected by an anion exchange chromatography-electrochemical detector, and the existence of the oligosaccharides with more than pentasaccharide is used as a judgment index of the maltose syrup in the honey. However, the pretreatment steps of the method are complicated, and the fructose and glucose need to be removed by a gel column chromatography method, so that the detection and judgment efficiency is influenced, experimental errors are easily caused, and the detection limit of the method is influenced (the standard detection limit is 5% of syrup). When the mixing ratio of the maltose syrup is less than 5%, the judgment cannot be made accurately.
The invention mainly establishes a novel, convenient and quick method for analyzing maltose and malto polysaccharide in honey by ion chromatography, and provides a new basis for judging whether the honey is mixed with maltose syrup. The method can simultaneously determine the reducing polysaccharides such as glucose, fructose and the like, so that gel column chromatography removal is not required, the experimental steps can be simplified, and the detection efficiency can be improved. In addition, the method has better separation degree and sensitivity to oligosaccharides with more than pentasaccharide, and can obviously improve the detection limit of the method.
Disclosure of Invention
In order to solve the problems, the invention provides a method for detecting the maltose syrup mixed in honey, and whether the maltose syrup is mixed in the honey is judged.
The technical scheme for solving the problem is to provide a method for detecting the maltose syrup mixed in honey, which comprises the following steps:
s1, adding a mixture of vitamin E and ethanol into a honey sample, extracting and oscillating at-10-4 ℃, and filtering to obtain a filtrate;
s2, adding propyl p-hydroxybenzoate and sodium sulfite into the filtrate, diluting with a saturated sodium chloride aqueous solution, and filtering with a water-phase filter membrane to obtain a detection solution;
and S3, obtaining an ion chromatogram of the detection solution by using ion chromatographic analysis, and qualitatively analyzing whether the honey sample contains pentasaccharide and more than malto-polysaccharide according to retention time, wherein if the honey sample contains pentasaccharide and more than malto-polysaccharide, the honey sample is mixed with maltose syrup, and if the honey sample does not contain pentasaccharide and more than malto-polysaccharide, the honey sample is not mixed with maltose syrup.
Preferably, the step of qualifying according to retention time is: and analyzing the peak-out time of each component in the mixed standard by ion chromatography, and qualitatively judging whether the mixed standard contains the pentasaccharide and the maltopolysaccharide above by contrasting the peak-out time of each component in the detection solution, wherein the mixed standard comprises maltose, maltotriose, maltotetraose, maltopentaose, maltohexaose and maltoheptaose.
Preferably, in step S1, the mass ratio of the ethanol to the honey sample is 50-100: 1.
preferably, in step S1, the mass ratio of the ethanol to the vitamin E is 100: 1-8.
Preferably, the mass ratio of the propyl hydroxybenzoate in the step S2 to the ethanol in the step S1 is 0.5-1:100, and the mass ratio of the sodium sulfite in the step S2 to the ethanol in the step S1 is 0.5-1: 100.
Preferably, the ion chromatography conditions are such that the column: CarboPac PA20 Analytical Column (3X 150 mm); mobile phase: deionized water is phase C, sodium hydroxide solution is phase A, sodium acetate solution is buffer solution, gradient elution is carried out, and the gradient elution procedure is as follows: mobile phase A: 250mmol/L sodium hydroxide aqueous solution; mobile phase C: ultrapure water; mobile phase D: 100mmNaAc, the proportion of the mobile phase A is maintained at 25 percent, the proportion of the mobile phase D is maintained at 0 percent, and the proportion of the mobile phase C is maintained at 75 percent in 0-18 minutes; in 18-95 minutes, the proportion of the mobile phase A is maintained at 25%, the proportion of the mobile phase D is maintained at 10%, and the proportion of the mobile phase C is maintained at 65%; at 95-110 minutes, the proportion of mobile phase a was maintained at 25%, the proportion of mobile phase D was maintained at 0%, and the proportion of mobile phase C was maintained at 75%, and the entire gradient run was completed at the end of the 110 th minute.
Preferably, the malto-polysaccharide of pentasaccharide or more includes maltopentasaccharide, maltohexasaccharide, and maltoheptasaccharide.
The invention has the beneficial effects that:
1. according to the method, gel column passing removal is not required, the experiment steps are simplified, and the detection efficiency is improved;
2. the stability of maltose and malto-polysaccharide in the analysis process is inconsistent, the added vitamin E has an antioxidant effect, and can prevent the maltose and malto-polysaccharide from being degraded, so that the stability of an analysis object is improved;
3. propyl p-hydroxybenzoate and saturated sodium chloride aqueous solution have been added to this application to improve the stability that contains sugar and detect the liquid, avoid it to take place chemical change and lead to the detection accuracy to reduce.
Drawings
FIG. 1 is a chromatogram of a honey sample measured in example 1 of the present application.
Detailed Description
In order to make the present invention more comprehensible, the technical solutions of the present invention are further described below with reference to specific embodiments, but the present invention is not limited thereto.
Example 1
Weighing 1g of sample, adding 100g of ethanol and 1g of vitamin E, magnetically stirring and extracting at-10 ℃ for 1 hour, filtering with a water-phase filter membrane, taking 1ml of filtrate, adding 0.5g of propyl p-hydroxybenzoate and 0.5g of sodium sulfite, diluting 100 times with saturated sodium chloride aqueous solution to obtain a detection solution, and carrying out chromatographic analysis according to the conditions of ion chromatographic analysis, wherein the conditions of ion chromatographic analysis are as follows: a chromatographic column: CarboPac PA20 Analytical Column (3X 150 mm); mobile phase: deionized water is phase C, sodium hydroxide solution is phase A, sodium acetate solution is phase D, gradient elution is carried out, and the gradient elution procedure is as follows: mobile phase A: 250mmol/L sodium hydroxide aqueous solution; mobile phase C: ultrapure water; mobile phase D: 100mmNaAc, the proportion of the mobile phase A is maintained at 25 percent, the proportion of the mobile phase D is maintained at 0 percent, and the proportion of the mobile phase C is maintained at 75 percent in 0-18 minutes; in 18-95 minutes, the proportion of the mobile phase A is maintained at 25%, the proportion of the mobile phase D is maintained at 10%, and the proportion of the mobile phase C is maintained at 65%; at 95-110 minutes, the proportion of mobile phase a was maintained at 25%, the proportion of mobile phase D was maintained at 0%, and the proportion of mobile phase C was maintained at 75%, and the entire gradient run was completed at the end of the 110 th minute.
Respectively weighing 1mg of arabinose, glucose, xylose, maltose, maltotriose, maltotetraose, maltopentaose, maltohexaose and maltoheptaose standard substances, adding 100g of ethanol, 1g of vitamin E, 0.5g of propyl p-hydroxybenzoate and 0.5g of sodium sulfite, diluting 100 times by saturated sodium chloride aqueous solution to obtain mixed standard reference substances, and carrying out chromatographic analysis according to the conditions of ion chromatographic analysis, wherein the conditions of the ion chromatographic analysis are as follows: a chromatographic column: CarboPac PA20 Analytical Column (3X 150 mm); mobile phase: deionized water is phase C, sodium hydroxide solution is phase A, sodium acetate solution is phase D, gradient elution is carried out, and the gradient elution procedure is as follows: mobile phase A: 250mmol/L sodium hydroxide aqueous solution; mobile phase C: ultrapure water; mobile phase D: 100mmNaAc, the proportion of the mobile phase A is maintained at 25 percent, the proportion of the mobile phase D is maintained at 0 percent, and the proportion of the mobile phase C is maintained at 75 percent in 0-18 minutes; in 18-95 minutes, the proportion of the mobile phase A is maintained at 25%, the proportion of the mobile phase D is maintained at 10%, and the proportion of the mobile phase C is maintained at 65%; at 95-110 minutes, the proportion of mobile phase a was maintained at 25%, the proportion of mobile phase D was maintained at 0%, and the proportion of mobile phase C was maintained at 75%, and the entire gradient run was completed at the end of the 110 th minute.
The chromatogram of the control sample and the mixed standard are shown in figure 1. From the peak time of fig. 1, it can be seen that the sample contains a trace amount of maltohexaose, which is the honey ginseng.
Example 2
Weighing 2g of sample, adding 100g of ethanol and 8g of vitamin E, magnetically stirring and extracting at 4 ℃ for 1 hour, filtering with a water-phase filter membrane, taking 1ml of filtrate, adding 1g of propyl p-hydroxybenzoate and 1g of sodium sulfite, diluting 100 times with saturated sodium chloride aqueous solution to obtain a detection solution, and carrying out chromatographic analysis according to the conditions of ion chromatographic analysis, wherein the conditions of ion chromatographic analysis are as follows: a chromatographic column: CarboPac PA20 Analytical Column (3X 150 mm); mobile phase: deionized water is phase C, sodium hydroxide solution is phase A, sodium acetate solution is phase D, gradient elution is carried out, and the gradient elution procedure is as follows: mobile phase A: 250mmol/L sodium hydroxide aqueous solution; mobile phase C: ultrapure water; mobile phase D: 100mmNaAc, the proportion of the mobile phase A is maintained at 25 percent, the proportion of the mobile phase D is maintained at 0 percent, and the proportion of the mobile phase C is maintained at 75 percent in 0-18 minutes; in 18-95 minutes, the proportion of the mobile phase A is maintained at 25%, the proportion of the mobile phase D is maintained at 10%, and the proportion of the mobile phase C is maintained at 65%; at 95-110 minutes, the proportion of mobile phase a was maintained at 25%, the proportion of mobile phase D was maintained at 0%, and the proportion of mobile phase C was maintained at 75%, and the entire gradient run was completed at the end of the 110 th minute.
Respectively weighing 1mg of arabinose, glucose, xylose, maltose, maltotriose, maltotetraose, maltopentaose, maltohexaose and maltoheptaose standard substances, adding 100g of ethanol, 8g of vitamin E, 1g of propyl p-hydroxybenzoate and 1g of sodium sulfite, diluting 100 times by saturated sodium chloride aqueous solution to obtain a mixed standard reference substance, and carrying out chromatographic analysis according to the conditions of ion chromatographic analysis, wherein the conditions of the ion chromatographic analysis are as follows: a chromatographic column: CarboPac PA20 Analytical Column (3X 150 mm); mobile phase: deionized water is phase C, sodium hydroxide solution is phase A, sodium acetate solution is phase D, gradient elution is carried out, and the gradient elution procedure is as follows: mobile phase A: 250mmol/L sodium hydroxide aqueous solution; mobile phase C: ultrapure water; mobile phase D: 100mmNaAc, the proportion of the mobile phase A is maintained at 25 percent, the proportion of the mobile phase D is maintained at 0 percent, and the proportion of the mobile phase C is maintained at 75 percent in 0-18 minutes; in 18-95 minutes, the proportion of the mobile phase A is maintained at 25%, the proportion of the mobile phase D is maintained at 10%, and the proportion of the mobile phase C is maintained at 65%; at 95-110 minutes, the proportion of mobile phase a was maintained at 25%, the proportion of mobile phase D was maintained at 0%, and the proportion of mobile phase C was maintained at 75%, and the entire gradient run was completed at the end of the 110 th minute.
And analyzing the peak-out time of each component and the sample in the mixed standard by ion chromatography, and qualitatively determining whether the mixed standard contains the pentasaccharide or more by contrasting the peak-out time of each component in the detection solution.
Example 3
Weighing 1g of sample, adding 100g of ethanol and 3g of vitamin E, magnetically stirring and extracting at 0 ℃ for 1 hour, filtering with a water-phase filter membrane, taking 1ml of filtrate, adding 0.8g of propyl p-hydroxybenzoate and 0.5g of sodium sulfite, diluting 100 times with saturated sodium chloride aqueous solution to obtain a detection solution, and carrying out chromatographic analysis according to the conditions of ion chromatographic analysis, wherein the conditions of ion chromatographic analysis are as follows: a chromatographic column: CarboPac PA20 Analytical Column (3X 150 mm); mobile phase: deionized water is phase C, sodium hydroxide solution is phase A, sodium acetate solution is phase D, gradient elution is carried out, and the gradient elution procedure is as follows: mobile phase A: 250mmol/L sodium hydroxide aqueous solution; mobile phase C: ultrapure water; mobile phase D: 100mmNaAc, the proportion of the mobile phase A is maintained at 25 percent, the proportion of the mobile phase D is maintained at 0 percent, and the proportion of the mobile phase C is maintained at 75 percent in 0-18 minutes; in 18-95 minutes, the proportion of the mobile phase A is maintained at 25%, the proportion of the mobile phase D is maintained at 10%, and the proportion of the mobile phase C is maintained at 65%; at 95-110 minutes, the proportion of mobile phase a was maintained at 25%, the proportion of mobile phase D was maintained at 0%, and the proportion of mobile phase C was maintained at 75%, and the entire gradient run was completed at the end of the 110 th minute.
Respectively weighing 1mg of arabinose, glucose, xylose, maltose, maltotriose, maltotetraose, maltopentaose, maltohexaose and maltoheptaose standard substances, adding 100g of ethanol, 3g of vitamin E, 0.8g of propyl p-hydroxybenzoate and 0.5g of sodium sulfite, diluting 100 times by saturated sodium chloride aqueous solution to obtain mixed standard reference substances, and carrying out chromatographic analysis according to the conditions of ion chromatographic analysis, wherein the conditions of the ion chromatographic analysis are as follows: a chromatographic column: CarboPac PA20 Analytical Column (3X 150 mm); mobile phase: deionized water is phase C, sodium hydroxide solution is phase A, sodium acetate solution is phase D, gradient elution is carried out, and the gradient elution procedure is as follows: mobile phase A: 250mmol/L sodium hydroxide aqueous solution; mobile phase C: ultrapure water; mobile phase D: 100mmNaAc, the proportion of the mobile phase A is maintained at 25 percent, the proportion of the mobile phase D is maintained at 0 percent, and the proportion of the mobile phase C is maintained at 75 percent in 0-18 minutes; in 18-95 minutes, the proportion of the mobile phase A is maintained at 25%, the proportion of the mobile phase D is maintained at 10%, and the proportion of the mobile phase C is maintained at 65%; at 95-110 minutes, the proportion of mobile phase a was maintained at 25%, the proportion of mobile phase D was maintained at 0%, and the proportion of mobile phase C was maintained at 75%, and the entire gradient run was completed at the end of the 110 th minute.
And analyzing the peak-out time of each component and the sample in the mixed standard by ion chromatography, and qualitatively determining whether the mixed standard contains the pentasaccharide or more by contrasting the peak-out time of each component in the detection solution. Example 4
Weighing 1.2g of sample, adding 100g of ethanol and 5g of vitamin E, magnetically stirring and extracting at-10 ℃ for 1 hour, filtering by using an aqueous phase filter membrane, taking 1ml of filtrate, adding 0.6g of propyl p-hydroxybenzoate and 0.6g of sodium sulfite, diluting 100 times by using a saturated sodium chloride aqueous solution to obtain a detection solution, and carrying out chromatographic analysis according to the conditions of ion chromatographic analysis, wherein the conditions of ion chromatographic analysis are as follows: a chromatographic column: CarboPac PA20 Analytical Column (3X 150 mm); mobile phase: deionized water is phase C, sodium hydroxide solution is phase A, sodium acetate solution is phase D, gradient elution is carried out, and the gradient elution procedure is as follows: mobile phase A: 250mmol/L sodium hydroxide aqueous solution; mobile phase C: ultrapure water; mobile phase D: 100mmNaAc, the proportion of the mobile phase A is maintained at 25 percent, the proportion of the mobile phase D is maintained at 0 percent, and the proportion of the mobile phase C is maintained at 75 percent in 0-18 minutes; in 18-95 minutes, the proportion of the mobile phase A is maintained at 25%, the proportion of the mobile phase D is maintained at 10%, and the proportion of the mobile phase C is maintained at 65%; at 95-110 minutes, the proportion of mobile phase a was maintained at 25%, the proportion of mobile phase D was maintained at 0%, and the proportion of mobile phase C was maintained at 75%, and the entire gradient run was completed at the end of the 110 th minute.
Respectively weighing 1mg of arabinose, glucose, xylose, maltose, maltotriose, maltotetraose, maltopentaose, maltohexaose and maltoheptaose standard substances, adding 10g of ethanol, 1g of vitamin E, 0.5g of propyl p-hydroxybenzoate and 0.5g of sodium sulfite, diluting 100 times by saturated sodium chloride aqueous solution to obtain mixed standard reference substances, and carrying out chromatographic analysis according to the conditions of ion chromatographic analysis, wherein the conditions of the ion chromatographic analysis are as follows: a chromatographic column: CarboPac PA20 Analytical Column (3X 150 mm); mobile phase: deionized water is phase C, sodium hydroxide solution is phase A, sodium acetate solution is phase D, gradient elution is carried out, and the gradient elution procedure is as follows: mobile phase A: 250mmol/L sodium hydroxide aqueous solution; mobile phase C: ultrapure water; mobile phase D: 100mmNaAc, the proportion of the mobile phase A is maintained at 25 percent, the proportion of the mobile phase D is maintained at 0 percent, and the proportion of the mobile phase C is maintained at 75 percent in 0-18 minutes; in 18-95 minutes, the proportion of the mobile phase A is maintained at 25%, the proportion of the mobile phase D is maintained at 10%, and the proportion of the mobile phase C is maintained at 65%; at 95-110 minutes, the proportion of mobile phase a was maintained at 25%, the proportion of mobile phase D was maintained at 0%, and the proportion of mobile phase C was maintained at 75%, and the entire gradient run was completed at the end of the 110 th minute.
And analyzing the peak-out time of each component and the sample in the mixed standard by ion chromatography, and qualitatively determining whether the mixed standard contains the pentasaccharide or more by contrasting the peak-out time of each component in the detection solution.
Specific embodiments of the present invention have been described above in detail.
It should be understood that numerous modifications and variations could be devised by those skilled in the art in light of the present teachings without departing from the inventive concepts. Therefore, any technical solutions that can be obtained by a person skilled in the art through logical analysis, reasoning or limited experiments in the prior art based on the inventive concept should be within the scope of protection defined by the claims.
Claims (8)
1. A method for detecting maltose syrup doped in honey is characterized by comprising the following steps:
s1, adding a mixture of vitamin E and ethanol into a honey sample, extracting and oscillating at-10-4 ℃, and filtering to obtain a filtrate;
s2, adding propyl p-hydroxybenzoate and sodium sulfite into the filtrate, diluting with a saturated sodium chloride aqueous solution, and filtering with a water-phase filter membrane to obtain a detection solution;
and S3, obtaining an ion chromatogram of the detection solution by using ion chromatographic analysis, and qualitatively analyzing whether the honey sample contains pentasaccharide and more than malto-polysaccharide according to retention time, wherein if the honey sample contains pentasaccharide and more than malto-polysaccharide, the honey sample is mixed with maltose syrup, and if the honey sample does not contain pentasaccharide and more than malto-polysaccharide, the honey sample is not mixed with maltose syrup.
2. The method for detecting the maltose syrup doped in the honey as claimed in claim 1, wherein the step of qualitative according to the retention time is as follows: and analyzing the peak time of each component in the mixed standard by ion chromatography, and qualitatively judging whether the mixed standard contains the pentasaccharide and the maltopolysaccharose by contrasting the peak time of each component in the detection solution, wherein the mixed standard comprises arabinose, glucose, xylose, maltose, maltotriose, maltotetraose, maltopentaose, maltohexaose and maltoheptaose.
3. The method for detecting the maltose syrup doped in the honey as claimed in claim 1, wherein in the step S1, the mass ratio of the ethanol to the honey sample is 50-100: 1.
4. the method for detecting the maltose syrup doped with honey as claimed in claim 1, wherein in step S1, the mass ratio of the ethanol to the vitamin E is 100: 1-8.
5. The method for detecting the addition of maltose syrup into honey as claimed in claim 1, wherein the mass ratio of the propyl hydroxybenzoate in the step S2 to the ethanol in the step S1 is 0.5-1:100, and the mass ratio of the sodium sulfite in the step S2 to the ethanol in the step S1 is 0.5-1: 100.
6. The method for detecting the maltose syrup doped in the honey as claimed in claim 1, wherein the ion chromatography conditions are that the chromatographic column: CarboPac PA20 Analytical Column (3X 150 mm); mobile phase: deionized water as phase C, sodium hydroxide solution as phase A, and sodium acetate buffer solution as phase D, and performing gradient elution.
7. The method as claimed in claim 1, wherein the malto-polysaccharide is selected from the group consisting of maltopentaose, maltohexaose, and maltoheptaose.
8. A method for detecting the incorporation of maltose syrup into honey as claimed in claim 6 wherein the gradient elution procedure is: mobile phase A: 250mmol/L sodium hydroxide aqueous solution; mobile phase C: ultrapure water; mobile phase D: 100mmNaAc, the proportion of the mobile phase A is maintained at 25 percent, the proportion of the mobile phase D is maintained at 0 percent, and the proportion of the mobile phase C is maintained at 75 percent in 0-18 minutes; in 18-95 minutes, the proportion of the mobile phase A is maintained at 25%, the proportion of the mobile phase D is maintained at 10%, and the proportion of the mobile phase C is maintained at 65%; at 95-110 minutes, the proportion of mobile phase a was maintained at 25%, the proportion of mobile phase D was maintained at 0%, and the proportion of mobile phase C was maintained at 75%, and the entire gradient run was completed at the end of the 110 th minute.
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