CN104849395A - Method for detecting isomaltulose in food - Google Patents
Method for detecting isomaltulose in food Download PDFInfo
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- CN104849395A CN104849395A CN201510319842.8A CN201510319842A CN104849395A CN 104849395 A CN104849395 A CN 104849395A CN 201510319842 A CN201510319842 A CN 201510319842A CN 104849395 A CN104849395 A CN 104849395A
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- isomaltoketose
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- deionized water
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Abstract
The invention relates to a method for detecting isomaltulose in food. The detection method has the following chromatographic conditions: a chromatographic column Hypersil APS-2(NH2) is adopted and has the specification of 250mm*4.6mm*5mu m; a mobile phase comprises acetonitrile and deionized water, the volume ratio of the acetonitrile to the deionized water is (88: 12)-(85: 15), and the flow rate of the mobile phase is 1mL/ minute; the column temperature is 40 DEG C; the injection volume is 10mu L; a steaming detection light scattering detector is used as a detector, the nitrogen flow rate is 1.6L/ minute, the temperature of an atomizer is 50 DEG C, and the temperature of a steaming pipe is 80 DEG C. The method has the advantages of being simple in pretreatment, easy to operate, low in limit of detection, good in linearity in a linear range, high in correlation coefficient and high in accuracy of measuring results, and is an effective method for detecting the isomaltulose in the food.
Description
Technical field
The present invention relates to food analysis field, be specifically related to the detection method of isomaltoketose in a kind of food.
Background technology
Isomaltoketose (Isomaltulose, C
12h
22o
11h
2o) have another name called palatinose, formal name used at school α-D-glycopyranosyl-1,6-D-fructose, to be naturally present in honey, sugar-cane juice.It is that sucrose isomerase changes α-1,2 glycosidic bond of sucrose a kind of reducing disaccharides of α-1,6 glycosidic bond into.It is a kind of isomeride of sucrose, has the sweet taste characteristic similar to sucrose, is a kind of low sweet sweetener, has extraordinary mouthfeel, can be applicable in candy, low sweet drink and food.It can be used alone or uses together with sucrose.It also has extraordinaryly covers peculiar smell effect.Such as: the fish oil taste of DHA, the peculiar smell of fruit and vegetable juice and the beany flavor etc. of soymilk have good effect.Due to the isomeride that isomaltoketose is sucrose, be difficult to be separated, sucrose is the important interference detecting isomaltoketose in food; Not yet find the method being separated sucrose and isomaltoketose in food at present.Therefore, what liquid phase chromatography detected the main solution of detection of the isomaltoketose content in food is how to make sucrose and the complete separation problem of isomaltoketose in food.
Summary of the invention
First object of the present invention is to provide the method for separating and detecting of sucrose and isomaltoketose in a kind of food.
Second object of the present invention is to provide the high performance liquid chromatography evaporative light-scattering detector approach of isomaltoketose content in a kind of convenience, accurate, sensitive mensuration food.
In order to reach object of the present invention, the invention provides following technical scheme:
A method for separating and detecting for sucrose and isomaltoketose in food, it comprises employing Hypersil APS-2 (NH
2) chromatographic column, its specification is 250mm × 4.6mm × 5 μm, and mobile phase is the volume ratio 88: 12 ~ 85: 15 of acetonitrile and deionized water, and flow velocity is 1mL/min, and column temperature is 40; Evaporative light-scattering detector selected by detecting device, and nitrogen flow rate is 1.6L/min, and atomizer temperature is 50 DEG C, and evaporation tube temperature is 80 DEG C.
An assay method for the content of isomaltoketose in food, it comprises the following steps:
(1) pre-treatment of sample;
(2) setting device:
The chromatographic condition of high performance liquid chromatograph workstation is set: adopt Hypersil APS-2 (NH
2) chromatographic column, its specification is 250mm × 4.6mm × 5 μm, and mobile phase is the volume ratio 88: 12 ~ 85: 15 of acetonitrile and deionized water, and flow velocity is 1mL/min, and column temperature is 40; Evaporative light-scattering detector selected by detecting device, and nitrogen flow rate is 1.6L/min, and atomizer temperature is 50 DEG C, and evaporation tube temperature is 80 DEG C;
(3) drawing standard curve, obtains typical curve equation;
(4) content of isomaltoketose in sample is detected: what arranged by the sample implantation step (2) after step (1) pre-treatment enters duty performance liquid chromatographic column, sample size is 10 microlitres, calculate the peak area under same retention time, substitute into the typical curve equation of step (3) gained, calculate the content of isomaltoketose in testing sample.
Wherein, the pre-treatment of step (1) described sample comprises the following steps: transfer in volumetric flask by after sample deionized water dissolving, slowly add potassium ferrocyanide solution, acid acetic acid zinc solution, add deionized water constant volume to scale, mixing, leave standstill 30min, filter with dry filter paper, discard just filtrate, with 0.22 μm of filtering with microporous membrane to sample bottle, treat that machine measures.
Wherein, the method for drafting of described typical curve is as follows:
Standard working solution is measured under the chromatographic condition of step (2), by concentration (mg.L
-1) and chromatographic peak area between relation drawing standard curve, curvilinear equation is chromatographic peak area=0.399Amt
1.330, r2 is 0.9991;
The preparation of described isomaltoketose standard working solution: pipette above-mentioned isomaltoketose standard reserving solution 0.5mL, 1mL, 3mL, 8mL respectively in 10mL volumetric flask with liquid-transfering gun, more namely obtain to scale the typical curve working fluid that concentration is 0.0550mg/mL, 0.1100g/mL, 0.3300mg/mL, 0.8800mg/mL and 1.0002mg/mL with deionized water constant volume respectively.
The preparation of described isomaltoketose standard reserving solution: accurately weigh 0.0501g isomaltoketose in small beaker, to transfer to after adding deionized water dissolving in 50ml volumetric flask and constant volume, obtains the isomaltoketose standard reserving solution that concentration is 1.002mg/mL.
The high performance liquid chromatography evaporative light-scattering detector approach of isomaltoketose assay in food provided by the present invention, sucrose in food can be separated with isomaltoketose by described method completely; The detection limit of isomaltoketose is low to moderate 30ug.mL
-1; In 80 ~ 1200 μ g.mL-1, working curve is linearly good, and correlation coefficient r=0.9991, the recovery is between 85.6% ~ 98.1%.Precision (n=6) is 2.78% ~ 2.92%.And method pre-treatment provided by the present invention is simple, be easy to operation, detectability is low, and good in range of linearity internal linear, related coefficient is high, and measurement result degree of accuracy is high, is the effective ways detecting isomaltoketose in food.
Accompanying drawing explanation
What Fig. 1 showed isomaltoketose and sucrose in embodiment 3 sample is separated the high-efficient liquid phase chromatogram detected.
Fig. 2 shows the typical curve of embodiment 4 isomaltoketose.
Embodiment
In order to make those skilled in the art understand technical scheme of the present invention better, below disclose further some non-limiting embodiments the present invention is described in further detail.
Material, reagent and instrument:
1, sample and reagent
Sample: candy (comprehensive yogurt taste fruit drops, manufacturer: Jun Xuan international marketing company limited; The tasty and refreshing happy vanilla thoat-soothing drops sugar-free of Switzerland Ricola, manufacturer: the tasty and refreshing happy company limited of Switzerland Ricola)
Reagent: isomaltoketose, deionized water, it is pure that potassium ferrocyanide, zinc acetate are analysis.
2, key instrument equipment
High performance liquid chromatography cascade evaporation light scattering detector: 1260 type high performance liquid chromatographs, 380-LC evaporative light-scattering detector manufacture by Anjelen Sci. & Tech. Inc.
Balance: GH-252 type analysis balance, sensibility reciprocal 0.0001g, range 0-250g, Japanese AND company.
Numerical control supersonic cleaning apparatus: KQ-500DE type, Ultrasonic Cleaning instrument company limited of city of Kunshan.
Liquid-transfering gun:
p1000 type, range: 200-1000 μ L, Gilson company;
f2 type, range: 0.5-5mL, Thermo Scientific Inc.
The preparation of the various solution of embodiment 1
The preparation of isomaltoketose standard reserving solution: accurately weigh 0.0501g (being accurate to 0.0001g) isomaltoketose in small beaker, to transfer to after adding deionized water dissolving in 50ml volumetric flask and constant volume, obtain the isomaltoketose standard reserving solution that concentration is 1.002mg/mL.
The preparation of acetic acid zinc solution: weigh 22.00g zinc acetate [Zn (CH
3cOO)
22H
2o] be dissolved in a small amount of water, add 3mL glacial acetic acid, thin up is to 100mL.
The preparation of potassium ferrocyanide solution: weigh 10.60g potassium ferrocyanide [K
4fe (CN)
63H
2o] water-soluble after be diluted to 100mL.
The preparation of isomaltoketose standard working solution: pipette above-mentioned isomaltoketose standard reserving solution 0.5mL, 1mL, 3mL, 8mL respectively in 10mL volumetric flask with liquid-transfering gun, more namely obtain to scale the typical curve working fluid that concentration is 0.0550mg/mL, 0.1100g/mL, 0.3300mg/mL, 0.8800mg/mL and 1.0002mg/mL with deionization constant volume respectively.
The selection of embodiment 2 instrument condition
Chromatographic column: Hypersil APS-2 (NH2), 5 μm, 250 × 4.6mm; Mobile phase is acetonitrile/deionized water (V/V) is 88: 12; Flow velocity: 1mL/min; Column temperature: 40 DEG C; Sampling volume: 10 μ L; Evaporative light-scattering detector nitrogen flow rate: 1.6L/min; Atomizer temperature: 50 DEG C; Evaporation tube temperature: 80 DEG C.
In embodiment 3 sample, isomaltoketose and sucrose is separated detection
Sample pre-treatments: weigh about 0.2g sample in small beaker, transfer in 25mL volumetric flask after being about 10mL ultrasonic dissolution sample with deionized water, slowly add potassium ferrocyanide solution, each 1mL of acid acetic acid zinc solution, add deionized water constant volume to scale, mixing, leave standstill 30min, filter with dry filter paper, discard just filtrate and be about 2mL, with 0.22 μm of filtering with microporous membrane to sample bottle, treat that machine measures;
Upper machine is separated and detects: setting chromatographic condition, chromatographic column: Hypersil APS-2 (NH2), 5 μm, 250 × 4.6mm; Mobile phase: acetonitrile: deionized water=88: 12; Flow velocity: 1mL/min; Column temperature: 40 DEG C; Sampling volume: 10 μ L; Evaporative light-scattering detector nitrogen flow rate: 1.6L/min; Atomizer temperature: 50 DEG C; Evaporation tube temperature: 80 DEG C, injects the high performance liquid chromatography cascade evaporation light scattering detector set and carries out separation detection by the above-mentioned sample handled well.Testing result is as shown in Figure 1: isomaltoketose is separated completely with sucrose.
Implement the mensuration of isomaltoketose content in 4 samples
1, production standard curve
The chromatographic condition that standard embodiment 1 prepared uses solution to optimize in embodiment 2 measures, by concentration (mg.L
-1) and peak area between relation drawing standard curve, curvilinear equation is Area=0.399Amt
1.330, r2 is 0.9991, and typical curve is shown in Fig. 2;
2, the range of linearity, detection limit
In blank sample, add a certain amount of standard solution, make the ratio of the peak height of isomaltoketose and noise close to 3 (i.e. S/N=3).This concentration is minimal detectable concentration.From experiment, the minimal detectable concentration of this method is 30ug.mL
-1.
Embodiment 5 recovery and precision
Get blank candy, each three of beverage, adds the isomaltoketose of high, medium and low three concentration levels respectively, and each level does three parallel laboratory tests, analyzes the recovery and precision, the results are shown in Table 1.
Table 1 recovery and precision
Embodiment 6 sample determination
By the sample that embodiment 1 is handled well, under embodiment 2 chromatographic condition, the list of ingredients of 3 kinds of samples is indicated that representative sample solution containing isomaltoketose and standard solution carry out stratographic analysis.Result shows, in surveyed 3 kinds of samples, all contain sucrose, the sample that isomaltoketose has contains, and what have does not detect, and its result is as shown in table 2:
Table 2
| Project | Candy 1 | Candy 2 | Beverage |
| Isomaltoketose | 28.37 | Do not detect | Do not detect |
。
The foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, for a person skilled in the art, the present invention can have various modifications and variations.Within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (6)
1. the method for separating and detecting of sucrose and isomaltoketose in food, is characterized in that adopting Hypersil APS-2 (NH
2) chromatographic column, its specification is 250mm × 4.6mm × 5 μm, and mobile phase is the volume ratio 88 of acetonitrile and deionized water
:12 ~ 85
:15, flow velocity is 1mL/min, and column temperature is 40; Evaporative light-scattering detector selected by detecting device, and nitrogen flow rate is 1.6L/min, and atomizer temperature is 50 DEG C, and evaporation tube temperature is 80 DEG C.
2. the assay method of the content of isomaltoketose in food, is characterized in that comprising the following steps:
(1) pre-treatment of sample;
(2) setting device:
The chromatographic condition of high performance liquid chromatograph workstation is set: adopt Hypersil APS-2 (NH
2) chromatographic column, its specification is 250mm × 4.6mm × 5 μm, and mobile phase is the volume ratio 88: 12 ~ 85: 15 of acetonitrile and deionized water, and flow velocity is 1mL/min, and column temperature is 40; Evaporative light-scattering detector selected by detecting device, and nitrogen flow rate is 1.6L/min, and atomizer temperature is 50 DEG C, and evaporation tube temperature is 80 DEG C;
(3) drawing standard curve, obtains typical curve equation;
(4) content of isomaltoketose in sample is detected: what arranged by the sample implantation step (2) after step (1) pre-treatment enters duty performance liquid chromatographic column, sample size is 10 microlitres, calculate the peak area under same retention time, substitute into the typical curve equation of step (3) gained, calculate the content of isomaltoketose in testing sample.
3. assay method as claimed in claim 2, it is characterized in that the pre-treatment of step (1) described sample comprises the following steps: transfer in volumetric flask by after sample deionized water dissolving, slowly add potassium ferrocyanide solution, acid acetic acid zinc solution, add deionized water constant volume to scale, mixing, leave standstill 30min, filter with dry filter paper, discard just filtrate, with 0.22 μm of filtering with microporous membrane to sample bottle, treat that machine measures.
4. assay method as claimed in claim 2, is characterized in that the method for drafting of step (3) described typical curve is as follows:
Measured under the chromatographic condition of step (2) by isomaltoketose standard working solution, by the relation drawing standard curve between concentration and chromatographic peak area, curvilinear equation is chromatographic peak area=0.399Amt
1.330, r2 is 0.9991.
5. assay method as claimed in claim 4, it is characterized in that the compound method of described isomaltoketose standard working solution is as follows: pipette above-mentioned isomaltoketose standard reserving solution 0.5mL, 1mL, 3mL, 8mL respectively in 10mL volumetric flask with liquid-transfering gun, more namely obtain to scale the typical curve working fluid that concentration is 0.0550mg/mL, 0.1100g/mL, 0.3300mg/mL, 0.8800mg/mL and 1.0002mg/mL with deionized water constant volume respectively.
6. assay method as claimed in claim 5, it is characterized in that the compound method of described isomaltoketose standard reserving solution is as follows: accurately weigh 0.0501g isomaltoketose in small beaker, to transfer to after adding deionized water dissolving in 50ml volumetric flask and constant volume, obtain the isomaltoketose standard reserving solution that concentration is 1.002mg/mL.
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| CN201510319842.8A CN104849395A (en) | 2015-06-11 | 2015-06-11 | Method for detecting isomaltulose in food |
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|---|---|---|---|
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111796034A (en) * | 2020-06-18 | 2020-10-20 | 广东省生物工程研究所（广州甘蔗糖业研究所） | Method for separating and detecting isomaltulose in food |
| CN111812231A (en) * | 2020-06-29 | 2020-10-23 | 广东省生物工程研究所（广州甘蔗糖业研究所） | Method for separating and detecting sucrose/isomaltulose in food |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111796034A (en) * | 2020-06-18 | 2020-10-20 | 广东省生物工程研究所（广州甘蔗糖业研究所） | Method for separating and detecting isomaltulose in food |
| CN111812231A (en) * | 2020-06-29 | 2020-10-23 | 广东省生物工程研究所（广州甘蔗糖业研究所） | Method for separating and detecting sucrose/isomaltulose in food |
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