CN110174361A - A kind of measurement method of total reducing sugar - Google Patents
A kind of measurement method of total reducing sugar Download PDFInfo
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- CN110174361A CN110174361A CN201910369015.8A CN201910369015A CN110174361A CN 110174361 A CN110174361 A CN 110174361A CN 201910369015 A CN201910369015 A CN 201910369015A CN 110174361 A CN110174361 A CN 110174361A
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- 239000012488 sample solution Substances 0.000 claims abstract description 36
- 238000012360 testing method Methods 0.000 claims abstract description 20
- 238000010521 absorption reaction Methods 0.000 claims abstract description 17
- 238000002835 absorbance Methods 0.000 claims abstract description 16
- 239000007787 solid Substances 0.000 claims abstract description 16
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- 238000005259 measurement Methods 0.000 claims abstract description 13
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- 150000001720 carbohydrates Chemical class 0.000 claims abstract description 9
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- 238000000034 method Methods 0.000 claims description 26
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- 239000008103 glucose Substances 0.000 claims description 11
- 239000012086 standard solution Substances 0.000 claims description 8
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- 230000008033 biological extinction Effects 0.000 claims description 2
- 239000008121 dextrose Substances 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 238000001514 detection method Methods 0.000 abstract description 10
- 239000003795 chemical substances by application Substances 0.000 abstract description 8
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- 231100000206 health hazard Toxicity 0.000 abstract 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 7
- 230000000052 comparative effect Effects 0.000 description 5
- RJGDLRCDCYRQOQ-UHFFFAOYSA-N anthrone Chemical compound C1=CC=C2C(=O)C3=CC=CC=C3CC2=C1 RJGDLRCDCYRQOQ-UHFFFAOYSA-N 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
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- 230000007935 neutral effect Effects 0.000 description 4
- 239000013558 reference substance Substances 0.000 description 4
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 4
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- 238000004737 colorimetric analysis Methods 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
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- 241000196324 Embryophyta Species 0.000 description 2
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- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 2
- 235000003140 Panax quinquefolius Nutrition 0.000 description 2
- 229920002230 Pectic acid Polymers 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 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 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- MWPLVEDNUUSJAV-UHFFFAOYSA-N anthracene Chemical compound C1=CC=CC2=CC3=CC=CC=C3C=C21 MWPLVEDNUUSJAV-UHFFFAOYSA-N 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 235000008434 ginseng Nutrition 0.000 description 2
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- 239000000463 material Substances 0.000 description 2
- CTYRPMDGLDAWRQ-UHFFFAOYSA-N phenyl hydrogen sulfate Chemical compound OS(=O)(=O)OC1=CC=CC=C1 CTYRPMDGLDAWRQ-UHFFFAOYSA-N 0.000 description 2
- 239000010318 polygalacturonic acid Substances 0.000 description 2
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- -1 saccharide compound Chemical class 0.000 description 2
- 210000003491 skin Anatomy 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
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- 239000005720 sucrose Substances 0.000 description 2
- 239000001117 sulphuric acid Substances 0.000 description 2
- 235000011149 sulphuric acid Nutrition 0.000 description 2
- 239000012085 test solution Substances 0.000 description 2
- 238000002371 ultraviolet--visible spectrum Methods 0.000 description 2
- 229920001285 xanthan gum Polymers 0.000 description 2
- 229940082509 xanthan gum Drugs 0.000 description 2
- 235000010493 xanthan gum Nutrition 0.000 description 2
- 239000000230 xanthan gum Substances 0.000 description 2
- 206010037423 Pulmonary oedema Diseases 0.000 description 1
- 235000009754 Vitis X bourquina Nutrition 0.000 description 1
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- 240000006365 Vitis vinifera Species 0.000 description 1
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- 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 description 1
- VQLYBLABXAHUDN-UHFFFAOYSA-N bis(4-fluorophenyl)-methyl-(1,2,4-triazol-1-ylmethyl)silane;methyl n-(1h-benzimidazol-2-yl)carbamate Chemical compound C1=CC=C2NC(NC(=O)OC)=NC2=C1.C=1C=C(F)C=CC=1[Si](C=1C=CC(F)=CC=1)(C)CN1C=NC=N1 VQLYBLABXAHUDN-UHFFFAOYSA-N 0.000 description 1
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- 238000007689 inspection Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000001225 nuclear magnetic resonance method Methods 0.000 description 1
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
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- 239000000376 reactant Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 239000009452 shenxiong glucose Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/314—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry with comparison of measurements at specific and non-specific wavelengths
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/314—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry with comparison of measurements at specific and non-specific wavelengths
- G01N2021/3148—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry with comparison of measurements at specific and non-specific wavelengths using three or more wavelengths
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/314—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry with comparison of measurements at specific and non-specific wavelengths
- G01N2021/3155—Measuring in two spectral ranges, e.g. UV and visible
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- Physics & Mathematics (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
Abstract
The present invention relates to total reducing sugar detection technique fields, in particular to a kind of measurement method of total reducing sugar includes the following steps: the preparation of 1) unknown sugared sample solution: soluble solids carbohydrate sample being taken to be configured to solution of the concentration in the section 0.01-0.07g/L with ultrapure water;2) unknown sugared sample solution, acid solution the total sugar content measurement of unknown sugared sample solution: are sequentially added in test tube, it is placed in ice water bath environment after mixing, it is placed in room temperature environment, then measures its absorbance at maximum absorption band with ultraviolet-visual spectrometer;3) total sugar content of unknown sugared sample solution calculates: the total sugar content in soluble solids sugar-like sheet is calculated using the configuration proportion of standard curve regression equation and unknown sugared sample solution;Solve the problems, such as that accuracy is low, the reaction time is long for color developing agent healthhazard and detection.
Description
Technical field
The present invention relates to total reducing sugar detection technique field, in particular to a kind of measurement method of total reducing sugar.
Background technique
The concentration of total reducing sugar in solution is detected in each research field, such as petrochemical industry, medicine, food field are all particularly significant
's.At present there are many kinds of the analysis methods of detection total reducing sugar, such as chromatography, capillary electrophoresis, infra-red sepectrometry, nuclear magnetic resonance
Method, evaporative light-scattering spectroscopic methodology, colorimetric method etc..But these methods have cost more and greatly, excessively rely on expensive instrument, time-consuming, inspection
The disadvantages of surveying high operation requirements.
Colorimetric method becomes detection always because of the advantages that its operation is relatively easy, testing cost is cheap, detecting instrument is relatively cheap
The common method of sugar;The basic principle is that the product and the reaction of specific color developing agent after saccharide compound is hydrolyzed generate to have and add lustre to
The derivative compound of group, this derivative can generate absorption at the Vis scan wave band of uv-vis spectra scanner
Peak.Phenol-sulphate acid method is hydrolyzed dewatered in concentrated sulfuric acid as the most widely used colorimetric method using saccharide compound
Product can generate a kind of coloured compound with phenol reactant, and this compound has maximum absorption band, but the party at 490nm
Method still has following problems: 1) color developing agent phenol used in has very big harm to the health of human body, and phenol steam is to skin
Skin, eyes and respiratory system are all corrosive, and contact phenol will cause scytitis or second level very to skin repeatedly or for a long time
To burning for three-level;Same sucking phenol will cause pulmonary edema for a long time or repeatedly, influence central nervous system.2) reaction time
Too long, the time for generally requiring 30min or more could fully reacting.3) accuracy is low;Make standard curve with glucose, and it is practical
Carbohydrate complicated composition in the sample of life, not only contain glucose, also contain acid sugar, due to the phenol sulfuric acid of various carbohydrates
The molar absorptivity rate difference of the substance generated after derivatization is larger, and especially neutral sugar and acid sugar is the most significant, so cause
The resultant error of detection is larger.
Patent publication No. CN105510257A discloses a kind of method for measuring total sugar content in shenxiong glucose injection,
1) reference substance solution preparation step: precision weighs glucose control product 10mg, sets in 100ml measuring bottle, adds water to make to dissolve and dilutes
To scale, the reference substance solution to get 0.1mg/ml is shaken up;2) preparation step of test solution: precision measures ginseng rhizome of chuanxiong grape
Sugared injection 0.5ml, sets in 25ml volumetric flask, adds water 5.0ml, then plus 2.0ml hydrochloric acid, shake up;It hydrolyzes 1 hour, puts at 95 DEG C
It is cold, scale is added water to, is shaken up;The accurate 1.0ml that measures is set in 10ml measuring bottle again, is diluted with water to scale, is shaken up to get test sample
Solution;3) sulfuric acid solution preparation steps: 760ml sulfuric acid is diluted with water 1000ml to obtain the final product;4) Anthrone Sulphuric acid solution prepares step
It is rapid: 0.2g anthrone being taken to be dissolved in 100mL sulfuric acid solution.The same day, which prepares, to be used;5) determination step: precision measures test solution
1.0mL is set in 10mL tool plug test tube, is added Anthrone Sulphuric acid solution 8.0mL prepared by step 4) after adding water 1ml, is dipped in rapidly
It is cooling in ice-water bath, then in 100 DEG C of water-baths, accurately boil l0min taking-up, be cooled to room temperature to obtain measurement test sample with ice-water bath
Solution;Precision is measured for reference substance solution 1.0mL, is set in 10mL tool plug test tube, adds anthracene prepared by step 4) after adding water 1ml
Ketone-sulfuric acid solution 8.0mL, is dipped in rapidly in ice-water bath cooling, then in 100 DEG C of water-baths, accurately boils l0min taking-up, uses ice
Water-bath cooling to room temperature must measure reference substance solution;Its absorption value is measured in 626nm with ultraviolet-uisible spectrophotometer, calculates ginseng
Total sugar content in rhizome of chuanxiong glucose injection.But it also belongs to poisonous substance using anthrone, still without solve it is toxic harm and
The problem of multiple types sugar part influences the accuracy of measurement.
(Su Yushun et al. is in 2011 for document " application of the UV-VIS spectrophotometry in plant polyose assay "
Year deliver) and document " progress of uronic acid and neutral sugar analyzing detecting method in plant polyose " (Chen Hui et al. in
Deliver within 2011) all illustrate that the middle resulting result difference of different color developing agents is larger, and material molecule is to different wave length and spy
The radiation absorption degree of given wavelength is different and different material molecule is also different to the radiation absorption degree of certain wave strong point,
Thus illustrate that the use of color developing agent and sample multicomponent will affect the accuracy that total sugar content detects.
Summary of the invention
In order to solve the above technical problems, The present invention provides a kind of measurement methods of total reducing sugar.
It is realized especially by following technical scheme:
A kind of measurement method of total reducing sugar, includes the following steps:
1) preparation of unknown sugared sample solution: soluble solids carbohydrate sample is taken to be configured to concentration in 0.01- with ultrapure water
Solution in the section 0.07g/L;
2) the total sugar content measurement of unknown sugared sample solution: sequentially adding unknown sugared sample solution, acid solution in test tube,
It places in ice water bath environment, is placed in room temperature environment after mixing, then inhaled with ultraviolet-visual spectrometer in maximum
It receives and records its absorbance at peak;
3) total sugar content of unknown sugared sample solution calculates: utilizing standard curve regression equation and unknown sugared sample solution
Configuration proportion calculate soluble solids sugar-like sheet in total sugar content.
The work of the ultraviolet-visual spectrometer is from wavelength be 200 to wavelength be 800nm carry out full scan, in maximum inhale
It receives and records its absorbance at peak.
The wavelength of the maximum absorption band is 315nm.
The determination method of the maximum absorption band, includes the following steps:
1. glucose standards solution is prepared: taking dextrose standard sample dry to constant weight, be first configured to mother liquor with ultrapure water, so
Afterwards with mother liquor at the standard solution of more concentration gradients;
2. the determination of maximum absorption band: sequentially adding glucose standards solution, acid solution in test tube, exist after mixing
It places in ice water bath environment, is placed in room temperature environment, is then scanned with ultraviolet-visual spectrometer, in maximum absorption band
Place's record absorbance.
The standard curve regression equation is to calculate gained according to absorbance and corresponding concentration of standard solution.
The standard curve regression equation: Y=0.0762X-0.0017.
The concentration of the mother liquor is 0.2g/L.
The concentration of standard solution of more concentration gradients is followed successively by 0.01,0.03,0.05,0.07g/L.
The temperature of the drying is 103-107 DEG C.
The standing time is 4-6min.
The utility model has the advantages that
Using the method for the present invention, the reaction time is shortened under conditions of not using color developing agent, reduces the detection of total reducing sugar
Error, the present invention do not use color developing agent, improve detection safety and health, are reacted merely with acid and sugar, are contracted significantly
In the short reaction time, influence of the color developing agent to accuracy of detection is also reduced, and overcomes neutral sugar and acid sugar is mutually dry
The problem of disturbing and then increasing detection error.
Detailed description of the invention
Fig. 1: sugar juice concentration comparison chart (a is 1 measurement result of comparative example, and b is 1 measurement result of embodiment);
Fig. 2: reaction time comparison diagram (solid expression control methods is as a result, hollow expression the method for the present invention result).
Specific embodiment
Specific embodiments of the present invention will be described in further detail below.For those of skill in the art
For member, from detailed description of the invention, the features and advantages of the present invention be will be evident.
Embodiment 1
A kind of measurement method of total reducing sugar, includes the following steps:
1) preparation of unknown sugared sample solution: soluble solids carbohydrate sample is taken to be configured to concentration in 0.01- with ultrapure water
Solution in the section 0.07g/L;
2) 1mL unknown sugared sample solution, 5mL the total sugar content measurement of unknown sugared sample solution: are sequentially added in test tube
Concentrated sulfuric acid solution places 5min in ice water bath environment after mixing, and 5min is placed in room temperature environment, then with ultraviolet
Visible spectrophotometer from wavelength be 200 to wavelength be 800nm carry out full scan, its absorbance is recorded at 315nm;
3) total sugar content of unknown sugared sample solution calculates: utilizing standard curve regression equation and unknown sugared sample solution
Configuration proportion calculate soluble solids sugar-like sheet in total sugar content.
Embodiment 2
A kind of measurement method of total reducing sugar, includes the following steps:
1) preparation of unknown sugared sample solution: soluble solids carbohydrate sample is taken to be configured to concentration in 0.01- with ultrapure water
Solution in the section 0.07g/L;
2) 1mL unknown sugared sample solution, 5mL the total sugar content measurement of unknown sugared sample solution: are sequentially added in test tube
Concentrated sulfuric acid solution places 4min in ice water bath environment after mixing, and 6min is placed in room temperature environment, then with ultraviolet
Visible spectrophotometer from wavelength be 200 to wavelength be 800nm carry out full scan, its absorbance is recorded at 350nm;
3) total sugar content of unknown sugared sample solution calculates: utilizing standard curve regression equation and unknown sugared sample solution
Configuration proportion calculate soluble solids sugar-like sheet in total sugar content.
Embodiment 3
A kind of measurement method of total reducing sugar, includes the following steps:
1) preparation of unknown sugared sample solution: soluble solids carbohydrate sample is taken to be configured to concentration in 0.01- with ultrapure water
Solution in the section 0.07g/L;
2) 1mL unknown sugared sample solution, 5mL the total sugar content measurement of unknown sugared sample solution: are sequentially added in test tube
Concentrated sulfuric acid solution places 4min in ice water bath environment after mixing, and 4min is placed in room temperature environment, then with ultraviolet
Visible spectrophotometer from wavelength be 200 to wavelength be 800nm carry out full scan, its absorbance is recorded at 300nm;
3) total sugar content of unknown sugared sample solution calculates: utilizing standard curve regression equation and unknown sugared sample solution
Configuration proportion calculate soluble solids sugar-like sheet in total sugar content.
Comparative example 1
A kind of measurement method of total reducing sugar, includes the following steps:
1) preparation of unknown sugared sample solution: soluble solids carbohydrate sample is taken to be configured to concentration in 0.01- with ultrapure water
Solution in the section 0.07g/L;
2) 1mL unknown sugared sample solution, 1mL the total sugar content measurement of unknown sugared sample solution: are sequentially added in test tube
After the phenol solution of 5% concentration mixes, 5mL concentrated sulfuric acid solution is added, 10min vortex 30s is placed after mixing, in ice water
20min is placed in bath environment, be then 200 from wavelength with ultraviolet-visual spectrometer to wavelength is that 800nm carries out full scan, in
Its absorbance is recorded at 490nm;
3) total sugar content of unknown sugared sample solution calculates: utilizing standard curve regression equation and unknown sugared sample solution
Configuration proportion calculate soluble solids sugar-like sheet in total sugar content;
Test example 1
In order to check the error condition of the method for the present invention, glucose that the present inventor's compound concentration is 0.03g/L, fructose,
Sucrose, starch, glucan 1 (dextran), glucan 2 (actigum), xanthan gum, polygalacturonic acid standard solution, so
Embodiment 1 is respectively adopted in various sugar juices afterwards and the method for comparative example 1 measures its absorbance value, the relative error of two methods
Mean value comparing result is as shown in table 1:
Table 1
Sugar type | True value (g/L) | Phenol-sulphate acid method relative error mean value (%) | This method relative error mean value (%) |
Neutral sugar | 0.03 | 6.5 | 2.8 |
Acid sugar | 0.03 | 14.4 | 7.5 |
Meanwhile the present inventor also prepares glucose, fructose, sucrose, the starch, Portugal of 0.01g/L, 0.05g/L, 0.07g/L
Glycan 1 (dextran), glucan 2 (actigum), xanthan gum, polygalacturonic acid standard solution, be respectively adopted embodiment 1 with
And the method for comparative example 1 records its absorbance value, draws sugar juice concentration comparison chart, as (a is 1 measurement result of comparative example, b to Fig. 1
For 1 measurement result of embodiment) shown in.
The acquisition of method veracity and precision data: pass through extinction of the test various concentration known sample under this method
Angle value is brought into, and the standard curve that this method is established calculates measured value.And by true value and measured value be calculated standard deviation,
The results are shown in Table 2 for the rate of recovery:
Table 2
Test example 2
In order to check influence of the reaction time to test result after acid adding, the present inventor prepares the Portugal 0.01g/L and 0.07g/L
Grape sugar juice, respectively as follows:
Control methods: sequentially adding the phenol solution of 1mL glucose standards solution, 5% concentration of 1mL in test tube, mixes
After add 5mL concentrated sulfuric acid solution, place after mixing, then be placed in water bath and place, then use uv-vis spectra
Instrument is scanned, and absorbance is recorded at maximum absorption band;
The method of the present invention: 1mL glucose standards solution, 5mL concentrated sulfuric acid solution are sequentially added in test tube, after mixing
It places in ice water bath environment, is placed in room temperature environment, is then scanned with ultraviolet-visual spectrometer, in absorption maximum
Absorbance is recorded at peak;
Wherein, after acid adding the reaction time be 5min, 15min, 30min, 45min, 75min, 105min, 135min,
225min, and reaction time comparison diagram is drawn according to this, as (solid expression control methods is as a result, hollow expression the method for the present invention by Fig. 2
As a result shown in).
It should be appreciated that although the present invention has carried out clear explanation by above embodiments, without departing substantially from the present invention
Spirit and its essence in the case where, person of ordinary skill in the field make in accordance with the present invention it is various it is corresponding variation and
Amendment, but these corresponding variations and modifications all should belong to scope of protection of the claims of the invention.
Claims (9)
1. a kind of measurement method of total reducing sugar, which comprises the steps of:
1) preparation of unknown sugared sample solution: soluble solids carbohydrate sample is taken to be configured to concentration in 0.01- with ultrapure water
Solution in the section 0.07g/L;
2) unknown sugared sample solution, acid solution, mixing the total sugar content measurement of unknown sugared sample solution: are sequentially added in test tube
It places in ice water bath environment after uniformly, is placed in room temperature environment, then with ultraviolet-visual spectrometer in maximum absorption band
Place records its absorbance;
3) total sugar content of unknown sugared sample solution calculates: utilizing matching for standard curve regression equation and unknown sugared sample solution
Ratio processed calculates the total sugar content in soluble solids sugar-like sheet.
2. the measurement method of total reducing sugar as described in claim 1, which is characterized in that the work of the ultraviolet-visual spectrometer is from wave
A length of 200 carry out full scan to wavelength for 800nm, its absorbance is recorded at maximum absorption band.
3. the measurement method of total reducing sugar as described in claim 1, which is characterized in that the determination method of the maximum absorption band, including
Following steps:
1. glucose standards solution is prepared: taking dextrose standard sample dry to constant weight, be first configured to mother liquor with ultrapure water, then use
Mother liquor at more concentration gradients standard solution;
2. the determination of maximum absorption band: glucose standards solution, acid solution are sequentially added in test tube, after mixing in ice water
It is placed in bath environment, places in room temperature environment, be then scanned with ultraviolet-visual spectrometer, remembered at maximum absorption band
Record absorbance.
4. the measurement method of total reducing sugar as described in claim 1, which is characterized in that the standard curve regression equation is according to extinction
Degree and corresponding concentration of standard solution calculate gained.
5. the measurement method of total reducing sugar as described in claim 1, which is characterized in that the standard curve regression equation: Y=
0.0762X-0.0017。
6. the measurement method of total reducing sugar as described in claim 1, which is characterized in that the concentration of the mother liquor is 0.2g/L.
7. the measurement method of total reducing sugar as claimed in claim 3, which is characterized in that the concentration of standard solution of more concentration gradients according to
It is secondary be 0.01,0.03,0.05,0.07g/L.
8. the measurement method of total reducing sugar as claimed in claim 3, which is characterized in that the temperature of the drying is 103-107 DEG C.
9. the measurement method of total reducing sugar as claimed in claim 1 or 3, which is characterized in that the standing time is 4-6min.
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