CN108362691A - The method for measuring sweet broomcorn straw dry sample soluble sugar content - Google Patents
The method for measuring sweet broomcorn straw dry sample soluble sugar content Download PDFInfo
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- CN108362691A CN108362691A CN201810030850.4A CN201810030850A CN108362691A CN 108362691 A CN108362691 A CN 108362691A CN 201810030850 A CN201810030850 A CN 201810030850A CN 108362691 A CN108362691 A CN 108362691A
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Abstract
The present invention provides a kind of method measuring sweet broomcorn straw dry sample soluble sugar content, and step includes：The configuration of reagent；Sample measures；The sugar content of sweet broomcorn straw is judged using the light absorption value of colorimetric method for determining compound.Phenol sulfuric acid procedure measurement total Soluble Sugar method is simple, and reagent is cheap, and laboratory apparatus used is less, and sensitivity is higher, is not influenced substantially by existing for protein in experimentation, and the color generated can stablize 3h or so, is convenient for test operation.
Description
Technical field
The present invention relates to sweet broomcorn straw dry sample soluble sugar content detection technique field more particularly to a kind of measurement sweet tea are high
The method of fine strain of millet cane dry sample soluble sugar content.
Background technology
The purposes of sugar grass is very extensive, it not only produces grain, and also production sugar, syrup, can also cook wine, alcohol and monosodium glutamate,
Fiber can for a long time be planted with papermaking, sugar grass as feeding and sugar crop.Sugar grass harvest time storage capacity is big, it is difficult to
It is all processed in short time, and fresh sample is stored inconveniently, it is easy to cause the loss of sugar grass.Therefore, sweet broomcorn straw is fresh
It can then be stored for a long time after sample drying.But after sample drying, what the sugar content inside cane has change, it is necessary to build
Found a kind of assay method of reliable and stable sweet broomcorn straw dry sample soluble sugar content.
Invention content
The purpose of the present invention is to provide a kind of methods measuring sweet broomcorn straw dry sample soluble sugar content, in solution
State technical problem.
The present invention using following technical scheme in order to solve the above technical problems, realized：
A method of measuring sweet broomcorn straw dry sample soluble sugar content, which is characterized in that include the following steps：
One, the configuration of reagent
A, 9g/ml phenol solutions：By phenol crystal, density 1.017g/ml dissolves in 60 DEG C of water-baths, takes 88.496ml
Liquid phenol is added in the volumetric flask containing 10ml distilled water；
B, 10mg/ml sucrose standards liquid：Sucrose 1g is weighed, in 100ml volumetric flasks, after adding 5ml distilled water to be pre-dissolved, slowly
The 0.25ml concentrated sulfuric acids are added, fully shakes up immediately, is placed at room temperature for 30s, continue to be slowly added to the 0.25ml concentrated sulfuric acids, fully shake immediately
It is even, it is placed at room temperature for 1min, with distilled water constant volume 100ml；
C, 100 μ g/ml sucrose solutions：The sucrose standard liquid 1ml for drawing 10mg/ml, is settled to 100ml；
Two, sample measures
A, standard curve making：
A1 substrate reactions liquids prepare：Prepare 20ml scale test tubes 7, number consecutively：0-7, wherein No. 0 is control tube), 1-
No. 6 are sample cell, according to the form below configuration testing sample, total volume 2ml：
A2 reaction solutions configure：9g/ml phenol solution 1ml are added in 7 test tubes successively, after mixing well, are tried respectively to 7
It manages and is rapidly added the 5ml concentrated sulfuric acids successively, within time interval 10s；
A3 samples measure：Reaction solution places 30min at room temperature, and absorbance is measured under 485nm wavelength, draws standard curve,
Find out standard straight-line equation；
B, sweet broomcorn straw dry sample soluble sugar measures
B1 sample treatments
One cane of sugar grass is taken, drying to constant weight；Cane dry sample is crushed using plant sample crushing machine, the sieving of 50 mesh
It is loaded on afterwards spare in polybag；
The extraction of B2 total Soluble Sugars
3 parts of sweet broomcorn straw powder is weighed, is respectively placed in three scale test tubes, 0.3g/ parts, is divided in three scale test tubes
Not Jia Ru 8ml distilled water, cork closure boils 30min extraction total reducing sugars in boiling water, and filter paper filters extracting solution to 25ml volumetric flasks
In, continue to rinse test tube 2-3 times, constant volume is spare；
B3 is measured
1. drawing in above extracting solution 0.5ml to 3 scale test tube, respectively plus after distilled water 1.5ml；
2. being rapidly added the phenol solution 1ml of 9g/ml, mixing is rocked immediately, is placed in 20s on rack for test tube；
3. being slowly added to the 5ml concentrated sulfuric acids, immediately slight wobble mixing, liquid splash is prevented；
4. 10mim is placed in 4 DEG C of refrigerators, colour developing,
5. 485nm wavelength measures absorbance and acquires sugar content according to standard curve；
Three, result calculates
The sugared content of sample is calculated as follows
Soluble sugar content
In formula, m：The sugared content checked in from standard curve, unit μ g；Vt is extracting liquid volume, units/ml；Vs is when measuring
The sample volume taken, units/ml；N is extension rate；m0For sample quality.
Further, in the B2 small steps of step 2, to ensure that test tube is immersed in boiling water during heating, test tube cannot paste
Nearly walls of beaker, it is ensured that be heated evenly, and the time of each test tube heating will be consistent.
Soluble sugar in sugar grass body is primarily referred to as that the monosaccharide and oligosaccharide of water can be dissolved in.Sugar the concentrated sulfuric acid effect under,
It is dehydrated the furfural or methyl furfural generated, easily with phenol condensation at a kind of orange red compound (within the scope of 10-100mg),
The depth of color is directly proportional to the content of sugar.Therefore, can utilizing colorimetric method, (485nm wavelength, the compound have absorption maximum
Peak) light absorption value that measures compound judges the sugar content of sweet broomcorn straw.
The beneficial effects of the invention are as follows：
Phenol sulfuric acid procedure provided by the invention measures total Soluble Sugar, and method is simple, and reagent is cheap, laboratory apparatus used compared with
Few, sensitivity is higher, is not influenced substantially by existing for protein in experimentation, and the color generated can stablize 3h or so,
Convenient for test operation.Storage when can be produced and processed for sweet sorghum stalk provides Important Theoretic Foundation.
Description of the drawings
Fig. 1 is the absorbance standard curve schematic diagram of invention；
Fig. 2 is the sugar grass different growing soluble sugar content schematic diagram of invention.
Specific implementation mode
In order to make the technical means, the creative features, the aims and the efficiencies achieved by the present invention be easy to understand, tie below
Specific embodiment is closed, the present invention is further explained, but following embodiments are only the preferred embodiment of the present invention, and not all.
Based on the implementation example in the implementation mode, those skilled in the art's obtained other realities without making creative work
Example is applied, protection scope of the present invention is belonged to.Experimental method in following embodiments is unless otherwise specified conventional method,
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
Embodiment 1
Instrument and equipment
Spectrophotometer, water-bath, scale test tube, measuring pipette, 10ml acid burets, scissors, rack for test tube, brandreth
Platform, funnel, small beaker, 80 μm of filter paper.
The configuration of reagent
1,9g/ml phenol solutions：Phenol crystal (density 1.017g/ml) is dissolved in 60 DEG C of water-baths, takes 88.496ml
Liquid phenol is added in the volumetric flask containing 10ml distilled water, can save 45 days at room temperature.
2,10mg/ml sucrose standards liquid：Weigh sucrose (analysis is pure) 1g adds 5ml distilled water pre- in 100ml volumetric flasks
After molten, the 0.25ml concentrated sulfuric acids are slowly added to, are fully shaken up immediately, 30s is placed at room temperature for, continues to be slowly added to the 0.25ml concentrated sulfuric acids,
It fully shakes up immediately, is placed at room temperature for 1min, with distilled water constant volume 100ml.
3,100 μ g/ml sucrose solutions：Sucrose standard liquid (10mg/ml) 1ml is drawn, 100ml is settled to.
Sample measures
1, standard curve making：
1.1 substrate reactions liquids prepare：Prepare 20ml scale test tubes 7, number consecutively：0 (control tube), 1-6 (sample cell,
Repeat twice), according to the form below configuration testing sample (total volume 2ml)：
1.2 reaction solutions configure：9g/ml phenol solution 1ml are added in 7 test tubes successively, after mixing well, respectively to 7
Test tube is rapidly added the 5ml concentrated sulfuric acids successively (10s or so, time interval can not be too long).
1.3 samples measure：Reaction solution places 30min at room temperature, and absorbance is measured under 485nm wavelength, draws standard curve,
Find out standard straight-line equation (Fig. 1).
2, sweet broomcorn straw dry sample soluble sugar measures
2.1 sample treatment
One cane of sugar grass is taken, drying to constant weight.Cane dry sample is crushed using plant sample crushing machine, the sieving of 50 mesh
It is loaded on afterwards spare in polybag.
The extraction of 2.2 total Soluble Sugars
3 parts of sweet broomcorn straw powder (0.3g/ parts) is weighed, (3 repetitions) is placed in three scale test tubes, is separately added into
8ml distilled water, cork closure boil 30min extraction total reducing sugars in boiling water, and filter paper filters in extracting solution to 25ml volumetric flasks, after
Continuous rinsing test tube 2-3 times, constant volume is spare.
Remarks：To ensure that test tube is immersed in boiling water during heating, test tube cannot be close to walls of beaker, it is ensured that heated equal
It is even, and heat time to be consistent.
2.3 measuring
1. drawing in extracting solution 0.5ml to 3 scale test tube of the above processing (3 repetitions), add distilled water 1.5ml respectively
Afterwards；
2. being rapidly added 1ml phenol solutions (9g/ml), mixing is rocked immediately, is placed in 20s on rack for test tube；
3. being slowly added to the 5ml concentrated sulfuric acids, immediately slight wobble mixing, liquid splash is prevented；
4. 4 DEG C of refrigerators place 10mim, develop the color,
5. measuring absorbance (485nm wavelength), according to standard curve, sugar content is acquired.
As a result it calculates
The sugared content of sample is calculated as follows
Soluble sugar content
In formula, m：The sugared content (μ g) checked in from standard curve；Vt is extracting liquid volume (ml)；Vs takes when measuring
Sample volume (ml)；N is extension rate；M0 is sample quality.
Data result is analyzed
As shown in Fig. 2, this experiment has chosen the cane sample of three period sugar grasses altogether, fresh sample is respectively adopted and does
Sample measures soluble sugar content in sweet broomcorn straw using improved Phenol sulfuric acid procedure.As a result (Fig. 2) is shown：In this measurement
Under method, the soluble sugar content index of sweet broomcorn straw dry sample and fresh sample is very close, and explanation can utilize this improvement
Phenol sulfuric acid procedure measure the soluble sugar in sweet broomcorn straw dry sample, storage when this is produced and processed for sweet sorghum stalk provides
Important Theoretic Foundation.
The basic principles, main features and advantages of the present invention have been shown and described above.The technology of the industry
For personnel it should be appreciated that the present invention is not limited to the above embodiments, described in the above embodiment and specification is only the present invention
Preference, be not intended to limit the present invention, without departing from the spirit and scope of the present invention, the present invention also has various
Changes and improvements, these changes and improvements all fall within the protetion scope of the claimed invention.The claimed scope of the invention is by institute
Attached claims and its equivalent thereof.
Claims (2)
1. a kind of method measuring sweet broomcorn straw dry sample soluble sugar content, which is characterized in that include the following steps：
One, the configuration of reagent
A, 9g/ml phenol solutions：By phenol crystal, density 1.017g/ml dissolves in 60 DEG C of water-baths, takes 88.496ml phenol
Liquid is added in the volumetric flask containing 10ml distilled water；
B, 10mg/ml sucrose standards liquid：Sucrose 1g is weighed, in 100ml volumetric flasks, after adding 5ml distilled water to be pre-dissolved, is slowly added to
The 0.25ml concentrated sulfuric acids, fully shake up immediately, are placed at room temperature for 30s, continue to be slowly added to the 0.25ml concentrated sulfuric acids, fully shake up immediately,
It is placed at room temperature for 1min, with distilled water constant volume 100ml；
C, 100 μ g/ml sucrose solutions：The sucrose standard liquid 1ml for drawing 10mg/ml, is settled to 100ml；
Two, sample measures
A, standard curve making：
A1 substrate reactions liquids prepare：Prepare 20ml scale test tubes 7, number consecutively：0-7, wherein No. 0 is control tube), No. 1-6
For sample cell, according to the form below configuration testing sample, total volume 2ml：
A2 reaction solutions configure：9g/ml phenol solution 1ml are added in 7 test tubes successively, after mixing well, respectively to 7 test tubes according to
It is secondary to be rapidly added the 5ml concentrated sulfuric acids, within time interval 10s；
A3 samples measure：Reaction solution places 30min at room temperature, and absorbance is measured under 485nm wavelength, draws standard curve, finds out
Standard straight-line equation；
B, sweet broomcorn straw dry sample soluble sugar measures
B1 sample treatments
One cane of sugar grass is taken, drying to constant weight；Cane dry sample is crushed using plant sample crushing machine, is filled after the sieving of 50 mesh
It is spare in polybag；
The extraction of B2 total Soluble Sugars
3 parts of sweet broomcorn straw powder is weighed, is respectively placed in three scale test tubes, 0.3g/ parts, is added respectively in three scale test tubes
Entering 8ml distilled water, cork closure boils 30min extraction total reducing sugars in boiling water, and filter paper filters in extracting solution to 25ml volumetric flasks,
Continue to rinse test tube 2-3 times, constant volume is spare；
B3 is measured
1. drawing in above extracting solution 0.5ml to 3 scale test tube, respectively plus after distilled water 1.5ml；
2. being rapidly added the phenol solution 1ml of 9g/ml, mixing is rocked immediately, is placed in 20s on rack for test tube；
3. being slowly added to the 5ml concentrated sulfuric acids, immediately slight wobble mixing, liquid splash is prevented；
4. 10mim is placed in 4 DEG C of refrigerators, colour developing,
5. 485nm wavelength measures absorbance and acquires sugar content according to standard curve；
Three, result calculates
The sugared content of sample is calculated as follows
In formula, m：The sugared content checked in from standard curve, unit μ g；Vt is extracting liquid volume, units/ml；Vs is to be taken when measuring
Sample volume, units/ml；N is extension rate；m0For sample quality.
2. the method according to claim 1 for measuring sweet broomcorn straw dry sample soluble sugar content, it is characterised in that：Step
In two B2 small steps, to ensure that test tube is immersed in boiling water during heating, test tube cannot be close to walls of beaker, it is ensured that heated equal
It is even, and each test tube heating time to be consistent.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110174361A (en) * | 2019-05-05 | 2019-08-27 | 贵州中烟工业有限责任公司 | A kind of measurement method of total reducing sugar |
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| CN105548055A (en) * | 2015-12-11 | 2016-05-04 | 中国农业科学院棉花研究所 | Determination method of soluble sugar in cotton fiber |
| CN107314986A (en) * | 2017-06-12 | 2017-11-03 | 华中农业大学 | A kind of method that utilization near infrared spectrum detects rape stem soluble sugar content |
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2018
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Patent Citations (5)
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| CN102879347A (en) * | 2012-10-18 | 2013-01-16 | 中国科学院西北高原生物研究所 | Detection method of polysaccharide content in medlar polysaccharide extract doped with saccharide chaff interferent |
| CN103604763A (en) * | 2013-12-03 | 2014-02-26 | 广东中测食品化妆品安全评价中心有限公司 | Detection method for health-care food functional components |
| CN103837384A (en) * | 2014-01-21 | 2014-06-04 | 吉林大学 | Measuring method of soluble saccharide |
| CN105548055A (en) * | 2015-12-11 | 2016-05-04 | 中国农业科学院棉花研究所 | Determination method of soluble sugar in cotton fiber |
| CN107314986A (en) * | 2017-06-12 | 2017-11-03 | 华中农业大学 | A kind of method that utilization near infrared spectrum detects rape stem soluble sugar content |
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