CN108956499A - A method of using glucose in DNS method measurement soil - Google Patents
A method of using glucose in DNS method measurement soil Download PDFInfo
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- CN108956499A CN108956499A CN201810891591.4A CN201810891591A CN108956499A CN 108956499 A CN108956499 A CN 108956499A CN 201810891591 A CN201810891591 A CN 201810891591A CN 108956499 A CN108956499 A CN 108956499A
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- glucose
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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
Abstract
The invention belongs to Soil Testing, specifically a kind of method using glucose in DNS method measurement soil, comprising the following steps: step A, extract the glucose in soil；Step B, glucose leaching liquor is filtered；Step C, DNS reagent is produced；Step D, sample measures: passing through absorbance of the solution after measurement constant volume at λ=540nm, to calculate the surplus of glucose in soil in measurement incubation, in the soil incubation experiment using glucose, the method that can not have to isotope labelling can measure the glucose surplus retained in soil, and it is more accurate, the rate of recovery is high.
Description
Technical field
It is specifically a kind of to use glucose in DNS method measurement soil the invention belongs to Soil K+adsorption technical field
Method.
Background technique
Soil environment monitoring refers to through the measurement to the typical value for influencing soil environment quality factor, determines environmental quality
(or pollution level) and its variation tendency.We refer to soil environment monitoring at usually said soil monitoring, generally comprise cloth
The technology contents such as point sampling, sample preparation, analysis method, result characterization, statistics and quality evaluation, the glucose in soil
Nutrition can be provided to the bacterium in soil, therefore, the measurement of glucose content has the existence of bacterium in soil in soil
There is not replaceable effect.
Glucose culture can be used in the culture experiment of many soil at present, but microorganism is to grape during the cultivation process
The consumption of sugar can only often use in the test using the glucose of isotope labelling, and in the training using common glucose sugar
Support in test to be detected later, although this method comparison of isotope labelling is accurate, at high cost, test period
Long measuring accuracy is low, and detection efficiency is low, in consideration of it, the present invention provides a kind of side using glucose in DNS method measurement soil
Method has the following characteristics that
(1) present invention is detected by surplus of the DNS method to the glucose in the culture experiment of soil, can be accurate
Ground detects the surplus of glucose in soil, accelerates detection efficiency, saves the time.
(2) in the soil incubation experiment using glucose, the method that can not have to isotope labelling can measure soil
In the glucose surplus that retains, and more accurate, the rate of recovery is high.
Summary of the invention
In order to make up for the deficiencies of the prior art, a kind of side using glucose in DNS method measurement soil proposed by the present invention
Method, the present invention are detected by surplus of the DNS method to the glucose in the culture experiment of soil, can be accurately detected
The surplus of glucose in soil accelerates detection efficiency, saves the time；It, can be in the soil incubation experiment using glucose
The glucose surplus retained in soil can be measured without the method for isotope labelling, and more accurate, the rate of recovery is high.
The technical solution adopted by the present invention to solve the technical problems is: of the present invention a kind of to be measured using DNS method
The method of glucose in soil, comprising the following steps:
Step A, it extracts the glucose in soil: weighing the fresh earth sample of 15g with electronic balance and be fitted into the centrifuge tube of 30mL,
15mL distilled water is added, is placed in 25 DEG C of concussion instrument after shaking 30min with 150r/min, is then placed on 3000r/min's
10min is centrifuged in centrifuge；
Step B, it filters glucose leaching liquor: being centrifuged resulting supernatant with the syringe aspiration step A of 15mL, then with 0.22
The water-based syringe filters of μ filter, and filtrate are collected to be measured in the 10mL centrifuge tube of sterilizing；
Step C, DNS reagent is produced: by 5.0g 3,5- dinitrosalicylic acid is dissolved in 200mL 2mol/L NaOH solution,
The solution of 500mL sodium potassium tartrate tetrahydrate containing 130g is added, 5g crystalline phenol and 5g sodium sulfite, stirring and dissolving is added after slowly mixing
Afterwards, 1000mL placement is settled to save backup in the dark；
Step D, sample measures: the addition filtered glucose leaching liquor 1mL of step B in the colorimetric cylinder of 25ml, then plus
Enter the 2mL DNS reagent that 1mL distilled water and step C are produced and is placed on boiling water bath in water-bath after liquid blending in test tube
5min is settled to 25ml with distilled water after cooling, absorbance of the solution after measuring constant volume at λ=540nm, to calculate
Measure the surplus of glucose in soil in incubation.
Specifically, the step C3,5- dinitrosalicylic acid, which is dissolved in 200mL 2mol/L NaOH solution, does not use high temperature
Dissolution mode is handled.
Specifically, in the step D calculate soil in glucose content specific method the following steps are included:
1) it, draws standard curve: the colorimetric cylinder of 6 25mL is taken, wherein being packed into a certain amount of distilled water conduct in 1 colorimetric cylinder
In addition control is packed into the glucose standards solution of different capabilities, then adds distilled water and be settled to a constant volume in 5 colorimetric cylinders
Amount；2mL DNS reagent is added, after liquid blending in test tube, is placed on boiling water bath 5min in water-bath, after cooling with steaming
Distilled water is settled to 25ml, absorbance of the solution after measuring constant volume at λ=540nm, finally using concentration of glucose as abscissa,
Absorbance is ordinate, draws standard curve；
2) concentration C of glucose in glucose leaching liquor, is calculated by the standard curve that step 1) is drawnGLU；
3) the glucose content C, being calculated by the following formula in Unit Weight dry groundGLU-soil:
CGLU-soil=(Wwater+15)×CGLU/Wdry soil
Wherein: Wwater--- the weight of moisture in the fresh soil of 15g；
Wdry soil--- the weight of dry ground in the fresh soil of 15g.
Specifically, the distilled water in the step 1) is 2ml, distilled water is settled to 2ml, glucose mark in 5 colorimetric cylinders
Quasi- solution is respectively 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0mL.
Specifically, the glucose standard is placed in 80 DEG C of baking ovens the preparation method is as follows: will analyze pure glucose in advance
11-12h accurately weighs 500mg glucose in beaker, after being dissolved with distilled water, moves in 500mL volumetric flask, constant volume shakes up
Being placed on temperature is that 4 DEG C of storages are spare.
Specifically, the Glucose standards liquor occurs muddy and floccule phenomenon occurs and need to prepare again, prevent
The glucose solution of configuration goes bad, and influences experimental result.
Beneficial effects of the present invention are as follows:
(1) present invention is detected by surplus of the DNS method to the glucose in the culture experiment of soil, can be accurate
Ground detects the surplus of glucose in soil, accelerates detection efficiency, saves the time.
(2) in the soil incubation experiment using glucose, the method that can not have to isotope labelling can measure soil
In the glucose surplus that retains, and more accurate, the rate of recovery is high.
Detailed description of the invention
The present invention will be further explained below with reference to the attached drawings.
Fig. 1 is flow chart of the invention；
Fig. 2 is soil concentration of glucose variation diagram in continuous seven days incubations in the embodiment of the present invention.
Specific embodiment
In order to be easy to understand the technical means, the creative features, the aims and the efficiencies achieved by the present invention, tie below
Specific embodiment is closed, the present invention is further explained.
As shown in Figure 1 to Figure 2, a kind of method using glucose in DNS method measurement soil of the present invention, including with
Lower step:
Step A, it extracts the glucose in soil: weighing the fresh earth sample of 15g with electronic balance and be fitted into the centrifuge tube of 30mL,
15mL distilled water is added, is placed in 25 DEG C of concussion instrument after shaking 30min with 150r/min, is then placed on 3000r/min's
10min is centrifuged in centrifuge；
Step B, it filters glucose leaching liquor: being centrifuged resulting supernatant with the syringe aspiration step A of 15mL, then with 0.22
The water-based syringe filters of μ filter, and filtrate are collected to be measured in the 10mL centrifuge tube of sterilizing；
Step C, DNS reagent is produced: by 5.0g 3,5- dinitrosalicylic acid is dissolved in 200mL 2mol/L NaOH solution,
The solution of 500mL sodium potassium tartrate tetrahydrate containing 130g is added, 5g crystalline phenol and 5g sodium sulfite, stirring and dissolving is added after slowly mixing
Afterwards, 1000mL placement is settled to save backup in the dark；
Step D, sample measures: the addition filtered glucose leaching liquor 1mL of step B in the colorimetric cylinder of 25ml, then plus
Enter the 2mL DNS reagent that 1mL distilled water and step C are produced and is placed on boiling water bath in water-bath after liquid blending in test tube
5min is settled to 25ml with distilled water after cooling, absorbance of the solution after measuring constant volume at λ=540nm, to calculate
Measure the surplus of glucose in soil in incubation.
Specifically, the step C3,5- dinitrosalicylic acid, which is dissolved in 200mL 2mol/L NaOH solution, does not use high temperature
Dissolution mode is handled.
Specifically, in the step D calculate soil in glucose content specific method the following steps are included:
1) it, draws standard curve: the colorimetric cylinder of 6 25mL is taken, wherein being packed into a certain amount of distilled water conduct in 1 colorimetric cylinder
In addition control is packed into the glucose standards solution of different capabilities, then adds distilled water and be settled to a constant volume in 5 colorimetric cylinders
Amount；2mL DNS reagent is added, after liquid blending in test tube, is placed on boiling water bath 5min in water-bath, after cooling with steaming
Distilled water is settled to 25ml, absorbance of the solution after measuring constant volume at λ=540nm, finally using concentration of glucose as abscissa,
Absorbance is ordinate, draws standard curve；
2) concentration C of glucose in glucose leaching liquor, is calculated by the standard curve that step 1) is drawnGLU；
3) the glucose content C, being calculated by the following formula in Unit Weight dry groundGLU-soil:
CGLU-soil=(Wwater+15)×CGLU/Wdry soil
Wherein: Wwater--- the weight of moisture in the fresh soil of 15g；
Wdry soil--- the weight of dry ground in the fresh soil of 15g.
Specifically, the distilled water in the step 1) is 2ml, distilled water is settled to 2ml, glucose mark in 5 colorimetric cylinders
Quasi- solution is respectively 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0mL.
Specifically, the glucose standard is placed in 80 DEG C of baking ovens the preparation method is as follows: will analyze pure glucose in advance
11-12h accurately weighs 500mg glucose in beaker, after being dissolved with distilled water, moves in 500mL volumetric flask, constant volume shakes up
Being placed on temperature is that 4 DEG C of storages are spare.
Specifically, the Glucose standards liquor occurs muddy and floccule phenomenon occurs and need to prepare again.
Embodiment:
50g dry ground is added in the jar of 1L, adjusts soil moisture content to the 40% of maximum water-holding capacity, then again into soil
The glucose solution 2ml of various concentration is added, the glucose content in solution is 1,2.5 and 4mg/g dry weight soil,
Using 0h, 12h, 1d of the above method after culture starts, 2d, 3d, 7d extracts remaining glucose in soil, as shown in Figure 2.
In addition, it is as shown in table 1 to extract the glucose sugar rate of recovery measured in the case where 0d known glucose concentrations.
By Fig. 2 it can be seen that it can be seen that glucose additive amount be 1 and 2.5mg/g dry weight soil when, 3
Breakdown of glucose totally, and adds the glucose of 4mg/g dry weight soil within it, because additive amount is higher, at 3 days
It is interior not decomposed, but decompose and complete within the scope of 3 days to 7 days, meet the rule of general breakdown of glucose experiment.In addition,
After glucose addition, pedotheque is obtained immediately and is extracted, find the rate of recovery under three kinds of concentration on 100% left side
It is right, it is seen that this method is extracted to the glucose in soil and detection efficiency is reliable, is suitble to the soil incubation in addition glucose real
Test middle use.
The basic principles, main features and advantages of the invention have been shown and described above.The technical staff of the industry should
Understand, the present invention is not limited to the above embodiments, and the above embodiments and description only describe originals of the invention
Reason, without departing from the spirit and scope of the present invention, various changes and improvements may be made to the invention, these changes and improvements
It all fall within the protetion scope of the claimed invention.The claimed scope of the invention is by appended claims and its equivalent circle
It is fixed.
Claims (6)
1. a kind of method using glucose in DNS method measurement soil, it is characterised in that: the following steps are included:
Step A, it extracts the glucose in soil: weighing the fresh earth sample of 15g with electronic balance and be fitted into the centrifuge tube of 30mL, then plus
Enter 15mL distilled water, is placed in 25 DEG C of concussion instrument after shaking 30min with 150r/min, is then placed on the speed of 3000r/min
10min is centrifuged in centrifuge；
Step B, it filters glucose leaching liquor: being centrifuged resulting supernatant with the syringe aspiration step A of 15mL, then with 0.22 μ's
Water-based syringe filters filtering, filtrate is collected to be measured in the 10mL centrifuge tube of sterilizing；
Step C, DNS reagent is produced: by 5.0g 3,5- dinitrosalicylic acid is dissolved in 200mL 2mol/L NaOH solution, then plus
Enter the solution of 500mL sodium potassium tartrate tetrahydrate containing 130g, slowly addition 5g crystalline phenol and 5g sodium sulfite after mixing, after stirring and dissolving,
1000mL placement is settled to save backup in the dark；
Step D, sample measures: the filtered glucose leaching liquor 1mL of step B being added in the colorimetric cylinder of 25ml, adds 1mL
The 2mL DNS reagent that distilled water and step C are produced is placed on boiling water bath 5min in water-bath after liquid blending in test tube, to
It is settled to 25ml with distilled water after cooling, absorbance of the solution after measuring constant volume at λ=540nm, to calculate measurement training
During supporting in soil glucose surplus.
2. a kind of method using glucose in DNS method measurement soil according to claim 1, it is characterised in that: described
Step C3,5- dinitrosalicylic acid, which is dissolved in 200mL 2mol/L NaOH solution, does not use high temperature dissolution mode to be handled.
3. a kind of method using glucose in DNS method measurement soil according to claim 1, it is characterised in that: described
In step D calculate soil in glucose content specific method the following steps are included:
1) it, draws standard curve: taking the colorimetric cylinder of 6 25mL, wherein being packed into a certain amount of distilled water conduct pair in 1 colorimetric cylinder
According to, in addition in 5 colorimetric cylinders be packed into different capabilities glucose standards solution, then add distilled water and be settled to a constant volume
Amount；2mL DNS reagent is added, after liquid blending in test tube, is placed on boiling water bath 5min in water-bath, after cooling with steaming
Distilled water is settled to 25ml, absorbance of the solution after measuring constant volume at λ=540nm, finally using concentration of glucose as abscissa,
Absorbance is ordinate, draws standard curve；
2) concentration C of glucose in glucose leaching liquor, is calculated by the standard curve that step 1) is drawnGLU；
3) the glucose content C, being calculated by the following formula in Unit Weight dry groundGLU-soil:
CGLU-soil=(Wwater+15)×CGLU/Wdry soil
Wherein: Wwater--- the weight of moisture in the fresh soil of 15g；
Wdry soil--- the weight of dry ground in the fresh soil of 15g.
4. a kind of method using glucose in DNS method measurement soil according to claim 3, it is characterised in that: described
Distilled water in step 1) is 2ml, and distilled water is settled to 2ml, in 5 colorimetric cylinders glucose standards solution be respectively 0.2ml,
0.4ml、0.6ml、0.8ml、1.0mL。
5. a kind of method using glucose in DNS method measurement soil according to claim 3, it is characterised in that: described
Glucose standard is placed in 11-12h in 80 DEG C of baking ovens the preparation method is as follows: will analyze pure glucose in advance, accurately weighs 500mg
Glucose is in beaker, after being dissolved with distilled water, moves in 500mL volumetric flask, constant volume, and shaking up and being placed on temperature is that 4 DEG C of storages are standby
With.
6. a kind of method using glucose in DNS method measurement soil according to claim 5, it is characterised in that: described
Glucose standards liquor occur muddy and floccule phenomenon occur and need to prepare again.
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| CN107066823A (en) * | 2017-04-18 | 2017-08-18 | 中山大学 | Based on plant, soil, microorganism heavy-metal contaminated soil repairing effect integrated evaluating method |
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2018
- 2018-08-07 CN CN201810891591.4A patent/CN108956499A/en active Pending
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| CN103300132A (en) * | 2013-07-05 | 2013-09-18 | 中国农业科学院农产品加工研究所 | Method for manufacturing Chinese date pieces by utilizing medium short wave infra-red drying method |
| CN104614223A (en) * | 2015-02-03 | 2015-05-13 | 新疆农业科学院土壤肥料与农业节水研究所 | Method for extracting soluble organic carbon in soil |
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| CN106248664A (en) * | 2016-07-18 | 2016-12-21 | 中国科学院东北地理与农业生态研究所 | The assay method of soil saccharase |
| CN106770606A (en) * | 2016-11-16 | 2017-05-31 | 中国科学院遗传与发育生物学研究所 | A kind of method for determining Soil dissolved orgnic carbon isotope |
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Application publication date: 20181207 |