CN106290339A - The assay method of the content of reducing sugar of pesticide infringement Nicotiana tabacum L. - Google Patents
The assay method of the content of reducing sugar of pesticide infringement Nicotiana tabacum L. Download PDFInfo
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- CN106290339A CN106290339A CN201610824345.8A CN201610824345A CN106290339A CN 106290339 A CN106290339 A CN 106290339A CN 201610824345 A CN201610824345 A CN 201610824345A CN 106290339 A CN106290339 A CN 106290339A
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- G—PHYSICS
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- 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
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
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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
- G01N21/3103—Atomic absorption analysis
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Abstract
The present invention proposes the assay method of a kind of content of reducing sugar by pesticide infringement Nicotiana tabacum L., comprises the following steps: (1) reagent preparation, including preparation 1mg/mL glucose standard and 3.5-edlefsen's reagent；(2) reducing sugar standard curve is made；(3) reducing sugar is extracted: clip Nicotiana tabacum L., totally three parts, add water grinding, constant volume 10mL distilled water, and 50 DEG C of tepidarium 20min make reducing sugar leach, and centrifugal filtration, as reducing sugar test liquid；(4) mensuration of reducing sugar, (5) move into dichloro quinolinic acid to add concentration Nicotiana tabacum L. 5～6 leaf periods is 1.04 × 10‑3mg/kg、2.08×10‑3mg/kg、4.17×10‑3mg/kg、8.33×10‑3mg/kg、1.67×10‑ 2In mg/kg medicine soil, using step (1)～(4) to measure the content of reducing sugar of Nicotiana tabacum L. at 7d, 14d, 21d, 28d, 35d respectively, present method solves Nicotiana tabacum L. in prior art is affected the problem inadequate with change detection accuracy by reducing sugar after chloro-quinolinic acid infringement.
Description
Technical field
The present invention relates to chemical analysis detection field, particularly relate to the mensuration side of the content of reducing sugar of pesticide infringement Nicotiana tabacum L.
Method.
Background technology
Dichloro quinolinic acid is the specially good effect selective herbicide preventing and kill off barnyard grass in paddy field, belongs to hormone-type quinoline carboxylic acid herbicide,
Weeds poisoning symptom is similar to auxins effect, is mainly used in preventing and treating barnyard grass and working life is the longest, and the 1-7 leaf phase is the most effective.Water
Rice safety is good.
Relevant research shows that dichloro quinolinic acid is as hormone herbicide, it is possible to stimulate ethylene at plant interior accumulation.Two
The target site of chloro-quinolinic acid is positioned in the biosynthesis of cell wall, plays Cell wall synthesis inhibitor in grass
Effect, the different grass action site sensitivity to dichloro quinolinic acid.The activity of ethylene wall capable of inhibiting cell synzyme
Thus reducing the synthesis of cell wall, the raising of ethylene contents causes the accumulation of ABA, and ABA declines at coordinate plant growth, blade
Always, stomatal opening plays an important role.
Dichloro quinolinic acid has summation in soil, and tobacco leaf production can be produced obvious poisoning.China's part vega
Once large area Nicotiana tabacum L. teratogenesis phenomenon occurred because of chloro-quinolinic acid, symptom is that young leaves irregular growth, leaf margin last volume, leaf first occurs
Sheet is supported or opposed shrinkage, causes blade long and narrow, and severe patient is wire, has a strong impact on yield and the quality of Nicotiana tabacum L..
Due to the particularity of this crop of Nicotiana tabacum L., it is affected will be directly affected receiving of tobacco grower by chloro-quinolinic acid
Become, and the quality of Nicotiana tabacum L., therefore, for the chloro-quinolinic acid impact on Nicotiana tabacum L., need to carry out many, accurately, reliably
Analysis, although the analysis currently for pesticide infringement plant has been reported that and general analysis method, but for chloroquinoline
Nicotiana tabacum L. infringement is researched and analysed the most relatively fewer by acid, the change impact especially encroached on the reducing sugar of Nicotiana tabacum L., causes at chlorine
The quinolinic acid degree to the infringement of Nicotiana tabacum L., the rare deep understanding such as impact, and analysis result on there is also bigger standard
Exactness difference.
Summary of the invention
The present invention proposes the assay method of the content of reducing sugar of a kind of pesticide infringement Nicotiana tabacum L., solves Nicotiana tabacum L. in prior art
After being encroached on by chloro-quinolinic acid, reducing sugar is affected the problem inadequate with change detection accuracy.
The technical scheme is that and be achieved in that: by the assay method of the content of reducing sugar of pesticide infringement Nicotiana tabacum L., bag
Include following steps:
(1) reagent preparation, including preparation 1mg/mL glucose standard and 3.5-edlefsen's reagent；
(2) reducing sugar standard curve is made；
(3) reducing sugar is extracted: clip Nicotiana tabacum L., totally three parts, add water grinding, constant volume 10mL distilled water, 50 DEG C of tepidariums
20min, makes reducing sugar leach, centrifugal filtration, as reducing sugar test liquid；
(4) mensuration of reducing sugar:
Take 3 25mL scale test tubes, numbering, it is separately added into reducing sugar liquid to be measured 1mL, 3.5-edlefsen's reagent
1.5mL, measures the extinction value of each pipe；On standard curve, find corresponding reducing sugar milligram number respectively, press examination and calculate reducing sugar
Percentage composition:
The reducing sugar amount that in formula, C standard curve equation is tried to achieve；
V extracting liquid volume；
During a colour developing, pipette samples liquid amasss；
W sample weight；
(5) move into dichloro quinolinic acid adding concentration Nicotiana tabacum L. 5～6 leaf periods is 1.04 × 10-3mg/kg、2.08×10-3mg/kg、4.17×10-3mg/kg、8.33×10-3mg/kg、1.67×10-2In mg/kg medicine soil, respectively 7d, 14d, 21d,
28d, 35d use step (1)～(4) to measure the reducing sugar total amount of Nicotiana tabacum L..
Further, in step (1), the preparation following proportioning of employing of reagent and parameter:
1mg/mL glucose standard: accurately weigh 100mg analytical pure glucose, be placed in small beaker, with a small amount of distillation
After water dissolution, quantitatively transfer to, in the volumetric flask of 100mL, be settled to scale with distilled water, shake up, put in refrigerator and save backup；
3.5-edlefsen's reagent: 3.5-dinitrosalicylic acid 0.63g, NaOH solution 26.2mL of 2mol/L,
It is added in the hydrothermal solution that 50mL contains 18.5g sodium potassium tartrate tetrahydrate, then adds 0.5g phenol and 0.5g sodium sulfite, stirring and dissolving,
Add distilled water after cooling and be settled to 100mL, now with the current.
Further, in step (3), clip the 4th launches fresh tobacco leaf 0.3g.
The measuring method of the content of reducing sugar of the described pesticide infringement Nicotiana tabacum L. of the present invention, for the characteristic of Nicotiana tabacum L., in conjunction with right
The infringement study mechanism of chloro-quinolinic acid, the change of content of reducing sugar during pesticide is encroached on, the apolegamy of reagent, operating environment,
Parameter etc. carried out the strictest adjust with control so as to get the measurement of content of reducing sugar of chloro-quinolinic acid infringement Nicotiana tabacum L.
More accurate, reliably, for reasonable employment chloro-quinolinic acid and eliminate the effects of the act, it is provided that the foundation of science.
Accompanying drawing explanation
Fig. 1 is reducing sugar standard curve；
Fig. 2 is the dichloro quinolinic acid dynamic analysis to tobacco leaf content of reducing sugar.
Detailed description of the invention
Below in conjunction with embodiments of the invention, the technical scheme in the embodiment of the present invention is clearly and completely retouched
State, it is clear that described embodiment is only a part of embodiment of the present invention rather than whole embodiments.Based on the present invention
In embodiment, the every other enforcement that those of ordinary skill in the art are obtained under not making creative work premise
Example, broadly falls into the scope of protection of the invention.
Embodiment
Tobacco seed uses kind to be K326.Greenhouse is sowed, nursery, transplants in five to six leaf periods, carries out pesticide and invade
The mensuration of the content of reducing sugar of evil Nicotiana tabacum L.
1., preparation of reagents
1mg/mL glucose standard: accurately weigh 100mg analytical pure glucose (drying to constant weight at 80 DEG C in advance), be placed in
In small beaker, after dissolving with a small amount of distilled water, quantitatively transfer to, in the volumetric flask of 100mL, be settled to scale with distilled water, shake
Even, put in refrigerator and save backup.
3.5-edlefsen's reagent: 3.5-dinitrosalicylic acid 0.63g, NaOH solution 26.2mL of 2mol/L,
It is added in the hydrothermal solution that 50mL contains 18.5g sodium potassium tartrate tetrahydrate, then adds 0.5g phenol and 0.5g sodium sulfite, stirring and dissolving.
Add distilled water after cooling and be settled to 100mL, now with the current.
2., the making of standard curve
Take 7 braces and have blood sugar tube or the scale test tube of 25mL scale, numbering, the amount as shown in table 3-3, accurately add concentration
Glucose standard and 3.5-edlefsen's reagent for 1mg/mL.
Table 5-3 reducing sugar standard curve
Each pipe is shaken up, boiling water heats 5min, is immediately placed in after taking-up in the beaker of cold water and is cooled to room temperature, then
Being settled to, at 25mL scale, clog the mouth of pipe with rubber closure with distilled water, reverse mixing is (as with Boiling tube, then added to often pipe
21.5mL distilled water, mixing).Under 540nm wavelength, with No. 0 pipe zeroing, read the extinction value of 1～No. 6 pipe respectively.With delustring
Value is vertical coordinate, draws standard curve, tries to achieve linear equation, with glucose content as abscissa, with the extinction density at 540nm
Value is vertical coordinate, obtains the standard curve of glucose as shown in Figure 1.
3., the extraction of reducing sugar
Clip the 4th launches fresh tobacco leaf 0.3g, totally three parts, adds a small amount of water and grinds, constant volume 10mL distilled water, 50 DEG C of temperature
Water-bath 20min, makes reducing sugar leach, centrifugal filtration, as reducing sugar test liquid.
4., the mensuration of reducing sugar
Take 3 25mL scale test tubes, numbering, it is separately added into reducing sugar liquid to be measured 1mL, 3.5-edlefsen's reagent
1.5mL, remaining operation is all identical with making standard curve, measures the extinction value of each pipe.Find corresponding respectively on standard curve
Reducing sugar milligram number, presses and tries to calculate reducing sugar percentage composition:
The reducing sugar amount (mg) that in formula, C standard curve equation is tried to achieve；
V extracting liquid volume (mL)；
During a colour developing, pipette samples liquid is long-pending (mL)；
W sample weight (g).
(5) move into dichloro quinolinic acid adding concentration Nicotiana tabacum L. 5～6 leaf periods is 1.04 × 10-3mg/kg、2.08×10-3mg/kg、4.17×10-3mg/kg、8.33×10-3mg/kg、1.67×10-2In mg/kg medicine soil, respectively 7d, 14d, 21d,
28d, 35d use step (1)～(4) to measure the content of reducing sugar of Nicotiana tabacum L..
According to said method measure the chloro-quinolinic acid impact on tobacco leaf content of reducing sugar, measurement result as in figure 2 it is shown,
It can be seen that content of reducing sugar is above comparison from 7d to 28d in dichloroquinoline acid treatment, cigarette strain blade.7d after transplanting, two
Chloro-quinolinic acid concentration is 1.04 × 10-3mg/kg、2.08×10-3mg/kg、4.17×10-3mg/kg、8.33×10-3mg/kg、
1.67×10-2Mg/kg process content of reducing sugar in tobacco leaf be respectively 5.06%, 5.06%, 5.97%, 6.85%,
7.99%, process relatively comparison raising 87.72%, 87.60%, 121.33%, 154.11%, 196.31%.35d after transplanting, two
Chloro-quinolinic acid concentration is 4.17 × 10-3mg/kg、8.33×10-3mg/kg、1.67×10-2Mg/kg, reducing sugar in tobacco leaf
After content is relatively transplanted, 28d measures reduction.It is probably Nicotiana tabacum L. and has produced obvious deformity, make unit fresh weight content of reducing sugar drop
Low.
The foregoing is only presently preferred embodiments of the present invention, not in order to limit the present invention, all essences in the present invention
Within god and principle, any modification, equivalent substitution and improvement etc. made, should be included within the scope of the present invention.
Claims (3)
1. encroached on the assay method of the content of reducing sugar of Nicotiana tabacum L. by pesticide, it is characterised in that comprise the following steps:
(1) reagent preparation, including preparation 1mg/mL glucose standard and 3.5-edlefsen's reagent；
(2) reducing sugar standard curve is made；
(3) reducing sugar is extracted: clip Nicotiana tabacum L., totally three parts, add water grinding, constant volume 10mL distilled water, and 50 DEG C of tepidarium 20min make
Reducing sugar leaches, and centrifugal filtration, as reducing sugar test liquid；
(4) mensuration of reducing sugar:
Take 3 25mL scale test tubes, numbering, it is separately added into reducing sugar liquid to be measured 1mL, 3.5-edlefsen's reagent
1.5mL, measures the extinction value of each pipe；On standard curve, find corresponding reducing sugar milligram number respectively, press examination and calculate reducing sugar
Percentage composition:
The reducing sugar amount that in formula, C standard curve equation is tried to achieve；
V extracting liquid volume；
During a colour developing, pipette samples liquid amasss；
W sample weight；
(5) move into dichloro quinolinic acid adding concentration Nicotiana tabacum L. 5～6 leaf periods is 1.04 × 10-3mg/kg、2.08×10-3mg/
kg、4.17×10-3mg/kg、8.33×10-3mg/kg、1.67×10-2In mg/kg medicine soil, respectively 7d, 14d, 21d, 28d,
35d uses step (1)～(4) to measure the content of reducing sugar of Nicotiana tabacum L..
The assay method of the content of reducing sugar of Nicotiana tabacum L. is encroached on the most as described in the appended claim 1 by pesticide, it is characterised in that: reagent
The preparation following proportioning of employing and parameter:
1mg/mL glucose standard: accurately weigh 100mg analytical pure glucose, be placed in small beaker, water-soluble with a small amount of distillation
Xie Hou, quantitatively transfers to, in the volumetric flask of 100mL, be settled to scale with distilled water, shake up, put in refrigerator and save backup；
3.5-edlefsen's reagent: 3.5-dinitrosalicylic acid 0.63g, NaOH solution 26.2mL of 2mol/L, is added to
50mL contains in the hydrothermal solution of 18.5g sodium potassium tartrate tetrahydrate, then adds 0.5g phenol and 0.5g sodium sulfite, stirring and dissolving, cooling
After add distilled water and be settled to 100mL, now with the current.
The assay method of the content of reducing sugar of Nicotiana tabacum L. is encroached on the most as described in the appended claim 1 by pesticide, it is characterised in that: step
(3), in, clip the 4th launches fresh tobacco leaf 0.3g.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610824345.8A CN106290339A (en) | 2016-09-14 | 2016-09-14 | The assay method of the content of reducing sugar of pesticide infringement Nicotiana tabacum L. |
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| CN201610824345.8A CN106290339A (en) | 2016-09-14 | 2016-09-14 | The assay method of the content of reducing sugar of pesticide infringement Nicotiana tabacum L. |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101701911A (en) * | 2009-11-30 | 2010-05-05 | 吉林燃料乙醇有限责任公司 | Method for detecting content of reducing sugar in fermentation materials |
| CN101963578A (en) * | 2010-09-11 | 2011-02-02 | 大连工业大学 | Method for determining reducing sugar content of pulping black liquor |
| CN104390958A (en) * | 2014-10-24 | 2015-03-04 | 西安莹朴生物科技股份有限公司 | Method for measuring content of reducing sugar in xylitol |
-
2016
- 2016-09-14 CN CN201610824345.8A patent/CN106290339A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101701911A (en) * | 2009-11-30 | 2010-05-05 | 吉林燃料乙醇有限责任公司 | Method for detecting content of reducing sugar in fermentation materials |
| CN101963578A (en) * | 2010-09-11 | 2011-02-02 | 大连工业大学 | Method for determining reducing sugar content of pulping black liquor |
| CN104390958A (en) * | 2014-10-24 | 2015-03-04 | 西安莹朴生物科技股份有限公司 | Method for measuring content of reducing sugar in xylitol |
Non-Patent Citations (2)
| Title |
|---|
| 李晶新: "二氯喹啉酸对烤烟生理特性影响的研究", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
| 金青: "《生物化学实验指导》", 31 August 2014, 中国农业大学出版社，第1版 * |
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Application publication date: 20170104 |