CN101900680A - Method for quickly detecting soluble protein content of rape leaves - Google Patents
Method for quickly detecting soluble protein content of rape leaves Download PDFInfo
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- CN101900680A CN101900680A CN 201010219042 CN201010219042A CN101900680A CN 101900680 A CN101900680 A CN 101900680A CN 201010219042 CN201010219042 CN 201010219042 CN 201010219042 A CN201010219042 A CN 201010219042A CN 101900680 A CN101900680 A CN 101900680A
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- 238000000034 method Methods 0.000 title claims abstract description 21
- 238000012417 linear regression Methods 0.000 claims abstract description 11
- 238000002310 reflectometry Methods 0.000 claims abstract description 11
- 239000000843 powder Substances 0.000 claims abstract description 10
- 102000004169 proteins and genes Human genes 0.000 claims description 19
- 108090000623 proteins and genes Proteins 0.000 claims description 19
- 238000006467 substitution reaction Methods 0.000 claims description 2
- 238000001514 detection method Methods 0.000 abstract description 7
- 238000004458 analytical method Methods 0.000 abstract description 2
- 238000001035 drying Methods 0.000 abstract description 2
- 238000003912 environmental pollution Methods 0.000 abstract description 2
- 229920003266 Leaf® Polymers 0.000 description 31
- 239000004009 herbicide Substances 0.000 description 5
- 230000002363 herbicidal effect Effects 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 240000002791 Brassica napus Species 0.000 description 3
- 235000004977 Brassica sinapistrum Nutrition 0.000 description 3
- QOSMNYMQXIVWKY-UHFFFAOYSA-N Propyl levulinate Chemical compound CCCOC(=O)CCC(C)=O QOSMNYMQXIVWKY-UHFFFAOYSA-N 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 238000009825 accumulation Methods 0.000 description 2
- 230000035508 accumulation Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000003921 oil Substances 0.000 description 2
- 235000019198 oils Nutrition 0.000 description 2
- 239000005416 organic matter Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- 238000009010 Bradford assay Methods 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 208000035126 Facies Diseases 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000003570 biosynthesizing effect Effects 0.000 description 1
- NKLPQNGYXWVELD-UHFFFAOYSA-M coomassie brilliant blue Chemical compound [Na+].C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=C1 NKLPQNGYXWVELD-UHFFFAOYSA-M 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000000875 corresponding effect Effects 0.000 description 1
- 239000008157 edible vegetable oil Substances 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 244000037666 field crops Species 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000003966 growth inhibitor Substances 0.000 description 1
- 238000002329 infrared spectrum Methods 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 238000002331 protein detection Methods 0.000 description 1
- 239000011546 protein dye Substances 0.000 description 1
- 229940121649 protein inhibitor Drugs 0.000 description 1
- 239000012268 protein inhibitor Substances 0.000 description 1
- 238000001814 protein method Methods 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 238000009333 weeding Methods 0.000 description 1
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- Investigating Or Analysing Materials By Optical Means (AREA)
Abstract
The invention discloses a method for quickly detecting soluble protein content of rape leaves. The method comprises the following steps of: crushing the rape leaves after fixation and drying to obtain powder of the leaves; acquiring reflectivities of the rape leaves corresponding to light waves with wavelengths of 2,208nm, 2,174nm, 2,236nm, 1,398nm, 1,422nm, 2,258nm, 2,282nm, 1,730nm, 1,890nm, 1,684nm, 2,366nm, 1,250nm, 1,658nm, 1,602nm, 1,462nm, 2,304nm, 1,910nm, 2,418nm, 1,858nm and 2,048nm respectively; and substituting each reflectivity into a multiple linear regression equation to calculate the soluble protein content of the rape leaves respectively. By the method, multiple linear regression analysis is performed by using a few characteristic wavelengths, so the soluble protein content of the rape leaves is quickly and accurately detected, the detection time is greatly shortened, and the environmental pollution and the detection cost are reduced.
Description
Technical field
The present invention relates to the rape detection range, relate in particular to a kind of method for quickly detecting soluble protein content of rape leaves.
Background technology
Rape is one of the main oil crops of China and nectar source crop, and the cultivated area of rape accounts for more than 40% of the national oil crops total area, and output accounts for more than 60% of national edible vegetable oil total production, occupies first place in the world.The upgrowth situation of rape different times is related to the output and the quality of rapeseed.Content of soluble protein in the rape leaf is very important nutrient and physical signs in the growth of rape process, and is significant to physiology courses such as rape leaf nutrient and organic matter accumulations.Therefore can obtain the upgrowth situation of rape by detecting the content of soluble protein in the rape leaf.In recent years, because a large amount of herbicides has been used in rural laborer's minimizing in the rapeseed cultivation.Some herbicide is a kind of growth inhibitor, and for example propyl-ester nitorfen can stop the amino acid whose biosynthesizing of straight chain, has rape safety, kills that the grass spectrum is wide, activity of weeding is high, the lasting period is long, and low toxicity, low-residual, with characteristics such as environmentally compatible.Detection to content of soluble protein in the rape leaf, can reflect of the influence of some herbicide to rape nutrient situation and organic matter accumulation, the land for growing field crops of realizing the growth of rape situation is detected and continuous monitoring, and the output and the quality that improve rapeseed are significant.
Traditional soluble protein content of rape leaves is measured according to Bradford method (Bradford MM.1976.A rapid and sensitive method for the quantitative of microgramquantities of protein utilizing the principle of protein-dye binding.Analyt.Biochem.72,248-54), drawing enzyme liquid measures with the Coomassie brilliant blue method.This method measuring process is time-consuming, effort, chemical depletion product expensive, and environment is caused certain pollution.Therefore, be badly in need of research a kind of simple, quick, harmless rape leaf content of soluble protein detection method and technology.
As seen/near infrared spectrum can reflect that the frequency multiplication of groups such as the organic principle of material inside, particularly C-H, O-H, N-H and sum of fundamental frequencies absorb, and can be used for the content of quantitative measurement material organic substance.Spectral technique has been widely used in industries such as agricultural, food, petrochemical complex, pharmacy, feed.In the research of plant life information fast detecting, has application potential widely.
Summary of the invention
The invention provides a kind of rape leaf content of soluble protein method for quick, solved problems such as existing detection method is time-consuming, effort, complicated operation, contaminated environment.
A kind of method for quickly detecting soluble protein content of rape leaves may further comprise the steps:
(1) rape leaf completed, dries the back and pulverize, obtain the blade powder;
(2) gather the reflectivity of wavelength 2208nm, 2174nm, 2236nm, 1398nm, 1422nm, 2258nm, 2282nm, 1730nm, 1890nm, 1684nm, 2366nm, 1250nm, 1658nm, 1602nm, 1462nm, 2304nm, 1910nm, 2418nm, 1858nm and the pairing rape leaf of 2048nm light wave respectively;
(3) with each reflectivity difference substitution multiple linear regression equations: y=10000* (53.15 λ
1-43.39 λ
2-32.77 λ
3-52.50 λ
4+ 54.04 λ
5+ 24.95 λ
6-14.24 λ
7-43.73 λ
8-29.69 λ
9+ 47.87 λ
10+ 16.31 λ
11+ 14.64 λ
12-30.90 λ
13+ 24.20 λ
14-24.52 λ
15-1.159 λ
16+ 6.542 λ
17-7.164 λ
18+ 26.40 λ
19+ 11.21 λ
20Content of soluble protein in the rape leaf is calculated in)+1779.728;
Y is the rape leaf content of soluble protein in the equation, and unit is μ g/g FW; λ
1～λ
20Be respectively the reflectivity of wavelength 2208nm, 2174nm, 2236nm, 1398nm, 1422nm, 2258nm, 2282nm, 1730nm, 1890nm, 1684nm, 2366nm, 1250nm, 1658nm, 1602nm, 1462nm, 2304nm, 1910nm, 2418nm, 1858nm and the pairing rape leaf Powder samples of 2048nm light wave.
In the step (1), the blade powder is 40～80 orders, and the temperature that completes is 95～115 ℃, and fixation time is 20～40min.This complete temperature and fixation time are suitable, help the reservation of major component in the rape leaf, particularly soluble protein.In the step (1), bake out temperature is 70～90 ℃, and blade is dried to constant weight, and this bake out temperature is suitable, can burned blade.
Because the rape leaf Powder samples is a non-transparent medium, when light-wave irradiation in its when surface, except that part is absorbed, all the other all reflect, and what (perhaps reflectivity) that light wave absorbs are to be correlated with the inner content of chemical substances of rape leaf, and only follow the reflectivity of some characteristic wavelength light wave of minority relevant at the vigor of the concrete material of this class of content of soluble protein.
Use the successive projection algorithm and in 1100～2500nm wavelength coverage, carry out choosing of characteristic wavelength, obtain these characteristic wavelengths and be respectively 2208nm, 2174nm, 2236nm, 1398nm, 1422nm, 2258nm, 2282nm, 1730nm, 1890nm, 1684nm, 2366nm, 1250nm, 1658nm, 1602nm, 1462nm, 2304nm, 1910nm, 2418nm, 1858nm and 2048nm.With the reflectance value of the above-mentioned characteristic wavelength input variable as multiple linear regression model, the soluble protein content of actual measurement calculates above-mentioned multiple linear regression equations as output variable.
The present invention carries out multiple linear regression analysis by a few features wavelength, quick and precisely detects content of soluble protein in the rape leaf, has shortened the time of detecting greatly, has reduced environmental pollution, has reduced the detection cost.
Description of drawings
Fig. 1 is a diffusing some distribution plan for modeling collection sample of the present invention predicts the outcome;
Fig. 2 is a diffusing some distribution plan for forecast set sample of the present invention predicts the outcome.
Embodiment
248 rape leafs of random collecting, the rape leaf sample of gathering for making has more representativeness and diversity, not only comprise the rape leaf (totally 62) under the normal growth state in 248 blades being gathered, also comprise and sprayed variable concentrations (100,500,1000mg/L) rape leaf of propyl-ester nitorfen herbicide is 186 (100,500, each concentration gradient of 1000mg/L is gathered 62 of rape leafs), spraying time is one heart stages of five leaves, spray pattern is a foliage-spray.The propyl-ester nitorfen herbicide is a kind of soluble protein inhibitor, inhibited to the rape leaf soluble protein, can make the content of soluble protein in the rape leaf have bigger scope, make institute's established model have better adaptability and robustness.
248 rape leafs are placed under 80 ℃, dry after completing 30 minutes in 105 ℃ the baking oven to constant weight.Rape leaf sample mill powdered after will drying with mortar again, and mistake 60 mesh sieves, at room temperature measure the reflectance value (Foss NIR Systems 5000) of rape leaf Powder samples, utilize the mentioned classic method of background technology to measure the content of soluble protein of all blades simultaneously at wavelength 2208nm, 2174nm, 2236nm, 1398nm, 1422nm, 2258nm, 2282nm, 1730nm, 1890nm, 1684nm, 2366nm, 1250nm, 1658nm, 1602nm, 1462nm, 2304nm, 1910nm, 2418nm, 1858nm and 2048nm place.Select 186 samples as modeling collection sample then at random, all the other 62 as the forecast set sample, with the reflectance value of each characteristic wavelength of 186 rape leafs of modeling collection sample as input variable, with the content of soluble protein of corresponding measurement as output variable, utilize multiple linear regression method (Multiple linear regression MLR) obtains following multiple linear regression equations:
y＝10000*(53.15λ
1-43.39λ
2-32.77λ
3-52.50λ
4+54.04λ
5+24.95λ
6-14.24λ
7-43.73λ
8-29.69λ
9+47.87λ
10+16.31λ
11+14.64λ
12-30.90λ
13+24.20λ
14-24.52λ
15-1.159λ
16+6.542λ
17-7.164λ
18+26.40λ
19+11.21λ
20)+1779.728。
Y is the rape leaf content of soluble protein in the equation, and unit is μ g/g FW; λ
1～λ
20Be respectively the reflectivity of wavelength 2208nm, 2174nm, 2236nm, 1398nm, 1422nm, 2258nm, 2282nm, 1730nm, 1890nm, 1684nm, 2366nm, 1250nm, 1658nm, 1602nm, 1462nm, 2304nm, 1910nm, 2418nm, 1858nm and the pairing rape leaf Powder samples of 2048nm light wave.
Utilize above-mentioned multiple linear regression equations that the content of soluble protein of modeling collection and forecast set rape leaf sample is predicted then, and estimate predicting the outcome.Related coefficient and slope approach 1 more in the evaluation index, and the absolute value of root-mean-square error, deviation and the absolute value of intercept are more little, illustrate that the model prediction performance is good more.It is as shown in the table to predict the outcome:
Modeling collection and forecast set are predicted the facies relationship number average between the predicted value of rape leaf content of soluble protein and the true chemical method measured value has obtained satisfied precision of prediction greater than 0.95.The diffusing some distribution plan that modeling collection and forecast set sample are predicted the outcome as depicted in figs. 1 and 2, sample has good linear prediction effect near being distributed in the regression straight line both sides among the figure.The detection that method of the present invention can realize the rape leaf content of soluble protein is rapidly and accurately used in the The above results explanation.
Claims (4)
1. method for quickly detecting soluble protein content of rape leaves may further comprise the steps:
(1) rape leaf completed, dries the back and pulverize, obtain the blade powder;
(2) gather the reflectivity of wavelength 2208nm, 2174nm, 2236nm, 1398nm, 1422nm, 2258nm, 2282nm, 1730nm, 1890nm, 1684nm, 2366nm, 1250nm, 1658nm, 1602nm, 1462nm, 2304nm, 1910nm, 2418nm, 1858nm and the pairing rape leaf of 2048nm light wave respectively;
(3) with each reflectivity difference substitution multiple linear regression equations: y=10000* (53.15 λ
1-43.39 λ
2-32.77 λ
3-52.50 λ
4+ 54.04 λ
5+ 24.95 λ
6-14.24 λ
7-43.73 λ
8-29.69 λ
9+ 47.87 λ
10+ 16.31 λ
11+ 14.64 λ
12-30.90 λ
13+ 24.20 λ
14-24.52 λ
15-1.159 λ
16+ 6.542 λ
17-7.164 λ
18+ 26.40 λ
19+ 11.21 λ
20Content of soluble protein in the rape leaf is calculated in)+1779.728;
Y is the rape leaf content of soluble protein in the equation; λ
1～λ
20Be respectively the reflectivity of wavelength 2208nm, 2174nm, 2236nm, 1398nm, 1422nm, 2258nm, 2282nm, 1730nm, 1890nm, 1684nm, 2366nm, 1250nm, 1658nm, 1602nm, 1462nm, 2304nm, 1910nm, 2418nm, 1858nm and the pairing rape leaf Powder samples of 2048nm light wave.
2. method for quickly detecting soluble protein content of rape leaves according to claim 1 is characterized in that: described blade powder is 40～80 orders.
3. method for quickly detecting soluble protein content of rape leaves according to claim 1 is characterized in that: in the step (1), the temperature that completes is 95～115 ℃, and fixation time is 20～40min.
4. method for quickly detecting soluble protein content of rape leaves according to claim 1 is characterized in that: in the step (1), bake out temperature is 70～90 ℃.
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|---|---|---|---|
| CN 201010219042 CN101900680B (en) | 2010-07-05 | 2010-07-05 | Method for quickly detecting soluble protein content of rape leaves |
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| CN101900680B CN101900680B (en) | 2012-06-27 |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0487733A1 (en) * | 1990-06-15 | 1992-06-03 | Snow Brand Milk Products Co., Ltd. | Method and apparatus for measuring the contents of components of skim milk, milk, cream and cheese using near infrared ray |
| WO2001046678A2 (en) * | 1999-12-23 | 2001-06-28 | Textron Systems, Corp. | Near infrared analysis of constituents |
| JP2005083901A (en) * | 2003-09-09 | 2005-03-31 | Pola Chem Ind Inc | Noninvasive assay of collagen |
| CN101706421A (en) * | 2009-11-19 | 2010-05-12 | 浙江大学 | Characteristic wave bands based method and device for rapidly detecting content of proteins in black fungi |
| CN101711347A (en) * | 2007-04-26 | 2010-05-19 | Ese公司 | Chemical constituent analyzer |
-
2010
- 2010-07-05 CN CN 201010219042 patent/CN101900680B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0487733A1 (en) * | 1990-06-15 | 1992-06-03 | Snow Brand Milk Products Co., Ltd. | Method and apparatus for measuring the contents of components of skim milk, milk, cream and cheese using near infrared ray |
| WO2001046678A2 (en) * | 1999-12-23 | 2001-06-28 | Textron Systems, Corp. | Near infrared analysis of constituents |
| JP2005083901A (en) * | 2003-09-09 | 2005-03-31 | Pola Chem Ind Inc | Noninvasive assay of collagen |
| CN101711347A (en) * | 2007-04-26 | 2010-05-19 | Ese公司 | Chemical constituent analyzer |
| CN101706421A (en) * | 2009-11-19 | 2010-05-12 | 浙江大学 | Characteristic wave bands based method and device for rapidly detecting content of proteins in black fungi |
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| CN101900680B (en) | 2012-06-27 |
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Inventor after: Liu Fei Inventor after: He Yong Inventor after: Kong Menmen Inventor after: Bao Yidan Inventor before: He Yong Inventor before: Liu Fei Inventor before: Kong Menmen |
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Free format text: CORRECT: INVENTOR; FROM: HE YONG LIU FEI KONG WENWEN TO: LIU FEI HE YONG KONG WENWEN BAO YIDAN |
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