CN101900678A - Method for quickly detecting leucine content in rape leaves - Google Patents
Method for quickly detecting leucine content in rape leaves Download PDFInfo
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- CN101900678A CN101900678A CN 201010219028 CN201010219028A CN101900678A CN 101900678 A CN101900678 A CN 101900678A CN 201010219028 CN201010219028 CN 201010219028 CN 201010219028 A CN201010219028 A CN 201010219028A CN 101900678 A CN101900678 A CN 101900678A
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- optical wave
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- leucine content
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Abstract
The invention discloses a method for quickly detecting leucine content in rape leaves. The method comprises the following steps of: fixing green and drying the rape leaves and crushing the rape leaves to obtain leaf powder; acquiring the reflectivity of the rape leaves corresponding to 2,360nm optical wave, 2,386nm optical wave, 2,306nm optical wave, 2,314nm optical wave, 2,330nm optical wave, 1,194nm optical wave, 2,198nm optical wave, 1,420nm optical wave, 1,462nm optical wave, 2,106nm optical wave, 1,602nm optical wave, 1,878nm optical wave, 2,426nm optical wave, 1,814nm optical wave, 2,274nm optical wave, 1,676nm optical wave, 1,910nm optical wave and 2,052nm optical wave respectively; and substituting the reflectivity into a multiple linear regression respectively to calculate the leucine content in the rape leaves. In the method, multiple linear regression analysis is performed through minor characteristic wavelengths; and thus, the leucine content in the rape leaves is quickly and accurately detected, the detection time is greatly shortened, the environmental pollution is reduced, and the detection cost is reduced.
Description
Technical field
The present invention relates to the rape detection range, relate in particular to a kind of method for quickly detecting leucine content in 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.Amino acid content in the rape leaf is very important physical signs in the growth of rape process, and is significant to physiology courses such as rape leaf physiology and organic matter accumulations.Therefore can obtain the upgrowth situation of rape by detecting the amino acid content 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.To the detection of leucine content in the rape leaf, can reflect of the influence of some herbicide to rape physiology and organic matter accumulation, the land for growing field crops of realizing the growth of rape situation is detected and continuous monitoring, the output and the quality that improve rapeseed are significant.
Traditional leucine content in rape leaves detection method is according to GB/T5009.124-2003, adopt ten thousand/balance accurately to take by weighing the ampere bottle that powder sample 0.05g places the 10ml volume, add 5ml6M HCL, vacuumize and on alcohol blast burner, seal to be placed in 110 ℃ of baking ovens with the circulating vacuum pump and clear up 24h; Cooled digestion solution is transferred to constant volume in the 50ml volumetric flask fully, shaken up; Draw the 5ml supernatant and place round bottom ground conical flask, (60～70 ℃) vacuumize 15min left and right sides evaporate to dryness hydrochloric acid on Rotary Evaporators; Again add 0.02M HCl (pH 2.2) 10ml, ultrasound wave fully dissolves back one-shot injector sucking-off, disposable filter through 0.22 μ m is filtered in the special-purpose sample bottle of amino acid analysis, 4 ℃ of dark preservations are with the L-8900 of Hitachi automatic amino acid analyser (Japan) assay determination leucine content.This method sample preprocessing process and measuring process is time-consuming, effort, chemical depletion product expensive, and environment has been caused certain pollution.Therefore, be badly in need of research a kind of simple, quick, harmless leucine content in rape leaves 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 method for quickly detecting leucine content in rape leaves, solved problems such as existing detection method is time-consuming, effort, complicated operation, cost height, contaminated environment.
A kind of method for quickly detecting leucine content in 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 2360nm, 2386nm, 2306nm, 2314nm, 2330nm, 1194nm, 2198nm, 1420nm, 1462nm, 2106nm, 1602nm, 1878nm, 2426nm, 1814nm, 2274nm, 1676nm, 1910nm and the pairing rape leaf of 2052nm light wave respectively;
(3) with each reflectivity difference substitution multiple linear regression equations: y=199.490 λ
1-205.134 λ
2-157.082 λ
3+ 158.636 λ
4-98.514 λ
5-40.158 λ
6+ 74.919 λ
7+ 79.434 λ
8-79.398 λ
9-76.208 λ
10+ 142.368 λ
11+ 45.322 λ
12+ 47.134 λ
13-45.866 λ
14+ 27.124 λ
15-99.757 λ
16-19.316 λ
17+ 44.704 λ
18+ 2.5554, calculate leucine content in the rape leaf;
Y is a leucine content in rape leaves in the equation, and unit is mg/100mg DW, λ
1～λ
18Be respectively the reflectivity of wavelength 2360nm, 2386nm, 2306nm, 2314nm, 2330nm, 1194nm, 2198nm, 1420nm, 1462nm, 2106nm, 1602nm, 1878nm, 2426nm, 1814nm, 2274nm, 1676nm, 1910nm and the pairing rape leaf Powder samples of 2052nm 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 leucine.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 the reflectivity with some characteristic wavelength light wave of minority is relevant at the concrete material of this class of leucine content.
Use the successive projection algorithm and in 1100～2500nm wavelength coverage, carry out choosing of characteristic wavelength, obtain these characteristic wavelengths and be respectively 2360nm, 2386nm, 2306nm, 2314nm, 2330nm, 1194nm, 2198hm, 1420nm, 1462nm, 2106nm, 1602nm, 1878nm, 2426nm, 1814nm, 2274nm, 1676nm, 1910nm and 2052nm.With the reflectance value of the above-mentioned wavelength input variable as multiple linear regression model, the leucine 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 leucine content 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 leucine inhibitor, and is inhibited to the rape leaf leucine, can make the leucine content in the rape leaf have bigger scope, makes 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 rape leaf Powder samples at wavelength 2360nm, 2386nm, 2306nm, 2314nm, 2330nm, 1194nm, 2198nm, 1420nm, 1462nm, 2106nm, 1602nm, 1878nm, 2426nm, 1814nm, 2274nm, 1676nm, the reflectance value at 1910nm and 2052nm place (Foss NIR Systems 5000) utilizes the mentioned classic method of background technology to measure the leucine content of all blades simultaneously.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 leucine content of corresponding measurement as output variable, utilize multiple linear regression method (Multiple linear regression MLR) obtains following multiple linear regression equations:
Y (mg/100mg DW)=199.490 λ
1-205.134 λ
2-157.082 λ
3+ 158.636 λ
4-98.514 λ
5-40.158 λ
6+ 74.919 λ
7+ 79.434 λ
8-79.398 λ
9-76.208 λ
10+ 142.368 λ
11+ 45.322 λ
12+ 47.134 λ
13-45.866 λ
14+ 27.124 λ
15-99.757 λ
16-19.316 λ
17+ 44.704 λ
18+ 2.5554, calculate leucine content in rape leaves.
Y is a leucine content in rape leaves in the equation, and unit is mg/100mg DW; λ
1～λ
18Be respectively the reflectivity of wavelength 2360nm, 2386nm, 2306nm, 2314nm, 2330nm, 1194nm, 2198nm, 1420nm, 1462nm, 2106nm, 1602nm, 1878nm, 2426nm, 1814nm, 2274nm, 1676nm, 1910nm and the pairing rape leaf Powder samples of 2052nm light wave.
Utilize above-mentioned multiple linear regression equations that the leucine content 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 leucine content in rape leaves 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 leucine content in rape leaves is rapidly and accurately used in the The above results explanation.
Claims (4)
1. method for quickly detecting leucine content in 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 2360nm, 2386nm, 2306nm, 2314nm, 2330nm, 1194nm, 2198nm, 1420nm, 1462nm, 2106nm, 1602nm, 1878nm, 2426nm, 1814nm, 2274nm, 1676nm, 1910nm and the pairing rape leaf of 2052nm light wave respectively;
(3) with each reflectivity difference substitution multiple linear regression equations: y=199.490 λ
1-205.134 λ
2-157.082 λ
3+ 158.636 λ
4-98.514 λ
5-40.158 λ
6+ 74.919 λ
7+ 79.434 λ
8-79.398 λ
9-76.208 λ
10+ 142.368 λ
11+ 45.322 λ
12+ 47.134 λ
13-45.866 λ
14+ 27.124 λ
15-99.757 λ
16-19.316 λ
17+ 44.704 λ
18+ 2.5554, calculate leucine content in the rape leaf;
Y is a leucine content in rape leaves in the equation; λ
1～λ
18Be respectively the reflectivity of wavelength 2360nm, 2386nm, 2306nm, 2314nm, 2330nm, 1194nm, 2198nm, 1420nm, 1462nm, 2106nm, 1602nm, 1878nm, 2426nm, 1814nm, 2274nm, 1676nm, 1910nm and the pairing rape leaf Powder samples of 2052nm light wave.
2. method for quickly detecting leucine content in rape leaves according to claim 1 is characterized in that: described blade powder is 40～80 orders.
3. method for quickly detecting leucine content in 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 leucine content in 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 201010219028 CN101900678B (en) | 2010-07-05 | 2010-07-05 | Method for quickly detecting leucine content in rape leaves |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1180839A (en) * | 1996-08-01 | 1998-05-06 | 株式会社佐竹制作所 | Content measuring apparatus for plant leaf |
| JP2005257676A (en) * | 2004-02-09 | 2005-09-22 | National Agriculture & Bio-Oriented Research Organization | Method for determining quantity of chemical component in tea leaves |
| WO2005111583A1 (en) * | 2004-05-17 | 2005-11-24 | The New Industry Research Organization | Method for nondestructively examining component of vegetable or the like by near-infrared spectroscopy and its device |
| CN1715880A (en) * | 2005-08-03 | 2006-01-04 | 北京农业信息技术研究中心 | Non-destructive detecting method and detecting instrument for portable plant nitrogen and water content |
| CN1789980A (en) * | 2005-12-16 | 2006-06-21 | 浙江大学 | Plant growth information acquisition device based on near infrared spectrum |
-
2010
- 2010-07-05 CN CN 201010219028 patent/CN101900678B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1180839A (en) * | 1996-08-01 | 1998-05-06 | 株式会社佐竹制作所 | Content measuring apparatus for plant leaf |
| JP2005257676A (en) * | 2004-02-09 | 2005-09-22 | National Agriculture & Bio-Oriented Research Organization | Method for determining quantity of chemical component in tea leaves |
| WO2005111583A1 (en) * | 2004-05-17 | 2005-11-24 | The New Industry Research Organization | Method for nondestructively examining component of vegetable or the like by near-infrared spectroscopy and its device |
| CN1715880A (en) * | 2005-08-03 | 2006-01-04 | 北京农业信息技术研究中心 | Non-destructive detecting method and detecting instrument for portable plant nitrogen and water content |
| CN1789980A (en) * | 2005-12-16 | 2006-06-21 | 浙江大学 | Plant growth information acquisition device based on near infrared spectrum |
Non-Patent Citations (2)
| Title |
|---|
| 《光学学报》 20100430 孙光明 等 基于近红外光谱技术检测除草剂胁迫下油菜叶片中脯氨酸含量的方法 1192-1196 1-4 第30卷, 第4期 2 * |
| 《广西工学院学报》 20040331 余炜 等 纸色谱-色斑洗脱比色法定量测定L-亮氨酸的研究 51-54 1-4 第15卷, 第1期 2 * |
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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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