Centre wallon de Recherches agronomiques

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Centre wallon de Recherches agronomiques

Centre wallon de Recherches agronomiques -Département Qualité des productions agricoles Chaussée de Namur, 24 - B – 5030 GEMBLOUX - Tél : ++ 32 (0)81 62 03 50 - Fax : ++ 32 (0)81 62 03 88

dptqual@cra.wallonie.be - http://cra.wallonie.be

«Un autre regard sur votre process »

La spectrométrie dans le proche infrarouge appliquée au suivi de processus de transformation

CRA-W – Gembloux

Département Qualité des Productions agricoles

Laboratoire de Spectrométrie

Après-midi d’étude

Jeudi 16 mars 2006

asbl

Ministère de la Région Wallonne.

Centre wallon de Recherches agronomiques

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Centre wallon de Recherches agronomiques

¾ PRESENTATION OF CRA-W

¾ NIR theory

¾ HISTORY & NIRS APPLICATIONS AT CRA-W

¾ NETWORKING

¾ CHEMOMETRICS

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Centre wallon de Recherches agronomiques

Walloon Agricultural Research Centre

(Gembloux)

¾ Founded in 1872

¾Public Interest Organisation (OIP)

¾ Under the Regional Government of Wallonia.

http://www.cra.wallonie.be/

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Centre wallon de Recherches agronomiques

Walloon Agricultural Research Centre 7 Departments:

1. Biotechnology 2. Crop production

3. Biological Control and Plant Genetic Resources 4. Pesticide research

5. Agricultural Engineering

6. Animal Production and Nutrition

7. Quality of Agricultural Products

+ 2 units

1. Agricultural Systems (in Libramont)

2. Biometry, Data Management and Agrometeorology

Personnel: ~450, Scientists: ~120

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Centre wallon de Recherches agronomiques

Quality of Agricultural Products Department

3 laboratories :

• Microbiology & Molecular biology Dr Gilbert Berben

• Analytical chemistry & Technology Dr Georges Sinnaeve

• Physical methods

Dr Vincent Baeten

50 employees - 15 scientists

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Centre wallon de Recherches agronomiques

Quality of Agricultural Products Department

Physical methods

•Near Infrared Spectroscopy

Near Infrared Micro-spectroscopy Hyperspectral Imaging Spectroscopy

• Mid Infrared Spectroscopy

• Raman Spectroscopy

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Centre wallon de Recherches agronomiques

Near Infrared Spectroscopy

Why NIR ?

- Fast

- < 2 min/spl (Æ sec)

- multiple analyses (cheap) - Simple to operate

- short training - on-line

- no sample prep. or reduced - Simple to maintain

- lamps, …

Drawbacks:

- Need of calibrations - Lack of accuracy

- LOD >>

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Centre wallon de Recherches agronomiques

Near Infrared Spectroscopy

How does it work?

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Centre wallon de Recherches agronomiques

Discovery of Near InfraRed Radiation

• 17 March 1800

• William Herschel,

Astronomer Royal attempts to find out the spectral

region responsible for heat formation in his telescope.

•The NIR is discovered.

•Philosophical Transactions

of the Royal Society 90:255-

83

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Centre wallon de Recherches agronomiques

The Electromagnetic Spectrum

Micro- Far Mid NIR Visible Ultra- Vacuum X-Ray

10 5000 2500 800 400 170 20 nm

10 200 4000 12500 25000 60000

6 **5*10 5 cm -1**

Overtones of Molecular vibrations

Electronic transitions

rotation vibrations Inner shell

electrons

wave IR IR violet UV

λ

stretch bend v

Valence

electrons ionisation Molecular bond

HIGH PHOTON ENERGY LOW

PHOTON ENERGY

λν = c = 3 x 10 ⁸ m s ^-1 E _photon = hν h=6.6 x 10 ^-34 Js

ENERGY

- +

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Centre wallon de Recherches agronomiques

WAVENUMBER (cm ^-1 ) =

FREQUENCY (Hz)/100. C

λν=c

Wavelength(nm)=

10,000,000/wavenumber(cm ^-1 )

1 000 nm = 10 000 cm-1

2 500 nm = 4 000 cm-1

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Centre wallon de Recherches agronomiques

NATURE OF THE COVALENT BOND

C H

•Covalent bonds SHARE electrons between atoms in a molecule

1.09 Å

•Bonds have length, strength & direction unique to each pair of atoms

•Bonds act like springs joining atoms

•Bonds vibrate at unique frequencies due to atomic masses & ‘stiffness’

_ +

•If vibration changes the dipole moment then bond can absorb INFRARED photons

•A photon of EXACTLY the right

frequency is absorbed & excites the bond to a higher vibrational state

•Frequency = qualitative analysis:IDENTITY

•Amplitude = quantitative analysis:AMOUNT

Courtesy Ian Murray

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Centre wallon de Recherches agronomiques

Courtesy Ian Murray

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Centre wallon de Recherches agronomiques

Spectres de produits purs

1928 2100 2055 2182

1722 2306 2342

1 200 1 400 1 600 1 800 2 000 2 200 2 400 0

0.2 0.4 0.6 0.8 1

Longueurs d ’onde (nm)

Densité optique: LOG(1/R)

Amidon Gluten

Huile

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Centre wallon de Recherches agronomiques

Instruments

Source Detector

Grating

Sample

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Centre wallon de Recherches agronomiques

Instruments

SAMPLE PRESENTATION MODE

INTERACTANCE Fibre optic probe

T R I

I ₀ I _t

I ₀

TRANSMISSION REFLECTANCE

or transflectance 0-45°optics or integrating sphere

I _r

+ -

I ₀

∞

I _t =0

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Centre wallon de Recherches agronomiques

Instruments

I _r R=

I ₀

I _a I ₀

I _t

I _r

REFLECTION REFLECTION

I _t T=

I ₀ TRANSMISSION

TRANSMISSION

ABSORBANCE or OPTICAL DENSITY ABSORBANCE or OPTICAL DENSITY

 

 



= 

 

 



= 

T Log 1 R

Log 1

A

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Centre wallon de Recherches agronomiques

L OG (1/R)

0 0.5 1 1.5 2 2.5 3 3.5 4

0 0.1 0.2

0.3 0.4

0.5 0.6

0.7 0.8

0.9 1

R EF L EC T IO N

OP TIC A L D E N S ITY or ABS O R BANCE

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Centre wallon de Recherches agronomiques

Instruments

- Filters (interferential filters)

- Monochromator : dispersive system (tilting grating)

- AOTF (Acoustic Optical Tunable Filter) - LCTF (Liquid Crystal Tunable Filter)

- Interferometers (Michelson or polarization) --- - -

- Diode Array (fixed grating)

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Centre wallon de Recherches agronomiques

INSTRUMENT IMPROVEMENTS:

- Sources

- Optical components (laser) - Detectors

- Electronics (communication)

Sample presentation:

- large cups - slurries - liquids

- on-line (belts, pipes, …) - fibre optics

- remote scanning

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Centre wallon de Recherches agronomiques

THE PAST

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Centre wallon de Recherches agronomiques

NIR INSTRUMENTS – CRA-W

•1976 R.Biston au CRA – Station de Haute Belgique

•1979 Neotec FQA51 (Feed Quality Analyser)

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Centre wallon de Recherches agronomiques

NIR INSTRUMENTS – CRA-W

•1981 Neotec 6350 (Research Analyser) + Nova III

New Instrumentations

>65 companies Better noise, More flexible,

Better sample presentation

…

Portable (miniaturization) But

Almost the same spectral

information as 25 years ago

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Centre wallon de Recherches agronomiques

APPLICATIONS

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Centre wallon de Recherches agronomiques

APPLICATIONS developed at CRA-W

•Fertilizers (N, P, K, Humic acids)

•Soils (N, C, CEC, …)

•Seeds & Phyto-sanitary Protection (seed/seed)

•Crop monitoring ^(N)

•Precision Agriculture

•Nutritive value (feed & forages)

•Technology (flour, baking quality,…)

•Authenticity (orange juice, wine, olive oil, meat, honey,…)

•Fruits

•Bio-fermentation monitoring

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Centre wallon de Recherches agronomiques

•NUTRITIVE VALUE OF FEED

CHEMICAL COMPOSITION & DIGESTIBILITY

Forages:

Fresh grass Hay

Grass Silage

Fresh corn silage Corn silage

Tropical Forages Cereals

High-protein crops

(soya, pea, …

High fat crops

(rape, sunflower,..)

Feed Ingredients

Cereals & by-products Wheat bran

Soyameal

Sugarbeet pulp

Animal protein (MBM)

……

Complete feed Cattle

Swine Poultry Pet food

•Moisture – DM

•Ashes – OM

+ P, Ca, K, Mg

•Fat

+ FA profile

•Proteins (N) + AA profile

•Fibres

(cellulose, NDF,ADF,ADL)

•Starch

+ amylose - amylopectin

•Total Sugar

+ sugar profile

•OMD

in vivo, in vitro, enzymatic

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Centre wallon de Recherches agronomiques

NIRS MICROSCOPY &

IMAGING

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Centre wallon de Recherches agronomiques

•NIR INFRARED MICROSCOPE

Meat and Bone Meal DETECTION

PerkinElmer FT-NIR

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Centre wallon de Recherches agronomiques

- Camera InGaAs - 900 - 1700 / 10 nm - 240 x 320 pixels

- **Each pixel: 70 µm*70 µm - Analyzed surface : 5 cm² - 76 800 spectra 24 MB - 300 - 350 particles**

- Analysis time : +/- 5 min

NIR Camera

(x)

(y)

(Wavelength nm)

Matrix NIR,

Spectral Dimensions, Inc.

CRA-W – DQPA 23-02-2006

Community Reference Laboratory for Animal Proteins

ISO 17025

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Centre wallon de Recherches agronomiques

NETWORKING

STANDARDISATION OF INSTRUMENTS Same models on all the instruments

Make the instruments alike

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Centre wallon de Recherches agronomiques

Asbl REQUASUD

Cellule de coordination Chaussée de Namur, 24 5030 GEMBLOUX

081/62.03.71

requasud@cra.wallonie.be

Michamps

CRA

La Hulpe Ath

Soignies Tinlot

Libramont Waremme

Ciney

FUSAGx

1987

9 laboratoires interconnectés

en région wallonne

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ABERDEEN DUBLIN CAMPDEN LELYSTAD ARNHEM ETTELBRUCK LILLE

REIMS NANCY

NAMSHEIM PARIS

NANTES

CLERMONT-F RODEZ

MONTPELLIER TOULOUSE PAU

PADOVA CORDOBA MADRID

EUROPEAN CRAGx CONTACTS

ROTTERDAM VARSOVIE BUCAREST PARIS

RENNES FRAGA MADRID LISBONNE

EUROPEAN PROVIMI NETWORK

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Centre wallon de Recherches agronomiques

WORLD NIRS CONTACTS

ARGENTINE

AFRIQUE DU SUD AUSTRALIE

ARGENTINE BRESIL CHILI

ETATS-UNIS SENEGAL TUNISIE INDONESIE VIETNAM

Nouvelle ZELANDE

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Centre wallon de Recherches agronomiques

dptqual@cra.wallonie.be - http://cra.wallonie.be 0

0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1

40 0 50 0

60 0 70 0

80 0 90 0

10 00 11 00

12 00 13 00

14 00 15 00

16 00 17 00

18 00 19 00

20 00 21 00

22 00 23 00

24 00 25 00

NIRSystems 5000

FT-Buchi NF 500

FT-Antaris Nicolet FT-Brucker MPA

Unity Spectrastar Foss – Infraxact

Perten DA7200

CALIBRATION TRANSFER

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Centre wallon de Recherches agronomiques

•Networks of instruments (2000)

RINA ^® : Remote Instrument Near Analysis (Foss ISI)

NIRS

400-2500nm

INTERNET

NIRS

400-2500nm

CRA SERVER:

-equations -data bases Spectra

Values

User 1

User 2

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Centre wallon de Recherches agronomiques

CHEMOMETRICS

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Centre wallon de Recherches agronomiques

Calibration : Multiple Linear Regression

Y X

∑

+

= b b _i x _i

Y ˆ ₀

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Centre wallon de Recherches agronomiques

the wet chemistry cost X

Æ Y Minimising

Spectral variability

Varieties, Years, Locations, Origins, ….

Samples prep., Grinders,

Temperatures, Operators, … Batches, …

Experimental Designs Reference

Wet Chemistry SEL ??

r (repeatability)

R (reproducibility)

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Centre wallon de Recherches agronomiques

¾CHEMOMETRICS

Many Packages (manufacturers)

+ MatLab

Quantitative

MLR, PCR, PLS, ANN, SVM,….

Qualitative

LDA, QDA, KNN, SVM, SIMCA,..

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Centre wallon de Recherches agronomiques

Chemometrics

Uncertainty of the NIR analyses

( y ^ˆ i y i ) ( [ ¹ h i ) ^. SEC ² S ref ² ] ¹ ^/ ²

s − = + −

Fernandez Pierna & al., Chemom., Intell. Lab. Syst. 65,281 (2003)

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Centre wallon de Recherches agronomiques

CALIBRATION

y = 91.516x - 45.342 R ² = 0.9055

3 3.5 4 4.5 5 5.5 6 6.5 7

0.53 0.535 0.54 0.545 0.55 0.555 0.56 0.565 0.57

Absorbances

Y re fe re n ce

Least Squares

NIRS: Inverse regression Yref= b.X+ b0 + e

2 ) ,

cov(

s X

y b = x

Chemometrics : uncertainty

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Centre wallon de Recherches agronomiques

Chemometrics : uncertainty

PREDICTION

y = 91.516x - 45.342 R ² = 0.9055

3 3.5 4 4.5 5 5.5 6 6.5 7

0.53 0.535 0.54 0.545 0.55 0.555 0.56 0.565 0.57

Absorbances

Y ref e ren c e

y _est = bx+b0

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Centre wallon de Recherches agronomiques

N=20

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Centre wallon de Recherches agronomiques

N=500

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Centre wallon de Recherches agronomiques

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Centre wallon de Recherches agronomiques

http://www.nirpublications.com/

Near infrared on-line measurement of degree of cook in extrusion processing of wheat flour

On-line monitoring of rumen fluid in milking cows by fibre optics in transmittance mode using the longer NIR region

Non-invasive fermentation analysis using an artificial nerual network algorithm for processing near infrared spectra (ethanol)

Monitoring carotenoid and chlorophyll pigments in virgin olive oil by visible-near infrared transmittance spectroscopy. On-line application On-line monitoring of melt-extrusion transesterification of ethylene

vinylacetate copolymers by near infrared spectroscopy and chemometrics On-line monitoring of kinetic and sensory parameters in acetic