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Copyright 2010 Iowa State University

The New Guidance Method for


Calibration, Validation and Use
of Near Infrared Analyzers
Dr. Charles R. Hurburgh, Jr.
Agricultural and Biosystems Engineering
Iowa State University
Dr. Benoit Igne
Mylan School of Pharmacy
Duquesne University
IOWA STATE UNIVERSITY
w w w. g r a i n l a b. o r g
Copyright 2010 Iowa State University
NIR Light
Electromagnetic spectrum
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What is so special about the waves in NIR range?
NIR light is absorbed by molecules containing C-H, N-H, and O-H
groups (fats, proteins, carbohydrates, organic acids, alcohol, water)
NIR range:
750 - 2500 nm
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Transmittance
3
Source Wavelength
selector
SAMPLE Detector Readout
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Reflectance
4
Source
Wavelength
selector
SAMPLE
Detector
Readout
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NIRS Instruments
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NIRS and AOCS
Near Infrared Spectroscopy (NIRS)is a
rapid, indirect, analytical method with
expanding use in oilseed marketing.
The Soybean Quality Traits (SQT) has
four programs to promote uniformity
across NIRS users and platforms.
Core sample set
Proof of application testing
Standard Calibration Method
Proficiency (check sample) testing
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AOCS Analytical Guidelines Am 1a-09
Near Infrared Spectroscopy Instrument
Management and Prediction Model Development
Main Topics
Performance
Sample Sets
Lab Values
Preprocessing
Validation/Calibration Statistics
Calibration Transfer/Standardization
Moisture Basis
Correlated Y Variables
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NIRS Calibration
Steps required for estimation of a mathematical
relationship between optical properties of the
sample and its known chemical composition.
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Spectral Data
X
Spectral Data
X
Constituent concentration
Y
(Obtained by standard wet
chemistry methods)
Constituent concentration
Y
(Obtained by standard wet
chemistry methods)
Regression
algorithm
Regression
algorithm
Mathematical
relationship
(calibration model)
Y=f (X)
Mathematical
relationship
(calibration model)
Y=f (X)
Multiple Linear Regression (MLR)
Principal Component Regression (PCR)
Partial Least Squares (PLS)
Artificial Neural Networks (ANN)
Locally Weighted Regression (LWR)
Support Vector Machines (SVM)
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Spectral Preprocessing
- Remove noise
- Enhance useful information
Example using multiplicative
scatter correction
Function of derivatives
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NIRS Prediction
Calculation of composition by using spectral data
(transmittance or reflectance) in calibration model
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Spectral Data
X
Spectral Data
X
Constituent concentration
Y
Constituent concentration
Y
Mathematical
relationship
(calibration model)
Y=f (X)
Mathematical
relationship
(calibration model)
Y=f (X)
Copyright 2010 Iowa State University
NIRS Validation
Proof that a calibration is maintaining accuracy
with standard, accepted chemical methods over
the useful life of the calibration.
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NEW SAMPLES Not in Calibration Population
Spectral Data
X
Spectral Data
X
Constituent
concentration
Y
(Obtained by standard wet
chemistry methods)
Constituent
concentration
Y
(Obtained by standard wet
chemistry methods)
Comparison of
calculated values to
reference data
SEP, RPD, etc
Comparison of
calculated values to
reference data
SEP, RPD, etc
Mathematical
relationship
(calibration model)
Y=f (X)
Mathematical
relationship
(calibration model)
Y=f (X)
Copyright 2010 Iowa State University
Evaluation Statistic - RPD
RPD Class Application
0.02.3 Very poor Not recommended
2.43.0 Poor Rough screening
3.14.9 Fair Screening
5.06.4 Good Quality control
6.58.0 Very good Process control
8.1+ Excellent Any application
RPD = (Std. Dev. of Reference Data)/(Std. Error of Prediction)
Copyright 2010 Iowa State University
SBM Protein Calibration
Figure 2. Protein in soybean meal
Calibration Cross Validation
2006-2008 Samples
SECV=0.65; RPD = 6.2
Validation Set
2009 Samples
SEP = 0.36; RPD = 2.6
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Sample Set Size is Important
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Reproducibility of Reference Data
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Reproducibility of Reference Data
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Inter-Lab Reproducibility
See Also: Hartwig, R.A. and Charles R. Hurburgh, Jr. 1991. Interlaboratory comparison of soybean
protein and oil determinations. JAOCS 68(12):949-957.
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Calibration Transfer Basic
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Calibration Transfer Difficult
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Moisture Basis
Terminology
Native moisture basis = moisture basis as
developed. Choices:
As is (at the moisture as scanned)
Direct (Reference values adjusted to an MB)
Reported moisture basis = moisture basis as
converted and reported (if different)
These are all the same sample:
35% Pro@10% M=33.8% Pro@13%M
=38.9% Pro@ 0% M
Copyright 2010 Iowa State University
Correlated Y - Amino Acids
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Correlated Y:
Amino Acids
Comparison of NIR
calibration models
(average r
2
) with
linear regressions of
reference amino
acids to reference
protein
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
LEU LYS HIS PHE ASP TYR ARG GLY ALA ISO PRO VAL THR GLU MET SER CYS TRY
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The best of NIR
calibrations
Linear regression
to protein
Regression vectors
of 18 AA PLS
models for FOSS
Infratec 1241 Grain
Analyzer; most of
the curves follow
the same pattern,
which indicates that
calibrations predict
one constituent.
Copyright 2010 Iowa State University
Summary
AOCS Analytical Guidelines Am 1a-09
Near Infrared Spectroscopy Instrument Management and
Prediction Model Development
A guideline to assist in NIRS calibration efforts.
Covers the main issues of NIRS Operation.
Users need to be able to do their own calibration.
Guidelines are not static; expect updates!
Provide input!
Copyright 2010 Iowa State University
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