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 Copyright (c)2010 Iowa State University NIRS and AOCS Near Infrared Spectroscopy (NIRS)is a rapid, indirect, analytical method with expanding use in oilseed marketing.
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 Copyright (c)2010 Iowa State University NIRS and AOCS Near Infrared Spectroscopy (NIRS)is a rapid, indirect, analytical method with expanding use in oilseed marketing.
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 Copyright (c)2010 Iowa State University NIRS and AOCS Near Infrared Spectroscopy (NIRS)is a rapid, indirect, analytical method with expanding use in oilseed marketing.
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 2 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 Copyright 2010 Iowa State University Transmittance 3 Source Wavelength selector SAMPLE Detector Readout Copyright 2010 Iowa State University Reflectance 4 Source Wavelength selector SAMPLE Detector Readout Copyright 2010 Iowa State University NIRS Instruments Copyright 2010 Iowa State University 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 Copyright 2010 Iowa State University 7 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 Copyright 2010 Iowa State University NIRS Calibration Steps required for estimation of a mathematical relationship between optical properties of the sample and its known chemical composition. 8 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) Copyright 2010 Iowa State University 9 Spectral Preprocessing - Remove noise - Enhance useful information Example using multiplicative scatter correction Function of derivatives Copyright 2010 Iowa State University NIRS Prediction Calculation of composition by using spectral data (transmittance or reflectance) in calibration model 10 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. 11 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 Copyright 2010 Iowa State University Sample Set Size is Important Copyright 2010 Iowa State University Reproducibility of Reference Data Copyright 2010 Iowa State University Reproducibility of Reference Data Copyright 2010 Iowa State University 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. Copyright 2010 Iowa State University Calibration Transfer Basic Copyright 2010 Iowa State University Calibration Transfer Difficult Copyright 2010 Iowa State University 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 Copyright 2010 Iowa State University 22 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 C o e f f i c i e n t
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D e t e r m i n a t i o n 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 Where To Find Us