Time-Domain

Based Reduction
of Complex NMR
Data

Dave Russell
FoodMR 2014
I know what I like and I like what I know.
Dr. J. Josephs, Sr.

Courtesy: Lydia Park

Overview - CRAFT analysis of NMR data

Introduction to the problems of frequency-

based analyses

Comparison of CRAFT to standard

techniques

Real-world application soy bean extracts

Quantitation in frequency domain
This is easy when the spectra are simple
CRAFT example
Complex overlapped spectra are the rule in real samples

Zinfandel Wine

100x
Spectral Deconvolution
What do we mean by deconvolution ?
Spectral Deconvolution
What do we really mean by deconvolution ?
Quantitation in frequency domain
Quantitation in frequency domain
Spectra from the same FID

Three spectra differ in linewidth only!

Time vs. Frequency domain: the good and the bad

! x y presentation of frequency vs. intensity

peak height = f(amplitude, linewidth, phase)
integral = f(amplitude, linewidth, phase)

! Baseline issues
A few corrupted data points in the fid
translate to several corrupted data points in
frequency axis
Time vs. Frequency domain: the good and the bad

! Sum of several sinusoids ! x y presentation of frequency vs. intensity

! Each sinusoid represented by independent peak height = f(amplitude, linewidth, phase)
NMR properties integral = f(amplitude, linewidth, phase)
Frequency
Amplitude
! Baseline issues
Decay rate
A few corrupted data points in the fid
Phase
translate to several corrupted data points in
frequency axis
Spectral Deconvolution
What do we really, really want by deconvolution ?

Quantitative analysis in time

domain
"Minimize quantitation issues
induced by phase and baseline
correction errors.
? Non-intuitive (i.e., not visual)
and abstract.
CRAFT workflow
Krishnamurthy, K., Mag. Reson. Chem., 2013
doi: 10.1002/mrc.4022

Cluster of
FIDs

Define ROI
Array of spectra

Alignment table

Relative
Statistical amplitudes
Analysis (components)
Fingerprint chemical
shift for chemicals of
interest
Quantitation in frequency domain
Spectra from the same FID

Three spectra differ in linewidth only!

CRAFT example
Total CRAFTing time: 3.5 minutes

Models

Residual

Reconstructed

Experimental
Overview

Introduction to the problems of frequency-

based analyses

Comparison of CRAFT to standard

techniques

Real-world application soy bean extracts

 CRAFT Results: Fingerprint peaks

17& SA Bradley, TA Smitka, DJ Russell and K Krishnamurthy. Current Metabolomics, submitted March&2014&
 CRAFT Results: Comparison

18& SA Bradley, TA Smitka, DJ Russell and K Krishnamurthy. Current Metabolomics, submitted March&2014&
 CRAFT Results: Comparison

19& SA Bradley, TA Smitka, DJ Russell and K Krishnamurthy. Current Metabolomics, submitted March&2014&
Overview

Introduction to the problems of frequency-

based analyses

Comparison of CRAFT to standard

techniques

Real-world application soy bean extracts

CRAFT applications
Food sciences soy dietary supplement analysis

! Over-the-counter soy dietary

supplements
! 3 different soy supplements
! 15 (replicate) capsules from
each group
! Extracted, filtered, ., 1H
NMR collected
Soy Extract - Aromatics
Isoflavone peaks of interest

RO O
8
7 9 2
A 10
B
6 3
5 4
2'
1' 3'
C 4'
OH O
OH
Soy Extract - Aromatics
Isoflavone peaks of interest
Soy Extract Analyte Table
Isoflavone peaks of interest
CRAFT example
Soy extract analysis

3 groups; 15 samples/each; ~2 min/spectrum CRAFTing time

CRAFT example
Soy extract analysis

3 groups; 15 samples/each; ~2 min/spectrum CRAFTing time

CRAFT example
Soy extract analysis

3 groups; 10 samples/each; 30 sec/spectrum CRAFTing time

CRAFT applications
Pure Shift 1D Homonuclear broadband decoupled 1H spectrum
K. Zangger, H. Sterk, J. Magn. Reson. 124. (1997) 486. (Original PS1D)
J.A. Aguliar, S. Faulkner, M. Nilsson, G.A. Morris, Angew. Chem. Int. Ed. 49. (2010) 3901.

sucrose in d2o
Normal PROTON

Pure Shift 1D

5.0 4.5 4.0 3.5 3.0

CRAFT example
Soy extract analysis

4 groups; 10 samples/each; 30 sec/spectrum CRAFTing time

CRAFT
Conclusions

Time-domain analysis yields accurate and precise results on

even the most complex spectra.
CRAFT allows fast, automated conversion of NMR data into
the desired tabular format.
Our concept of overlap in NMR data needs to be revisited.
NMR is a fantastic technique for use in many types of Food
Science investigations.
You still cant cheat physics.
CRAFT - Acknowledgements
Scott Bradley (Eli Lilly)
Andreas Kaerner (Eli Lilly) Agilent
Jonus Buser (Eli Lilly)
Allen Kline (ex Eli Lilly) Krish Krishnamurthy
Ross Johnson (Eli Lilly) Frank Delaglio
Juel DeHoniesto (Eli Lilly) Dan Iverson
He Liu
Daina Avizonis (McGill University) Mauro Cremonini
Greg Helms (Washington State U) Dimitris Argyropoulos
Heiko Schill
Scott Baggett (Acorn) Ron Crouch

Joe Ackerman (WashU, St. Louis)

Larry Bretthorst (WashU, St. Louis)
Thank you !