Nutritional Supplement and Diesel Fuel Application Development for Benchtop NMR Systems Operating at 42, 60, and

80 MHz – Equivalency with Supercon NMR

John C. Edwards – Process NMR Associates, LLC Danbury CT
150 4
Abstract: Benchtop high-resolution NMR systems are available at a number of field strengths and probe configurations. However beyond the obvious academic instruction market for these instruments very few applications have been
demonstrated across all available platforms and thus proving the general applicability of benchtop NMR technology to industrial quality control. 300 MHz 1H NMR EPA/DHA Analysis 3

Q Residuals (0.11%)

Y Stdnt Residual 1
We present two chemometric-based applications that have been developed at 4 different field strengths utilizing Varian Mercury 300 MHz, Magritek Spinsolve 42 MHz, Aspect AI 60 MHz, and Thermo Picospin 80 MHz NMR systems. 2
100
Partial-least-squares (PLS) regression correlations were obtained on all 4 platforms relating to: Experimental 1
1) Omega-3 fatty acid composition of samples taken from various points in a nutritional supplement manufacturing process. Excellent correlations were obtained on all 4 NMR instruments proving that NMR technology is applicable Varian Mercury - 300 MHz NMR Supercon Spectrometer
0
to in-lab, at-line. or on-line analysis of fish oil derived omega-3 fatty acid supplements. The 40 second NMR analysis effectively replaces a 60+ minute GC analysis. 4 pulse on pure sample in 5 mm tube, Run Unlocked
50 -1
In MNova 8.1.2 SPC Files Imported, Stacked, Binned at 3 Hz interval, EPA
2) Physical and chemical property determination of diesel fuels where excellent correlations were obtained between 1H NMR variability and parameters such as density, aromatic content by GC, hydrogen content by 1H TD-NMR Area Normalized to 100, Saved as Transposed Ascii Matrix -2
(ASTM D7171 method), and sulfur content. Many more physical and chemical properties can be correlated to the 1H NMR spectrum allowing a single 40 second NMR experiment to predict 10-15 parameters that each require For Peak Integrals Used Advanced Feature – Create Integral Graph
-3
dedicated analyzers. from Stacked Plot 0
0 5 10 15 20 25 30 0 0.2 0.4 0.6 0.8
Finally, we will present the concept and initial results from an independent server-based NMR application software that can be utilized in conjunction with the NMR software of the current benchtop NMR systems, or alternatively as a PLS Regression Performed Thermo Grams IQ and Eigenvector Solo Hotelling T^2 (99.89%) Leverage
stand-alone application platform. This software would effectively make chemometric and direct measurement NMR application ubiquitous across all NMR platforms.
EPA 80 40

R2 = 0.993

Scores on LV 2 (0.45%)
DHA 60 20

Y CV Predicted 1
8 Latent Variables
RMSEC=1.20
Eicosaoentaenoic Acid (EPA) 20:5(n-3) 40 RMSECV=1.68 0

20 -20

Docosahexaenoic Acid (DHA) 22:6(n-3) Expansion of Correlated Region

0 -40
0 20 40 60 80 -400 -200 0 200 400
Wt% EPA
Y Measured 1 Scores on LV 1 (95.40%)

Variables/Loadings Plot for 1H NMR - 55 Fish Oil Samples - Processed - 8-9-13 - Transposed.xlsx
0.04

0.03

300 MHZ 0.02

300 MHz NMR
Wt% DHA by GC Regression

Reg Vector for Y 1

0.01

Aspect AI – 60 MHz EPA/DHA Analysis EPA/DHA Analysis 300 MHz NMR

0
Vector
Wt% DHA by GC
4 pulse on pure sample in 5 mm tube Regression Vector
Wt% DHA by GC -0.01

Locked on 1H NMR signal Wt% DHA by GC

In MNova 8.1.2 Magritek Spinsolve – 42.5 MHz -0.02

SPC Files Imported, Stacked, Binned at 3 Hz interval, Area Normalized to 1000 4 pulse on pure samples in 5mm tube
-0.03
20 40 60
Variable
80 100 120

Saved as Transposed Ascii Matrix Data Processed in Mestrelab Mnova 9

For Peak Integrals Used Advanced Feature – Create Integral Graph from Stacked Plot
PLS Regression Performed Thermo Grams IQ and Eigenvector Solo
Data converted to 0.05 ppm integration bins
Spectral area Normalized to 1000
Physical and Chemical Properties of Diesel Fuel
1H benchtop NMR has great potential to increase the throughput of both routine and emergency fuel sample analysis in refinery laboratories. Currently fuel samples must be
Saved as transposed matrix
passed through multiple dedicated analyzers to obtain information such as density, H-Content, aromatics, olefins, saturates, benzene,
Octane numbers, cetane index, cetane number, distillation curves, vapor pressure, flash point, pour point, freeze point, cloud point, etc. Correlation of the 1H NMR spectra of
300 MHz 1H NMR Spectra of Samples at Different Points in the Process these refinery fuel samples to these primary test results will allow all parameters to be predicted in about 40 seconds from the 4 pulse spectrum of the pure fuel. Here we have
a few examples obtained on some diesel fuels that were submitted to our lab for ASTM D7171 – Hydrogen Content by TD-NMR. We had density, H-content, and aromatics wt%
NMR Processing for Multivariate Analysis by GC. Below are three example correlation obtained on the Picospin 80 system (that requires 32 pulses per sample due to the capillary sample size). The results were very
NMR ID
FO3h026
EPA (Area %) DHA (Area %)
44.55 19.30
Sample Description
Clathration similar for the 300, 60, and 42 MHz data obtained on the three other NMR system in our laboratory. The comparative results are shown in Table II. The results are very similar
NMR ID
FO3h001
EPA (Area %)
0.64
DHA (Area %)
0.01
Sample Description
First Esterification
50 sample initial data set from FO3h027
FO3h028
16.69
6.87
9.89
4.53
First Esterification
Crude Pollock Oil independent of the field strength of the NMR system. The data from all 4 NMR systems is provided in this section.
FO3h002
FO3h003
21.55
62.97
13.34
15.66
First Esterification
Clathration all points in manufacturing process. FO3h029
FO3h030
62.79
37.29
19.43
22.03
Separator
PolyUnsat Ester
FO3h004 29.43 18.16 Mol Dist
FO3h005
FO3h006
14.21
52.74
9.54
28.90
Pollock Oil
Separator
40 Samples used for 80 MHz dataset FO3h031
FO3h032
9.71
32.00
0.38
15.10
First Esterification
First Esterification
FO3h033 38.79 26.75 Clathration
FO3h007 15.21 10.51 PolyUnsat Ester
FO3h034 41.87 23.25 Mol Dist
FO3h008
FO3h009
7.18
16.95
0.23
10.04
First Esterification
First Esterification First round of PLS regression analysis FO3h035
FO3h036
35.49
0.30
43.99
0.00
Separator
First Esterification
FO3h010 36.35 16.47 Clathration
FO3h011
FO3h012
61.09
13.32
21.26
5.95
Mol Dist
MSC Pollock Oil
revealed concentration outliers that FO3h037
FO3h038
34.09
15.44
8.09
9.82
Clathration
MonoUnsat Ester
FO3h013 71.78 7.43 Separator
were linked to limitations in the GC FO3h039 60.08 24.52 PolyUnsat Ester
30 Diesel Fuels - 80 MHz
FO3h014
FO3h015
41.40
1.19
25.91
0.06
PolyUnsat Ester
First Esterification
method. This study was performed on
FO3h040
FO3h041
8.77
12.41
5.75
57.59
Crude Salmon Oil
Separator 3 Diesel Fuels - 60 MHz
FO3h016 11.73 12.23 First Esterification FO3h042 36.36 21.49 PolyUnsat Ester 32 Pulse Average
FO3h017
FO3h018
43.38
6.07
19.30
2.78
Clathration
Clath Raffinate improved GC data values.
FO3h043
FO3h044
3.79
29.23
0.15
13.78
First Esterification
Clath Raffinate
25 2 4 Pulse Average

Q Residuals (0.09%)
FO3h045 45.99 33.51 PolyUnsat Ester

Y Stdnt Residual 1
FO3h019 9.77 0.72 First Esterification
FO3h020 58.93 23.41 Mol Dist FO3h046 12.10 5.69 MSC Pollock Oil
FO3h021 10.62 5.18 MSC Pollock Oil
Final 80 Sample models were utilized FO3h047 24.57 14.32 Separator 20 1
FO3h022 43.91 21.52 Separator FO3h048 45.86 33.61 PolyUnsat Ester
FO3h023 54.05 28.18 PolyUnsat Ester FO3h049 6.39 3.68 First Esterification
FO3h024 0.00 0.00 First Esterification to validate the calibrations on a 24 FO3h050 58.13 24.66 Clathration
FO3h025 26.97 12.82 First Esterification 15 0
sample validation set (below).
10 -1

5 -2

-3
0
0 5 10 15 20 0 0.2 0.4 0.6 0.8
Hotelling T^2 (99.91%) Leverage

80 20 80 15
60 MHz 60 MHz - EPA 42.5 MHz
Wt% EPA
Wt% EPA

Scores on LV 2 (0.64%)
10
60 10 60

Y CV Predicted 1
30 4

Scores on LV 2 (2.48%)
5

Y CV Predicted 1
25
40

Q Residuals (0.09%)
40 02

Y Stdnt Residual 1
0
20
R2=0.993 -5
R2=0.988 Diesel Fuels – 42 MHz
20 15 SECV=2.13 wt% -100 20
SECV=2.20 wt%
Diesel Fuels - 300 MHz
8 Latent Variables
10 8 Latent Variables -10 4 Pulse Average 4 Pulse Average
-2 0
0 5 -20 -15
0 20 40 60 80 -200 -100 0 100 200
42 MHz - EPA
0 Y Measured 1 -4 Scores on LV 1 (98.04%)
0 5 10 15 20 25 30 0 0.2 0.4 0.6 0.8 -20 -20
Hotelling T^2 (99.91%) Leverage 0 20 40 60 80 -100 -50 0 50
Y Measured 1 Scores on LV 1 (95.78%)

60 20 80 20
60 MHz 60 MHz - DHA
50 42.5 MHz
Integration of Peaks to

Scores on LV 2 (0.36%)
Wt% DHA
10 Wt% DHA
Y CV Predicted 1

60
Produce Multivariate 40
10
Integration Spectra

Scores on LV 2 (2.23%)
30 0

Y CV Predicted 1
R2=0.992 40
20
SECV=1.13 wt%
-10 0
600 7 Latent Variables
10 R^2 = 0.989
20
500 8 Latent Variables
0 -20
Omega-3 Ethyl Ester Sample obtained at 4 field strengths – ppm scale 0 10 20 30 40 50 60 -200 -100 0 100 200 RMSEC = 0.75363
400 Y Measured 1 Scores on LV 1 (97.83%) -10
0 RMSECV = 1.2463

300 Calibration Bias = 0

42.5 MHz - DHA
Data

Chemometric Software Utilized in this study: CV Bias = 0.098796

200
1) Eigenvector Research Solo 7.0.3 -20 -20
2) LatentiX 2.12 0 10 20 30 40 50 60 -100 -50 0 50
100
Y Measured 1 Scores on LV 1 (95.78%)

0
EPA/DHA Analysis
-100
1 2 3 4 5 6
Variables
7 8 9 10 11 Thermo Picospin80 – 82.3 MHz 80 MHz
Wt% EPA
32 pulse average on pure sample
Data Processed in Mestrelab Mnova 9 Density Aromatics
The 1H NMR PLS correlations to EPA and DHA content in the various manufacturing streams and products are Data converted to 0.05 ppm integration bins
provided in Table I and demonstrate that all 4 NMR systems produce excellent correlations. Good correlations (kg/L) (wt%) Hydrogen Content
Spectral area Normalized to 1000
can also be obtained from gross integration region “spectra” indicating that model robustness may be improved Saved as transposed ascii matrix (%H)
by data reduction of NMR spectra into wider chemically relevant integration bins.

Table I: Regression Results for Wt% DHA/EPA Content at 4 Field Strengths Table II: PLS Regression Results for Diesel Quality Parameters at 4 Field Strengths
Conclusion
1H
NMR and Diesel Parameter Correlation – PLS Regression A wide range of PLS correlation models can be readily built
EPA Content (Wt%) R2, SECV DHA Content (Wt%) R2, SECV based on NMR data obtained on both superconducting and
Hydrogen Content (%H) Aromatic Content
PLS Regression Data (Wt%) (Wt%) 80 MHz benchtop permanent magnet NMR systems. Currently models
ASTM D7171 TD-NMR (Wt% GC) Density (Kg/L)
Wt% DHA require that data be obtained on each individual spectrometer
42 MHz NMR 0.988, 2.20 0.989, 1.25 system but it may be possible that various spectral ‘de-resolution’
NMR Freq R2 SECV LV R2 SECV LV R2 SECV LV
60 MHz NMR 0.993, 2.13 0.992, 1.13 techniques may make models obtained on one system transferable
between NMR systems at varying magnetic field strengths.
82 MHz NMR 0.989, 2.17 0.992, 1.11 42 MHz 0.951 0.10 3 0.959 0.54 4 0.944 0.0021 5 At-line and in-line permanent magnet NMR systems can
300 MHz NMR 0.988, 1.68 0.991, 1.09 yield the same high quality correlations as data obtained on
60 MHz 0.952 0.09 3 0.962 0.53 2 0.951 0.0020 2 much higher field superconducting NMR systems. Very little
Peak Integral Data 0.981, 2.71 0.964, 2.24 difference is observed in the quality of the correlations and errors
Fused - NMR-FTIR 0.993, 1.62 0.992, 1.08 80 MHz 0.934 0.10 3 0.953 0.57 3 0.937 0.0022 3 of prediction on models developed on the 4 systems in our
Omega-3 Ethyl Ester Sample obtained at 4 field strengths – frequency scale laboratory.
300 MHz 0.974 0.07 3 0.953 0.57 5 0.937 0.0021 5