Industrial qNMR Using Low-Field NMR Instrumentation:

Perspectives and Progress

John C. Edwards
Process NMR Associates, LLC, 87A Sand Pit Rd, Danbury, CT 06810 USA

132nd AOAC Annual Meeting and Exposition

August 26-29, 2018
Toronto, Canada

www.process-nmr.com www.nmrblog.com Tel: 203-744-5905

 Pulsar
60 MHz NMR
3-5 mm Samples Fourier 60
Lab Only 60 MHz NMR Picospin 45 and 80
3-5 mm Samples 45 MHz and 82 MHz NMR
42 & 60 MHz NMR Lab Only 300 micron capillary injection
3-5 mm Samples Lab Only
Lab Only
Cryogen Free
Permanent Magnet
High-Resolution
NMReady 60 Benchtop NMR 60 and 90 MHz
60 MHz NMR
5 mm Samples
3-5 mm Samples
Lab Only
Lab Only
aspectimaging
60 MHz NMR 60 MHz NMR
3-10 mm Samples 3-10 mm Samples
Lab or On-Line Lab or On-Line
 Magritek 43 MHz

Purity of Lidocaine
By Internal Standard

Six independent samples were created by co-

dissolving lidocaine and maleic acid in DMSO-d6 at
a concentration of approximately 50 mg/mL. A
mean purity of 99.0 ±0.7 % was measured from the
six samples, confirming the manufacturer’s purity
specification of >98%.
43 MHz NMR – L-Lysine in D2O + MA Internal Standard
60 MHz NMR – L-Lysine in D2O + MA Internal Standard
300 MHz NMR – L-Lysine in D2O + MA Internal Standard
 Lysine in D2O - Concentration Linearity - Lysine in D2O - Concentration Linearity - 60 MHz
300 MHz 12.0
12.0
10.0
y = 0.9985x
10.0 R² = 0.9969
y = 0.9718x 8.0
R² = 0.9999
8.0
6.0

6.0
4.0

4.0
2.0

2.0
0.0
0.00 2.00 4.00 6.00 8.00 10.00 12.00
0.0
0.00 2.00 4.00 6.00 8.00 10.00 12.00 Lysine in D2O - Concentration Linearity - 43 MHz
12.0

WLys (%w/v) = 100*(WMA * (MWLys / MWMA ) * ( ILys / IMA )) / Vtotal

10.0
WMA = weight of Maleic Anhydride Internal Standard (10mg)
MWLys = 182.65 amu - Molecular weight of L-Lysine.HCl 8.0

MWMA = 116.07 amu – Molecular Weight of Maleic Acid

6.0
y = 1.0155x
ILys = Molar Integral of L-Lysine Signal (adjusted for number of protons) R² = 0.998

IMA = Molar Integral of Maleic Acid Signal (adjusted for number of protons) 4.0

Vtotal – total volume of solvent (1000 l)

2.0

0.0
0.00 2.00 4.00 6.00 8.00 10.00 12.00
50 sample initial data set from all points in manufacturing process.
First round of PLS regression analysis revealed concentration outliers that were
linked to limitations in the GC method. This study was performed on improved
GC data values.
Final 50 Sample models were utilized to validate the calibrations on a 24 sample
validation set
Experimental
Varian Mercury - 300 MHz NMR Supercon Spectrometer 300 MHz 1H NMR In chain
4 pulse on pure sample in 5 mm tube, Run Unlocked CH2
In MNova 8.1.2 SPC Files Imported, Stacked, Binned at 3 Hz interval,
Area Normalized to 100, Saved as Transposed Ascii Matrix Ethyl
PLS Regression Performed Thermo Grams IQ and Eigenvector Solo
CH3
EPA-a

EPA-b
DHA w3
CH3

Olefins
w6/ w9
CH3-CH2-O-OC-CH2-R
=C-CH2-C= EtOOC-CH2-R CH3
EPA
R-CH2-C=

DHA
 DHA EPA
EPA

EPA
EPA

DHA

Comparison of High EPA and High DHA PUFA-Ester after Separation

www.process-nmr.com www.nmrblog.com Tel: 203-744-5905
www.process-nmr.com www.nmrblog.com Tel: 203-744-5905
Spinsolve 1H NMR data at 42.5 MHz of a series of fish oils and esterified omega-3 fatty acids
3 Hz binning interval

0.5 Hz binning interval

Picospin-80 NMR Data – 82.3 MHz

DHA
 Regression Results for Various NMR Approaches

EPA Content (Wt%) DHA Content (Wt%)

R2, SEP (Wt%) R2, SEP (Wt%)
43 MHz NMR 0.988, 2.20 0.989, 1.25
60 MHz NMR 0.993, 2.13 0.992, 1.13
82 MHz NMR 0.984, 2.53 0.990, 1.20
300 MHz NMR 0.988, 1.68 0.991, 1.09

www.process-nmr.com www.nmrblog.com Tel: 845-240-1177

Question:
Why is this competitors product so good?

Ingredients : Oyster, Barberry,

Dextrose, Snow Lotus, Bombyx Mori L,
Ginger Root, Salfron Crocus.

Sample preparation:
Extract with 20:80 D2O:CD3CN
Centrifuge or Settle
Supernatant analyzed.

Answer:
Also contained ~10 wt% Sulfoaildenafil
A Viagra Analog
60 MHz

300 MHz
Tadalafil
 43 MHz

60 MHz
Eugenol
 Anethol

Fenchone

EO % Pinene Fenchone Limonene Anethol

0.98 2.1% 15.6% 3.5% 74.5%
 Acknowledgements

Anton Bzhelyansky, Kristie Adams - USP

Tal Cohen, Aspect AI – 60 MHz NMR Systems

Paul Giammatteo, Cosa-Xentaur – 60 MHz NMR Systems
Cortney Phillips, Leap Technologies – 43 MHz Magritek Spinsolve
Mark Dixon, Thermo – Picospin-80