The Complex Fingerprint Analysis of Alcoholic Beverages

by Quantitative NMR (qNMR)

John Edwards, Ph.D.,

Process NMR Associates, LLC, Danbury, CT

Presented at TTB Beverage Alcohol Laboratory, Beltsville MD

February 2, 2017
NMR Spectrometer
Beer
 CH3-CH2-OH
CH3

CH2

OH
Precision of Measurement = 0.5%
 N=3

HO-CH2-CH3

Maleic Acid
Internal
Standard

N=2
 Sigma-Aldrich Organic TraceCERT®
products are characterized by:
•Manufactured according to
ISO/IEC 17025 and ISO Guide 34
•Certified Content by quantitative
NMR (qNMR)

•Superior level of accuracy,

calculated uncertainties and
lot-specific values
•Traceability to NIST
•Comprehensive
documentation delivered
with the product
(certification according to ISO
Guide 31)
 d, 5.32 ppm

Glucose d, 4.1 ppm

d, 5.21 ppm
Sucrose d, 5.4 ppm
Fructose Galacuronic Acid

Lactic Acid
Malic Acid d, 1.41 ppm Acetic Acid Succinic Acid Citric m, 2.94 ppm
m, 2.89 ppm s, 2.08 ppm s, 2.67 ppm

d, 0.88 ppm Ethanol

s, 3.32 ppm 1,3-propandiol

Glycerol Isobutanol Isoamyl alcohol t, 1.17 ppm p 1.76 ppm
Methanol m, 3.61 ppm d, 0.87 ppm
d 1.14 ppm
d, 1.56 ppm
s=singlet
Examples of Major Protons Utilized d=doublet
for 1H Quantitative (qNMR) Analysis t=triplet
p=pentet
of Alcoholic Beverages m=multiplet 2,3-butandiol
Alanine
Quinic Acid
2 x d 1.9 ppm
Calculations:
Standard Solution = 10mg Maleic Acid (MA) per 100ml of D2O Solution
Original Sample: The following volatile components were always obtained on a 175 ml beer sample as the
freeze drying process would compromise the amount present: ethanol, acetic acid, fusel alcohols
(isobutanol, isopentanol (isoamyl alcohol), and 1-propanol) ethyl acetate, methanol. The calculation
utilized to quantify these components was:

Component mg/L = 0.995 * 10 mg * ((Icomp / Ncomp) / 50) * (Mwcomp / 116.1) * (1,000,000 / 175)

Where, wt of maleic acid (MA) = 10mg, Icomp = Integration of component resonance, Ncomp = number of
protons integrated, IMA/NMA = 50 (MA integral set as 100), Mwcomp = molecular weight of component
molecule, MW of MA = 116.1 amu, the 1,000,000/175 factor rectifies the volumetric component of the
calculation to allow mg/L to be calculated. 0.995 represents the 99.5% purity of the maleic acid standard.
Freeze Dried Sample: The following non-volatile components were always obtained on a 1000 ml freeze
dried sample: lactic acid, succinic acid, malic acid, citrate, various amino acids, glucose, malt and
carbohydrates, glycerol. The calculation utilized to quantify these components was:

Component mg/L = 0.995 * 10 mg * ((Icomp / Ncomp) / 50) * (Mwcomp/ 116.1) * (1,000,000 / 1,000)

Where, wt of MA = 10 mg, Icomp = Integration of component resonance, Ncomp = number of protons

integrated, IMA/NMA = 50 (MA integral set at 100), Mwcomp = molecular weight of component molecule,
MW of MA = 116.1 amu, the 1,000,000/1,000 factor rectifies the volumetric component of the calculation
to allow mg/L to be calculated. 0.995 represents the 99.5% purity of the maleic acid standard.
Kombucha
Liquor, Liquers, Wine, Sake, Mead
Vinegars
 qNMR Developments
• Bruker had developed the NMR WineScreener – which can provide fingerprint chemistry
analysis, grape variety, vintage and geographical origin.

• Various Summits are being held to encourage the use of qNMR in quality control, purity and
authentication analysis and standard test method development
*1st qNMR Summit - US Pharmacopoeial Convention – Rockville MD October 2016
* 2nd qNMR Summit – BAM, Berlin, Germany, March 15-17, 2017

One Reference of Many:

Quantitative 1H NMR Analysis of Egg Yolk, Alcohol, and Total Sugar Content in Egg Liqueurs
• Monika Hohmann, Verena Koospal, Claudia Bauer-Christoph, Norbert Christoph, Helmut Wachter,
Bernd Diehl, and Ulrike Holzgrabe
• J. Agric. Food Chem., 2015, 63 (16), pp 4112–4119
• DOI: 10.1021/acs.jafc.5b00940

A comprehensive list of NMR articles on alcoholic beverage analysis can be provided upon request
 Thank You for Your Attention

Questions ?

References and Presentations can be found on PNA Website http://www.process-nmr.com

Contact John Edwards - email: john@process-nmr.com Tel: (203) 744-5905