13C/12C IRMS Analyses of Alcohol Beverages at the Alcohol and

Tobacco Tax and Trade Bureau (TTB)

Amanda Smith, Alcohol and Tobacco Tax and Trade Bureau

June, 2014

Alcohol and Tobacco Tax and Trade Bureau

TTB Mission

• TTB’s mission is to collect the taxes on alcohol, tobacco, firearms, and ammunition; protect the consumer by ensuring the integrity of alcohol products; and prevent unfair and unlawful market activity for alcohol and tobacco products.

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IRMS Systems

IRMS #1 • CE Elantech EA • Trace Ultra GC • LEAP Autosampler • GCCIII and ConFlo IV • Delta V Plus IRMS IRMS #2 • Gasbench II • Trace Ultra GC • TriPlus Autosampler • GC Isolink • ConFlo IV • Delta V Plus IRMS

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13C/12C IRMS Analyses of Alcohol Beverages

• Probing fermented versus distilled character or markers of beverage alcohol

• Determining agricultural sources of alcohol in sakés

• Differentiating between bourbons and other types of whiskeys

• Determining natural versus artificial carbonation in wine

Probing Fermented Versus Distilled Character or Markers of Beverage

Alcohol

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Fermentation Products

• Naturally produced during fermentation: – Methanol,

acetaldehyde, ethyl acetate

– Fusel oils • n-propanol • iso-butanol • n-butanol • amyl alcohol

• Detected using GC – DB-WAX column, 30m

x 0.53 mm, 1 µm film thickness

– Hydrogen carrier gas

2.5 5 7.5 10 12.5

pA

100

200

300

400

500

600

FID1 A, (ALS031109\2009-03-11ALS\003B0401.D)

4.1

77 -

Eth

yl A

ceta

te 4

.392

- M

etha

nol

7.9

35 -

n-P

ropa

nol

9.1

79 -

iso-

Buta

nol

10.

086

- n-

Buta

nol

11.

111

- Ac

tive

Amyl

11.

791

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Distilled Scotch

935.48 sec -30.88 + 0.13‰ not detected not detected 698.27 sec -31.81 + 0.20‰ 556.36 sec -24.85 + 0.19‰ 485.09 sec = backflush bump. Headspace ethanol = -27.34 + 0.06‰

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Neat Wine

n-Propanol (37 ppm)

Isobutanol? Unknown? Co-elution?

n-Butanol?

Amyl Alcohol (122 ppm)

Unknowns

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Grape Wine Results

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Repeatability Issue

Determining Agricultural Sources of Alcohol in

Sakés

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Collect the Revenue

• Saké is taxed as a beer – $18 per barrel (31 US liquid gallons)

• Saké which contains alcohol produced by distillation is taxed as a distilled spirit – $13.50 per proof gallon

• 7 times higher tax rate per US liquid gallon for a 15% ABV saké

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Plant Type δ 13C values (‰)* Examples

C3 (95% of plants

are C3) -30 to -23

rice, barley, grape, sugar beet, molasses made from sugar beet,

coconut, palm, peanut, plum, potatoes, rye, soybean, sunflower, wheat

C4 (1% of plants

are C4) -10 sugar cane, corn, millet, sorghum,

molasses made from sorghum

CAM -18 to -12** vanilla, agave, cactus, pineapple

Photosynthesis and Carbon Isotopes

* These ranges are approximations. **Results in this range can also indicate a mixture of C3 and C4 plant origin alcohols.

• Different photosynthetic pathways lead to different 13C/12C ratios detectable by IRMS

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HS-GC-IRMS 13C/12C of Ethanol

• Standards – Rum and Vodka standardized in relation to

Vienna-Pee Dee Belemnite (VPDB) • Rum: -10.98 ‰, C4 • Vodka: -27.53‰, C3

• Samples – Alcohol beverages, above ~20% ABV are

diluted to ~20% with water – 200 µL in a 2 mL autosampler vial, triplicate

headspace injections (standards and samples)

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Example Chromatogram

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Sake Results

-30.00

-28.00

-26.00

-24.00

-22.00

-20.00

-18.00

-16.00

-14.00

-12.00

-10.00

0 100 200 300 400 500

δ13C

(‰) v

s. V

PDB

Sample Number

Pure SakéSaké with Added AlcoholSuspect SakéSaké with Other IngredientsRum Standard ValueVodka Standard ValuePure Saké/Added Alcohol Cutoff Line

Differentiating between Bourbons and Other Types of Whiskeys

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Bourbon Whiskey

Standard of Identity Bourbon whiskey is whisky produced at not exceeding 160° proof from a fermented mash of not less than 51 percent corn and stored at not more than 125° proof in charred new oak containers.

27 CFR § 5.22

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Whiskey Authentication

0

100

200

300

400

500

600

700

-30 -25 -20 -15 -10 -5

Amyl Alcohols (g/100L) abs. EtO

H

δ13C(‰)

Straight Bourbon and Tennesse

Canadian

Irish Blended Scotch

Single Malt Scotch

Determining Natural Versus Artificial

Carbonation in Wine

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Tax Implications

• Sparkling Wines – > 0.392 g CO2 / 100 mL

• Naturally Sparkling – $3.40 / wine gallon

• Artificially Carbonated – $3.30 / wine gallon

• Hard Cider – Still wine

• $0.226 / wine gallon

– Naturally Sparkling • $3.40 / wine gallon

– Artificially Carbonated • $3.30 / wine gallon

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Ring Study

Sponsor • MOU Partner DGCCRF

France’s General Directorate for Competition Policy, Consumer Affairs, and Fraud Control – Francois Guyon,

coordinator

• Inter-Laboratory ring test on CO2 ᵟ13C Ratio Determination In Sparkling Beverages, published Feb. 2013

Participants

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Ring Study Results

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Database Results

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Thank You

• www.ttb.gov • Any questions? • Thank you!

Acknowledgements: Armen Mirzoian whiskey authentication Abdul Mabud and Jeffrey Ammann resources and support