The world leader in serving science

Lars Lindqvist and Per Nilsson

Thermo Fisher Scientific

New Technologies Applied to the Brewing Industry

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Analytical Instruments for Beer Analysis

• Discrete Analyser – Beermaster

• Chromatography

• Ion Chromatography (IC) - Instruments and Application overview

• High Performance Liquid Chromatography (HPLC) Example application – Isohumulones

• Gas Chromatography (HPLC) Example application diketones and profiling studies

• Resources

Gallery Plus Beermaster – automated efficiency Outline

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• Automated, discrete photometric system for

beer, wort and malt analysis

• Superior platform for beer, wort and malt

quality control and testing

• Highly specific colorimetric and enzymatic

measurements

• Automated, effective bitterness measurement

• Compact, easy-to-use system

• Wide range of system reagents

Gallery Plus Beermaster – automated efficiency Gallery Plus Beermaster – automated efficiency

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• Automates all necessary analysis steps

• Bitterness and other photometric tests, like beta- glucan, pH, SO2 etc.,

can be run parallel

• Up to 350 photometric tests/h when running one-reagent methods

• Typical capacity 8 bitterness tests/h

• First bitterness result in 10 minutes

Time and cost savings

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• Easy to use – once loaded all the steps of analysis are performed automatically

• Fully automated pretreatment and measurement for bitterness simultaneously with other tests

• Contamination free analysis –disposable cuvettes

• Cost-effective – low reagent usage

• Reduced environmental impact –low water consumption

• Easy to install and maintain

Key Benefits of Gallery Plus Beermaster

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• Acids

• Acetic acid (ACS)

• Acetic acid

• Citric acid

• D-Lactic acid

• L-Lactic acid

• L-Malic acid

• Oxalic acid

• Total Acids

• Metals

• Calcium

• Total Iron

• Magnesium

Thermo ScientificTM System Reagents for Beer Samples

Over 30 optimized system products for brewing analysis

• Sugars• D-Fructose

• D-Glucose

• D-Glucose + D-Fructose

• D-Glucose + D-Fructose + Sucrose

• Sucrose (Total Glucose)

• Lactose

• Alcohols• Glycerol

• Ethanol

• Others• Acetaldehyde

• Ammonia

• SO2 Total

• Alpha-amino nitrogen (NOPA)

• pH (ECM or Colorimetric)

• Bitterness

• Beta-Glucan (High MW)

• Total protein

• Calibrators• Acetaldehyde standard

• Acid combination standard

• Glycerol standard

• NOPA standard

• Sugar combination standard

• Beta-Glucan standard

• Oxalic acid standard

• Lactose standard

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Key benefits of Thermo Scientific system reagents

• Optimized system solution

• System applications for beer analysis

• Continuously expanding menu

• Electronically loadable application data

• Optimized kit sizes and on-board stability

• Wide range of standards

• Productivity and efficiency

• Ready-to-use liquid reagents eliminate

reagent preparation

• Minimal reagent waste

• Bar-coded reagent vials provide easy and

reliable identification

• lot, exp date, vial size

• real-time reagent monitoring

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Key benefits of Thermo Scientific system reagents

• Optimized system solution

• System applications for beer analysis

• Continuously expanding menu

• Electronically loadable application data

• Optimized kit sizes and on-board stability

• Wide range of standards

• Productivity and efficiency

• Ready-to-use liquid reagents eliminate

reagent preparation

• Minimal reagent waste

• Bar-coded reagent vials provide easy and

reliable identification

• lot, exp date, vial size

• real-time reagent monitoring

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• Reports available

• Through LIS

• As printouts

• Electronically as PDF files

• Spreadsheet reporting for further calculations

Various reporting options

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Gallery Plus Beermaster - Summary

• Fully automated system for beer

analysis

• Quick and accurate results

securing quality and consistency

during the brewing process

The world leader in serving science

Chromatography Instrumentation- High Performance Liquid Chromatography (HPLC)

- Ion Chromatography (IC)- Gas Chromatography

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Ion Chromatography Portfolio: For every Beer Analysis Challenge

Aquion

Integrion

ICS-4000

ICS-5000+

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• Multi Analyte Detection and Quantification in a Single Run

• Applications Available for:

Carbohydrates, Alcohols, Organic Acids, Inorganic Anions, and Cations

IC Application Summary

Download Technical Note 126:

Determination of Organic Acids in Beer Samples

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• The power to analyze Fermentable Sugars and Malto-Oligosaccharides in Beer

IC Application Examples: Carbohydrates

Download Application Note 46: Ion Chromatography: A Versatile Technique for the Analysis of Beer

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HPLC: Versatile and Powerful Technique

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Example Application: Isohumulones

• Different contributions to beer properties

• Iso-n-humulones provide balanced bitterness

• Isocohumulones considered to provide harsher bitterness

• Different contributions to beer flavor, odor and aroma

• Isoadhumulones stabilize beer foam the most, isocohumulones the least

Source: Handbook of Brewing: Processes, Technology, Markets. H.M. Eßlinger, ed. Wiley-VCH, Weinheim. 2009.

Isohumulone Foam / Beer Ratio

Isocohumulone 1.3

Iso-n-humulone 2.5

Isoadhumulone 3.3

Total Isohumulones 2.2

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• Derived from hops during wort boiling

• Foam stability, Bitterness and Anti-microbial preservation chemistry

Isohumulones (Iso-α-acids) Chemistry

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Brewers need to know the exact isohumulone patternto be innovative and successful!

Example Solution to the market

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Ready, Steady, Go! One-Click Total Solutions

Instrument Columns, Reagents and Standards Method

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Get the Result in Less than 10 Minutes

Brewers can create new innovative brands and flavors

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Information Resources

• Brochure: Total Solution for Isohumulones in Beer

• Webpage: Thermo Scientific Isohumulones in Beer Application Kit

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Gas Chromatography Solutions: Diketone analysis

Trace 1300/1310 Gas Chromatograph

with automated head-space sampler

Head-space analysis

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Static head-space

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Calibration curves 2,3 – butanedione and 2,3 - pentanedione

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Qualitative analysis: Olfactometry + chromatography = true

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The selectivity of the nose

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Experimental - Samples

• 9 whisky extracts in ethyl acetate analysed 4 times in random order.

• Pool sample prepared from 20 µl each whisky.

• Run at 60,000 resolution on Q Exactive GC

Sample ID Type Age Country of Origin Region

2263 Single 12 Scotland Lowlands

2264 Single 18 Scotland Lowlands

2265* Single NAS Scotland Lowlands

2281 Single 10 Scotland Campbeltown

2282 Single 15 Scotland Campbeltown

2283 Single 15 Scotland Campbeltown

2284 Single 12 Scotland Highland

2285 Single 18 Scotland Highland

2295 Bourbon - USA Kentucky

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Analytical approach in this proof of concept study

• Are there any chemical differences between whisky samples?

• Type of whisky ie Bourbon or Scotch Whisky

• Geographical origin ie USA or Scotland, Highland or Lowland

• Age of whisky ie 10 or 15 Year aging

• Approach was to run a non-targeted analysis under generic conditions

and use software tools to identify:

• If there are differences? Using statistical software ie Sieve 2.2

• What the differences are? Using software tools including NIST libraries,

deconvolution software and elemental composition and fragment matching

software.

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TIC whisky sample

F:\Work\...\8April_60K_Whisky_025 04/10/15 06:10:59

RT: 3.45 - 34.00 SM: 7B

4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34

Time (min)

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90

95

100

Re

lative

Ab

un

da

nce

12.45

14.93

15.698.53

18.90

12.55

12.2814.62 23.9020.7812.19 26.02

18.999.38 9.5122.1817.15

29.1410.54 27.94 31.9824.897.42

NL:2.46E10

TIC MS 8April_60K_Whisky_025

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Software Algorithm for Unknowns Analysis

Deconvolve TIC Create “clean” spectrum

Acquired

Library

Library Search

Proposed Compounds

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PCA from Sieve 2.2

2282

2283

2281

22652295

Pool 22852263

2264

2284

Technical replicates in good agreement and clear differences between samples

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PCA from Sieve 2.2

Single distillery

Bourbon

3 wood aged

Pool

Bourbon and 3 wood aged clearly different from other whiskies.

Single distillery whiskies also show clear differences.

10 year

15 year

15 year

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Taking a closer look at the data

• Initial approach is to investigate the differences in the bourbon and 3

wood aged from other whiskies

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Peak at 13.61 min. m/z 177.1274Elevated in Bourbon

• From the PCA plot and the table of the detected peaks the following peak was

identified as being significantly elevated in bourbon.

• The peak was investigated by peak extraction and spectrum assessment.

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NIST Library hit for Trans β Ionone

Acquired spectrum

NIST spectrum

Trans β Ionone

NIST hits filtered

based on high resolution

filtering

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Literature search for beta ionone supports

• Beta-ionone is a breakdown product of beta-carotene and expected to be

found in whisky.

• Bourbon brewed from corn; Scotch Whisky from barley.

• Corn is higher in beta-carotene (53 µg/200g) compared to barley (7

µg/200g (Source: Nutritiondata.com)

• This would support the higher levels of beta-ionone found in our data.

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Resources

thermofisher.com/BeverageTesting

Dedicated Beer Analysis Compendium

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Many More Resources

• Dedicated Website and Learning Centre

thermofisher.com/BeverageTesting

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Thank You For Your Time – Any Questions…………