105th Brewing and Engineering Congress 2018

07/03/2018, Dortmund

Producing a gluten-free beer using a special

barley variety–
Challenges from the perspective of Quality
Assurance

Madeleine Adolf
Radeberger Gruppe

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 What is this presentation about?

• Introduction
What is gluten?
What gluten-related disorders are there?
What is the legal definition of “gluten-free”?

• What possibilities of producing gluten-free beer are there?

• How does the Radeberger Gruppe produce its gluten-free

beer PIONIER?

• What challenges arise from gluten testing?

• What challenges exist in the production of PIONIER?

 What is gluten?

Fractionation of wheat proteins by T. B. Osborne 1907

Extraction with Protein fraction Wheat Barley
Water Albumins
Ratio depends on
Sodium-chloride solution, Globulins
type of grain
e.g. 0.5M NaCl
70 % ethanol Prolamines Gliadins Hordeins
Residue Glutelins Glutenins Hordenines

Gluten = a protein fraction from wheat, rye, barley, oats or their crossbred varieties and
derivatives thereof, to which some persons are intolerant and that is insoluble in water and 0.5M
NaCl.

(AOECS definition)

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 Gluten as the cause of various diseases

• Coeliac Disease

− chronic autoimmune disease of the small intestine

− about 1% of the population affected (800,000 Germans /
corresponds to slightly more than the population of
Frankfurt a.M.)
− genetic disposition
− therapy requires a strict gluten-free diet
• Dermatitis herpetiformis

• Gluten ataxia

• Wheat sensitivity

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 What happens in the case of Coeliac Disease? (simplified)

Tabelle entnommen aus: E. Arranz et al: Advances in the Understanding of Gluten Related Pathology and
Relevant (immunogenic) epitopes of
prolamins and glutelins from wheat,
rye, barley, partially oats

Sensitivity
varies across
those affected

Inflammation of intestinal mucosa

Regression of villi

the Evolution of Gluten-Free Foods

Reduced nutrient uptake

Dyspepsia
Weight loss
Malnutrition
Other symptoms

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Two essential legal regulations have to be observed

Commission Implementing Regulation (EU) Regulation (EU)

No 828/2014 No 1169/2011 (food labelling)
(old: Commission Regulation (EC) No 41/2009) Cereals containing gluten (wheat, rye,
Basis: Codex Alimentarius Standard for barley, oats) and products thereof shall be
food for special dietary use for persons stated in the list of ingredients according
intolerant to gluten (Codex Stan 118-1979) to their legal name.
DZG / AOECS standard (licensing of the The name shall be emphasised through a
Crossed Grain symbol) typeset that clearly distinguishes it from
the rest of the list of ingredients, for
Gluten-free Max. 20 mg/kg gluten example by means of the font, style or
background colour.
Very low Max. 100 mg/kg gluten
gluten
e.g. barley malt
content

Comments:
• daily intake of gluten should not exceed 20 mg for coeliacs
• sensitivity varies across those affected
• beverages are consumed in larger quantities than solids

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 Beer according to the “Reinheitsgebot” is produced from grains
containing gluten

0 10 20 30 40 50 60 70 80 90 100
Barley
Barley
malt
Wheat

g gluten / kg raw material Literature values

> 270 (testing limit)

approx. 1,000

< 20 - 60

mg gluten / l beer

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 Possibilities of producing gluten-free beers by removing gluten

Removing gluten using enzymes

e.g. transglutaminase, endopeptidase
Advantages: Disadvantages:
• taste close to that of traditional beers • acc. to Article 9 of Vorl. BierG (provisional beer law),
only permissible if obtained within the scope of the
“Reinheitsgebot”, e. g. during malting, technical
enzymes not permissible
• doubtful whether all relevant epitopes are removed

Removing gluten using protein-binding filter aids

e.g. tannin, silica gel
Advantages: Disadvantages:
• taste close to that of traditional beers • unclear whether permissible acc. to Article 9 of Vorl.
BierG (provisional beer law)
• doubtful whether all relevant epitopes are removed

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 Possibilities of producing gluten-free beer by replacing glutenous raw
materials

Replacing glutenous raw materials with gluten-free raw materials

e. g. millet, rice, maize, buckwheat
Advantages: Disadvantage:
• definitely no gluten content • taste differs from that of traditional beers
• not permissible to be labelled “beer“ according to
Article 9 of Vorl. BierG (provisional beer law)

Use of a barley variety in which gluten was largely removed by breeding

Ultra-Low Gluten (ULG) Barley (Kebari® from CSIRO + GRDC /Australia)
Kebari® is protected by various international patents
Advantages: Disadvantages:
• beer is definitely gluten-free • necessary to ensure the whole supply chain
• taste close to that of traditional beers remains contamination-free
• meets Article 9 of Vorl. BierG (provisional
beer law)

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 Producing gluten-free beer using a barley variety for which gluten has been
largely removed by breeding

0 10 20 30 40 50 60 70 80 90 100
ULG
Barley
barley Barley
malt
3.0
g gluten / kg raw material Literature and analysis values

< 20 - 60

< 10

mg gluten / l beer

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Producing gluten-free beer using a barley variety for which gluten has been
largely removed by breeding

ULG Barley 3.0 was developed by crossing

different barley mutations, i.e. a barley
chromosome with a deactivated hordein locus

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Producing gluten-free beer using a barley variety from which gluten has
been largely removed by breeding

Determining hordein using MS:

ULG 3.0 compared to other barley varieties and
earlier stages of development

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 Radeberger Gruppe’s choice – PIONIER

GLUTEN-FREE STRATEGY: ULG barley

BEER CATEGORY: Vollbier
(original gravity between 11 – 14 %)
TYPE: Pilsener
ORIGINAL WORT: 11.6 %
VOLUME PERCENTAGE : 4.5% vol
CARBONATION PERCENTAGE: 0.55%
BREWING PROCESS: bottom-fermented
BITTERNESS UNITS: 24 EBC
COLOUR: 12 EBC
pH VALUE: 4.6

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PRODUCING PIONIER

Diagram source: http://www.brauer-bund.de/bier-ist-rein/so-wird-bier-gebraut/der-

brauprozess.html

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Gluten analysis using ELISA

ELISA (enzyme-linked immunosorbent assay / enzyme immunoassay)

most commonly used simple application, required detection limit:

method to detect gluten fast results <10mg gluten / kg

Sandwich ELISA to detect prolamins

of rye, barley and wheat in natural and Competitive ELISA to detect
heat-treated foods hydrolysed prolamins e.g. in fermented
products

Source: http://archiv.aktuelle-wochenschau.de/2009/w30/w30.html

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 Today’s standard analytics have weaknesses – further development is
required here!

Factors ELISA method Evaluation

• there are various antibodies with varying sensitivity to Further epitopes may be
specific epitopes present in the sample!
Antibody used • acc. to Codex and AOECS Standards:
monoclonal R5 antibodies (Mendez) against secalin Coeliacs’ reactions vary
• epitopes detected by R5: QQPFP, QQQFP, LQPFP, QLPFP across epitopes!

• no certified reference material available

Standard The sample may have a
• differences between ELISA kits
different composition than
material • PWG gliadin (wheat) partly used in Sandwich ELISA; the standard material!
hydrolysate from barley, wheat, rye used in Comp. ELISA

• prolamines dissolved with ethanol

Complete extraction needs
Extraction • reduction agents used can simultaneously change the to be ensured!
solubility of glutelins

Calculation • gluten = prolamin x 2

Compositions of grains
factor differ!

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Gluten analysis using a mass spectrometric method

specific analysis with the

detection of different +
relevant epitopes is
possible

no true quantification
possible yet,
sophisticated -
analytics

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The greatest challenge in the production of Pioneer is to avoid
contamination with glutenous material.

Partner companies Radeberger Gruppe

Cultivation Malthouse Brewery

Sample Sample Clearance

Storage Storage high risk of contamination

Sample Sample
medium risk of contamination
Transport Transport low risk of contamination
Sample

• cooperation with reliable, experienced partners

• very high cleaning requirements, beyond the common standard of
hygiene incl. corresponding inspections
• more difficult to implement in dry areas than in wet areas
• executing control tests where possible (contamination with foreign
grains, gluten analysis)
• integration into food safety systems

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Further challenges in the production of Pionier arise from the
processability of ULG barley.

Size and • lower extract yield, brewhouse

yield and final degree of
shape of fermentation
the grain • requires malt mill adjustments

• larger volumes during

transport and production
Density • requires storage adjustments
• influences soaking

common barley malt ULG barley malt

Protein • influences foam
composition • inclination towards turbidity

common barley malt ULG barley malt

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 Additional challenges exist in securing the supply of raw materials

Supply risk
• raw materials are in limited supply
• reasonable risk management using
− various growing regions
− various transport routes
− various warehouses
− various batches of malt

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Conclusion

Using ultra-low-gluten barley leads to a beer that

is definitely gluten-free, which meets Article 9 of
Vorl. BierG (provisional beer law) and the taste
expectations for a beer.

From our The greatest challenge is to ensure that

experience in contamination with glutenous material is avoided
throughout the supply chain.
producing Pioneer
from ULG barley Due to the shape of the grains and the different
(Kebari®), we can composition as compared to normal barley,
adjustments have to be made in its processing.
conclude:
Today’s standard analytics have weaknesses – for
better results in gluten detection, analytical
options have to be developed further.

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