identification of molecules linked to low filtration … of molecules linked to low filtration...

Identification of molecules linked to

low filtration performances.

Examples of malting barleys tested at

pilot plant level.

EBC Brewing Science Group 10th Technical meeting

Vienna – September 2014

Marc SCHMITT

IFBM / EBC BSG WIEN / September 2014 2

Context of the study

Justification : Mash & beer filtration problems are of importance for barley to beer chain

Aims : Identification of molecules linked with low filtration performances

Study of behaviour during malting & brewing

Industrial strategy : Improve filtration conditions

Reduce enzymes utilization

Expected results : Optimised mash and beer filtration conditions

Looking for tools to improve barley selection


Introduction : Beer production

Possible filtration

problems


Outline

Focus on malting barley homologation (France)

Molecules of interest : Presentation

Materials & methods used

Samples production, analytical plan & results Some examples of samples used

Impact of process

Conclusions & perspectives

Malting barley varieties homologation

• Registering on french catalog

• TAV, DHS, techno barley

• Malting barley List

• Quality index (malt)

• Pre-validation technological

• Functional index

Yield Sieving ; proteins ; SW

Malt : Extract ; enzymes ; proteins

Malt : Fermentability ; Functionalities

Extract, friability, diastasic power, viscosity, beta-glucans, etc…

Modification, attenuation, FAN, a-amylase, tepral filtration, etc…



• Technological validation

• Pilot tests Tests CBMO

• Commercial and industrial observation

• Maltsters & brewers

Barley to beer : Looking for big troubles

Agronomical development

Industrial productions

Industrial validations

Preferred variety


BRASSEURS DE FRANCE - 25, boulevard Malesherbes - 75008 PARIS - TEL. 01 42 66 29 27 - FAX. 01 42 66 07 66 - [email protected]

MALTEURS DE FRANCE- 66, rue de La Boétie - 75008 PARIS - TEL. 01 43 59 44 93 - FAX. : 01 43 59 46 02 – [email protected]

GESINE / RGT PLANET / PRUNELLA

Step 2

Juin

2014

Step 1 AMISTAR / VOYEL

BEATRIX / SEBASTIAN VANESSA ESTEREL / PASSEREL

CALYPSO

ODYSSEY / OVERTURE / TRAVELER

KWS AURELIA / PIONIER / SANETTE / STYLE

BELLINI / CONCERTO /NFC TIPPLE PRESTIGE/SUNSHINE

EXPLORER /GRACE / KWS IRINA / ZEPPELIN SALAMANDRE

ARTURIO / AZUREL

CASINO / ETINCEL / ISOCEL

More than 15 000 ha

Less than15 000 ha

Limited use

In commercial and industrial observation: Step 1 : Varieties which have successfully passed the IFBM pilot tests and subjected to industrial tests in order to check that all functional Malting and Brewing expectations are fulfilled. This period should allow commercial development of the variety Step 2 :Varieties under industrial tests in order to check that all functional Malting and Brewing expectations are fulfilled. They must be multiplied over 150 hectares and must have an interest for a maltster and a brewer. Admitted in technological validation : New registered varieties on Brewery orientation list of CTPS and proposed by CBMO to IFBM pilot tests. Limited use : Variety adapted to certain specifications which commercial output has to be secured.

LIST OF PREFERRED VARIETIES of MALTEURS DE FRANCE and BRASSEURS DE FRANCE

MALTING BARLEY - CROP 2015

SPRING BARLEY WINTER BARLEY

2 ROWS 6 ROWS

VARIETIES IN COMMERCIAL AND INDUSTRIAL OBSERVATION

VARIETIES ADMITTED IN TECHNOLOGICAL VALIDATION

PREFERRED VARIETIES

SY TEPEE



System is made to keep only the better lines…

Important to detect possible problems linked to a cultivar

Filtration problems are only detected at pilot plant level


Functional qualities of malt Looking for relevant tools to improve knowledge &

database collection

Barley breeders are looking for tests to improve barley selection Detection of “problems” as early as possible

Providing more specific definitions of required specifications

EBC congress mash : Mashing profile…

Increasing gap with results observed in current industrial practices

=> Lack of informations in EBC congress wort IFBM / EBC BSG WIEN / September 2014 9

Non starchy polysaccharides :

linked with filtration problems…

Aleurone layer

Husks

Arabinoxylans

6 to 10 % dm

Starchy endosperm

b-glucans

3 to 5 % dm


β-Glucans

Arabinoxylans

Introduction


Water retention

Hydrogen bonds

Formation of viscous solutions & gels

Wort viscosity

Wort turbidity

Filtration problems

Consequences

Properties

Introduction


Principle :

Colorimetric reaction with phloroglucinol

Reaction with hexoses & pentoses

Quantification of xylose after acid hydrolysis

Measurement :

Differences of absorbancy

(510 & 552 nm)

ARAX content determination

Wort or extract

Wort hydrolysis

Colorimetric

reaction

Xylose content

ARAX content

Revision of the method for better accuracy IFBM / EBC BSG WIEN / September 2014 13

//upload.wikimedia.org/wikipedia/commons/8/87/Alpha-D-Xylose_.svg

Why ?

Need to have more informations

How ?

Precipitation of HMW arax

Principle :

Precipitation with 1-propanol

Quantification

HMW-ARAX content determination

Precipitation with

1-propanol

Centrifugation

Concentration of

precipitate

Hydrolysis

Quantification of

arabinoxylans

(Adapted from

Novozymes, J. Obrecht, 2012

&

INRA Nantes, L. Saulnier, 2012)


//upload.wikimedia.org/wikipedia/commons/e/e3/Propan-1-ol-3D-balls.png

Why ?

Need to have more informations

Principle : (MEBAK 2.5.5)

Thermal treatment

Quantification

Beta-glucan “gel” content

10 mL

wort

10 mL

wort

E1 E2

Water bath 15 min x 80°C

20°C

Centrifuge, 20 min, 3500g

Supernatent

Quantification of soluble β-Glucans

(EBC 8.13.2)

Procedure Calculation E1 [mg/L] - E2 [mg/L] = β-GG [mg/L]


Steric exclusion chromatography

Injection

Small

molecules

Separation

according size of

molecules

Solvant

Sample

Cavités

poreuses

Detection

Retention time

Big

molecules


HPSEC : research &

identification of molecules linked

with viscosity & filtration

problems

Steric exclusion chromatography with triple detection

Equipment

1 2 3

Light diffusion signal

Viscosity signal

Refractometric signal

Study

Study of interesting samples from IFBM pilot plant

With & without filtration problems

Study of brewing (& malting) conditions

« Tepral » brewing & filtration system

Analyses :

HPSEC

Wort viscosity

b-glucans & arabinoxylans

… IFBM / EBC BSG WIEN / September 2014 18

Malting pilot plant test

Steeping : 600 kg of inox cylindro vessel with aeration and temperature management

Germination : 2 saladins vessels, 70 cm

Kilning : simple desk electric energy and air flow variation


Pilot plant brewing

20 hl & automatised

cereal cooker & mash tun

Lauter tun

Brew Kettle

Whirlpool

Fermenter of 20 hl

Propagator

Beer filtration: Filtrox frame

Bottling : CARBOFILL IFBM / EBC BSG WIEN / September 2014 20

Brewing conditions

Malt throw = 300 kg (± 5)

Water = 850 kg

pH = 5.4+/- 0.1

Pilot plant brewing

40

45

50

55

60

65

70

75

80

0 15 30 45 60 75 90

Time (min)

Tem

pera

ture

(°C

)

50°C

64°C

74°C

78°C

Wort gravity :

12°P end of boiling

Lauter tun load :

170 kg/m2

Fermentation :

Pitching : 17 +/- 2 M – O2 : 8 ppm

T° : 12 – 14°C

P : 0.5 – 1.0 bar

Boiling : 60’

Evaporation : 6%


Computerized mashing system

57 g fine gring & 200 ml water

Mash-in temperature : 50°C (15 min)

Rests temperatures : 63 & 75°C (15 min)

Mash filtration speed (1 bar pressure)

Sparging with 200 ml water at 75°C

Sparging filtration speed (1.5 bar pressure)

40

45

50

55

60

65

70

75

80

0 20 40 60 80

Time (min)

Te

mp

era

ture

(°C

)

Filtration

Computerized mashing system

57 g fine gring & 200 ml water

Mash-in temperature : 50°C (15 min)

Rests temperatures : 63 & 75°C (15 min)

Mash filtration speed (1 bar pressure)

Sparging with 200 ml water at 75°C

Sparging filtration speed (1.5 bar pressure)

40

45

50

55

60

65

70

75

80

0 20 40 60 80

Time (min)

Te

mp

era

ture

(°C

)

Filtration

40

45

50

55

60

65

70

75

80

0 20 40 60 80

Time (min)

Te

mp

era

ture

(°C

)

FiltrationFiltration

Laboratory scale :Tepral filtration system

Mash-in : 45, 50, 55 & 60°C


Tepral filtration system

Tepral filtration :

Registered parameters : • Filtration speed

• Time to filter 350 g of wort

• Filtration duration

• Sparging speed

Test used in Fonctional Index calculation IFBM / EBC BSG WIEN / September 2014 23

Study : analytical plan

Malt (600 kg pilot plant)

Pilot brew (20 hl)

Process control (all steps)

- Malt analyses

- Tepral filtration - ≠ mash-in t°

- …

Analyses on tepral wort

bGL

- ARAX

- FAN

- Soluble N

- etc IFBM / EBC BSG WIEN / September 2014 24

Study

13/09/2014 25

Interesting available samples :

CBMO malts :

Batches Mash filtration Beer filtration

Cargo 1 LL K

Cargo 2 K K

Cargo 3 LL L

CARGO

ATLANTICK Atlantick 2 K LL

SY TEPEE SY Tepee 2 J K

EXPLORER Explorer 3 J K


Example of pilot brew

CARGO : 6RW Observations CARGO 3

Brewing :

Saccharification (min) 15

Wort + sparging filtration duration (min) 189

Time to filter 450 kg (min) 50

Weight filtered before raking (Kg) 1002

Raking during filtration 0

Raking during sparging 0-6-9

Wort aspect hazy + floaters

Yield end boiling / throw (%DM) 74,0

Sediments - Whirlpool (ml/l) 70

Fermentation :

% attenuation / FA at 7 days (%) 100,0

Diketons at cooling (mg/l) 0,42

Cooling day 8

Observations CARGO 3

Beer filtration :

Nomber of filtrations 1

DP end filtration 1 (kg/cm2) 1,0

Flow end filtration 1 (hl/h) 10,0

Volume filtered (hl) 15,5

Filtration duration (min) 90

Yeast before filtration (millions cell/ml) 5,5

Temperature before filtration (°C) -1,8



ATLANTICK : 6RW Observations ATLANTICK 2

Brewing :










Fermentation :



Cooling day 8

Observations ATLANTICK 2

Beer filtration :










SY TEPEE : 2RW

Observations SY TEPEE 2

Brewing :










Fermentation :



Cooling day 8

Observations SY TEPEE 2

Beer filtration :










EXPLORER : 2RS Observations EXPLORER 3

Brewing :










Fermentation :



Cooling day 8

Observations EXPLORER 3

Beer filtration :









Study – Results Tepral filtration with ≠ mash-in temperatures :

Friabilité

Beta

glucanesViscosité

Protéines

solublesIK

CARGO1 67 362 1,75 3,29 30

CARGO2 68 303 1,70 3,70 35

CARGO3 67 330 1,71 4,40 40

Filtration speed

Beta-glucans



Friabilité

Beta

glucanesViscosité

Protéines

solublesIK

CARGO1 67 362 1,75 3,29 30

CARGO2 68 303 1,70 3,70 35

CARGO3 67 330 1,71 4,40 40

Filtration speed

Viscosity



Solubles & HMW Arax

Filtration speed

Arax


Study – Results

13/09/2014 Titre 33

Tepral filtration with ≠ mash-in temperatures :

Filtration speed

Viscosity


Solubles & HMW Arax

Filtration speed



b-GG…

Only CARGO worts present significant content of b-GG… b-glucans content with mash-in temperature

b-GG content with mash-in temperature

Mash-in t° CARGO1 CARGO2 CARGO3 ATLANTICK2 SY TEPPE2 EXPLORER3

45°C 26 22 32 3 1 16

50°C 27 15 28 0 2 2

55°C 24 19 22 0 2 0

60°C 15 9 12 2 2 10

Beta glucans gel (mg/l)


Study – Results Relation between some parameters



Study – Results

13/09/2014 38

Calculated R2 :

0,93


Study – Results

13/09/2014 39

Calculated R2 :

-0,85

Study – Results

13/09/2014 40

Calculated R2 :

0,94


Malt batches tested showed big differences for their filtration performances

Malt batches (varieties) tested didn’t have the same sensitivity to the mash-in temperature

Filtrations properties are linked to solubles polysaccharides

b-GG seems to be related with mash filtration problems

content with mash-in temperature

Solubles arax of HMW seems to be related with beer filtration problems

no clear relation with mash-in temperature (ARAX)

Great correlation between filtration parameters & HMW NSPS

Study : First Conclusions


Study – HPSEC Results



CARGO wort



ATLANTICK wort



Teprals worts T 45°C

Cargo 1

Cargo 3

Atlantick 2

Explorer 3




Cargo 1

Cargo 3

Atlantick 2

Explorer 3




Cargo 1

Cargo 3

Explorer 3



Cargo 1

Atlantick 2


HPSEC profile in accordance with analyses & previous tests :

peak at 26-27 min : arax

peak at 30-32 min : beta-glucans IFBM / EBC BSG WIEN / September 2014 48


Barley EXPLORER

Malt EXPLORER

LRGP - CNRS

Wort EXPLORER



Interpretation : Comparison of profiles

Integration of peaks with significant informations

Calculation of some parameters :

Average size of molecules

Average intrinsic viscosity of molecules

Average concentration of molecules

LRGP - CNRS



Atlantick Cargo Explorer

Mean molecular weight (kDa) 54,7 49,7 43,9

Intrisec viscosity 0,29 0,39 0,14

Concentration (g.L-1

) 2,19 3,31 2,49

Mean molecular weight (kDa) 103,2 74,0 137,0

Intrisec viscosity 0,20 0,21 0,12

Concentration (g.L-1) 2,48 2,06 1,63

Barley

Malt

Summary of results :


Conclusions : Barleys, Malts & worts showed different profiles

Varieties analysed showed different profiles

Molecules produced during malting process showed various properties with HPSEC (MW, IV, CC)

HPSEC Conclusions

Perspectives : Identification of concerned molecules with help of enzymes


Conclusions

Conclusions & hypotheses

Atlantick Cargo Explorer SY Tepee

Beta glucansSolubles ++ +++ + -

beta-GG - + - -

Solubles ARAXTotal ++ +++ + ++

Arax HPM + + - -



Conclusions : Developped technics are of interest

Tepral filtration give interesting informations (vs EBC wort)

Filtrations properties are linked to solubles polysaccharides (in the observed samples) :

b-GG seems to be related with mash filtration problems

Solubles arax of HMW seems to be related with beer filtration problems (same observation in recent samples)

HMW NSPS are related to filtration performances



Perspectives : Collection of informations to go on

Study of samples with “other” problems :

Linked to proteins (data not shown)

Study of enzymes & their activities…

Are differences observed due to :

Different or more / less active enzymes ?

Fine barley grain structure…

Are differences observed due to different barley grain composition & structure ?


Contact : IFBM

Marc SCHMITT [email protected]


Thanks to the support of :

Malteurs de France

&

Brasseurs de France

Thank you for your attention

mailto:[email protected]



identification of molecules linked to low filtration … of molecules linked to low filtration...

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