Index of Beer & Malt Methods

Acetaldehyde Diacetyl Polyphenols

Amino nitrogen (free) Diastatic power Protein

Amylase Ethanol Protein (soluble)

Anthocyanogen Glucan (ß) Reducing Sugars (total)

Bicarbonate Glucanase (ß) Sulphite

Bitterness Hydroxymethylfurfural Sulphur dioxide (total)

Carbohydrate Nitrogen (total) Thiobarbituric acid

Carbonate Nitrate + Nitrite Turbidity

Color Nitrate + Nitrite Viscosity

Density pH
 This method is not available in this presentation.

For details please contact Skalar specifying sample matrix and range.

Skalar’s Headquarters
P.O. Box 3237
4800 DE Breda
The Netherlands

Tel. +31 (0)76 5486 486

Fax. +31 (0)76 5486 400
Email: [email protected]
Internet: http://www.skalar.com
 SKALAR METHODS
ANALYSIS : ACETALDEHYDE

Catnr. 101-000

PRINCIPLE

The automated procedure for the determination of Acetaldehyde is based on an enzyme catalysed reaction; in
the presence of aldehyde dehydrogenase (AT-DH), acetaldehyde is oxidised quantitatively by NAD to acetic
acid and NADH. The amount of NADH formed in the reaction is stoichiometric with the amount of
acetaldehyde and is measured at 340 nm.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA.

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer , typical range 0.8 – 40 ppm Acetaldehyde.

For sample pretreatment see 2.1.1. and 2.1.2.

REAGENTS

A. Buffer

Required chemicals: tetra Potassium pyrophosphate (K4 P2 O7)

Hydrochloric acid (HCl (1N))
Distilled water (H2 O)
Brij 35 (30%)

B. Buffer NAD

Required chemicals: Buffer solution A

Nicotinamide adenine dinucleotide (NAD)

C. Aldehyde dehydrogenase solution

Required chemicals: Aldehyde dehydrogenase (AL –DH)

Distilled water (H2 O)

D. Rinsing liquid sampler

Required chemicals: Distilled water (H2 O)

STANDARDS

Stock standard Acetaldehyde

Required chemicals: Acetaldehyde (C2 H4O)

Distilled water (H2 O)

Working standards

Dilute standards according to the range.

GENERAL REMARKS

1. The analysis needs blank analysis to eliminate own color interference. This can be analyzed
simultaneously using two analysis modules, one for the analysis and the second for the blank. It is also
possible to analyse on the same module, separate batch of analysis and blank after each other, that
requires only one module. The Skalar software can subsract automatically the blank value from the
sample value.
2. Micro inlet connectors, coils and flow cell are required.

REFERENCES

1. Lazaro, F., Luque de Castro, M.D., Valcarcel, M., “Individual and simultaneous determination of ethanol
and acetaldehyde in wines”, Anal. Chem. 59, 1987, page 1859-1863.
2. Methods of biochemical and food analysis, using single reagents,Boehringer Mannheim, page 6-7, 1989.

FLOW DIAGRAM
waste
ml/min

Flow cell
Filter 340 nm
Cor. Filter 420 nm
37°C

Aldehyde dehydrogenase solution

Air ****
Flow cell 10 mm
Filter 340 nm
Buffered NAD solution

Air
Buffer solution pH 9.00

Sample
waste

waste

Sampler

**** bubble gating flow cell catnr. 6441

 SKALAR METHODS
ANALYSIS : AMINO NITROGEN (FREE)

Catnr. 149-000

PRINCIPLE

The automated procedure for the spectrophotometric determination of free amino nitrogen (FAN) is based on
the ninhydrin method; amino acids are decarboxylised with ninhydrin, ammonia will be released as well. The
reduced ninhydrin reacts with the unreduced ninhydrin and ammonia to a blue-yellow dye and is measured at
570 nm. Potassium iodate addition stops the reaction and avoids interfering reactions.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA.

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.
3. N2 gas.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer, typical range 6-300 ppm Amino nitrogen.

For sample pretreatment see 2.1.2. and 2.1.5.

B. Wort and malt, typical range 6-300 ppm Amino nitrogen.

For sample pretreatment see 1.1.2. and 1.1.3.

REAGENTS

A. Distilled water + Brij 35

Required chemicals: Distilled water (H2 O)

Brij 35 (30%)

B. Hydrazinium sulfate solution

Required chemicals: Hydrazinium sulfate (N2H6 SO4)

Distilled water (H2 O)
Brij 35 (30%)

C. Ninhydrin solution

Required chemicals: Ninhydrin (C9 H6 O4 )

Sodium acetate (CH3COONa)
Acetic acid (CH3 COOH (100%))
Distilled water (H2 O)
D. Potassium iodate solution

Required chemicals: Potassium iodate (KIO3 )

Distilled water (H2 O)

E. Rinsing liquid sampler

Required chemicals: Distilled water (H2 O)

STANDARDS

Stock standard Amino nitrogen

Required chemicals: Glycine (C2 H5 O2N)

Distilled water (H2 O)

Working standards

Dilute standards according to the range

GENERAL REMARKS

1. EBC method 141, 8.8.1 (manual method)

2. Standardization on glycine.
3. Non specific method.
4. These standards are only used to check linearity. The factor between glycine standards and beer samples
is ± 3. To check the factor analyse manually the beer and watery standard and compare with the automatic
gained absorbances.

REFERENCES

1. Prof. Dr. F., Drawert, “Brautechnische analysenmethoden”, Mitteleuropaischen Brautechnischen

Analysenkommission (MEBAK), D-8050, 1979, 261-265.
2. European Brewery Convention, Analytica EBC, fourth edition, 1987, method E 141, 8.8.1 (manual
method).
3. Baier, B.,Jödicke, T., Harms, J., Lück, M., Müke, O., Krüger, E., "Anwendung der Continuous Flow
Analysentechnik zur Qualitätskontrolle in der Brauindustrie, Teil IV : Bestimmung des Gehaltes an Freier
Amino-Stickstoff in Würze und Bier", Monatsschrift für Brauwissenschaft, 46, Heft 6, 1993,
page 223-229. (English translation available)
FLOW DIAGRAM
waste
ml/min

Flow cell
Filter 570 nm
Cor. Filter 620 nm

Potassium iodate solution

Ninhydrin solution

Nitrogen gas
Hydrazinium sulfate solution

Resample waste 90°C

Distilled water + Brij 35

Nitrogen gas
Distilled water + Brij 35

Sample

waste

Sampler
 SKALAR METHODS
ANALYSIS: AMMONIA NITROGEN (TKN)

Catnr. 155-000

PRINCIPLE

The automated procedure for the determination of Ammonia Nitrogen is based on the modified Berthelot
reaction; ammonia is chlorinated to monochloramine which reacts with salicylate to 5-aminosalicylate. After
oxidation and oxidative coupling a green coloured complex is formed. The absorption of the formed complex
is measured at 660 nm.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA. Check voltage at the
back of the instrument before installation.
2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Malt, typical range 3-150 ppm Nitrogen.

For sample pretreatment see 1.1.4.

REAGENTS

A. Buffer solution

Required chemicals: Potassium sodium tartrate (C4 H4O6KNa.4H 2O)

Sodium citrate (C6 H5 O7Na3 .2H 2O)
Distilled water (H2O)
Brij 35 (30%)

B. Sodium salicylate solution

Required chemicals: Sodium hydroxide (NaOH)

Sodium salicylate (C7H5 NaO3 )
Distilled water (H2O)

C. Sodium nitroprusside solution

Required chemicals: Sodium nitroprusside (Na2 [Fe(CN)5NO].2H 2 O)

Distilled water (H2O)

catnr. 155
D. Sodium dichlororisocyanurate solution

Required chemicals: Sodium dichloroisocyanurate (C3 N3 O3Cl2 Na.2H2 O)

Distilled water (H2O)

E. Rinsing liquid sampler

Required chemicals: Sulfuric acid (H2 SO4 )

Distilled water (H2O)

STANDARDS

Stock solution Nitrogen

Required chemicals: Ammonium chloride (NH4 Cl)

Distilled water (H2O)

Working standards

Dilute standards according to the range.

GENERAL REMARKS

1. % Protein = 6.25 x % N

module numbers:

Total Kjeldahl Nitrogen, malt, typical range 3-150 ppm N requires chemistry module 473-316 (w/r)

SETTINGS

1. Sample time: 60 sec., wash time: 60 sec., air time: 0 sec.

REFERENCES

1. Krom, M.,"Spectrophotometric determination of ammonia; a study of modified Berthelot reaction using

salicylate and dichloroisocyanurate", The Analyst, April 1980, Vol. 105, page 305-316.
2. Searle, P.L.,"The Berthelot or indophenol reaction and its use in the analysis chemistry of nitrogen", The
Analyst, Vol. 109, May 1984, page 549-565.

catnr. 155
 FLOW DIAGRAM
waste
ml/min

Flow cell
Filter 660 nm
Cor. Filter 520 nm
40°C

Sodium dichloroisocyanurate sol.

Sodium nitroprusside solution

Sodium salicylate solution

Air
Buffer solution

Air
Buffer solution
Sample
waste

waste

Sampler

catnr. 155
 SKALAR METHODS
ANALYSIS: α -AMYLASE

Catnr. 164-000

PRINCIPLE

The automated procedure for the determination of α-Amylase is based on the following reaction; the malt
extract is diluted with a buffer solution, the diluted sample is mixed with a β-limit dextrin substrate and
incubated for 10 minutes at 35°C. The sample is then mixed with an iodine solution and the rate of breakdown
of the dextrin is measured colorimetrically. The colour is measured at 610 nm.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA. Check voltage at the
back of the instrument before installation.
2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

PROCEDURE SAMPLE PREPARATION

A. Malt and wort, typical range 2.6-130

Dextrinizing Units (D.U.).

For sample pretreatment see 1.1.1.

REAGENTS

A. Dilution buffer solution

Required chemicals: Sodium chloride (NaCl.)

Sodium acetate(CH3COONa.)
Acetic acid (CH3COOH)
Distilled water (H2O)

B. Stock iodine solution

Required chemicals: Iodine (I2)

Potassium iodine (KI)
Distilled water (H2O)
C. Iodine working solution

Required chemicals: Potassium iodide (KI)

Stock solution B
Distilled water (H2O)

D. Substrate solution β-Limit dextrin

Required chemicals: β-Limit dextrin .......................

Dilution Buffer sol. A.............

E. Rinsing liquid sampler

Required chemicals: Sodium chloride (NaCl)

Distilled water (H2O)

STANDARDS

Required chemicals: α-Amylase type VIII-A from Barley Malt

GENERAL REMARKS

1.M.U.G. Method

MODULE NUMBERS

a-amylase, typical range 2.6 – 130 D.U requires chemistry module 164-001.

SETTINGS

1. Sample time: 100 sec., wash time: 100 sec., air: 0 sec.
 FLOW DIAGRAM
waste
ml/min

Flow cell
Filter 610 nm

Air
Iodine solution

Resample
waste

Air

Substrate solution
Resample 35°C
waste

Dilution buffer solution

Air

Dilution buffer solution

Sample built up in glass

waste

Sampler
 SKALAR METHODS
ANALYSIS : ANTHOCYANOGEN

Catnr. 176-000

PRINCIPLE

The automated procedure for the determination of Anthocyanogen is based on the following reaction ;
leucoanthocyanide reacts, in contrast with sodium disulfite, in a buffered solution with sodium molybdate to a
yellow coloured complex. This complex is measured at 420 nm.

LABORATORY FACITITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA.

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer, typical range 3-150 ppm Anthocyanogen.

For sample pretreatment see 2.1.5.

REAGENTS

A. Distilled water + Brij 35

Required chemicals: Distilled water (H2O)

Brij 35 (30%)

B. Sodium disulfite solution

Required chemicals: Sodium disulfite (Na2S2O5)

Distilled water (H2O)
Brij 35 (30%)

C. Stock solution buffer 1

Required chemicals: di-Sodium hydrogen o-phosphate (Na2HPO4.2H 2O)

Distilled water (H2O)
D. Stock buffer solution 2

Required chemicals: Potassium dihydrogen o-phosphate (KH2PO4)

Distilled water (H2O)

E. Buffer solution (pH 7)

Required chemicals: Stock solution buffer

Stock solution buffer

F. Sodium molybdate solution

Required chemicals: Sodium molybdate (Na2MoO4.2H 2O)

Distilled water (H2O)
Brij 35 (30%)

G. Rinsing liquid sampler

Required chemicals: Distilled water (H2O)

STANDARDS

A standard is required that corresponds to the beer samples involved.

GENERAL REMARKS

1. Dark ales need a blank analysis to eliminate own color interference. This can be analyzed simultaneously
using two analysis modules, one for the analysis and one for the blank. It is possible to analyse on the
same module, separate batch of analysis and blank after each other, that requires only one module. The
Skalar software can substract automatically the blank value from the sample value.

REFERENCES

1. Amerine, M.A., Ough, C.S., Methods for Analysis of musts and wines, A Wilkey-Interscience publication,
page 175-195, 1980. (manual method)
2. Baier, B., (autor) “Anwendung der Continuous Flow Analysentechik zur Qualitätskontrolle in der
Brauindustrie”, Monatsschift für Brauwissenschaft Heft 4/1992, 116-121.
(English translation available)
FLOW DIAGRAM
waste
ml/min

Flow cell
Filter 420 nm
Cor. Filter 630 nm

Sodium molybdate solution

Buffer solution

Sodium disulfite solution

Air
Distilled water + Brij 35

Sample

waste

Sampler
 SKALAR METHODS
ANALYSIS : CARBON DIOXIDE

Catnr. 185-000

PRINCIPLE

The automated procedure for the determination of carbon dioxide is based on a pH shift; the released carbon
dioxide diffuses through a silicon membrane into a weak buffer solution containing a pH indicator
(phenolphthalein). The decrease in pH causes a decrease in the colour development of the phenolphthalein.
The colour is measured at 550 nm.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA.

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer, typical range 100- 5000 ppm Carbon dioxide.

For sample pretreatment see 2.1.3.

REAGENTS

A. Sulfuric acid solution

Required chemicals: Sulfuric acid (H2SO4 (97%))

Distilled water (H2O)
Brij 35 (30%)

B. Stock solution colour reagent

Required chemicals: Sodium carbonate (Na2CO3)

Sodium hydrogen carbonate (NaHCO3)
Distilled water (H2O)

C. Colour reagent

Required chemicals: Stock solution colour reagent

Phenolphthalein (C20H14O4 (1% in ethanol))
Distilled water (H2O)
Brij 35 (30%)

catnr. 185-000 1 of 3
D. Rinsing liquid sampler

Required chemicals: Distilled water (H2O)(CO2free)

STANDARDS

Stock standard carbon dioxide

Required chemicals: Sodium hydrogen carbonate (NaHCO3)

Distilled water (H2O)

Working standards

Dilute standards according to the range.

GENERAL REMARKS

1. Store the samples at 4°C in a closed bottle.

2. Dialysis step can be replaced by means of a gas separator.
3. The reaction determines total carbonates.
4. Segmentation of the flow must be made with carbon dioxide free gas: supply by pure nitrogen or scrubbed
air through an alkaline solution.

REFERENCES

1. Skeggs, L.T., Jr. and Hochstrasser, H., “Multiple Automatic Sequential Analysis”, Clin. Chem., 10, pp.
918-936.
2. Moll, M., Flayeux, R., Leheude, J., “Determination automatique du CO2 dans la biere”, extracts de bois.
No. 3, 75, 6 annee.

catnr. 185-000 2 of 3
FLOW DIAGRAM
waste
ml/min

Flow cell
Filter 550 nm
Cor. filter 620 nm

40°C

Nitrogen gas
Colour reagent
40°C

Nitrogen gas
Sulfuric acid solution

Sample 40°C
waste

waste

Sampler

catnr. 185-000 3 of 3
 SKALAR METHODS
ANALYSIS : BITTERNESS

Catnr. 191-000

PRINCIPLE

The automated procedure for the determination of Bitterness is based on an extraction method; after
acidification the sample is extracted in iso-octane. The absorption of the extract is measured at 275 nm.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA. Facilities for chemical
wastes.
2. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer, typical range 1 – 50 Bitterness Units.

For sample pretreatment see 2.1.5.

B. Wort, typical range 1 – 50 Bitterness Units

For sample pretreatment see 1.1.2.

REAGENTS

A. iso-Octane

Required chemicals: Iso-octane (2,2,4 Trimethyl-pentane)(C8 H18 (p.a.))

B. Hydrochloric acid 3 N solution

Required chemicals: Hydrochloric acid (HCl (32%))

Distilled water (H2 O)
Brij 35 (30%)

C. Rinsing liquid sampler

Required chemicals: Distilled water (H 2 O

STANDARDS

A standard is required that corresponds to the beer samples involved.

catnr. 191-000 1 of 3
GENERAL REMARKS

1. If nitrogen gas is not available, also compressed air can be used.

2. EBC method E 155, 9.6 (manual method)
3. UB photometer is required, catnr. SA 6700.
4. Ambient cooling is necessary.
5. Standardization of beer depends on the callibration of laboratory beer samples.
6. Standardization on beer, but iso-humulon can be used to check the linearity of the system.

REFERENCES

1. Cooper, A.H., Hudson, J.R., “Automated analysis applied to brewing”, Brewing industry research
foundation, 1961, page 436-438.
2. European Brewery Convention, Analytica EBC, fourth edition, method E 155, 9.6 (manual method).
3. Methods of Analysis of the ASBC, 1992.

Correlation with the manual method

Seq Beer type Manual Pabst B.U. Skalar B.U. Difference

1 HLTS 7B2 12.9 13.5 0.6

2 HLTS 8B2 13.0 13.8 0.8
3 HLTS 6B2 12.9 12.9 0.0
4 HOPS 4R2 12.4 12.3/11.9 0.1/0.5
5 HOPS 7R2 12.4 12.1/11.8 0.3/0.6
6 GOLR 4L2 12.0 12.6/12.0 0.6/0.0
7 GOLR 5L2 11.9 12.6/12.0 0.7/0.1
8 GOLR 1B3 11.7 12.4/12.0 0.7/0.3
9 GOLR 2B3 12.1 12.3/11.8 0.2/0.3

FLOW DIAGRAM
free waste
ml/min
polyethylene tube
Flow cell
Filter 275 nm

** phase separator

20°C

Hydrochloric acid solution

Nitrogen gas silicone tube
iso-Octane *
Sample 20°C

*
waste
* waste
Sampler

* acidflex tubing

catnr. 191-000 2 of 3
 SKALAR METHODS
ANALYSIS: CARBOHYDRATE

Catnr. 227-000

PRINCIPLE

The automated procedure for the determination of Carbohydrate is based on the following reaction; the
anthron reagent reacts with carbohydrate in the presence of sulfuric acid at 95°C, forming a blue dye. The
colour is measured at 630 nm.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA. Check voltage at the
back of the instrument before installation.
2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer, typical range 2000 –100,000 ppm Carbohydrate.

For sample pretreatment see 2.1.1. and 2.1.6.

REAGENTS

A. Distilled water + Triton X-100

Required chemicals: Distilled water (H2O)

Triton X-100

B. Colour reagent

Required chemicals: Anthron (C14H10O)

Sulfuric acid (H2SO4)
Distilled water (H2O)

C. Rinsing liquid sampler

Required chemicals: Distilled water (H2O)

catnr. 227
STANDARDS

Stock solution Carbohydrate

Required chemicals: Glucose monohydrate (C6H12O6.H2O)

Distilled water (H2O)

Working standards

Dilute standards according to the range.

GENERAL REMARKS

1. The colour reagent can not be used in combination with the rinsing valve.

module numbers:
Carbohydrate, typical range 2000-100,000 ppm requires chemistry module 227-101.

SETTINGS

1. Sample time: 80 sec., wash time: 100 sec., air time: 1 sec.

REFERENCES

1. Pinnegar, M.A., “Automated analysis in relation to fermentation problems”, Automation in Analytical

Chemistry, 1965, page 80-81.

FLOW DIAGRAM
waste
ml/min

Flow cell
Filter 630 nm
Cor. Filter 520 nm

Air
Colour reagent
Resample 95°C 20°C
waste

Air
Distilled water + Triton X-100
Resample
waste

Air

Distilled water + Triton X-100

Sample

waste

Sampler

catnr. 227
FLOW DIAGRAM
waste
ml/min

Flow cell 30 mm
Filter 600 nm
Cor. Filter 750 nm

Ammonia solution 0.42 5247

Ammonium ferric citrate * 0.16 5247

Air
5324 5324 5327
Buffer solution 0.60 5232
5221 5221
Sample 0.60

0.80
waste

Sampler

* for a blank analysis, the ammonium ferric citrate

is replaced by distilled water

catnr. 227
 SKALAR METHODS
ANALYSIS : COLOR

Catnr. 275-000

PRINCIPLE

The automated method for the determination of the Color is based on the following procedure; the color of the
samples is measured at 430 nm.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA.

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer, malt, wort, typical range 0.8 – 40 B.C.-units.

For sample pretreatment see 1.1.2. 1.1.3. 2.1.2. and 2.1.5.

GENERAL REMARKS

1. Standardization on known beer samples, according to references 1 and 2.

2. High turbidity or variable turbidity : a correction filter of 700 nm and a matrix photometer are required.
3. Malts have to be measured within 1 hour.
4. Filter the malt and wort extract over a membrane filter if the haze is mort than 1 EBC unit.

REFERENCES

1. van Strien, J., Drost, B., Journal of the American Society of brewing chemists, (1979), 37, 84.
2. European Brewery Convention, Analytica EBC, Chapter 9.4 and 8.3. (manual method)
3. Methods of Analysis of the ASBC, 1992, Beer-10

catnr. 275-000 1 of 2
 FLOW DIAGRAM
ml/min

length 30 cm
to waste
flow cell

A:
430 nm
detector head
B:
700 nm
sampler

length 55 cm
from sampler

Absorbance filter A = 430 nm for colour

Absorbance filter B = 700 nm for turbidity

Measurement for colour and turbidity; connect interface cable to channel A

Measurement for colour only; connect interface cable to A-B

catnr. 275-000 2 of 2
 SKALAR METHODS
ANALYSIS : DIACETYL

Catnr. 298-000

PRINCIPLE

The automated procedure for the determination of Diacetyl is based on the following reaction; the diacetyl is
distilled from the beer. The distillate is mixed with an acidified solution of o-phenylenediamine to form
derivatives of quinoline. The color is measured at 340 nm.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA.

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer, typical range 6 – 300 ppb Diacetyl

For sample pretreatment see 2.1.7.

REAGENTS

A. Sulfuric acid solution

Required chemicals: Sulfuric acid (H2 SO4 (97%))

Distilled water (H2O)

B. Stock solution FFD6

Required chemicals: FFD6

Distilled water (H2O)

C. Colour reagent

Required chemicals: Hydrochloric acid (HCl (32%))

1,2-Phenylene diamine (C6 H8 N2)
Distilled water (H2 O)
Stock solution FFD6

D. Distilled water + FFD6

Required chemicals: Distilled water (H2 O)

Stock solution FFD6

E. Rinsing liquid sampler

Required chemicals: Distilled water (H2 O)

catnr. 298-000 1 of 3
STANDARDS

As a standard one takes a manually analyzed beer.

GENERAL REMARKS

1. Requires matrix compensation on 370 nm.

2. Standards of diacetyl are used only to check linearity. Artificial diacetyl standards have a factor of ± 2.3
compared to beer samples.

REFERENCES

1. Vicinale Diketone, Brautechnische Analysen methoden, Mebak, 1979, page 340-342.

2. Amerine, M.A., and Ough, C.S., Acetoin and diacetyl, Wine and must analysis, page 60-62.
3. Merdock, D.I., “Methods employed by the citrus concentrate industry for detecting diacetyl and
acetylmethyl carbinol, Food technology, April 1967, page 157-161.
4. Buijten, J.C., Holm, B., “An automated distillation method for the determination of diacetyl in beer; a
comparison of analysis by autosampler and gas chromatography”, Jour. of. Autom. Ch. 1979, Vol. 1, No.
2, page 91-94.

FLOW DIAGRAM
waste
ml/min

Flow cell
Filter 340 nm
Cor. Filter 370 nm
Nitrogen gas
Resample

Distilled water + FFD6 BLACK

cooling
waste
88°C
Colour reagent

waste
waste

Nitrogen gas
Nitrogen gas

Sample
Sulfuric acid solution

waste

Sampler

catnr. 298-000 2 of 3
 SKALAR METHODS
ANALYSIS : DENSITY

Catnr. 305-000

PRINCIPLE

The sample is thermostated and measured with a density meter.

The output is connected with the analyser data system.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA.

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer, malt and wort, typical range 0.9000 – 1.1000 g/cm3.

For sample pretreatment see 1.1.2. 1.1.3. and 2.1.5.

STANDARDS

Calibration is done on known beer samples.

GENERAL REMARKS

1. Always contact Skalar for this application.

2. Typical throughput 40 samples per hour.
3. Calculation of the density is realized by the Skalar software package.
4. Cooling device ± 0.01°C is required.
5. Extract calculation in combination with alcohol results can be achieved by our software.

catnr. 305-000 1 of 2
FLOW DIAGRAM
ml/min

waste

Air
Sample
20°C PAAR

Sampler

waste

catnr. 305-000 2 of 2
 SKALAR METHODS
ANALYSIS : DIASTATIC POWER

Catnr. 308-000

PRINCIPLE

The automated procedure for the determination of Diastatic Power is based on the following reaction; the
sample is mixed with a starch solution. The starch is hydrolyzed to reducing sugars by the diastatic power of
the sample. The potassium ferricyanide solution is added which oxidises (at 95°C) the reducing sugars and to
be reduced to the ferrous cyanide ion. The absorbance decrease of the ferrous ferricyanide is measured at
420 nm.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA.

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Malt, wort, typical range 5.2 – 260 DP Units.

For sample pretreatment see 1.1.1.

REAGENTS

A. Sodium chloride solution

Required chemicals: Sodium chloride (NaCl)

Distilled water (H2O)
Brij 35 (30%)

B. Sodium acetate buffer

Required chemicals: Sodium acetate (CH3COONa)

Acetic acid (CH3COOH (100%))
Distilled water (H2O)

C. Starch solution

Required chemicals: Starch (Lintner).

Sodium acetate buffer.
Distilled water (H2O)

catnr. 308-000 1 of 3
D. Alkaline potassium ferricyanide solution

Required chemicals: Potassium ferricyanide (K3Fe(CN)6)

Sodium carbonate (Na2CO3)
Distilled water (H2O)
Brij 35 (30%)

E. Rinsing liquid sampler

Required chemicals: Sodium chloride (NaCl)

Distilled water (H2O)

STANDARDS

Required chemicals: α-Amylase type VIII-A from Barley Malt

Sodium chloride (NaCl (0.5%))

Working standards

Dilute standards according to the range.

GENERAL REMARKS

1. EBC method 4.12.1 (manual method).

REFERENCES

1. European Brewery Convention, Analytica EBC, fourth edition, 1987, method E 79, 4.12.2 (manual
method).
2. Methods of Analysis of the ASBC, 1992, Malt 6-A.
3. Banasik, O.J., "An automated Analysis of Malt Diastatic Power and Alpha Amylase Activity",
Wallerstein Laboratories Communications, Vol XXXIV, no 113, april 1971, page 45-46.

catnr. 308-000 2 of 3
FLOW DIAGRAM
waste
ml/min

Flow cell
Filter 420 nm
Cor. filter 500 nm

95°C

Air
Potassium ferricyanide solution

Air

Starch

Resample 35°C
waste
waste

Air

Sodium chloride solution

Sample

waste

Sampler

catnr. 308-000 3 of 3
 SKALAR METHODS
ANALYSIS : ETHANOL

Catnr. 329-000

PRINCIPLE

The automated procedure for the determination of Ethanol is based on the following reaction, the sample is
distilled at 96°C. The distilled ethanol is resampled and mixed with a potassium dichromate.
The chromium 6+ reduced to the green colored chromium 3+ by oxidation of the ethanol. The color is
measured at 590 nm

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyzer configuration, 2000 VA.

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer, typical range 0.1 – 5% Ethanol.

For sample pretreatment see 2.1.2. 2.1.4. and 2.1.5.

REAGENTS

A. Distillation reagent

Required chemicals: Distilled water (H 2 O)

Antifoam

B. Distilled water + Brij 35

Required chemicals: Distilled water (H 2 O)

Brij 35 (30%)

C. Colour reagent

Required chemicals: Potassium dichromate (K2 Cr2O7 )

Nitric acid (HNO3 (65%))
Distilled water (H2 O)

D. Rinsing liquid sampler

Required chemicals: Distilled water (H2 O)

catnr. 329-000 1 of 3
STANDARDS

Stock solution Ethanol

Required chemicals: Ethanol (CH3 CH2 OH (abs.))

Working standards

Dilute standards according to the range.

GENERAL REMARKS

1. Contact Skalar for the enzymatic method 323.

2. Volatile substances, which can be oxidized interfere.

CALCULATIONS
Alcohol w/w % * Density Beer
Alcohol v/v % =
Density Alcohol

Original wort extract = p (°Plato)

Real extract in % = ER
Apparent extract in % = EA
Real degree of fermentation in % = VR
Apparent degree of fermentation in % = VA
A = alcohol content (w/w)

From the table of Goldiner/Klemann, Kämpf can be calculated =

EA = - 460.234 + 662.649 .density filtered beer – 202.414 .density filtered beer2

ER = - 460.234 + 662.649 .density residue – 202.414 .density residue 2

A = 517.4.(1 – density distillate) + 5084 . (1 – density distillate)2 + 33503 (1 – density distillate)3

100 . (A . 2.0665 + E )
p= R
100 + 1.0665 . A

(Original wort extract – E ) . 100

VA = A
Original wort extract

(Original wort extract – E ) . 100

VR = R
Original wort extract

REFERENCES

1. Sawyer, R. Dixon, E.J. Analyst, Vol. 93, October 1968, page 680-687.
2. Hart & Fisher, Modern Food analysis, 1971, page 41-42.
3. Official journal of the European Communities, L 272, Vol. 33, October 1990, ISBN no. 0378-6978, page
187-189. (manual method)

catnr. 329-000 2 of 3
 FLOW DIAGRAM
waste
ml/min

Flow cell
Filter 590 nm
Cor. Filter 720 nm

Nitrogen gas
Colour reagent

Resample 40°C

waste

cooling
Distilled water + Brij 35 waste

waste

96°C
Nitrogen gas

Distillation reagent
X
Sample

waste

Sampler

flow meter
90 units

catnr. 329-000 3 of 3
 SKALAR METHODS
ANALYSIS : β-GLUCAN

Catnr. 349-000

PRINCIPLE

The automated procedure for the determination of β-Glucan is based on the following reaction; the sample is
diluted with distilled water and is resampled in a cellufluor reagent. Cellufluor reacts with the β-glucans.
The cellufluor-β-glucan complex fluorescence intensity is measured. The excitation wavelength is 365 nm
and the emission wavelength is 425 nm.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA.

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer, typical range 6 - 300 ppm β-glucan.

For sample pretreatment see 2.1.1.

B Malt, wort, typical range 6 – 300 ppm β-glucan.

For sample pretreatment see 1.1.1. and 1.1.2.

REAGENTS

A. Distilled water + FFD6

Required chemicals: Distilled water (H2 O)

FFD6

B. Stock glycine buffer (1M)

Required chemicals: Glycine (C2 H5 O2N)

Distilled water (H2O)

C. Cellufluor working reagent

Required chemicals: Cellufluor

Stock glycine buffer B
D. Rinsing liquid sampler

Required chemicals: Distilled water (H2 O)

STANDARDS

Required chemicals: Biocon β-Glucan

Distilled water (H2O)

Working standards

Dilute standards according to the range.

GENERAL REMARKS

1. A fluorimeter SA 6300 is required.

REFERENCES

1. Aastrup, S., and Jorgensen, K.G., "Application of the Calcofluor Flow Injection Analysis Method for
determination of β-Glucan in barley, malt, wort and beer", ASBC Journal, 1988, Vol.46, No.3,
page 76-81.
2. Analytical EBC (ed) Moll, M., fourth Ed. 1987, E85 method 4.1.3.
2. Jorgensen, K.G., Munck, J., Ruud-Hansen, J. and Hansen, K.T., "The EBC methods for determination of
high molecular weight β-Glucan in barley, malt, wort and beer", M., Journal of the institute of Brewing,
1989, vol. 95, page 79-82..

FLOW DIAGRAM
waste
ml/min

FLUORIMETER
Flow cell
Filter prim. 365 nm
Filter sec. 425 nm

Air
Cellufluor

Resample waste

Air
Distilled water + FFD6

Sample

waste

Sampler
 SKALAR METHODS
ANALYSIS : β-GLUCANASE

Catnr. 351-000

PRINCIPLE

The automated procedure for the determination of β-Glucanase is based on the following reaction; the
enzyme is reacted with β-glucan substrate for 10 minutes and the reducing end group is determinated at 540
nm by means of Dinitro Salicylic acid (D.N.S.).

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA.

Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Malt, wort, typical range 20 – 1000 ppm Maltose activity.

For sample pretreatment see 1.1.1.

REAGENTS

A. Distilled water + Teepol 610

Required chemicals: Distilled water (H2O)

Teepol 610

B. Stock acetate buffer solution

Required chemicals: Sodium acetate (CH3COONa)

Acetic acid (CH3COOH)

C. Colour reagent

Required chemicals: di-Nitro salicylic acid (C9H10O5)

Sodium hydroxide (NaOH)
Potassium sodium tartrate (C4H4KNaO6.4H 2O)
Distilled water (H2O)
Teepol 610.

D. Substrate

Required chemicals: β-Glucan

Stock acetate buffer solution (B)
Distilled water (H2O)

catnr. 351-000 1 of 3
E. Rinsing liquid sampler

Required chemicals: Sodium chloride (NaCl)

Distilled water (H2O)

STANDARDS

Stock solution Maltose

Required chemicals: Maltose (C12H22O11.H2O)

Distilled water (H2O)

Working standards

Dilute standards according to the range.

GENERAL REMARKS

1. A blank analysis (90°C, 5 min.) is required.

REFERENCES

1. Miller, G.L., Analytical Chemistry, Vol. 31, page 426-428 (1959).

2. Wainwright, T., Buckee, G.K., "Barley and Malt Analysis – A revieuw", Journal of the institute of
brewing, Vol. 83, 1977, page 325-347.

FLOW DIAGRAM
waste
ml/min

Flow cell
Filter 540 nm

Air

Distilled water + Teepol 610

Resample waste

Colour reagent

Sample

Air

Substrate
50°C cooling 95°C
25°C

waste

Sampler

catnr. 351-000 2 of 3
 SKALAR METHODS
ANALYSIS : NITRATE + NITRITE

Catnr. 455-000

PRINCIPLE

The automated procedure for the determination of Nitrate and Nitrite is based on the hydrazine reduction
method; nitrate is reduced to nitrite by hydrazinium sulfate. The nitrite reacts with sulfanilamide and
α-naphthyl-ethylenediamine dihydrochloride to form a highly coloured azo dye.
The colour is measured at 540 nm.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyzer configuration, 2000 VA.

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer, typical range 0.2 – 10 ppm Nitrate + Nitrite.

For sample pretreatment see 2.1.1.

REAGENTS

A. Buffer solution

Required chemicals: Sodium citrate (C6H5O7Na3.2H 2O)

Sodium hydroxide (NaOH)
Distilled water (H2O)
Brij 35 (30%)

B. Sodium hydroxide solution

Required chemicals: Sodium hydroxide (NaOH)

Distilled water (H2O)
Brij 35 (30%)

C. Stock solution cupric sulfate

Required chemicals: Cupric sulfate (CuSO 4.5H 2O)

Distilled water (H2O)
D. Hydrazinium sulfate solution

Required chemicals: Hydrazinium sulfate (N2H6SO4)

Stock solution cupric sulfate
Distilled water (H2O)

E. Colour reagent

Required chemicals: o-Phosphoric acid (H3PO4 (85%))

Sulfanilamide (C6H8N 2O2S)
α-Naphthylethylene di- amine dihydrochloride (C12H16Cl2N2)
Distilled water (H2O)

F. Rinsing liquid sampler

Required chemicals: Distilled water (H2O)

STANDARDS

Stock standard Nitrate

Required chemicals: Sodium nitrate (NaNO3)

Distilled water (H2O)

Working standards

Dilute standards according to the range .

GENERAL REMARKS

1. Procedure based on reduction with cadmium instead of hydrazinium sulfate is available.

2. Reduction equilibrium is critical and depends on the quality of the hydrazinium sulfate.
3. The nitrite, present in the sample will be analyzed as well. Separate determination can be made
by replacing the hydrazinium sulfate by water.

REFERENCES

1. Kamphake, L.J., Hannah, S.A., Cohen, J.M., "Automated analysis for nitrate by hydrazine reduction",
Water Research, 1, 1967, page 205-216.
2. Kempers, A.J., Luft, A.G., "Re-examination of the determination of environmental Nitrate as Nitrite by
reduction with hydrazine", Analyst, Vol. 113, July 1988, pp. 1117-1120.
FLOW DIAGRAM
waste
ml/min

Flow cell
Filter 540 nm
Cor. Filter 620 nm

Color reagent

37°C

Hydrazinium sulfate

Air

Buffer
Air
Buffer

Sample
waste

waste

Sampler
 SKALAR METHODS
ANALYSIS : NITRATE + NITRITE

Catnr. 461-000

PRINCIPLE

The automated procedure for the determination of Nitrate + Nitrite is based on the cadmium reduction
method; the sample is passed through a column containing granulated copper-cadmium to reduce the nitrate
to nitrite. The nitrite reacts with sulfanilamide and α-naphthylehylenediamine dihydrochloride to form a
highly colored diazo dye. The color is measured at 540 nm.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyzer configuration, 2000 VA.

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.
3. Cadmium granules sieve twice, first aperture 0.3 mm and secondly aperture 1.0 mm.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer, typical range 0.2 – 10 ppm nitrate + Nitrite.

For sample pretreatment see 2.1.1.

REAGENTS

A. Distilled water + Brij 35

Required chemicals: Distilled water (H 2O)

Brij 35 (30%)

B. Buffer
Required chemicals: Ammonium chloride (NH4Cl)
Ammonia solution (NH4 OH(25%))
Distilled water (H2O)
Brij 35 (30%)

C. Colour reagent

Required chemicals: o-Phosphoric acid (H3 PO4 (85%))

Sulfanilamide (C6 H8N 2O2 S)
α-Naphthylethylenediamine dihydrochloride (C12H14N2 .2HCL)
Distilled water (H2O)

D. Rinsing liquid sampler

Required chemicals: Distilled water (H2 O)

catno. 461-000 1 of 3
STANDARDS

Stock standard Nitrite

Required chemicals: Sodium nitrate (NaNO 3 )

Distilled water (H2O)

Working standards

Dilute standards according to the range.

GENERAL REMARKS

1. Method of choice for complex sample matrices and precision.

2. Hydrazinium reduction see method no. 455.

REFERENCES

1. Standard method for examination of water and waste water, 15th edition, 1980.
2. Navone, R., Proposed method for nitrate in potable waters, Amer. J., Water Works Ass. 56:781,
1964.
3. Walinga, I., van Vark, W., Houba, V.J.G., van der Lee, J.J., Plant Analysis Procedures, Part 7,
Department of Soil Science and Plant Nutrition, Wageningen Agricultural University, Syllabus 1989,
page 197-200.

FLOW DIAGRAM
waste
ml/min

Flow cell
Filter 540 nm
Cor. Filter 620 nm
Colour reagent

Air

Cadmium column

waste

Air

Buffer solution
Air
Buffer solution
Sample waste

waste
Sampler

catno. 461-000 2 of 3
 SKALAR METHODS
ANALYSIS : pH

Catnr. 485-000

PRINCIPLE

The automated procedure for the determination of the pH value is based on a potentiometrically
determination with a pH flow throughput electrode; the flow cell contains two combined electrodes. The
potential difference between the electrodes is amplified and related to a negative logarithm of the hydrogen
ions, expressed in pH.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA.

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer, typical range : pH 2-8

For sample pretreatment see 2.1.1. and 2.1.2.

B. Wort, malt, typical range : pH 2-8.

For sample pretreatment see 1.1.2. and 1.1.3.

REAGENTS

A. Potassium chloride solution (3M)

Required chemicals: Potassium chloride (KCl)

Distilled water (H2O)

B. Rinsing liquid sampler

Required chemicals: Sodium chloride (NaCl)

Hydrochloric acid (HCl)(32%))
Distilled water (H2O)

STANDARDS

Dilute commercial available pH buffers with almost the same ionic strength as the samples.

catnr. 485-000 1 of 2
GENERAL REMARKS

1. To avoid matrix difference, prepare standards and rinsing water of the sampler under the same conditions.

REFERENCES

1. Official Journal of the European Communities, L 272, Vol. 33, 3 October 1990, ISBN nr. .378-6978,
page 121-122, (manual method)

FLOW DIAGRAM
ml/min

waste

pH-meter

Air
Sample

Potassium chloride solution cooling

Sampler

catnr. 485-000 2 of 2
 SKALAR METHODS
ANALYSIS : POLYPHENOLS

Catnr. 521-000

PRINCIPLE

The automated procedure for the determination of Polyphenols is based on the following reaction; the sample
is mixed with a solution of carboxymethyl cellulose and ethylenediamine tetra acetic acid (EDTA). An
alkaline ferric solution is added with which the polyphenols react to yield a red coloured dye. This color is
measured at 600 nm.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA.

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer, typical range 3 – 150 ppm PP.

For sample pretreatment see 2.1.5.

REAGENTS

A. Buffer solution

Required chemicals: Carboxymethyl cellulose ((low viscosity))

Ethanol (CH3CH2OH)
Ethylene diamine tetra acetic acid 2Na (C10H14N2Na2O8.2H 2O)
Distilled water (H2O)
Brij 35 (30%)

B. Ammonia solution

Required chemicals: Ammonia solution (NH4OH (25%))

Distilled water (H2O)

C. Ammonium ferric citrate solution

Required chemicals: Ammonium ferric citrate (Fe (14-17%))

Distilled water (H2O)
D. Rinsing liquid sampler

Required chemicals: Distilled water (H2O)

STANDARDS

Standardisation is performed by absolute Beer standards of the laboratory.

GENERAL REMARKS

1. EBC method 9.9.1 (manual method).

2. The analysis needs blank analysis to eliminate own color interference. This can be analyzed simultaneously
using two analysis modules, one for the analysis and the second for the blank. It is possible to analyze on
the same module, separate batch of analysis and blank after each other, that requires only one module.
The Skalar software can substract automatically the blank value from the sample value.
3. Standardization is possible on Eomaric acid or Gallic acid, but only to check the linearity of the system.

REFERENCES

1. American Society of Brewing Chemists, “Subcommittee Report on Polyphenols in Beer:, Journal 36: 129,
1978.
2. European brewery convention, Analytica EBC, fourth edition, 1987, method 9.9.1 (manual method).
3. Baier, B., Jödicke, T., Harms, J., Lück, M., Müke, O., Krüger, E., "Anwendung der Continuous Flow
Analysentechnik zur Qualitätskontrolle in der Brauindustrie, Teil III : Einsatz der Continuous Flow Analytik
zur Bestimmung des Gesamtpolyphenolgehaltes in Würze und Bier", Monatsschrift für Brauwissenschaft,
45, Heft 6, 1992, page 204-210. (English translation available)

FLOW DIAGRAM
waste
ml/min

Flow cell
Filter 600 nm
Cor. Filter 750 nm

Ammonia solution

Ammonium ferric citrate *

Air
Buffer solution

Sample

waste

Sampler

* for a blank analysis, the ammonium ferric citrate

is replaced by distilled water
 SKALAR METHODS
ANALYSIS : SOLUBLE PROTEIN

Catnr. 530-000

PRINCIPLE

The automated procedure for the determination of Soluble Protein is based on the following method; the
sample is diluted in a sodium chloride solution. After resampling the sample is measured directly at 215 and
225 nm. The protein contents is calculated based on the difference in absorbance at 215 and 225 nm.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA.

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Malt, typical range 10 – 500 ppm Protein.

For sample pretreatment see 1.1.2.

REAGENTS

A. Sodium chloride solution (0.5%)

Required chemicals: Sodium chloride (NaCl)

Distilled water (H2O)
Brij 35 (30%)

B. Rinsing liquid sampler

Required chemicals: Sodium chloride (NaCl)

Distilled water (H2O)

STANDARDS

Worts calibrated by the Kjeldahl method.

GENERAL REMARKS

1. The spectrophotometer wavelength has to be set exact at the required wavelength.

catnr. 530-000 1 of 2
 FLOW DIAGRAM
ml/min

Filter 215 nm
Filter 225 nm
waste
X

waste

Air

0.5% NaCl solution

Resample
waste

Air

0.5% NaCl solution

Sample

Sampler

catnr. 530-000 2 of 2
 SKALAR METHODS
ANALYSIS : REDUCING SUGARS (TOTAL)

Catnr. 551-000

PRINCIPLE

The automated procedure for the determination of Total Reducing Sugars is based on the following reaction;
before dialysis the sample is hydrolyzed at 95°C or inverted enzymatically at 37°C to invert sucrose. After
addition of a copper neocuproin solution, the stream is heated to 97°C. The copper neocuproin chelate is
reduced by the sugars and forms a yellow cupro-neocuproin complex which is measured at 460 nm.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer, wort, typical range 2-100 g Glucose/liter.

For sample pretreatment see 1.1.3. and 2.1.1.

REAGENTS

A1. Distilled water + Brij 35

Required chemicals: Distilled water (H2O)

Brij 35 (30%)

A2. Hydrochloric acid (HCl inversion)

Required chemicals: Hydrochloric acid (HCl (32%))

Distilled water (H2O)
Brij 35 (30%)

B1. Sodium chloride (without inversion and HCl inversion)

Required chemicals: Sodium chloride (NaCl)

Distilled water (H2O)
Brij 35 (30%)

catnr. 551-000 1 of 4
B2. Sodium carbonate solution

Required chemicals: Sodium chloride (NaCl)

β-fructosidase
Distilled water (H2O)
Brij 35 (30%)

C. Sodium carbonate

Required chemicals: Sodium carbonate (Na2CO3)

Distilled water (H2O)
Brij 35 (30%)

D. Colour reagent

Required chemicals: Neocuproin hydrochloride (C14H13ClN2)

Cupric sulfate (CuSO 4.5H 2O)
Distilled water (H2O)

E. Rinsing liquid sampler

Required chemicals: Distilled water (H2O)

STANDARDS

Stock solution Glucose

Required chemicals: Glucose (C6H12O6)

Distilled water (H2O)

Working standards

Dilute standards according to the range.

REFERENCES

1. Sawyer, R. et Dixon, E.J., The Analyst, 1968, page 669-679.

2. Sawyer, R. et Dixon, E.J., The Analyst, 1968, page 680-687.
3. Moll, M., Flayeux, R., Lehueda, J.M., “Determination automatique des sucres reducteurs totaux dans le
moût et la biere apres hydrolyse”.

catnr. 551-000 2 of 4
FLOW DIAGRAM (without inversion)
waste
ml/min

Flow cell
Filter460 nm
Cor. Filter 620 nm
97°C
Colour reagent
Air

Sodium carbonate solution

Air

Sodium chloride solution

Resample
waste waste

Air
Distilled water + Brij 35

Resample waste

Air

Distilled water + Brij 35

Sample

waste

Sampler

FLOW DIAGRAM (acid hydrolysis)

waste
ml/min

Flow cell
Filter460 nm
Cor. Filter 620 nm
97°C
Colour reagent

Air
Sodium carbonate solution

Air
Sodium chloride solution
Resample
waste waste

Air
Hydrochloric acid 3 N

Resample 95°C
waste

Air

Distilled water + Brij 35

Sample

waste

Sampler

catnr. 551-000 3 of 4
FLOW DIAGRAM (enzymatic inversion)
waste
ml/min

Flow cell
Filter460 nm
Cor. Filter 620 nm
97°C
Colour reagent
Air

Sodium carbonate solution

Air 37°C
b-fructosidase

Resample 37°C
waste waste

Air

Distilled water + Brij 35

Resample waste

Air

Distilled water + Brij 35

Sample

waste

Sampler

catnr. 551-000 4 of 4
 SKALAR METHODS
ANALYSIS : TOTAL SULFUR DIOXIDE

Catnr. 593-000

PRINCIPLE

The automated procedure for the determination of Sulfur Dioxide is based on the following reaction; the
sample is acidified with sulfuric acid and heated at 95°C to release the bound sulfur dioxide. Gaseous sulfur
dioxide formed is dialysed into a formaldehyde solution. para-Rosaniline is added. A red coloured complex
is formed by the reaction of the sulfur dioxide and the para-rosaniline.
The absorption is measured at 560 nm.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA.

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer, typical range 0.4 – 20 ppm Sulfur dioxide.

For sample pretreatment see 2.1.1.

REAGENTS

A. Sulfuric acid solution 4N

Required chemicals: Sulfuric acid (H2 SO4 (96%))

Distilled water (H2 O)
Triton X-100

B. Sulfuric acid solution 1%

Required chemicals: Sulfuric acid (H2 SO4 (96%))

Distilled water (H2 O)
Brij 35 (30%)

C. Formaldehyde solution

Required chemicals: Formaldehyde (CH2 O (37%))

Distilled water (H2 O)
D. para-Rosaniline stock solution

Required chemicals: para-Rosaniline HCl (C19 H17N3 HCl)

Hydrochloric acid (HCl (32%))
Distilled water (H2 O)

E. Color reagent

Required chemicals: para-Rosaniline stock solution

Hydrochloric acid (HCl (32%))
Distilled water (H2 O)

E. Rinsing liquid sampler

Required chemicals: Distilled water (H 2O)

STANDARDS

Stock solution Sulfur dioxide

Required chemicals: Sodium bisulfite (Na2 S2 O5 )

Sodium hydroxide (NaOH)
Distilled water (H2 O)

Working standards

Dilute standards according to the range.

REFERENCES

1. West, P.W., Gaeke, G.C., “Fixation of Sulfur Dioxide as Disulfitomercurate (II) and subsequent
Colorimetric Estimation”, Anal. Chem., vol. 12, 1956, page 1816-1819.
2. Faria, A.H., Tavares, M.J., "Determination of Total Sulphur dioxide in wines, by Segmented Flow
Analysis with destillation", Instituto da Vinha e do Vinho, Direcçao de Serviços de Controle de
Qualidade, Lissabon, Portugal.
FLOW DIAGRAM
waste
ml/min

Flow cell
Filter 560 nm
Cor. Filter 700 nm
45°C

para-Rosaniline solution

Formaldehyde solution
Nitrogen gas

Sulfuric acid solution (1%)

Nitrogen gas
Sulfuric acid solution (4N)
Sample 95°C
waste

waste

Sampler
 SKALAR METHODS
ANALYSIS : THIOBARBITURIC ACID VALUE

Catnr. 606-000

PRINCIPLE

The automated procedure for the determination of Thiobarbituric acid value is based on the following
reaction between thiobarbituric acid and carbonyl complexes, after predilution the sample is resampled in an
acetic acid / barbituric acid solution and is heated till 95°C. The thiobarbituric acid reacts with the carbonyl
components and forms a methin dye. The color measured at 450 nm.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA.

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer, typical range 1 – 50 ppm TBZ.

For sample pretreatment see 2.1.1.

REAGENTS

A. Distilled water + MSD

Required chemicals: Distilled water (H 2 O)

MSD (20%)

B. Colour reagent

Required chemicals: Acetic acid (CH3COOH (100%))

2-Thiobarbituric acid (C4 H4 N2O2 S)
Distilled water (H2O)
MSD (20%)

C. Rinsing liquid sampler

Required chemicals: Distilled water (H2 O) .

STANDARDS

Standardisation is done by the absolute standards of the laboratory.

GENERAL REMARKS

1. Standardization on manual determined samples.

2. Matrix photometer 6250 or 6260, is required.

REFERENCES

1. Birkenstock, B., “Anwendung der Thiobarbiursäurezahl (TBZ) in der brautechnischen

Betriebsüberwachung”, Brauweit, Nr. 3 (1990), page 44-46.
2. Brickenstock, B., Thalacker, R., "Eine neue Kennzahl in der brautechnischen Analyse die
Thiobarbitursäurezahl (TBZ)", Brauwissenschaft, 35 (1982), Heft 6, page 133-137.
3. Baier, B., Jödicke, T., Harms, J., Lück, M., Müke, O., Krüger, E., "Anwendung der Continuous Flow
Analysentechnik zur Qualitätskontrolle in der Brauindustrie, Teil V : Bestimmung der
Thiobarbitursäurezahl in Betriebswürze und Bier", Monatsschrift für Brauwissenschaft, 46, 1993.
(Englisch translation available).

FLOW DIAGRAM
waste
ml/min

Flow cell
Filter 450 nm
Cor. Filter 600 nm

waste
Air

Colour reagent

Resample 95°C cooling

waste

Air

Distilled water + MSD

Sample

waste

Sampler
 SKALAR METHODS
ANALYSIS : VISCOSITY

Catnr. 615-000

PRINCIPLE

The automated procedure for the determination of Viscosity is based on the following procedure; the sample
is measured in a viscosity meter at 20°C.

LABORATORY FACILITIES

1. Maximum power consumption depending on the analyser configuration, 2000 VA.

2. Facilities for chemical wastes. Check environmental regulations for proper disposal of waste.

RANGE, SAMPLE MATRIX AND PRETREATMENT

A. Beer, malt, wort typical range 1.00 – 2.00 cP (centipoise)

For sample pretreatment see 1.1.2. and 2.1.5.

REAGENTS

A. Rinsing liquid sampler

Required chemicals: Distilled water (H2O)

STANDARDS

Stock solution Glucose

Required chemicals: D(+)Glucose monohydrate (C6H12O6.2H 2O)

Distilled water (H2O)

Working solutions

Dilute standards according to the range.

REFERENCES

1. Methods of Analysis of the ASBC, 1992, Wort-13.

catnr. 615 1 of 2
FLOW DIAGRAM

ml/min

X
waste

Air

Sample
20°C

Sampler waste

waste

catnr. 615 2 of 2