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Frank Fürle, Senior Application Specialist Industrial Solutions

Analysis of Sulfites and Sugars in Wine with Discrete Analyzer Technology

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Overview of the Content

• Introduction to automated system solutions

• Reasons to measure sugars and sulfites in wine

• Ready-to-use test kits for analysis

• Method principles and performance of analysis

• Benefits of automated discrete analysis

• System kit offering for food & beverage analysis

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Complete Thermo Scientific Assay Solution

Analyzers automating: • Dispensing • Mixing • Dilution • Incubation • Measuring

Ready-to-use reagents for food& beverage testing, like wine, juice, beer, malt, water, milk

Validated methods

System support: • Application support • Technical support

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Open Platform

Analyzers automating: • Dispensing • Mixing • Dilution • Incubation • Measuring

Ready-to-use reagents for food& beverage testing, like wine, juice, beer, malt, water, milk

Validated methods

System support: • Application support • Technical support

Customer-tailored applications, e.g., enzyme activity measurements up to 60°C

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Benefits of Discrete Analyzers System Solutions

• Efficiency – time and cost savings

• Consistency in sample processing – no manual errors, short turn-around times

• Ease-of-use – requires minimal training and skill level to run routine analysis

• Traceability of results - fully automated and validated methods giving confidence in the accuracy and precision of results

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Discrete Analysis Process

1. Cuvette entry point 2. Cuvette loader 3. Incubator 4. Sample racks 5. Sample disk 6. Reagents 7. Reagent disk 8. Barcode reader 9. Reagent dispenser 10. Sample dispenser 11. Mixer 12. Photometer unit

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Photometric Principle and Measurement

Light source

Filter disk

Optical fiber

Beam splitter

Reference detector

Signal detector

Absorbance range : 0 - 3,5A Resolution: 0.001A Reproducibility: 0.005 at 2A

Light beam Filtered beam

Reference beam

Signal beam

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Reasons for Wine Analysis

• Control of must and final product

• Control of production processes (malolactic fermentation)

• Detection of adulterations and false declaration

• Label claims /legal limits

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Sugar Analysis in Wine

• Yeast enzymes convert sugars, sucrose, glucose and fructose, to • Ethanol • Carbon dioxide

• Sucrose is first converted to glucose and fructose, then glucose is consumed followed by fructose

• Results reported either as • Total sugars (sum of glucose, fructose and sucrose) • Fermentable sugars (sum of glucose and fructose) • Fructose and glucose separately

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Ready-to-use Test Kits for Sugar Analysis

• Kits include all reagents needed for each assay • Enzymes • Buffers

• The photometric measurement is based on the formation of NADH • The amount of formed NADH is directly proportional to the

amount of assay

• Measurements at 340 nm, 37oC • The increase of NADH is measured by means of its light

absorbance at 340 nm

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Advantages of Enzymatic Tests

• Specificity • Enzyme reaction with its substrate is highly specific

• High precision and sensitive determination • Generally CV values below 2 %

• Typically no sample preparation needed

• Fast and reliable

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Performance of Sugar Analysis

For sugar analysis Measuring range,g/L Precision CV% D-Fructose 0.02 – 140 1.0 - 1.4 D-Glucose 0.02 – 140 1.6 – 1.7 D-Glucose + D-Fructose 0.04 – 200 1.0 - 1.4 D-Glucose + D-Fructose + Sucrose

0.24 – 200 1.6 - 2.7

• Wide measurement range with automated dilutions

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Example of Method Comparison Study for Glucose

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Example of Method Comparison Study for Glucose

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Analysis of Sulfites in Wine

• Free SO2 form • Active preservative • Acts as an antioxidant to increase physical stability • Used to control the fermentation process

• Free and bound SO2 • Sulphur dioxide can cause potentially

fatal allergic reactions • Total SO2 is regulated by legislation,

needs to be labelled

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SO2 Method Principle

• SO2 Free • The method is based on the reaction between sulfur dioxide,

pararosaniline and formaldehyde. • Method is performed at 37oC, using 575 nm filter and for side

wavelength 700 or 750 nm filter.

• SO2 Total

• The method is based on the reaction between sulfur dioxide and 5,5’-dithiobis nitrobenzoic acid (DTNB) in basic conditions.

• Method is performed at 37oC, using 405 nm filter and for side wavelength 700 or 750 nm filter.

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Performance of Sulfite Analysis

For sulfite analysis Measuring range,mg/L Precision CV% SO2 Free 2 - 150 1.5 – 4.6 SO2 Total 2 - 300 3.4 – 4.1

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Example of Correlation Study for SO2 Free

Gallery versus FIA

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System Reagents for Food and Beverage Analysis

D-Fructose D-Glucose D-Fructose+D-Glucose D-Fructose+D-Glucose+Sucrose Lactose (Glucose) Sucrose (Total) Acetic acid L-Ascorbic acid ß-Hydroxybutyric acid Citric acid D-Gluconic acid D-Isocitric Acid D-Lactic Acid L-Lactic Acid L-Malic acid Oxalic acid Total Acids

Cholesterol Food Ethanol Glycerol Polyphenols (total) Ammonia Acetaldehyde α-Amylase α-Amino Nitrogen (NOPA) Bitterness Beta-Glucan (High MW) Calcium (Ca) Copper Magnesium pH (Colorimetric) Potassium SO2 free and total Total Iron Total Protein Urea (Ammonia)

pH Alcohol std Acetaldehyde std Acid combination std Beta-Glucan Std Beta Hydroxybutyrate std Cholesterol std Copper std Glycerol std Lactose std NOPA std Oxalic acid std Sugar combination std Urea std pH standards

Sugars A

cid analysis

Alcohol analysis

Others

Calibrators

ECM

Over 50 optimized system products

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Benefits of Automated Discrete Analysis

• Flexible – various tests can be analyzed from each sample, there is no limitation of number of tests

• Fast – ready for immediate analysis, no reagent priming, no method changeover time

• Precise – specific measurements with high reproducibility, achieves low detection levels

• No carry-over – each reaction takes place in its own reaction cuvette

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Benefits of Thermo Scientific Discrete Analyzers

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

• Contamination-free analysis – disposable cuvettes

• Cost-effective – low reagent usage

• Adaptable – same system can be used for various applications

• Reliable – offering consistent performance and low maintenance cost

• Reduced environmental impact – low water consumption

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Key Benefits of Thermo Scientific System Reagents

• Optimized system solution • System applications • 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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Thermo Scientific™ Discrete Platforms

Aquakem™ Workcell

Aquakem™ 600 Aquakem™ 200 Aquakem™ 250

Arena™ 60

Arena™ 30

Arena™ 20 Arena™ 20XT

Gallery™

Gallery™ Plus

Environmental

Industrial

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Thank You!

For more information: Welcome to our Thermo Scientific booth to see the Thermo Scientific™ Gallery™ Plus automated discrete analyzer