Wine analysis: STANDARDS & READY-TO-USE REAGENTS For Discrete Analyzers

Transcription

1 AMS Alliance February Non contractual pictures. Specifications may be changed without prior notice. All rights reserved. Wine analysis: STANDARDS & READY-TO-USE REAGENTS For Discrete Analyzers NEW ORDER ONLINE reagents and consumables. AMS France 10, avenue Charles de Gaulle Frepillon FRANCE Tel. : (+33) Fax : (+33) info.fr@amsalliance.com

2 READY-TO-USE REAGENTS FOR WINECHEM RANGE SUMMARY We have created SELECTION, a full range of enzymatic and colorimetric reagents dedicated to our Smartchem (environment) and Winechem (wine) range of Discrete analyzers. Developed, tested and validated by our application laboratory, they will guarantee a perfect stability and reproducibility of your analyses. Very easy to use, they are supplied liquid, in specific bottles specifically made for Winechem, and automatically detected by the barcode reader (optional). Thus, this ready solution greatly simplifies the preparation of your analysis 2 ranges Ready-to-use reagents for Wine analysis. (see details here) and facilitates the use and storage of reagents. It also secures your analyzes with reagents adapted to your application and your instrument. A unique reagent inventory management is proposed when using our ready to use reagents. Fast Convenient Cost-effective Ready-to-use Ready-to-use reagents for Water analysis. Alkalinity (MO or BPB) - Ammonia - Chromium VI - Chlorides - Fluoride - Hardness - Nitrate - Nitrite - Phosphates - Silicate - Sulfates ALL THESE REAGENTS KITS CAN BE PURCHASED ON- LINE ON OUR WEBSHOP. STANDARDS & CONTROLS FOR DISCRETE ANALYZERS Multiparametric calibrator 04 Dedicated kits 05 READY-TO-USE REAGENTS KITS FOR DISCRETE ANALYZERS Acetaldehyde Acetic Acid Alpha Amminic Nitrogen Ammoniacal Nitrogen Anthocyans Biogenic Amines Calcium Ionic Catechins Citric Acid C02 Colour Copper D-Gluconic Acid D-Lactic Acid Free SO2 Glucose Glucose-Fructose Glycerol Iron Ionic L-Ascorbic Acid L-Lactic Acid L-Malic Acid Magnesium ph Potassium Pyruvic Acid Reduced Glutathione Sucrose Tartaric Acid Total Acidity Total Polyphenols Total SO2 Urea

3 Dedicated kits KIT COMPOSITION Acetaldehyde Standard 100 mg/l 10 x 10 ml EN009STD Alpha Amminic Nitrogen Standard 250 mg/l 10 x 10 ml EN010STD1 STANDARDS & CONTROLS FOR DISCRETE ANALYZERS Biogenic Amines Standard 100 mg/l Calcium Standard 200 mg/l 10 x 2 ml EN130BSTD 10 x 10 ml EN020STD1 Catechins Standard 500 mg/l 10 x 10 ml EN019STD Multiparametric kit Copper Standard 2,0-1,0-0,1 mg/l 6 x 10 ml EN013STD KIT COMPOSITION ENOCAL LD Multiparametric calibrator kit for: Acetic Acid (1,0 g/l) Citric Acid (1,0 g/l) D-Gluconic Acid (2,0 g/l) L-Lactic Acid (3,0 g/l) L-Malic Acid (5,0 g/l) Ammonia (250 mg/l) Glucose+Fructose (10 g/l) Glycerol (0,40 g/l) 10 x 10 ml EN050STD-LD D-lactique acid Standard 0,60 g/l Free SO 2 Standard 80 mg/l Free SO 2 To use with Free SO2 reagent Ref. EN022BM Standard 60 mg/l Glucose + Fructose Standard 20 g/l Iron Standard 20 mg/l L-Ascorbic Acid Standard 500 mg/l 10 x 10 ml EN006STD 10 x 10 ml EN022STD 10 x 10 ml EN022BSTD 10 x 10 ml EN014STD 10 x 10 ml EN016STD 10 x 500 mg EN131STD STANDARDS & CONTROLS FOR DISCRETE ANALYZERS 4 5

4 Dedicated kits KIT COMPOSITION Magnesium Standard 120 mg/l 10 x 10 ml EN025STD Polyphenols Standard 3 g/l 10 x 10 ml EN018STD Polyphenols To use with Total Polyphenols reagent Ref. EN18EM Standard 2,1 g/l Potassium Standard 1500 mg/l 10 x 10 ml EN018ESTD 10 x 10 ml EN026STD READY-TO-USE REAGENTS KITS FOR DISCRETE ANALYZERS Pyruvic Acid Standard 0,4 g/l 10 x 10 ml EN017STD STANDARDS & CONTROLS FOR DISCRETE ANALYZERS Reduced Glutathione Standard 2 g/l Sucrose Standard 2,5 g/l Tartaric Acid Standard 5 g/l Total SO 2 Standard 160 mg/l Total SO 2 To use with Total SO2 reagent Ref. EN023BM Standard 120 mg/l Urea Standard 500 mg/l 10 x 200 mg EN110STD 10 x 10 ml EN149STD 10 x 10 ml EN021STD 10 x 10 ml EN023STD 10 x 10 ml EN023BSTD 10 x 10 ml EN055STD Winechem 140 reagents /200 reagents reagents reagents Tested and validated by our applications laboratory, the SelectION Oeno reagents kits have been created specifically for use with both AMS Alliance Discrete analyzers and those of other manufacturers. They are supplied in ready-to-use liquid form, each one being formulated, tested and dedicated to its specific parameter. Their stability is guaranteed. These same kits are available either in a form compatible with all Discrete analyzers on the market, or in a form specific to our Winechem range analyzers; these references differ in the format of their bottles, designed to be directly inserted into the instrument s reagents compartment. Three ranges are available, the bottle shape differing according to the instrument: For Winechem 140 For or 200 For or 600 Caution : some parameters are available only for Winechem analyzers and vice versa. 6 7

5 Acetaldehyde REAGENT KIT FOR MEASUREMENT OF ACETALDEHYDE CONCENTRATION IN WINE Acetaldehyde (or ethanal, or acetic aldehyde), found naturally in wine, is a chemical compound produced by yeasts during alcoholic fermentation and by oxidation during ageing. This compound contributes to the aroma and complexity of wines. At high concentration however, it may have a negative impact on the wine. Acetic acid REAGENT KIT FOR MEASUREMENT OF ACETIC ACID CONCENTRATION IN WINE Acetic acid is a volatile acid produced by alcoholic fermentation. It is always present in wine, but in very small amounts. Measuring this compound provides valuable information concerning the sanitary status of the wine, as some contaminants cause a significant increase in volatile acidity, considering that beyond 0.80 g, the wine is considered unfit for sale. Acetaldehyde is converted to acetic acid by AIDH (Aldehyde dehydrogenase) in presence of NAD. The amount of NADH formed in the reactions is stoichiometric to the amount of Acetaldehyde in the sample. The reaction is determinate by means of its light absorbance at 340 nm. Acetate + ATP + CoA ACS Acetyl CoA + Oxaloacetate + H 2 O Acetate in the sample consumes NADH (by the following reaction), which can be measured by spectrophotometry. Acetyl CoA + AMP + Pyrophosphate L MHD Citrate + CoA L Malate + NAD + + H 2 O Oxaloacetate + NADH + H + CS R3 R4 1 x 100 ml 5 vials 5 vials 5 x 11 ml 333 tests EN-0092-A170/200 1 x 100 ml 5 vials 5 vials 5 x 11 ml 333 tests EN-N009-A450/600 R3 5 x 18 ml 2 x 10 ml 5 x 1,0 ml 447 tests EN001LM R3 Winechem x 11 ml 1 x 4 ml 1 x 1,5 ml 100 tests EN-001L-A140 5 x 19 ml 2 x 10 ml 5 x 1 ml 333 tests EN-001L-A170/200 5 x 19 ml 2 x 10 ml 5 x 1 ml 333 tests EN-001L-A450/

6 Alpha-Amminic Nitrogen COLORIMETRIC OPA UV METHOD REAGENT KIT FOR MEASUREMENT OF α-amminic NITROGEN CONCENTRATION IN WINE The nitrogen compounds in grape must and wine play a key role in fermentation and yeast growth. Measuring amino nitrogen provides a good indication of amino acid content. Ammoniacal Nitrogen REAGENT KIT FOR THE DETERMINATION OF AMMONIACAL NITROGEN IN WINE Grape must contains two forms of nitrogen available to yeasts: amino nitrogen and mineral nitrogen, or ammoniacal nitrogen. Nitrogen measurement is important for controlling fermentation and monitoring winemaking. At alkaline ph, the nitrogen of the primary amminic groups (NH2) react with ophthaldialdehyde (OPA) in presence of N-Acetyl-Cysteine to give a chromophor which absorbes at 340 nm. The amount of this chromophor is stoichiometrically related to the amount of α -amminic nitrogen present in the sample. α chetoglutarate + NADH + NH 4 Ammoniacal nitrogen reacts with oxoglutarate and reduced NADH in presence of GLDH enzyme to form L-Glutamate and NAD. The amount of NADH oxidised to NAD is stoichiometrically related to the amount of ammoniacal nitrogen present in the sample. GLDH L Glutamate + NAD + H 2 O OPA + NH 2 R ph=9,5 reducing agent OPA NH 2 R 2 x 45 ml 1 x 10 ml 410 tests EN010BM Winechem x 20 ml 1 x 2 ml 100 tests EN-010B-A140 4 x 32 ml 1 x 16 ml 410 tests EN-010B-A170/200 3 x 42 ml 1 x 16 ml 410 tests EN-010B-A450/600 2 x 45 ml 1 x 12 ml 410 tests EN008BM Winechem x 20 ml 1 x 3 ml 100 tests EN-008B-A140 4 x 32 ml 1 x 16 ml 410 tests EN-008B-A170/200 3 x 42 ml 1 x 16 ml 410 tests EN-008B-A450/

7 Anthocyans COLORIMETRIC METHOD REAGENT KIT FOR MEASUREMENT OF POLYPHENOLS ANTHOCYANIC COMPOUNDS IN WINE Biogenic Amines REAGENT KIT FOR MEASUREMENT OF BIOGENIC AMINES CONCENTRATION IN WINE Anthocyanins are purple or red pigments found in grape skin and hence in must. These anthocyanins give the wines their colour. Anthocyanins (from the Greek anthos meaning flour and kyanos meaning blue) constitute a large family of organic molecules found, in particular, in the red grape berry. Anthocyanins are notable primarily for their colouring strength. When combined with other polyphenols, they help improve wine colour stability, hence need to analyse them. Anthocyans Polyphenolics compounds are ionized at acid ph reacting with a buffer at a fixed ionic strength. The reagent is optimized to allow the complete ionization of the Anthocyans. In case of turbidity of the sample, due to proteic compounds, the chromogen reagent is able to eliminate them. This method measure the ionized and ionizables Anthocyans. The polymerized Anthocyans with the tannins are not analyzed from this method. α ketoglutarate + NADH + HN 4 Biogenic amines are by-products of fermentation by lactic acid bacteria. Some can impact health (e.g.: histamine can cause allergies and headaches) and, as such, must be monitored. The Diamino Oxidase enzyme catalyzes the oxidative deamination of biogenic amines with production of hydrogen peroxide and ammonium ion. The Ammonium ion reacts with oxoglutarate and reduced NADH in presence of glutamate dehydrogenase (GLDH) enzyme to form L-Glutamate and NAD. The amount of NADH oxidised to NAD is stoichiometrically related to the amount of Biogenic Amines present in the sample. GLDH L glutamate + NAD + H 2 O Winechem x 24 ml 100 tests EN-N011-A140 3 x 49 ml 565 tests EN011M 6 x 33 ml 565 tests EN-0112-A170/200 4 x 50 ml 565 tests EN-N011-A450/600 4 x 13,5 ml 1 x 13 ml 230 tests EN130BM A B Winechem x 12 ml 1 x 2,5 ml 1 x 5 ml 100 tests EN-130B-A140 2 x 27 ml 1 x 6,5 ml 1 x 13 ml 203 tests EN-130B-A170/200 2 x 27 ml 1 x 6,5 ml 1 x 13 ml 203 tests EN-130B-A450/

8 Calcium Ionic ARSENAZE III COLORIMETRIC METHOD REAGENT KIT FOR THE DETERMINATION OF CALCIUM IN WINE Catechins COLORIMETRIC METHOD REAGENT KIT FOR THE DETERMINATION OF CATECHINS IN WINE Calcium concentration in wine may vary and generate calcium tartrate, thus impacting wine acidity. If calcium concentration exceeds a given value, it may precipitate, giving rise to typical whitish deposits. In slightly acid ph, Calcium reacts with Arsenaze III. Arsenaze III is a metallochromic dye which is used in the determination of calcium in samples. Being a very sensitive method, is it possible to have positive interferences provoked by calcium presents in Glassware used for the analysis. Hence, It is necessary to use bidistilled water for dilution and to clean glassware. Catechins are member of the flavonoid family of polyphenols. Flavonoids give wine its colour: flavonols colour it yellow, anthocyanins colour it red and catechins, though colourless alone, when combined can give all shades from white to red, via rose. They also play a role in wine structure and stability. Under acid environment Cinnamaldehyde react with Catechins to give a coloured compound that can be measured spectrophotometrically at 620 nm. The amount of the coloured compound is directly proportional to the Catechins concentration in the sample. Catechins + DMACA [ Catechins DMACA ] 2 x 40 ml 1 x 19 ml 351 tests EN020BM Winechem x 20 ml 1 x 5 ml 100 tests EN-020B-A140 3 x 35 ml 1 x 27 ml 354 tests EN-020B-A170/200 3 x 36 ml 1 x 27 ml 354 tests EN-020B-A450/600 2 x 45 ml 2 x 15 ml 410 tests EN019LM Winechem x 20 ml 1 x 7 ml 100 tests EN-019L-A140 4 x 32 ml 2 x 21 ml 410 tests EN-019L-A170/200 3 x 42 ml 1 x 42 ml 410 tests EN-019L-A450/

9 Citric acid C02 KINETIC UV METHOD ENZYMATIC METHOD REAGENT KIT FOR THE MEASUREMENT OF CITRIC ACID CONCENTRATION IN WINE REAGENT KIT FOR THE DETERMINATION OF CARBON DIOXIDE IN WINE Citric acid is an organic acid found in grapes of all varieties. It is a metabolic by-product of Botyris cinerea, a fungus responsible for noble rot, which generates citric acid by sugar oxidation. During malolactic fermentation, nearly all of the citric acid is used by lactic acid bacteria, leading to the formation of acetic acid. Citric acid is also used preventively, as a treatment for white casse. It is also used to improve the taste properties of dry white wines. Many red wines have no citric acid. Measuring its concentration is essential to determine total wine acidity. The concentration of dissolved carbon dioxide (CO2) in wine has a major impact on this latter s organoleptic properties. CO2 generally emphasizes sensations of freshness and acidity, reduces sweetness, intensifies bitterness and astringency and may cause a tingling sensation. Depending on the wine, insufficient CO2 may render white wine flat. Conversely, excess CO2 can make red wines harsh and tannic. For this reason, managing CO2 content is an important factor. Citrate CL Citric acid presents in the sample is converted to oxaloacetate and acetate in the reaction catalyzed by the enzyme citrate lyase (CL). In the presence of L-LDH and L-MDH enzymes the oxaloacetate and its decarboxylation product pyruvate and reduced to L-malate and L-lactate by reduced NADH. The variation of light absorbance at 340 nm is stechiometric to the amount of citrate in the sample. Oxaloacetate + acetate L MDH Oxaloacetate + NADH + H + L malate + NAD + R3 R4 R3 R4 4 x 13,5 ml 4x4 U 4 x 1,5 ml 4 x 2,1 ml 278 tests EN003M 4 x 13,5 ml 4 x 4 U 4 x 1,5 ml 4 x 2,1 ml 278 tests EN-0032-A170/200 4 x 13,5 ml 4 x 4 U 4 x 1,5 ml 4 x 2,1 ml 278 tests EN-N003-A450/600 The assay for determination of CO2 is based on two different enzymatic reactions: The enzyme Phosphoenolpyruvate carboxylase (PEPC) catalyzes the first reaction which produces oxalacetate. In the second reaction, the cofactor (NADH analog), is oxidated by oxaloacetate in the presence of MDH. The amount of NADH analog oxidised to NAD is stoichiometrically related to the amount of carbon dioxide present in the sample. This results in a decrease absorbance at 405 nm. Phosphoenolpyruvate(PEP) + HCO 3 Oxaloacetate + H 2 PO 4 Oxaloacetate + NADH analog MDH Malate + NAD +ANALOG PEPC Winechem x 20 ml 100 tests EN-N100-A140 2 x 33,5 ml 330 tests EN-N100-A170/200 2 x 50 ml 330 tests EN-N100-A450/

10 Colour COLORIMETRIC METHOD REAGENT KIT FOR MEASUREMENT OF COLOR IN RED WINE This method is used to measure the chromatic characteristics of a wine, i.e. its brightness and chromacity. Brightness corresponds to transmittance. This value is inversely proportional to the wine s colour intensity. Chromacity corresponds to the dominant wavelength (characterising the shade) and purity. Many of the inputs used in wine-making will impact its colour, hence the need for regular measurements. The sample is suitable diluted with a buffer solution know and constant. Absorbance reading at 420 nm, 520 nm and 620 nm allows the chromatic characteristics to be calculated. Copper COLORIMETRIC DI-BR-PAESA METHOD REAGENT KIT FOR MEASUREMENT OF COPPER CONCENTRATION IN WINE The presence of copper in wine is due to the winegrowing methods used to protect the vine from diseases such as downy mildew. The copper content of wine must be measured as regulations impose a maximum concentration of 1 mg/l in finished wine. Under acid environment, copper present in the sample reacts with Di-Br-PAESA to form a coloured complex. The intensity of the coloured complex is proportional to the copper concentration in the sample. The method doesn t need deproteinization, neither sample blank. Standard included ph=4,7 Copper (Cu) + 2PAESA Cu(PAESA) reducing agent 2 R3 Winechem x 20 ml 1 x 20 ml 1 x 20 ml 300 tests EN-N024-A140 6 x 40 ml 1090 tests EN024M 4 x 28 ml 4 x 28 ml 4 x 28 ml 1090 tests EN-N024-A170/200 3 x 38 ml 3 x 38 ml 3 x 38 ml 1090 tests EN-N024-A450/600 R3 R4 Winechem x 10 ml 1 x 10 ml 1 x 5 ml 1 x 1,4 ml 100 tests EN-N013-A140 R3 R4 4 x 17,5 ml 4 x 17,5 ml 8 x 2 ml 8 x 0,45 ml 350 tests EN013BM 4 x 19,5 ml 4 x 19,5 ml 8 x 2,2 ml 8 x 0,5 ml 266 tests EN-N013-A170/200 4 x 19,5 ml 4 x 19,5 ml 8 x 2,2 ml 8 x 0,5 ml 266 tests EN-N013-A450/

11 D-Gluconic acid D-Lactic acid REAGENT KIT FOR MEASUREMENT OF D-GLUCONIC ACID CONCENTRATION IN WINE REAGENT KIT FOR THE D-LACTIC ACID DETERMINATION IN WINE Occurring naturally at trace concentrations in wine, gluconic acid is an indicator of grape degradation and sanitary status. Bacterial fermentation, which produces lactic acid, follows on from alcoholic fermentation. This is malolactic fermentation. The malic acid found naturally in wine is broken down to lactic acid by the bacteria. This fermentation is essential as it serves to reduce the wine s acidity. The measurement of D-lactic acid is necessary during the wine-making process. D gluconate + ATP GK 6 PGDH D-gluconic acid (D-gluconate) is phosphorylated by ATP in the presence of the enzyme gluconate kinase to D-gluconate-6-phosphate with the simultaneous formation of ADP. The next reaction results NADPH which can be measured by the instrument. D gluconate 6 P + ADP D gluconate 6 P + NADP + Ribulose 5 P + NADPH + CO 2 + H + D-Lactic acid is oxidised by NAD to pyruvate using D-Lactate dehydrogenase (D- LDH ) as a catalyst. The formation of pyruvate is removed using a second reaction catalyzed by GPT enzyme that converts the pyruvate to L-alanine in the presence of L-glutamate. The amount of NADH formed and measured is stoichiometrically related to the amount of D-Lactic acid (L-Lactate) present in the sample. 2 x 45 ml 1 x 10 ml 410 tests EN004BM Winechem x 20 ml 1 x 2 ml 100 tests EN-004B-A140 4 x 32 ml 1 x 13 ml 410 tests EN-004B-A170/200 3 x 42 ml 1 x 13 ml 410 tests EN-004B-A450/600 D lactate + NAD + D LDH Pyruvate + NADH 2 x 45 ml 1 x 10 ml 410 tests EN006BM Winechem x 20 ml 1 x 2 ml 100 tests EN-006B-A140 4 x 32 ml 1 x 16 ml 410 tests EN-006B-A170/200 3 x 42 ml 1 x 16 ml 410 tests EN-006B-A450/

12 Free SO2 COLORIMETRIC P-FUCSINE METHOD REAGENT KIT FOR MEASUREMENT OF FREE SULPHUR DIOXIDE IN WINE Glucose REAGENT KIT FOR MEASUREMENT OF GLUCOSE CONCENTRATION IN WINE SO2, or sulphur dioxide, is an essential wine input as, for the moment, there are no viable wine-making alternatives to its use. These are the famous sulphites. The fraction of SO2 that does not combine with the various other organic compounds is called free SO2. It is this fraction which possesses most antiseptic and antioxidant properties. In acid environment, «free sulphur dioxide» reacts with p-fucsine and formaldehyde to produce a chromophor of magenta colour which is measured at 575 nm. The interference of the polyphenols and the colour of the wines is eliminated by measuring of the Sample Blank. The grape s sugar content determines the wine s alcohol content. This value increases during ripening, at the expense of acidity, which will tend to decrease. During alcoholic fermentation, the bacteria and/or yeasts convert the glucose naturally contained in the grape to ethanol, i.e. the sugar is converted to alcohol. Measuring glucose serves, on the one hand, to determine the optimum moment for grape harvesting and, on the other hand, to monitor the wine-making process. Glucose react with adenosine triphosphate (ATP) in the presence of the enzyme Hexokinase (HK) to form glucose- 6-phosphate (G-6-P). G-6-P in the presence of the enzyme G6P-DH reduce NADP to form gluconate-6-phosphate. D Glucose + ATP G6P DH HK Glucose 6 phosphate + ADP G 6 P + NADP + Gluconate 6 phosphate + NADPH + H + Winechem x 20 ml 1 x 3 ml 100 tests EN-022B-A140 2 x 35,5 ml 1 x 9,5 ml 321 tests EN022BM 3 x 33 ml 1 x 13 ml 321 tests EN-022B-A170/200 2 x 49 ml 1 x 13 ml 321 tests EN-022B-A450/600 2 x 45 ml 1 x 12 ml 410 tests EN140M Winechem x 20 ml 1 x 2,5 ml 100 tests EN-N140-A140 4 x 32 ml 1 x 13 ml 410 tests EN-N140-A170/200 3 x 42 ml 1 x 13 ml 410 tests EN-N140-A450/

13 Glucose-Fructose Glycerol ENZYMATIC OIV-MA-AS METHOD REAGENT KIT FOR MEASUREMENT OF GLUCOSE-FRUCTOSE CONCENTRATION IN WINE REAGENT KIT FOR THE DETERMINATION OF GLYCEROL CONCENTRATION IN WINE The grape s sugar content determines the wine s alcohol content. It is measured during grape ripening and wine-making, when alcoholic fermentation converts the sugar to alcohol. Measuring glucose and fructose, or total sugars, is necessary to determine the optimum time to harvest the grapes and to monitor the wine-making process. Glycerol is a polyol, a compound with multiple alcohol groups (two primary alcohol groups and one secondary group), which give the wine its easy to drink, full-bodied and mellow nature. It is thus the third natural constituent of wine, after water and ethanol. It is essential for defining the wine s mouth-feel. Glucose and fructose react with ATP in presence of the enzyme Hexokinase (HK) to form glucose-6-phosphate (G6P) and fructose-6-phosphate (F6P). F6P is converted to G6P by the enzyme PGI. G6P, in the presence of the enzyme G6P-DH reduce NADP to form gluconate-6-phosphate. The glycerolkinase (GK) catalyses the phosphorylation of glycerol to glycerol-3- phosphate by ATP. The ADP is then converted into ATP by PEP in presence of pyruvate-kinase (PK) with pyruvate formation. Pyruvate is converted into lactate by reduced NADH in presence of LDH. The quantity of NAD+ formed during the reaction is proportional to the quantity of glycerol. The NADH oxidization is measured by the decrease of its extinction at wavelengths of 340 nm. D Glucose + ATP G-6-P ADP + H + D Fructose + ATP F-6-P ADP + H + F-6-P G6P DH PGI G-6-P G-6-P + NADP + Gluconate 6 phosphate + NADPH + H + HK HK Glycerol + ATP ATP + PEP GK PK Glycerol 3 phosphate + ADP LDH ATP + Pyruvate Pyruvate + NADH + H + Lactate + NAD + 3 x 44 ml 1 x 14 ml 410 tests EN014M Winechem x 30 ml 1 x 3 ml 100 tests EN-N014-A140 4 x 32 ml 1 x 13 ml 410 tests EN-N014-A170/200 3 x 42 ml 1 x 13 ml 410 tests EN-N014-A450/600 2 x 45 ml 1 x 20 ml 370 tests EN015M R3 Winechem x 11 ml 1 x 2 ml 1 x 3 ml 100 tests EN-N015-A140 5 x 12 ml 1 x 4,5 ml 1 x 7,7 ml 306 tests EN-N015-A170/200 5 x 12 ml 1 x 4,5 ml 1 x 7,7 ml 306 tests EN-N015-A450/

14 Iron ionic COLORIMETRIC FERENE S METHOD REAGENT KIT FOR MEASUREMENT OF IRON (IONIC) CONCENTRATION IN WINE Wine naturally contains very small amounts of iron (white casse). In wines kept in airtight containers in a reducing environment, the iron is in its ferrous state. If the wine contains dissolved oxygen following aeration, however, the iron oxidises and switches to ferric state, which can impact the colour and clarity of the wine. Under acid environment, iron is ionized. After reduction at ferrous state (Fe2+), iron react with Ferene S to form a stable coloured complex with a maximum of absorbance at 600 nm. The intensity of the colour measured is proportional at the quantity of iron presents in the sample. L-Ascorbic acid REAGENT KIT FOR MEASUREMENT OF L-ASCORBIC ACID CONCENTRATION IN WINE AND MUST L-ascorbic acid, or vitamin C, occurs naturally in grapes and grape must. This potent antioxidant plays a major role in the wine s aromas. Its content in wine is limited by regulation. The L-ascorbic acid in the presence of the enzyme ascorbate oxidase (AAO), is converted to acid L-Dehydroascorbic. L-Dehydroascorbic acid selectively reacts with the chromogen, producing a compound evaluated photometrically at 340 nm, the concentration of which is directly proportional to the concentration of L-ascorbic acid present in the sample. L ascorbic acid + 1 O 2 2 AAO L deshydroascorbic acid + H2O 2 x 42 ml 1 x 20 ml 370 tests EN016M Winechem x 20 ml 1 x 5 ml 100 tests EN-N016-A140 4 x 28 ml 1 x 28 ml 370 tests EN-N016-A170/200 3 x 38 ml 1 x 28 ml 370 tests EN-N016-A450/600 2 x 33,5 ml 1 x 16,5 ml 302 tests EN131BM Winechem x 20 ml 1 x 5 ml 100 tests EN-131B-A140 3 x 31 ml 1 x 23 ml 302 tests EN-131B-A170/200 2 x 46 ml 1 x 23 ml 302 tests EN-131B-A450/

15 L-Lactic acid REAGENT KIT FOR THE L-LACTIC ACID DETERMINATION IN WINE L-Malic acid REAGENT KIT FOR MEASUREMENT OF L-MALIC ACID CONCENTRATION IN WINE L lactate + NAD + L LDH L-Malic and L-Lactic acids are analyzed to monitor the malolactic fermentation process. L-lactic acid is oxidised by NAD to pyruvate using L-LDH as a catalyst. The amount of NADH formed and measured is stoichiometrically related to the amount of L-lactic acid (L-lactate) present in the sample. Pyruvate + NADH Malic acid is the main acid in wine. It is a natural organic acid. L-Malic and L-Lactic acids are analyzed to monitor the malolactic fermentation process. Measuring L-Malic acid allows the reduction of wine acidity to be monitored and modulated. L-Malic acid is oxidised by nicotinamide adenine dinucleotide ( NAD ) to oxaloacetate using L-Malate dehydrogenase (L- MDH ) as a catalyst. The formation of oxaloacetate is removed using a second reaction catalyzed by glutamate oxaloacetate transaminase enzyme (GOT) that converts the oxaloacetate to L-aspartate in the presence of L-glutamate. L Malate + NAD + Oxaloacetate + L glutamate L MDH GOT Oxaloacetate + NADH Aspartate + 2 oxoglutarate 2 x 45 ml 1 x 10 ml 410 tests EN005BM Winechem x 20 ml 1 x 2 ml 100 tests EN-005B-A140 4 x 31 ml 1 x 16 ml 410 tests EN-005B-A170/200 3 x 42 ml 1 x 16 ml 410 tests EN-005B-A450/600 2 x 45 ml 1 x 18,5 ml 410 tests EN007BM Winechem x 20 ml 1 x 4 ml 100 tests EN-007B-A140 4 x 32 ml 1 x 25 ml 410 tests EN-007B-A170/200 3 x 42 ml 1 x 25 ml 410 tests EN-007B-A450/

16 Magnesium COLORIMETRIC XYLIDYL BLUE METHOD REAGENT KIT FOR MEASUREMENT OF MAGNESIUM IN WINE Magnesium (Mg) is an essential vine trace element. Magnesium levels do not decrease during fermentation and storage as the magnesium salts of the wine s mineral or organic anions are soluble. The presence of magnesium contributes to the beneficial effects of wine. It normally contains between 60 and 150 mg/l. Magnesium REAGENT KIT FOR MEASUREMENT OF MAGNESIUM IN WINE Magnesium (Mg) is an essential vine trace element. Magnesium levels do not decrease during fermentation and storage as the magnesium salts of the wine s mineral or organic anions are soluble. The presence of magnesium contributes to the beneficial effects of wine. It normally contains between 60 and 150 mg/l. The Magnesium Reagent utilizes a direct method in which magnesium forms a coloured complex with xylidyl blue in a strongly basic solution, where calcium interference is eliminated by glycoletherdiamine-n,n,n N tetraacetic acid (EGTA). The colour produced is measured at 520 nm and is proportional to the magnesium concentration. In buffer solution, the magnesium content actives a specific enzyme. This enzymatic reaction produces a compound measurable by spectrophotometer. The increment of enzymatic activity is directly proportional to magnesium content. The enzymes used are specific to magnesium, so there is no interference caused by other metals. 2 x 40 ml 1 x 19 ml 350 tests EN025BM Winechem x 20 ml 1 x 5 ml 100 tests EN-025B-A140 3 x 35 ml 1 x 27 ml 350 tests EN-025B-A170/200 3 x 36 ml 1 x 27 ml 350 tests EN-025B-A450/600 2 x 40 ml 1 x 19 ml 350 tests EN025EM 30 31

17 ph Potassium COLORIMETRIC METHOD TURBIDIMETRIC TETRAPHENYLBORATE METHOD REAGENT KIT FOR MEASUREMENT OF PH IN WINE REAGENT KIT FOR MEASUREMENT OF POTASSIUM CONCENTRATION IN WINE Measuring a wine s ph is supplemental to that of total acidity; it serves to monitor the wine s acidity. If the ph is too high, 4.0 or higher, the wine becomes microbiologically unstable. A low ph, on the other hand, will inhibit microbial growth, thus contributing to the wine s stability over time. Potassium is an essential element for vine growth and is found (in compound form) in most soils. This micronutrient plays a non-negligible role in our diet. In the mineral composition of wine, potassium is found at levels of 0.7 to 2%. ph determination proceeds through a colorimetric reaction, exploiting the capability of a specific indicator to change color on base of ph of the sample measured. The analysis is carried out at 620 nm. The reaction product between potassium ion and tetraphenylborate originates a colloidal solution. The use of special stabilizers and masks allows the suspension of the colloid and the elimination of interference caused by other cations. The turbidity measured at 550 nm is directly proportional to the potassium concentration in the sample. The concentrations of the reagents are optimized in order to obtain a linear analytical response. Winechem x 20 ml 1 x 4 ml 100 tests EN-N150-A140 5 x 33 ml 1 x 28 ml 452 tests EN-N150-A170/200 4 x 41 ml 1 x 28 ml 452 tests EN-N150-A450/600 2 x 33,5 ml 2 x 33,5 ml 302 tests EN026M Winechem x 20 ml 1 x 20 ml 100 tests EN-N026-A140 3 x 31 ml 3 x 31 ml 302 tests EN-N026-A170/200 2 x 46 ml 2 x 46 ml 302 tests EN-N026-A450/

18 Pyruvic acid REAGENT KIT FOR MEASUREMENT OF PYRUVIC ACID CONCENTRATION IN WINE Pyruvic acid is a natural organic acid produced by malolactic fermentation. It contributes to the combinations impacting the wine s structure and texture. The enzyme D-Lactate dehydrogenase (D- LDH ) in presence of reduced NADH converts the Pyruvic acid to D- Lactic acid and NAD+. The amount of NAD+ measured is stoichiometrically related to the amount of Pyruvic acid present in the sample. Reduced Glutathione REAGENT KIT FOR DETERMINATION OF REDUCED GLUTATHIONE IN WINE Glutathione (GSH) is a significant component of wine as it is able to trap orthoquinones, which are the primary protagonists of browning and of loss of flavour due to oxidation processes. Indeed, glutathione is a potent antioxidant, considered to be one of the most precious supplements for our health. High levels of glutathione in wine are important for preserving its aroma and colour. Glutathione levels may vary in grape must insofar as they are determined by the grape variety and by the wine-growing and wine-making processes, hence the importance of measuring its content, particularly in its reduced form as this is its active antioxidant form. D lactate + NAD + + D LDH Pyruvate + NADH The enzyme glutathione-s-transferase (GST) catalyzes the reaction between the reduced glutathione (GSH) and the CDNB (1-chloro-2,4-dinitrobenzene), producing an adduct (S-(2,4 -dinitrophenyl) -glutathione) that absorbs ultraviolet (340 nm) and the formation of which is directly proportional to the amount of GSH present in the sample. 2 x 40 ml 1 x 19 ml 351 tests EN017BM A B 5 x 9 ml 5 x 1 ml 1 x 5 ml 236 tests EN110M A B Winechem x 10 ml 1 x 2 ml 1 x 2,4 ml 100 tests EN-N110-A140 3 x 24 ml 1 x 12,5 ml 1 x 6 ml 236 tests EN-N110-A170/200 3 x 15 ml 1 x 5,5 ml 1 x 12 ml 236 tests EN-N110-A450/

19 Sucrose REAGENT KIT FOR MEASUREMENT OF TOTAL SUGAR IN WINE D glucose + ATP G-6-P ADP + H + G-6-P + NADP + Gluconate 6 phosphate + NADPH + H + Sucrose G6P DH INVERTASE HK Saccharose (or sucrose) is simply the name of sugar. During chaptalisation, saccharose may be added to the grape must if the grape lacks in sugar. A precise analysis of sucrose (or total sugar) content is thus essential for the wine-maker. The analytical determination of total sugar proceed through the following separate and distinct reactions. D glucose + D fructose Tartaric acid COLORIMETRIC AMMONIUM VANADATE METHOD REAGENT KIT FOR MEASUREMENT OF TARTARIC ACID CONCENTRATION IN WINE Tartaric acid is the acid specific to grapes and wine. It has been known since Antiquity as potassium acid salt. Moreover, vine is the only plant to produce tartaric acid, where it accumulates in the fruit (grape berries). It plays a major role in ph, which provides a measure of a wine s acidity. It is the acid that is best able to resist bacterial action. Under acid environment, Tartaric acid (tartrate) produce a coloured complex (metapervanadyl tartrate) by the reaction with Vanadic acid. The amount of this chromophor is stoichometrically related to the amount of tartrate present in the sample. ph=2,5 Tartaric Acid (TART) + Vanadium salt (V) [ TART V ] A B 4 x 10 ml 4 x 0,5 ml 1 x 3 ml 190 tests EN149M A B Winechem x 10 ml 1 x 1,1 ml 1 x 1,1 ml 90 tests EN-N149-A140 4 x 10 ml 4 x 0,5 ml 1 x 2,5 ml 140 tests EN-1492-A170/200 4 x 10 ml 4 x 0,5 ml 1 x 2,5 ml 140 tests EN-N149-A450/600 2 x 45 ml 1 x 20 ml 370 tests EN021AM Winechem x 20 ml 1 x 5 ml 100 tests EN-021A-A140 3 x 35 ml 1 x 25 ml 360 tests EN-021A-A170/200 2 x 50 ml 1 x 25 ml 345 tests EN-021A-A450/

20 Total Acidity COLORIMETRIC METHOD REAGENT KIT FOR MEASUREMENT OF TOTAL ACIDITY IN WINE OR MUST Total Polyphenols COLORIMETRIC FOLIN-CIOCALTEUS METHOD REAGENT KIT FOR THE DETERMINATION OF TOTAL POLYPHENOLS CONCENTRATION IN WINE Acidity is a major sensory parameter of wine. An insufficiently acidic wine will taste flat and dull. Conversely, excessive acidity will make the wine tart, aggressive, or even sour. It also impact the wine s microbial stability. Determining the acidity allows it to be corrected during winemaking. The determination of total acidity takes place by a colorimetric reaction exploiting the acidbase properties of a specific indicator whose absorption value measured at 420 nm, is correlated to the total acidity value of the sample. Polyphenols + Folin Ciocalteu s reagent (FC) Red wine contains numerous polyphenols, including the famous resveratrol (a known anti-cancer agent). As these polyphenols are extracted by fermentation, their concentration in wine is higher than in grape juice. As they are derived from the skin and pips of the grapes, they are found mainly in red wine. Their fundamental value resides in their antioxidant properties. They also contribute to the colour and structure of red wine. Under alkaline condition, the Folin-Ciocalteus reagent oxidate the -OH groups of the polyphenolics compound into the sample. The reduction products have a maximum absorbance at 620 and at 720 nm. The intensity of the colour at these wavelengths is proportional to the concentration of Total Polyphenols in the sample. ph=10,9 Polyphenols FC Winechem x 35 ml 100 tests EN-N134-A140 6 x 32 ml 530 tests EN-N134-A170/200 4 x 47 ml 530 tests EN-N134-A450/600 Winechem x 20 ml 1 x 5 ml 100 tests EN-N018-A140 2 x 45 ml 1 x 12 ml 410 tests EN018M 4 x 32 ml 1 x 16 ml 410 tests EN-N018-A170/200 3 x 42 ml 1 x 16 ml 274 tests EN-N018-A450/

21 Total Polyphenols ENZYMATIC METHOD REAGENT KIT FOR THE DETERMINATION OF TOTAL POLYPHENOLS IN WINE Total SO2 COLORIMETRIC DTNB METHOD REAGENT KIT FOR MEASUREMENT OF TOTAL SULPHUR DIOXIDE IN WINE Red wine contains numerous polyphenols, including the famous resveratrol (a known anti-cancer agent). As these polyphenols are extracted by fermentation, their concentration in wine is higher than in grape juice. As they are derived from the skin and pips of the grapes, they are found mainly in red wine. Their fundamental value resides in their antioxidant properties. They also contribute to the colour and structure of red wine. SO2, or sulphur dioxide, thanks to its antiseptic and antioxidant properties, is the main preservative of wines and grape must. As sulphur dioxide is considered to be toxic, however, its addition is limited and regulated, hence the essential need to measure its concentration. In a buffered reaction environment, the total polyphenols of sample in presence of the enzyme Polyphenol Oxidase react with a selective chromogen, producing a cromophore whose intensity is proportional to the total amount of polyphenols. In alkaline environment, «free sulphur dioxide» and «combined sulphur dioxide» react with 5,5 -ditiobis-2- nitrobenzoic acid (DTNB) to produce a chromophor which can be measured at 416 nm ( nm). A B A B 4 x 2 ml 4 x 1,5 ml 1 x 13 ml 230 tests EN018EM 4 x 12 ml 4 x 1,5 ml 1 x 7 ml 180 tests EN-018E-A170/200 4 x 12 ml 1 x 6,5 ml 1 x 12 ml 180 tests EN-018E-A450/600 2 x 35,5 ml 321 tests EN023BM Winechem x 20 ml 100 tests EN-023B-A140 3 x 33 ml 321 tests EN-023B-A170/200 2 x 49 ml 321 tests EN-023B-A450/

22 Urea KINETIC UV METHOD REAGENT KIT FOR THE MEASUREMENT OF UREA CONCENTRATION IN WINE Urea is a substance found in variable quantities in some wines. Urea is involved in the formation of two thirds of ethyl carbamate, a substance found to be carcinogenic at high doses. It is estimated that 1 mg/l urea leads to the formation of 0.2 µg/l ethyl carbamate. The acceptable daily intake (ADI) of ethyl carbamate for humans is of 0.3 µg/l. In order to limit the formation of ethyl carbamate, urea may be eliminated through the use of urease, an enzyme capable of hydrolysing urea. AMS ALLIANCE DISCRETE ANALYZERS RANGE Winechem 170 Up to 170 tests/hour Winechem 140 Up to 140 tests/hour Urea is transformated from urease to CO2and NH3. In presence of glutamate dehydrogenase (GLDH), the produced NH3 reacts with the α -ketoglutaric acid and NADH producing NAD+ and glutamate. The decrease of NADH concentration can be measured by spectrophotometry and is proportional to the quantity of urea in the sample. Winechem 200 Up to 200 tests/hour WINECHEM Fully automated instruments Multi-parameters analysis Ready-to-use reagents New analytical software Lower running costs 42 2 x 40 ml 1 x 19 ml 350 tests EN055M Winechem x 20 ml 1 x 5 ml 100 tests EN-N055-A140 4 x 26,5 ml 1 x 27 ml 350 tests EN-N055-A170/200 3 x 35,5 ml 1 x 27 ml 350 tests EN-N055-A450/600 Winechem 450 Up to 450 tests/hour 2 ranges: Smartchem (Environment) & Winechem (Wine) Winechem 600 Up to 600 tests/hour