Diagnostic test for reductive wine characters (Cu/Cd test)

The following procedure (commonly referred to as the copper/cadmium [Cu/Cd] test) outlines a quick simple aroma only diagnostic test that can be used to determine what type of reductive compounds are causing a wine to have a reductive wine fault or character. Reductive wine characters can be attributed to hydrogen sulfide, mercaptans, disulfides or dimethyl sulfide, each which have distinct aromas and different winemaking treatment options to remove them.

Hydrogen sulfide (H2S)
H2S has the aroma of ‘rotten egg gas’. During alcoholic fermentation, yeast will excrete H2S into the fermenting juice when placed under stress, e.g. when the yeast starts to run out of nitrogen. Australian juices can be low in nitrogen and winemakers often supplement the juice with a soluble nitrogen source, such as diammonium phosphate (DAP) at the first sign of development during fermentation. To calculate an appropriate DAP addition, access the AWRIs winemaking calculator. The AWRI strongly recommends that yeast assimilable nitrogen (YAN) be analysed before making decisions on DAP additions.

Mercaptans
Mercaptan is a general term applied to the aroma of a range of compounds containing a terminal SH moiety (thiols) and are variously described as ‘cabbage’, ‘garlic’, ‘onion’ and ‘rubber’. It is thought that methyl mercaptan (methanethiol) might be formed by the direct chemical reaction between H2S and acetalaldehyde (and possibly methanol). However, it is known that methyl mercaptan is formed directly as a result of yeast metabolism, therefore, it is best to remove H2S before it reacts further to form thiols. Other mercaptans include ethyl mercaptan (ethanethiol) are thought to be formed by reaction of H2S with other wine components.

Disulfides
Mercaptans such as methanethiol (methyl mercaptan) and ethanethiol (ethyl mercaptan) can be rapidly oxidised to dimethyldisulfide (DMDS) and diethyldisulfide (DEDS), respectively. The aroma of DMDS is described as ‘onions’ and ‘cooked cabbage’. The aroma of DEDS is described as ‘burnt rubber’ and ‘garlic’.

Dimethyl sulfide
Dimethyl sulfide (DMS) is one of the major compounds found in aged wines and is formed during the maturation of wine in the bottle. However, the mechanism of formation of DMS is not clearly known. It is probably more important in red wines and can give a ‘blackcurrant’ or ‘jam’ character. At higher concentrations, the aroma of DMS is a fault and is described as ‘vegy’, ‘asparagus’, ‘cooked corn’, ‘cooked tomato’ or ‘molasses’.

Principle of Cu/Cd test
The principle of this test is that different reductive volatiles react with different fining agents, including copper and cadmium salts (Note that cadmium sulfate is not an allowable fining agent and is only to be used in this diagnostic test). A fining trial can be conducted using these agents to determination of the type of reductive fault present in a wine to determine the appropriate course of action to remove the fault.

Reagents
1) 1% w/v Copper (II) Sulfate solution (1 g of CuSO4.5H2O in 100 mL 10% Ethanol)
2) 1% w/v Cadmium (II) Sulfate solution (1 g of CdSO4.8 H2O in 100 mL 10% Ethanol)

Caution:
Cadmium is TOXIC.
Do not taste samples to which cadmium has been added. Assess by aroma only.
Cadmium sulfate is not an allowable wine additive or processing aid and is only to be used in this diagnostic test.

3) 10% w/v Ascorbic acid (10 g ascorbic acid in 100 ml of 10% Ethanol)

Procedure

  1. Place 50 mL of wine into four separate glasses.
  2. Label the four glasses: (1) control, (2) copper, (3) cadmium, and (4) ascorbic acid +copper
  3. Add 1 mL of reagent 1, the copper sulfate solution to glass 2 (‘copper’ glass),
  4. Add 1 mL of reagent 2, the cadmium sulfate solution to glass 3 (‘cadmium’ glass),
  5. Add 0.5 mL of reagent 3, the ascorbic acid solution to glass 4 (‘ascorbic + Cu’ glass), wait 2 minutes, then add 1 mL of reagent 1, the copper sulfate solution to the same glass 4.

Wines glasses

  1. Assess and compare the aroma of the control (glass 1) to the aroma of the other three glasses, noting down any differences observed. Note that this is an AROMA ONLY diagnostic test. DO NOT TASTE THE SAMPLES. A tasting sheet template can be found here.

Interpretation

Glass 1
(control)
Glass 2
(copper)
Glass 3
(cadmium)
Glass 4
(Ascorbic +
Cu)
Type of
reductive
compounds
Presence of
offensive odour
Odour gone Odour gone Odour gone H2S
Odour gone No change Odour gone Mercaptans
Odour gone Slight
improvement
Odour gone H2S and
mercaptans
No change No change Odour gone Disulfides
No change No change No change Dimethyl sulfide

Note: Have several people perform the assessment. A triangle test can be used to determine if real differences exist.

Reference: Adapted from Zoecklein, B.W.; Fugelsang, K.C.; Gump, B.H.; Nury, F.S. Wine analysis and production. New York: Chapman and Hall; 1995; pp428-429.

Treatment for removal

H2S:

  • During fermentation
    Excess H2S can be removed from red wines by aerating at the first racking, thus volatilising the H2S. Supplement the juice or must with a soluble nitrogen source, such as diammonium phosphate at the first sign of development during fermentation. Addition of DAP after the first half of the fermentation (the yeast growth phase) is less effective at removing reductive characters and can result in wines with high residual nitrogen content.Minimise the formation of excess H2S in white wines by either settling, centrifuging or filtering the must before fermentation, which removes high-density solids which might contain elemental sulfur. Note that excessive clarification at the juice stage however can also lead to stuck fermentation issues.
  • After fermentation
    Copper sulfate fining can be used to remove H2S in red and white wines. Copper sulfate reacts with H2S to form copper sulfide, which is highly insoluble. However, careful laboratory trials should precede any CuSO4 additions to bulk wine, as an instability can result if the copper concentration in the wine exceeds approximately 0.5 mg/L (even lower in some wines). In 2008, Australian food law (Australian and New Zealand Food Standards Code) removed the maximum copper concentration of 5 mg/L, as copper ions. There is now no specified maximum limit in Australia. Note that maximum allowable levels vary depending on the country e.g. USA 0.5 mg/L.

Mercaptans:
Removal of mercaptans in red and white wines can be achieved by fining with copper sulfate (CuSO4). Copper sulfate reacts with mercaptans to form highly insoluble salts i.e.:

CH3CH2SH + Cu2+ → Cu(CH3CH2S)2

It is important to note however, that the addition of copper can also cause by-product formation of an unreactive disulfide complex via the below reaction:

3CH3CH2SH + Cu2+ → Cu-S-CH2CH3 ↓ + CH3CH2S-SCH2CH3

Disulfides:
Removal of disulfides requires the creation of reducing conditions, with the addition of ascorbic acid and SO2, in order to reduce these compounds back to the reactive mercaptan species which may then be removed by treatment with copper. Note that copper is not as effective at removing these mercaptans as it is at removing H2S, as discussed above.

Dimethyl sulfide:
As dimethyl sulfide does not bind to copper it can be difficult to remove the off-aroma, however, removal might be possible by sparging with nitrogen or by using reverse osmosis.