Ough (1988) and Ough and Amerine (1988) gave very detailed reviews of methods for free and total sulfur dioxide determinations in grapes and wines. The official methods of analysis for agricultural and food products are specific in their recommendations for analysis of sulfur dioxide (Table 5.8). Sulfites can be described as free, reversibly bound, or irreversibly bound (Beck et al., 2000). Free sulfites are readily converted to sulfur dioxide upon acidification and can be quantita- tively analyzed following distillation. Reversibly bound sulfites are converted to sulfur dioxide only after heat and acid or alkali treatments. Those sulfites that bind food matrices and are not converted to sulfur dioxide with heating or acidic conditions are irreversibly bound. Thiosulfonates are irreversibly bound (Beck et al., 2000). One official method for measuring total sulfurous acid is the modified Monier-Williams procedure. A sample is placed in a distilling flask, acidified, and
TABLE 5.7
Application of Sulfites in Foods as Antimicrobials and the Concentrations Used (Gould, 2000)
Food Use Concentration (mg/kg SO2)a
Beer 10–30
Fresh fruits 100
Fresh vegetables (onion, garlic, horseradish) 50–1000
Fruit juices 10–100
Fruit-based sauces and related products 50–100 Fruit pulps, purees, and fillings 50–500
Jams and jellies 50–100
Nonalcoholic beverages 20–200
Sausage 450
Sugar confectionary 50
Vinegar 50–200
Wine 100–300
a Sulfites are allowed only in certain foods in different countries, and concentration varies by country.
TABLE 5.8
AOAC International Official Methods of Analysis for Sulfites (Warner et al., 2000) Method
Number Year Adopted Title Type of Analysis
892.02 1892 Sulfurous Acid (Free) in Meats Titrimetry
961.09 1961 Sulfites in Meats Qualitative Test
962.16 1962 Sulfurous Acid (Total) in Food Modified Monier-Williams 963.20 1963 Sulfurous Acid (Total) in Dried Fruit Colorimetry
975.32 1975 Sulfurous Acid in Food Qualitative Test
980.17 1980 Preservatives in Ground Beef Colorimetry
987.04 1987 Sulfites (Total) in Foods Differential Pulse Polarography
990.28a 1990 Sulfites in Foods Optimized Monier-Williams
990.29 1990 Sulfites (Total) in Foods and Beverages Flow Injection Analysis 990.30 1990 Sulfites (Free) in Wine Flow Injection Analysis 990.31 1990 Sulfites in Foods and Beverage Ion Exclusion Chromatography
Sulfur Dioxide and Sulfites 159
heated, and the volatile sulfur dioxide is removed by a stream of nitrogen through a reflux condenser. The reflux condenser must retain all volatile acids except the sulfurous acid. The sulfurous acid is trapped in hydrogen peroxide solution:
SO2 + H2O2 → SO4 2–
+ 2H+
The acid produced by the oxidation of sulfur dioxide to sulfate can be titrated with sodium hydroxide; in addition, the sulfate can be precipitated with barium and measured gravimetrically. This method is good for most products, except dried onions, leeks, and cabbage. Mitsuhashi et al. (1979) showed that sodium sulfide and allyl isothiocyanate gave positive results in the Monier- Williams tests. The optimized Monier-Williams method is used by the FDA to measure sulfites in official samples (Table 5.8) (Warner et al., 2000).
The direct iodine titration method (Ripper) for measurement of total sulfur dioxide is the method used for routine analysis in the wine industry (Ough and Amerine, 1988). Briefly, the sample is made basic to break the bisulfite addition products; it is then acidified, and the freed sulfurous acid is titrated directly with iodine to a starch end point:
I3 –
+ SO2 + H2O → SO3 + 3I– + 2H+
SO3 + H2O → SO42–+ 2H+
This is satisfactory for certain materials, but if other oxidizable substances are present, artifi- cially high results occur. Joslyn and Braverman (1954) reviewed the errors associated with iodine titrations. Vahl and Converse (1980) made a collaborative study of the Ripper method for the Association of Official Analytical Chemists and concluded that the poor precision and large systematic error precluded it for adoption as an official method.
The Ripper method for free sulfur dioxide is given by Ough and Amerine (1988). This is a direct iodine titration of the substance at acid pH. The same errors are associated with this procedure as are associated with the Ripper procedure for total sulfur dioxide. Schneyder and Vlcek (1977) reported that iodate was superior to an iodine standard solution. Excess iodide is added to the sample, and it is then titrated with the iodate solution to a starch end point. The reactions are as follows: 8I– + IO 3– + 6H+→ 3I3 – + 3H2O I3–+ SO2 + H2O → SO3 + 3I – + 2H+
The advantage is a stable standard solution that does not require daily standardization. The Monier-Williams method can be modified to determine free sulfurous acid. Kielhofer and Aumann (1957), Paul (1958), Burroughs and Sparks (1964b), Rankine and Pocock (1970), and Ough and Amerine (1988) described the method and equipment needed. Basically the sample is sparged with gas for 12 to 15 minutes, and the hydrogen ion produced in the peroxide solution is titrated. Free sulfur dioxide is easily measured in white wines by the Ripper method, but determi- nations on red wines are more accurately done by this variation of the Monier-Williams method.
Sulfite test strips for protection of people with asthma who are hypersensitive to recognize that the food contained sulfites were questioned by Nordlee et al. (1988) and Wanderer and Solomons (1987). False-negative and false-positive results were found by both groups. Schwedt (1986) considered the test strips satisfactory but noted that if dyes or natural pigments came in contact with the cellulose, false results were recorded.
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