COMPOSITIONS AND METHODS FOR INHIBITING BROWNING IN FOODS AND BEVERAGES

Abstract: Compositions and methods are described for preventing or inhibiting oxidative darkening of foods and beverages. The compositions comprise at least one substituted resorcinol derivative and at least one additive which when applied in combination with the resorcinol derivative prevents enzymatic browning of the food or beverage. The compositions inhibit the enzymatic browning of foods and beverages susceptible to browning, such as shrimp, potatoes, apples, avocados, fruit juices and wines.

Description: COMPOSITIONS AND METHODS FOR INHIBITING BROWNING IN FOODS AND BEVERAGES Background of the Invention Browning of foods is a major problem in the food and beverage industry. Browning, or oxidative darkening, can be the result of the action of an enzyme, such as polyphenol oxidase (PPO; also known as tyrosinase) , or the result of non-enzymatic chemical reactions, for example, due to polymerization of phenolic compounds which are present in some foods. High PPO activity is present in foods which are susceptible to browning, e.g., shrimp, bananas and mushrooms. Browning causes deleterious changes in the appearance and texture of foods and beverages. Both enzymatic and non-enzymatic browning constitute serious problems for the food industry and result in millions of pounds of wasted food products per year. Enzymatic browning, in particular, has been the subject of much research, particularly as the causative agent of shrimp melanosis, which is characterized by the formation of dark spots on shrimp. Faulkner et al. , Advanced Food Research, 3L :302-310 (1953). Enzymatic browning is the result of PPO-catalyzed oxidation of mono- and diphenols to o-quinones which polymerize spontaneously to form dark-colored, high molecular weight polymers, leading to the characteristic browning or formation of dark spots. Several methods have been developed to prevent browning, including heat inactivation of PPO and various chemical treatments, such as altering the pH of the food. Heat inactivation is not appropriate for fresh foods, such as fruits and seafood, as the high temperatures necessary to inactivate PPO change the quality and texture of the foods. Likewise, reducing the pH by adding an acid (e.g., citric acid or phosphoric acid) deleteriously affects the appearance and quality of some foods. The control of PPO-catalyzed enzymatic browning in mushrooms using citric acid was reported by McCord and Kilara in the Journal of Food Science, 48:1479-1483 (1983) . The inhibition of polyphenol oxidase activity in an extract of Jerusalem artichokes using various sulfite compounds was described in Zawistowski et al. , in Can. Inst. Food Sci. Tech. J. , 20(3) :162-164 (1987). The use of cinnamic acid, p-coumaric acid and ferulic acid to control enzymatic browning in fruit juices was described by J.R.L. Walker in Food Technology, 11:341-345 (1976). T. C. Wong et al. report in Plant. Physiol., 48:24-30 (1971) that phloroglucinol and resorcinol, and their derivatives d-catechin and orcinol react with 4-methyl-o-qu.inone which is formed by PPO in peaches, although these compounds are not substrates for PPO. R. Kuttner and H. Wagreich, Arch. Biochem. Biophys., £3:80-87 (1952) report that mushroom PPO (catecholase) is inhibited by benzoic acid and selected benzoic acid derivatives. None of these methods have proven entirely satisfactory, however, due to expense, lack of availability, or inferior performance. Labuza in Cereal Foods World, 34(4) :353 (1989) describes the use of proteases, especially ficin, in the control of enzymatic browning of certain foods. The author attributed this effect to attack on PPO by the protease. -3- Another method for reducing browning which has been prevalent in the food industry is adding sulfite salts to foods and beverages. Some forms of enzymatic browning, such as shrimp melanosis, have traditionally been treated by dipping or coating the shrimp or other food in a sulfite solution, such as sodium bisulfite. Sulfites are also added to wines to prevent oxidation. Sulfites reduce o-quinones to the mono- and/or diphenols, thereby inhibiting the browning reaction. However, the use of sulfite in foods has been restricted due to adverse health effects in certain individuals, and may be restricted further or even eliminated completely. Summary of the Invention Compositions and methods are described for preventing or inhibiting oxidative darkening of foods and beverages. The compositions comprise at least one substituted resorcinol derivative and at least one additive which when applied in combination with the resorcinol derivative prevents enzymatic browning of the food or beverage. The additives can be a sulfite-contain- ing compound such as sodium metabisulfite; a mild food grade reducing agent such as ascorbic or erythorbic acids; a food grade organic acid such as citric, fumaric, tartaric or malic acid; a chelating agent such as ethylenediaminetetraacetic acid or sodium acid pyrophosphate; or an inorganic salt such as calcium chloride, sodium chloride, zinc chloride or sodium bicarbonate. Components of the present compositions act in synergy to effectively inhibit browning in selected foods or beverages, without adversely affecting the appearance, taste, texture or quality of the food or beverage. Detailed Description of the Invention This invention pertains to compositions and methods for inhibiting oxidative browning of foods and beverages, particularly enzymatic browning caused by PPO activity. The invention involves subjecting a food or beverage to a composition comprising at least one substituted resorcinol derivative and at least one additional additive for a period of time sufficient to inhibit enzymatic browning. An additive is a compound which is known to inhibit oxidative browning of certain foods or beverages, or one which when admixed with the substituted resorcinol compounds inhibits enzymatic browning. Additives that can be used in the composition of this invention include sulfite-containing compounds such as sodium metabisulfite; mild food grade reducing agents such as ascorbic acid, erythorbic acids and derivatives of these such as ascorbyl phosphate and ascorbyl fatty acid esters; food grade acidulents such as citric, fumaric, tartaric, malic acid, etc.; chelating agents such as ethylenediaminetetraacetic acid (EDTA) or sodium acid pyrophosphate; or inorganic salts such as calcium chloride, sodium chloride, zinc chloride or sodium bicarbonate. One or more additives can be used in combination to achieve the desired effect. The compositions may optionally include conventional additives such as emulsifiers, dispersing agents, etc. It has now been shown that substituted resorcinol derivatives when used in combination with at least one additional additive, act in synergy to inhibit browning in foods and beverages. The degree of inhibition is enhanced when the components are used in combination, than that observed when each component is individually applied to the food or beverage. Complete prevention of enzymatic browning is difficult to achieve due to the presence of pigment precursors formed prior to the addition of inhibitor or due to br wning that is not initiated by PPO catalysis (nonenzy atic browning) . However, compositions of this invention can be used to reduce and minimize browning beyond that which is observed when reducing agents alone are used. Resorcinol derivatives and especially those substituted at the 4-position comprise a class of compounds which are highly effective in inhibiting the activity of the enzyme, polyphenol oxidase, from a number of sources. Preferably, the 4-substituted resorcinols are compounds having the general formula: I wherein R and R are independently selected from the group consisting of H, CH_, COR*', CR' , PO.R'R" and SO R'R"; wherein R' and R" are independently H or an alkyl group having from 1 to about 6 carbon atoms in a linear, branched or cyclic configuration or a substituted aromatic compound having from about 4 to about 10 carbon atoms; and R_ is an organic or inorganic substituent selected so that the resulting compound is an inhibitor of the enzyme PPO. For example, R_ can be a heteroatom or a group containing a heteroatom, a saturated or unsaturated alkyl group, a substituted aromatic compound or an organic functional group selected so that the compound has inhibitory activity. The heteroatom can include, for example, oxygen (0) , nitrogen (N) , sulfur (S) , phosphorus (P) or halogens such as chlorine (Cl) , bromine (Br) , iodine (I) or fluorine (F) . The saturated or unsaturated alkyl group can have from 1 to 30 carbon atoms in a linear, branched or cyclic configuration and can include a substituted aromatic compound. The alkyl substituents or organic functional groups can contain a heteroatom or heteroato s, for example, oxygen (0) , nitrogen (N) , sulfur (S) , phosphorus (P) or halogens such as chlorine (Cl) , bromine (Br) , iodine (I) or fluorine (F). 4-Alkylresorcinol compounds in combination with other additives are particularly effective for inhibiting browning in foods. For example, 4-hexylresorcinol, having the structure shown below (wherein R. and R_ are both H and R -_ is Co..H1-._5) has been shown to be highly effective for this purpose. Formula II In one embodiment, R_ has the general structure: O II —-CH,,— Cl^-C—Z wherein n is 1 or 2 and Z is an alkyl or other organic functional group selected so that the compound has inhibitory activity. Z can be an alkyl substituent containing at least one heteroatom, such as oxygen (O) , nitrogen (N) , sulfur (S) , phosphorus (P) or halogens such as chlorine (Cl) , bromine (Br) , iodine (I) or fluorine (F) . In a preferred embodiment, Z is selected from the group consisting of OH, NH 2 , 0(CH 2 ) χ CH 3 , NHC0 2 (CH 2 ) χ CH 3 , NH(CH ? ) CH_, amino acids, polyamine metabolites, such as NH(CH 2 ) χ NH 2 , NH(CH 2 ) χ NH(CH 2 ) y NH 2 , NH(CH 2 ) χ NHR 4 , NH(CH_) NH(CH ) NHR. wherein x and y independently can be any integer from 0 to 5; and higher polyamine oligomers or substituted oligomers conisting of at least three monomers, wherein the monomer is a l,ω-diaminoalkane and R. has the following formula: wherein n, R. and R_ are defined above. Compounds which are particularly effective inhibitors of oxidative browning are resorcinol derivatives having the general formula: Formula III wherein n is 1 or 2 and R and R_ and Z are as defined above. Particularly useful resorcinol derivatives for inhibiting enzymatic browning are obtained when n = 2, R. and R 2 are both H and Z is OH, NH(CH 2 ) 4 NH 2 , NH(CH 2 ) 4 NHR 4 (where x = 4) or H CH 2 ) 4 NH(CH 2 ) 3 NHR 4 (where x = 4, y - ** ■ * 3 and R is as defined above) ■ These compounds are shown as Formulae IV, V, VI and VII respectively: Formula V Formula VI Formula VII The invention includes functional equivalents of these formulae (Formulae I-VII) . The term "functional equivalents" means a chemical derivative or analog of the compound which has similar anti-browning activity. As used herein, a molecule is said to be a "derivative" of another molecule when it contains additional or different chemical moieties not normally part of the molecule. Specific resorcinol compounds and their synthesis are described in detail in U.S. Serial No. 07/537,361, filed June 13, 1990. Low concentrations of the resorcinol derivatives reduce the level of enzymatic browning but will not completely prevent the nonenzymatic browning of foods due to the presence of pigment precursors prior to the addition of the inhibitor. Increasing the concentration of the resorcinol derivative does not affect the nonenzymatic component of browning. Non-enzymatic browning can be prevented by the addition of high concentrations of reducing agents such as sulfite, ascorbic acid, erythorbic acid, etc., which are known to react chemically with the pigment precursors and the products of ongoing PPO catalysis; however, the effectiveness of the reductants diminishes with time as they are consumed. The 4-substituted resorcinol compounds have been found to be synergistic with other agents such as reducing agents, acidulants, etc., that is the combination of the two types of agents are more effective than either agent individually, even if used at much higher concentrations. The synergistic mixture is used in an amount or concentration sufficient to inhibit or prevent browning. The form of treatment will depend upon the food or beverage being treated and the results sought, and can include, e.g., dipping, spraying, dusting, sprinkling, immersing, mixing and/or soaking. The mixture can be added to an aqueous diluent, for example water, salt water or buffer, and applied to the food, or can be added neat, e.g., to fruit juice or wine. The amount needed will depend upon the susceptibility of the food or beverage, to browning, the condition of the food or beverage and the storage conditions. The amount sufficient to prevent or inhibit browning can be determined empirically by one skilled in the food art. Compositions comprising about 0.001% to 5% by weight of the additive and about 0.001 to 0.5% by weight of the resorcinol derivative will be particularly effective for this purpose. Foods which are susceptible to browning include certain shellfish, crustaceans, fruits (e.g., apples, avocados, bananas, peaches) vegetables (e.g., potatoes, lettuce) and beverages, such as fruit juices and wines are treated with a composition containing an amount of the present resorcinol derivatives in combination with at least one other anti-browning agent. The amount applied to the food should be sufficient to inhibit the browning reaction. Browning is "prevented" if it is completely eliminated. Browning is "significantly inhibited" if browning takes place at a significantly lower rate compared to untreated foods in the same time frame. Components of the present compositions are non-toxic in levels used and act in synergy to effectively inhibit or prevent browning in selected foods and beverages, without adversely affecting the appearance, taste, texture or quality of the food of beverage. The invention will be further illustrated by the following non-limiting examples: EXAMPLES Example 1 Inhibition of Browning of Apples Using Formula II and Bisulfite Fresh, whole, Mclntosh apples, held at ambient temperature (22 to 24°C) were sliced by cutting 1/4 inch (top to bottom) sections from several apples. The outermost and core sections were discarded. The freshly cut slices were halved (top to bottom) and placed (2-3 slices) in each test solution. The samples were agitated during the soak time to ensure complete contact. After one minute, the slices were removed and placed in clear plastic containers and allowed to stand at ambient temperature and were exposed to air. The apple slices were visually inspected after 1 and 24 hours. The results are shown in Table 1. Table 1 Test Solution (%, w/v) 1 Hour 24 Hours Water browning browned Formula II (0.05 or 0.1) white white with core area browning Formula II (0.05) + bisulfite white white (0.05) Formula II (0.01) white browning, veins noticeable Bisulfite (0.01) some browned browning Formula II (0.01) + bisulfite white slight (0.01) browning near edges Example 2 Inhibition of Browning of Apples Using Formula IV and Bisulfite Mclntosh apples were treated and evaluated as described in Example 1. The results are shown below in Table 2. Table 2 (continued) Test Solution (%, w/v) 1 Hour 24 Hours Bisulfite (0.01) some browned browning Formula IV (0.01) + bisulfite white slight (0.01) browning veins noticeable Example 3 Inhibition of Browning of Apples Using Formula IV and Acidulants and/or Reducing Agents Mclntosh apples were treated and evaluated as described in Example 1. The results are shown in Table 3. Test Solution (%, w/v) Water Formula IV (0.2) Formula IV (0.05) Formula IV (0.01) Ascorbic Acid (0.05 or 1.0) Formula IV (0.05) + ascorbic acid (0.5) Citric Acid (0.5) white browned Table 3 (continued) Test Solution (%, w/v) 1 Hour 24 Hours Formula IV (0.05) + citric white slight Acid (0.5) browning Erythorbic Acid (0.5) white moderate browning Formula IV (0.05) + erythorbic white white Acid (0.5) Malic Acid (0.5) Formula IV (0.05) + Malic Acid (0.5) Malonic Acid (0.5) Formula IV (0.05) + Malonic Acid (0.5) Example 4 Inhibition of Browning of Apples Using Formula IV, Sulfites and Ascorbic Acid Mclntosh apples were treated and evaluated as described in Example 1. The results are provided in Table 4. Table 4 Test Solution (%, w/v) 1 Hour 24 Hours Water browning browned Formula IV (0.01) slight core browning, browning veins noticeable Bisulfite (0.01) some browned browning Table 4 (continued) Test Solution (%, w/v) Ascorbic Acid (0.1) Ascorbic Acid (0.5) Formula IV (0.01) + Bisulfite (0.01) + Ascorbic Acid (0.1) Formula IV (0.01) + Bisulfite (0.01) + Ascorbic Acid (0.5) Formula IV (0.01) + Bisulfite (0.05) + Ascorbic Acid (0.5) Formula IV (0.05) + Bisulfite (0.05) + Ascorbic Acid (0.5) Example 5 Inhibition of Browning of Potatoes Using Formula II and Sulfites Whole brown russet potatoes were peeled under water. Slices were made by cutting 1/4 inch sections from several peeled potatoes. Slices from the ends of the potato were discarded. The remaining freshly cut slices were placed (2-3 slices) in each test solution. The samples were agitated during the soak time to ensure complete contact. After one minute, the slices were removed and placed in clear plastic containers and allowed to stand at ambient temperature and were exposed to air. The potato slices were visually inspected after 1 to 24 hours. The results are provided in Table 5. Table 5 Test Solution (%, w/v) 1 Hour 24 Hours Water reddening blackened Formula II (0.05) off colored slightly browned vascular ring Bisulfite (0.05) white slightly browned from edge Bisulfite (0.01) slight browned reddening Formula II (0.01) + Bisulfite white browned (0.01) near edge Example 6 Inhibition of Browning of Potatoes Using Formula IV and Bisulfite Brown russet potatoes were treated and evaluated as described in Example 5 and the results are shown in Table 6 below. Table 6 Test Solution (%, w/v) 1 Hour 24 Hours Water reddening blackened Formula IV (0.1) white browned Formula IV (0.05) slight browned reddening Bisulfite (0.05) white - browned vascular ring Table 6 (continued) Test Solution (%, w/v) 1 Hour 24 Hours Formula IV (0.05) + Bisulfite white white (0.05) Formula IV (0.01) slight browned browning Bisulfite (0.01) slight browned browning Formula IV (0.01) + Bisulfite white browned (0.01) near edges Example 7 Inhibition of Browning of Potatoes Using Formula IV and Acidulants and/or Reducing Agents Brown russet potatoes were treated and evaluated as described in Example 5. The results are shown in Table 7. Test Solution (%, w/v) Water Formula IV (0.2) Formula IV (0.05) Formula IV (0.01) Ascorbic Acid (0.5) Ascorbic Acid (0.1) Formula IV (0.05) + Ascorbi Acid (0.5) Table 7 (continued) Test Solution (%, w/v) Citric Acid (0.5 or 1.0) Formula IV (0.05) + Citric Acid (0.5) Erythorbic Acid (0.5 or 1.0) Formula IV (0.05) + Erythorbic Acid (0.5) Malic Acid (0.5) Formula IV (0.05) + Malic Acid (0.5) Malonic Acid (0.5) Formula IV (0.05) + Malonic Acid (0.5) Example 8 Inhibition of Browning of Potatoes Using Formula IV, Sulfites and Acidulants Brown russet potatoes were treated and evaluated as described in Example 5. The results are provided in Table 8. Table 8 Test Solution (%, w/v) 1 Hour 24 Hours Water reddening blackened Formula IV (0.01) very slight browned reddening Bisulfite (0.01) very slight browned reddening Table 8 (continued) Test Solution (%, w/v) 1 Hour 24 Hours Ascorbic Acid (0.01) reddening browned Ascorbic Acid (0.5) white slight browning Formula IV (0.01) + Bisulfite very slight some browning (0.01) + Ascorbic Acid (0.1) reddening Formula IV (0.01) + Bisulfite white some browning (0.01) + Ascorbic Acid (0.5) on edges Formula IV (0.01) + Bisulfite white white (0.05) + Ascorbic Acid (0.5) Formula IV (0.01) + Bisulfite white white (0.05) + Ascorbic Acid (0.5) Equivalents Those skilled in the art will recognize, or be able to ascertain, using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims:

Claims: CLAIMS 1. A method for inhibiting enzymatic browning of foods or beverages susceptible to such browning, comprising applying to the foods or beverages a composition comprising at least one substituted resorcinol derivative and at least one additive selected from the group consisting of a reducing agent, a chelating agent, an acidulant, an inorganic salt and combinations thereof, the amount of additive and resorcinol derivative being sufficient to inhibit browning of the foods, wherein the resorcinol derivative has the following formula: wherein R. and are independently selected from the group consisting of H, CH , COR*', CR', PO-R'R" and SO-R'R" wherein R Λ and R" are independently H or an alkyl group having from 1 to 6 carbon atoms in a linear branched or cyclic configuration or a substituted aromatic compound; and wherein R_ is selected so that the resorcinol derivative inhibits enzymatic browning. 2. A composition for inhibiting enzymatic browning of foods or beverages susceptible to such browning. comprising at least one substituted resorcinol derivative and at least one additive selected from the group consisting of a reducing agent, a sulfite, a chelating agent, an acidulant, an inorganic salt and combinations thereof, the amount of additive and resorcinol derivative being sufficient to inhibit browning of the foods, wherein the resorcinol derivative has the following formula: wherein R. and R_ are independently selected from the group consisting of H, CH 3 , COR-', CR' , P0 3 R'R" and SO-R'R" wherein R' and R" are independently H or an alkyl group having from 1 to 6 carbon atoms in a linear branched or cyclic configuration or a substituted aromatic compound; and wherein R_ is selected so that the resorcinol derivative inhibits enzymatic browning. 3. A method or composition according to any one of Claims 1 and 2, wherein the foods or beverages are selected from the group consisting of crustaceans, shellfish, fruits, vegetables, fruit juices and wines. 4. A method or composition according to any one of Claims 1 and 2, wherein the amount of additive is from about 0.001 to about 5 percent by weight and the amount of resorcinol derivative is from about 0.001 to 0.5 percent by weight. 5. A method or composition according to any one of Claims 1 and 2, wherein the additive is one of the following: a) a reducing agent selected from the group consisting of ascorbic acid, erythorbic acid, sulfiting agents and derivatives of these; b) an acidulant that is any food grade acid such as citric acid, fu aric acid, tartaric acid or malic acid; c) an inorganic salt selected from the group cosisting of calcium chloride, sodium chloride, zinc chloride and sodium bicarbonate; or d) a chelating agent that is ethylenedia inetetra- acetic acid or sodium acid pyrophosphate. 6. A method or composition according to any one of Claims 1 and 2, wherein R_ comprises a heteroatom (e.g., oxygen, nitrogen, sulfur and phosphorus), a saturated or unsaturated alkyl group, a substituted aromatic group or an organic functional group containing a heteroatom. 7. A method or composition according to any one of Claims 1 and 2, wherein R_ is a saturated or unsaturated alkyl group having from about 1 to about 30 carbon atoms in a linear, branched or cyclic configuration. 8. A method or composition according to any one of Claims 1 and 2, wherein R- and 2 are both H. 9. A method or composition according to any one of Claims 1 and 2, wherein the resorcinol derivative has the following formula: 10. A method or composition according to any one of Claims 1 and 2, wherein the resorcinol derivative has the following formula: wherein n is 1 or 2; R. and R_ are independently selected from the group consisting of H, CH 3 , C0R , CR', P0 3 R , R" and S0 3 R , R" wherein R' and R" are independently H, an alkyl group having from about 1 to about 6 carbon atoms in a linear, branched or cyclic configuration or a substituted aromatic compound; and Z is an OH group, an alkyl or organic functional group selected so that the resorcinol derivative inhibits enzymatic browning. 11. A method or composition according to any one of Claims 1 and 2, wherein the resorcinol derivative is selected from the group consisting of: and 12. A food or beverage susceptible to enzymatic browning which has been treated with the composition of Claim 2 in an amount sufficient to prevent or inhibit browning of the food or beverage. 13. A food or beverage susceptible to enzymatic browning of Claim 12, wherein the composition comprises a resorcinol derivative selected from the group and

Inventors: GROSS AKIVA, IYENGAR RADHA, MCEVILY ARTHUR J, GROSS, AKIVA, IYENGAR, RADHA, MCEVILY, ARTHUR, J.
Applicants: OPTA FOOD INGREDIENTS INC, OPTA FOOD INGREDIENTS, INC.
Application year: 1992
Main classification: A23B7
IPC codes: A23B7, A23L1, A23L2, A23L3, C12H1