WO2011045643A1

WO2011045643A1 - Preservation of non-alcoholic beverages and functional waters against spoilage by bacteria of the genus alicyclobacillus

Info

Publication number: WO2011045643A1
Application number: PCT/IB2010/002456
Authority: WO
Inventors: Dusanka Dimitrijevic; Gregor Perne; Matej Bornsek; Majda Hadolin Kolar
Original assignee: Vitiva D.D.
Priority date: 2009-10-14
Filing date: 2010-09-23
Publication date: 2011-04-21

Abstract

This invention is directed to the preservation of non-alcoholic beverages and functional waters against spoilage by bacteria belonging to the genus Alicyclobacillus. The preservation is achieved by the extracts from plants of the Lamiaceae family, mixtures thereof, as well as their mixtures with other extracts, such as citrus extract, green tea extract, carrot extract and/or olive extract. The Lamiaceae family comprises Rosmarinus, Salvia, Melissa, Origanum, Satureja, Mentha, Thimus, and others.

Description

PRESERVATION OF NON-ALCOHOLIC BEVERAGES AND FUNCTIONAL WATERS AGAINST SPOILAGE BY BACTERIA OF THE GENUS Alicyclobacillus

FIELD OF THE INVENTION

[001 ] This invention is directed to the preservation of non-alcoholic beverages and functional waters against spoilage by bacteria belonging to the genus Alicyclobacillus. The preservation is achieved by the extracts from plants of the Lamiaceae family, mixtures thereof, as well as their mixtures with other extracts, such as citrus extract, green tea extract, carrot extract or olive extract. The Lamiaceae family comprises Rosmarinus, Salvia, Melissa, Origanum, Satureja, Mentha, Thimus, and others.

BACKGROUND OF THE INVENTION

[002] Synthesized preservatives have been used for decades in the preservation of products against microbial spoilage. Recently, the use of naturally occurring substances, such as extracts from plant and animal materials, has become more common in the protection against the microorganisms resposible for food spoilage and pathogenic bacteria. This is mainly since synthetic preservatives tend to exert undesired side effects on human health.

[003] Owing to the increased production of non-alcoholic beverages and functional waters during the past years, the spoilage thereof gained importance, and was found to be in many cases due to spore-forming acidophilic bacteria of the genus Alicyclobacillus.

[004] Alicyclobacillus spp. grow at a temperature in the range of 20-70°C and a pH in the range of 2.0 to 6.0; their growth optimum is within the temperature range of 40 to 60°C at a pH of 3.5 to 4.5. They are strict aerobes, although they are able to survive at micro-aerophilic conditions. Their growth is inhibited in an anoxic environment. They are not pathogenic, however, they cause the spoilage of non-alcoholic beverages and functional waters. The spoiled products have an unpleasant taste and odor, and further, they may show turbidity. Generally, they do not display distinctive precipitates nor do they cause the containers holding the beverages to inflate. They also do not significantly reduce the pH value, however, a phenolic or medicinal odor can be noticed in the beverages.

[005] Alicyclobacillus spp. are naturally occurring in the environment, especially in the soil, wherefrom they are transmitted to fruit and vegetables and consequently to the fruit/vegetable processing equipment. They do not cause spoilage of concentrates having over 30° Brix; however, their spores survive in the concentrates and cause spoilage of the final product after dilution. Even the pasteurization temperature does not destroy the spores; on the contrary, they are activated at such a temperature and after the thermal treatment the bacteria begin to multiply. Characteristically, spoilage can occur if a single cell per mL comes into contact with the product. Spores are able to survive before growing for long periods of time, even for periods of over three months.

[006] Therefore, there is a need in the beverage industry for a method to inhibit the growth of Alicyclobacillus spp. in non-alcoholic beverages and functional waters by using naturally occurring substances.

SUMMARY OF THE INVENTION

In an embodiment of the invention, there is provided a preservative for non-alcoholic beverages and functional waters comprising extracts from plants of the Lamiaceae family and a method for preserving non-alcoholic beverages or functional waters including the addition of the preservative described herein.

DETAILED DESCRIPTION OF THE INVENTION

[007] In the following detailed description, numerous specific details are set forth in order to provide a thorough understanding of the invention. However, it will be understood by those skilled in the art that the present invention may be practiced without these specific details. In other instances, well-known methods, procedures, and components have not been described in detail so as not to obscure the present invention.

[008] The invention relates to the inhibition of the growth of bacteria of the genus Alicyclobacillus in non-alcoholic beverages and functional waters by means of extracts from plants of the Lamiaceae family and their mixtures. The Laminaceae family includes, inter alia, Rosmarinus, Salvia, Melissa, Origanum, Satureja, Mentha, and Thimus.

[009] The invention further relates to the preservation of non-alcoholic beverages and functional waters against spoilage by Alicyclobacillus spp. by means of water-soluble extracts from plants of the Lamiaceae family, oil-soluble extracts from plants of the Lamiaceae family, or mixtures thereof. The invention is directed also to the preparation of such preservatives. It was found that the combination of water-soluble and oil-soluble active substances confers to the product superior protection against microorganisms. [0010] The major active substances in the extracts from plants of the Lamiaceae family are carnosic acid, rosmarinic acid, and derivatives thereof. Carnosic acid is an oil-soluble active substance, while rosmarinic acid is a water-soluble active substance.

[001 1 ] The oil-soluble extract from plants of the Lamiaceae family contains up to 100 % by weight of carnosic acid. The water-soluble extract from plants of the Lamiaceae family contains up to 100 % by weight of rosmarinic acid.

[0012] According to certain embodiments of the invention, the extracts from plants of the Lamiaceae family and their mixtures are combined with any appropriate additive. When the additive dilutes the extract it allows the extract to be better distributed in the product that is to be preserved and eases the handling thereof. Such additives include propylene glycol, maltodextrin, sugar and any other additives approved for consumption, as well as mixtures thereof. According to further embodiments, an emulsifier approved for consumption may be added to the extract.

[0013] According to this invention, the Lamiaceae extracts as well as their mixtures are used in concentrations varying for each type of beverage so that they do not alter the organoleptic characteristics thereof. Further, thermal treatment (pasteurization) of the beverage does not affect their activity.

[0014] According to certain embodiments, the total concentration of the active ingredients (i.e., mainly the sum of the carnosic acid and the rosmarinic acid), added to the non-alcoholic beverage or functional waters is 1 -100 mg/kg. According to further embodiments, the total concentration of the active ingredients is 1-90 mg/kg. According to further embodiments, the total concentration of the active ingredients is 1-80 mg/kg. According to further embodiments, the total concentration of the active ingredients is 1 -70 mg/kg. According to further embodiments, the total concentration of the active ingredients is 1-60 mg/kg. According to further embodiments, the total concentration of the active ingredients is 1 -50 mg/kg. According to further embodiments, the total concentration of the active ingredients is 1-40 mg/kg. According to further embodiments, the total concentration of the active ingredients is 1 -30 mg/kg. According to further embodiments, the total concentration of the active ingredients is 1- 20 mg/kg. According to further embodiments, the total concentration of the active ingredients is 1 -10 mg/kg.

[0015] According to certain embodiments, the extract from plants of the Lamiaceae family is added to a fruit/vegetable concentrate or flavoring. According to further embodiments, the extract from plants of the Lamiaceae family is added to the final product, i.e., the non-alcoholic beverage or functional water. According to further embodiments, the extract from plants of the Lamiaceae family is combined with any appropriate additive approved for consumption, e.g., any type of carrier, vehicle or emulsifier, before the addition thereof to the concentrate, flavoring, beverage or functional water.

[0016] According to certain embodiments, additional plant extracts may be added to the beverages or functional waters together with extracts from plants of the Lamiaceae family. The various extracts may be added to the beverages in a single solution or in two separate solutions, simultaneously or separately. Additional plant extracts that may be used are citrus extracts, green tea extract, carrot extract, olive extract, or any mixture thereof, as well as any other appropriate extracts that may enhance the antimicrobial protection of the beverage. In an embodiment of the invention, there is a synergy in the antimicrobial protection of extracts from plants of the Lamiaceae family together with the citrus extract, green tea extract, carrot extract, olive extract, or any mixture thereof. This can be clearly seen from Examples 2 and 3.

[0017] Various aspects of the invention are described in greater detail in the following Examples, which represent embodiments of this invention, and are by no means to be interpreted as limiting the scope of this invention.

EXAMPLES

EXAMPLE 1

[0018] A solution of the rosemary extract (a plant belonging to the Lamiaceae family) in propylene glycol containing at least 10% of both rosmarinicc and carnosic acids together, was tested on a non-alcoholic beverage for activity against the Alicyclobacillus spp. (ACB). The solution of the rosemary extract was added in a concentration of 0.04 % to the non-alcoholic beverage, which had been inoculated with ACB cells. The following samples were prepared:

1. 100 mL of beverage + 10 mL of ACB cells suspension (cone. ~ 10³ cells/mL) + solution of the rosemary extract (0.04%)

2. 100 mL of BAT bouillon (liquid nutrient medium for ACB) + 10 mL of ACB cells

suspension (cone. ~ 10 cells/mL) + solution of the rosemary extract (0.04%)

3. 100 mL of beverage + 10 mL of ACB cells suspension (cone. ~ 10³ cells/mL) - positive control

4. 100 mL of BAT bouillon (liquid nutrient medium for ACB) + 10 mL of ACB cells

suspension (cone. ~ 10 cells/mL) - positive control

5. 100 mL of flavored water - negative control

6. 100 mL of BAT bouillon - negative control [0019] The samples were kept at 45°C for one month.

[0020] In both positive controls growth (visible turbidity) occurred within two days. On the contrary, all samples supplemented with the solution of the rosemary extract, as well as the negative controls remained clear even after one month.

EXAMPLE 2

[0021] A solution of rosemary extract in propylene glycol containing at least 10% of rosmarinic and carnosic acids, was tested on four different non-alchoholic beverages for activity against the Alicyclobacillus spp. (ACB).

[0022] In order to leave the organoleptic characteristics of the beverages unaltered after the addition of the rosemary extract solution, the following concentrations of the extract solution were used:

Beverage 1 - 0.02%

Beverage 2 - 0.015%

Beverage 3 - 0.015%

Beverage 4 - 0.01%

[0023] The testing of antimicrobial activity was repeated with appropriate concentrations for each product separately, and in a parallel test with ACB cells suspension of varying strength (cone. ~ 10³ cells/mL and cone. ~ 10⁶ cells/mL).

The following samples were prepared:

1. 100 mL of Beverage 1 + 2 mL of ACB cells suspension (cone. ~ 10⁶ cells/mL) +

solution of the rosemary extract (0.02%)

2. 100 mL of Beverage 2 + 2 mL of ACB cells suspension (cone. ~ 10⁶ cells/mL) +

solution of the rosemary extract (0.015%)

3. 100 mL of Beverage 3 + 2 mL of ACB cells suspension (cone. ~ 10⁶ cells/mL) +

solution of the rosemary extract (0.015%)

4. 100 mL of Beverage 4 + 2 mL of ACB cells suspension (cone. ~ 10⁶ cells/mL) +

solution of the rosemary extract (0.01 %)

5. 100 mL of Beverage 1 + 2 mL of ACB cells suspension (cone. ~ 10³ cells/mL) +

solution of the rosemary extract (0.02%)

6. 100 mL of Beverage 2 + 2 mL of ACB cells suspension (cone. ~ 10³ cells/mL) +

solution of the rosemary extract (0.015%)

7. 100 mL of Beverage 3 + 2 mL of ACB cells suspension (cone. ~ 10³ cells/mL) +

solution of the rosemary extract (0.015%) 8. 100 mL of Beverage 4 + 2 mL of ACB cells suspension (cone. ~ 10³ cells/mL) + solution of the rosemary extract (0.01 %)

9. 100 mL of each beverage + 2 mL of ACB cells suspension (cone. - 10⁶ cells/mL) - positive controls

10. 100 mL of each beverage + 2 mL of ACB cells suspension (cone. ~ 10³ cells/mL) - positive controls

1 1. 100 mL of each beverage - negative controls

[0024] The samples were kept at 45 °C for one month.

[0025] In the positive controls with a concentration of ~10⁶ cells/mL in ACB cells suspension, growth (visible turbidity) occurred within one day; in positive controls having a concentration of ~10³ cells/mL in ACB cells suspension, growth occurred within three days. All negative controls remained clear even after one month.

[0026] The concentration of ~10⁶cells/mL in ACB cells suspension caused growth (visible turbidity) in the Beverage 4 sample after 5 days; in the Beverage 3 sample after 7 days; in the Beverage 2 and Beverage 1 samples after 14 days.

[0027] The concentration of 10 cells/mL in ACB cells suspension caused growth (visible turbidity) in the Beverage 4 sample after 9 days; in the Beverage 3 sample after 14 days; in the Beverage 2 sample after 16 days, while the Beverage 1 sample remained clear even after one month.

[0028] Since the contamination of the products is not expected to be so extreme in real production and products are rarely exposed for longer period of time to the temperature of 45 °C (optimal for growth of Alicyclobacillus spp.) as simulated in the above experiments, the inhibition of the spoilage of the products is expected even at concentrations of 0.01% to 0.02% of the solution of the rosemary extract.

EXAMPLE 3

[0029] A solution of rosemary, citrus, and green tea extracts in propylene glycol, comprising all together at least 10% of rosmarinic acid, carnosic acid, naringin and epigallocatechin gallate, was tested on non-alcoholic beverage for activity against the Alicyclobacillus spp. (ACB). The solution of the above extracts was added in a concentration of 0.01 % to beverage that had been inoculated with ACB cells. The following samples were prepared:

1. 500 mL of beverage + 0.1 mL of ACB cells suspension (cone. ~ 10⁷ cells/mL) + the

above extract solution in propylene glycol (0.01%) 2. 500 mL of beverage + 0.1 mL of ACB cells suspension (cone. ~ 10⁷ cells/mL) - positive control

3. 500 mL of beverage - negative control

[0030] The samples were kept at 45°C for one month.

[0031] Growth (visible turbidity) occurred in the positive control within three days. On the contrary, the samples supplemented with the extract solution as well as the negative controls remained clear even after one month.

[0032] A similar synergistic antimicrobial effect was also achieved by the addition of olive and carrot extracts.

[0033] While certain features of the invention have been illustrated and described herein, many modifications, substitutions, changes, and equivalents will now occur to those of ordinary skill in the art. It is, therefore, to be understood that the appended claims are intended to cover all such modifications and changes as fall within the true spirit of the invention.

[0034]

The above examples relate also to antioxidative extracts of other plants of the Laminaceae family, such as Rosmarinus, Salvia, Melissa, Origanum, Satureja, Mentha, Thimus, and others.

Claims

What is claimed is

1. A preservative for non-alcoholic beverages and functional waters comprising extracts from plants of the Lamiaceae family.

2. The preservative according to claim 1 , wherein the extract is water-soluble, oil-soluble, or a mixture of water-soluble and oil-soluble materials.

3. The preservative according to claim 1 further comprising at least one additive approved for consumption.

4. The preservative according to claim 3, wherein the additive is a propylene glycol, maltodextrin, sugar, a carrier, an emulsifier, or any combination thereof.

5. The preservative according to claim 1 , wherein the extract from plants of the Lamiaceae family is incorporated into a non-alcoholic beverage or functional waters in an amount of 1-100 mg/kg carnosic acid or rosmary or a combination thereof .

6. The preservative according to claim 1 , wherein the extract from plants of the Lamiaceae family is incorporated into a non-alcoholic beverage or functional waters in an amount of 1-50 mg/kg carnosic acid or rosmary or a combination thereof.

7. The preservative according to claim 1 , wherein the extract from plants of the Lamiaceae family is incorporated into a non-alcoholic beverage or functional waters in an amount of 1-10 mg/kg active ingredients.

.

8. The preservative according to claim 1 , further comprising additional plant extracts.

9. The presenvative according to claim 1, wherein the preservative prevents the spoilage of non-alcoholic beverages or functional waters due to bacteria belonging to the genus

Alicyclobacillus .

10. The preservative according to claim 1 , wherein citrus extract, green tea extract, carrot extract or olive extract is added.

1 1. A method for preserving non-alcoholic beverages or functional waters including the addition of the preservative according to any one of claims 1-10 to said non-alcoholic beverages or functional waters.

12. The method according to claim 1 1 , wherein the preservative is added to a fruit/vegetable concentrate or flavoring, which is later diluted to prepare the non-alcoholic beverage or functional water.

13. The method according to claim 1 1, wherein the preservative is added to directly to the nonalcoholic beverage or functional water.

14. The method according to claim 1 1, wherein said method does not alter the organoleptic characteristics of the non-alcoholic beverages or functional waters.

15. The method according to claim 1 1 , wherein the thermal treatment of non-alcoholic beverages or functional waters does not affect the activity of the preservative.