Ambient stable beverage

The present invention relates to an ambient stable drink, in particular a tea based drink which is preserved with a preservative system comprising cinnamic acid, dimethyl bicarbonate and at least one essential oil. The use of such a system for producing a cold-fillable beverage is also shown.

Known state of the art

In recent years, consumers wishing to quench their thirst with ready-made drinks have been offered the choice of a constantly increasing range of ready-made drinks. Today, many are turning from well-known cold soft drinks to tea-based beverages, which can be carbonated or still, and offer "natural refreshment."

Tea contains a complex combination of enzymes, biochemical intermediates, and structural elements commonly associated with plant growth and photosynthesis. Therefore, it contains many natural substances that give tea a unique taste, tartness, aroma and color. Many of these arise from the oxidation reactions that take place during the so-called fermentation step in the production of black tea. The production of tea has long been carried out by traditional processing methods with only a basic understanding of the chemical processes involved. Consequently, manufacturers have found that producing ambient stable tea-based beverages in the volumes required to compete with more traditional soft drinks is not merely a matter of flavoring non-alcoholic beverages with tea.

The flavor of the tea-based beverages and their durability relate to that of the drink as a whole. Fungi, including yeasts and molds, which can grow in tea based beverages and other soft drinks, can be degraded by heat treatment or at least controlled by the use of preservatives. Certain tea-based beverages are pasteurized and then bottled in glass or special heat-resistant PET containers. This process is known as "hot filling." Unfortunately, this is an expensive operation that produces a large amount of environmentally harmful waste. It would be more attractive to manufacturers if they could pack tea-based products in standard PET containers that can have a capacity from a single unit to multiple packs and maintain the shelf life of the product with a tailored flavor and preservative system. This is known as "cold filling." It is also useful because a tea concentrate or powder can be readily used.

Unfortunately, the use of common preservatives can adversely affect the flavor of the tea based beverage. This is especially the case with sulphites and sorbates. By adding strong flavors such as lemon, you can counterbalance the aroma of the preservative. However, consumers are sensitive to experiencing other smells. In addition, some of those consumers who seek tea-based products as healthier and as a natural alternative to soft drinks sometimes view preservatives as a type of synthetic additive that should rather be avoided.

Many countries have laws that prohibit the use of certain food additives in food and beverages, including certain fungicides (fungicides) and preservatives. Such recipes can vary widely, but there is a clear tendency for food to contain only a small assortment and smaller amounts of chemical fungicides and preservatives, especially synthetics.

There is a need to provide flavorful, ambient stable tea based beverages having a low level of synthetic preservatives.

The present inventors have now developed an ambient stable tea based beverage that is preserved with a preservative system comprising cinnamic acid, dimethyl bicarbonate and at least one essential oil in response to this need. Non-tea based beverages, including fruit drinks and cold soft drinks, can be preserved in a similar manner.

Object of the invention

The present invention relates to an ambient stable beverage containing a preservative system characterized in that the preservative system is cinnamic acid, dimethyl dicarbonate and at least one essential oil.

PL 203 188 B1

Preferably the beverage contains 1 to 175 ppm, in particular 1 to 60 ppm, of cinnamic acid.

Preferably the beverage contains 1 to 500 ppm, in particular 1 to 250 ppm, of dimethyl bicarbonate.

Preferably the beverage of the invention comprises an essential oil selected from the group consisting of: Benzyl 4-hydroxybenzoate, 4-tert-butylcyclohexanone, carvone, cinnamaldehyde, citral, citral dimethyl acetal, citronellol, cumic alcohol, cyclohexanobutyric acid, 2-cyclohexyl ethyl acetate, transnal-decaniene, transnal-decanoate , dihydrocarveol, 3,7-dimethyl-1-octanol, ethyl cyclohexanepropionate, ethyl pyruvate, ethylvanillin, jasmon, o-methoxycinnamaldehyde, methyl anthranilate, α-methyl-trans-cinnamaldehyde, methyl eugenol, methyl nonanoate, 2-methyl-2-pentenal, 5-methyl-2-phenyl-2-hexenal, methyl salicylate, 4-methyl-5-thiazoleethyl acetate, myrtenol, neomenthol, nonanoic acid, γ-nonalactone, δ-octalactone, octanoic acid, 1-octanol, 1-phenyl-1,2-propadione, piperonyl acetate, propyl benzoate, pulgeon, sorbic aldehyde, terpinene-4-ol, toluic aldehyde, γ- undecalactone, undecanal, 1-undecanol and vanillin, in particular an essential oil selected from the group consisting of: citral, citral dimethyl acetal, cumic alcohol, trans, trans-2,4-decadienal, 3,7-dimethyl-1-octanol, ethyl pyruvate, myrtenol and piperonyl acetate.

Preferably, the preservative system comprises 1 to 100 ppm of one or more essential oils.

Preferably the beverage is a tea based beverage, especially 0.01 to 3% tea solids.

The invention also relates to the use of a preservative system comprising cinnamic acid, dimethyl bicarbonate and at least one essential oil as defined above for the preparation of an ambient stable tea based beverage suitable for cold fill.

The drink preferably contains 1 to 175 ppm cinnamic acid, 1 to 500 ppm dimethyl dicarbonate (DMDC) and 1 to 100 ppm of at least one essential oil. When the beverage is a tea based beverage it preferably contains 0.01 to 3% tea solids, especially about 0.14% tea solids.

The method of producing an ambient stable tea based beverage suitable for cold filling comprises adding a preservative system containing cinnamic acid, dimethyl bicarbonate and at least one essential oil to the tea extract.

"Beverage as defined in the present invention is any beverage other than water and includes soft cold drinks, fruit drinks, coffee based drinks and tea based drinks.

The term "essential oil" as used in the present invention includes any of the volatile vegetable oils which have a foul or pleasant odor in the plant from which it is extracted. It also includes one or more of the components of this oil which is, or is responsible for, or at least contributes to imparting a bad or pleasant smell to plants.

The term "tea in the present invention" denotes the leaf material of Camellia sinensis var. sinensis or Camellia sinensis var. assamica. The term "tea also includes the product of blending two or more of these teas."

In order to remove doubt as to the meaning of the word "comprising, it means that" includes but not necessarily "consists of or" consists of. In other words, the listed steps or options need not be exhaustive.

Except in embodiments and comparative examples, or where not expressly stated, all numbers in the specification indicating amounts or concentrations of materials should be read in conjunction with the modifying word "about.

Detailed description of the drawings

Figure 1 shows the results of a Saccharomyces cerevisiae X2180-1B yeast growth control experiment in a matrix of tubes with Ready-To-Drink Tea (RDT) containing 0.14% tea.

Figure 2 shows the combined effect of citral dimethyl acetal, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of Ready-To-Drink tea, 0.14% tea.

Figure 3 shows the combined effect of cumic alcohol, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of Ready-To-Drink tea, 0.14% tea.

PL 203 188 B1

Figure 4 shows the combined effect of citral, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of Ready to Drink tea containing 0.14% tea.

Figure 5 shows the combined effect of 3,7-dimethyloctanol, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of Ready-To-Drink tea, 0.14% tea.

Figure 6 shows the combined effect of myrtenol, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of Ready-To-Drink tea, 0.14% tea.

Figure 7 shows the combined effect of piperonyl acetate, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of Ready to Drink tea containing 0.14% tea.

Figure 8 shows the combined effect of trans, trans-2,4-decadienal, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes with Ready to Drink tea containing 0.14% tea.

Figure 9 shows the combined effect of δ-decanolactone, cinnamic acid and DMDC on the growth of yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of Ready-To-Drink tea containing 0.14% tea.

Figure 10 shows the combined effect of citral dimethyl acetal, cumic alcohol, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of Ready to Drink tea tubes containing 0.14% tea.

Figure 11 shows the results of a Saccharomyces cerevisiae X2180-1B yeast growth control experiment in a matrix tube with a cold soft drink containing 0% tea.

Figure 12 shows the combined effect of citral dimethyl acetal, cinnamic acid, and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of synthetic soft drink, 0% tea.

Figure 13 shows the combined effect of cumic alcohol, cinnamic acid, and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of test tubes of synthetic soft drink, 0% tea.

Figure 14 shows the combined effect of citral, cinnamic acid, and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of synthetic soft drink, 0% tea.

Figure 15 shows the combined effect of 3,7-dimethyloctanol, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of test tubes of a synthetic soft drink, 0% tea.

Figure 16 shows the combined effect of myrtenol, cinnamic acid, and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of synthetic soft drink, 0% tea.

Figure 17 shows the combined effect of piperonyl acetate, cinnamic acid, and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of synthetic soft drink, 0% tea.

Figure 18 shows the combined effect of trans, trans-2,4-decadienal, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of synthetic soft drink, 0% tea.

Figure 19 shows the combined effect of δ-decalactone, cinnamic acid, and DMDC on the growth of yeast Saccharomyces cerevisiae X2180-1B in a matrix of test tubes of synthetic soft drink, 0% tea.

Figure 20 shows the combined effect of citral dimethyl acetal, cumic alcohol, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of synthetic soft drink, 0% tea.

Figure 1 shows the effective concentrations of trans, trans-2,4-decadienal.

Figure 22 shows the effective concentrations of citral.

Detailed Description of the Invention

The ambient stable beverage of the present invention is preserved with a preservative system comprising cinnamic acid, dimethyl bicarbonate, and at least one essential oil. Cinnamic acid

PL 203 188 B1

Cinnamic acid (3-phenyl-2-propenoic acid) is a well-known flavoring agent used in cakes, drinks, chewing gum and ice cream. It is derived from cinnamon, which has long been added to food and is considered a useful and harmless flavoring in many countries. When it is dissolved in a tea-based drink, the cinnamic acid gives the drink a mild resinous scent, reminiscent of honey and flowers, along with a sweet and slightly spicy aftertaste. The odor effect is pronounced at concentrations above about 10 ppm. At concentrations above 30 ppm, the smell becomes particularly strong. An additional benefit is the suppression of unwanted odors from chemicals such as sorbic acid and benzoic acid. Of the two existing stereoisomers, the trans isomer is more often used for aromatization.

For cinnamic acid, FEMA (Flavoring Extract Manufacturers Associacion) granted GRAS status (i.e. Generally Recognized as Safe) in 1965. Although there are as yet no regulations in the European Union that inhibit or restrict use of cinnamic acid in food or beverages, the usual maximum use that has previously been used in industry is 31ppm. Recently, 174.9 ppm has been allowed for non-alcoholic beverages.

Many derivatives of cinnamic acid are known and used in industry. These include p-dimethylaminocinnamate, cinnamaldehyde, cinnamic acetate, cinnamic alcohol, cinnamon benzoate, cinnamon cinnamate, cinnamon formate, cinnamon isobutyrate, cinnamon isovalerate and cinnamic phenylacetate. For the purposes of the present invention, cinnamic acid may be substituted or combined with one or more of its derivatives, although its effects on flavor and taste should be taken into account when considering the concentration required to achieve the desired effect.

The preservative or antimicrobial activity of cinnamic acid in combination with an acidulant in low pH beverages is known from US Patent No. 6,042,861. However, there is no disclosure of a preservative system including cinnamic acid, methyl bicarbonate, and at least one essential oil.

Without wishing to be bound by theory, the inventors believe that cinnamic acid acts as an active compound in the cell membrane which, at low pH, increases the concentration of cinnamic acid dissolved in the cell membrane, i.e., it does not act as a classic weak acid preservative.

The drink according to the present invention preferably comprises 1 to 175 ppm cinnamic acid, more preferably 1 to 60 ppm and especially 1 to 30 ppm.

Dimethyl dicarbonate

Dimethyl dicarbonate is a well known sterilant for cold soft drinks. It is also known as dimethyl pyrocarbonate or DMDC and is sold by Bayer AG under the tradename VELCORIN ™. DMDC is a useful sterilant and kills microorganisms quickly when added. DMDC degrades quickly in the aquatic environment and therefore poses no risk to consumers. However, it does not provide a long-term preservative effect. DMDC was approved for use as a yeast inhibitor in wines at the bottling site by the United States FDA / US Department of Food and Drug Control on October 21, 1988. The European Union introduced DMDC as a cold sterilization agent in 1989.

DMDC is known to be ineffective against mold contamination.

For the purposes of the present invention, the preservative system should preferably contain between 1 and 500 ppm dimethyl dicarbonate, more preferably between 1 and 250 ppm dimethyl dicarbonate.

Essential oil

The present inventors have tested many antibacterial agents and found the following suitable for use in the preservative system of the invention. The minimum inhibitory concentration (MIC) of each compound is given.

Table 1

Beneficial essential oils

<td>MIC union</td><td>(ppm)</td>

<td> 1</td><td> 2</td>

<td>Benzyl 4-hydroxybenzoate</td><td> 68</td>

<td>4-t-butylcyclohexanone</td><td> 462</td>

Table 1

<td> 1</td><td> 2</td>

<td>carvone</td><td> 300</td>

<td>cinnamaldehyde</td><td> 66</td>

<td>lemon aldehyde</td><td> 228</td>

<td>citral dimethyl acetal</td><td> 198</td>

<td>citronellol</td><td> 125</td>

<td>cumic alcohol</td><td> 450</td>

<td>cyclohexanobutyric acid</td><td> 68</td>

<td>2-cyclohexylethyl acetate</td><td> 102</td>

<td>trans, trans-2,4-decadienal</td><td> 8</td>

<td>decanal</td><td> 47</td>

<td>decanol</td><td> 24</td>

<td>dihydrocarveol</td><td> 540</td>

<td>3,7-dimethyl-1-octanol</td><td> 15,8</td>

<td>ethyl cyclohexanepropionate</td><td> 184</td>

<td>ethyl pyruvate</td><td> 1392</td>

<td>ethylvanillin</td><td> 249</td>

<td>jasmon</td><td> 246</td>

<td>o-methoxycinnamaldehyde</td><td> 130</td>

<td>methyl anthranilate</td><td> 310</td>

<td>α-methyl-trans-cinnamic aldehyde</td><td> 58,4</td>

<td>methyl eugenol</td><td> 356</td>

<td>methyl nonanoate</td><td> 90</td>

<td>2-methyl-2-pentenal</td><td> 1274</td>

<td>5-methyl-2-phenyl-2-hexenal</td><td> 162</td>

<td>methyl salicylate</td><td> 152</td>

<td>4-methyl-5-thiazole ethanol acetate</td><td> 1110</td>

<td>myrtenol</td><td> 137</td>

<td>neomenthol</td><td> 156</td>

<td>nonanoic acid</td><td> 63</td>

<td>γ-nonalactone</td><td> 63</td>

<td>δ-octalactone</td><td> 568</td>

<td>octanoic (caprylic) acid</td><td> 115</td>

<td>1-octanol</td><td> 247</td>

<td>1-phenyl-1,2-propanedione</td><td> 222</td>

<td>piperonyl acetate</td><td> 242</td>

<td>propyl benzoate</td><td> 66</td>

<td>pulegon</td><td> 152</td>

PL 203 188 B1 cont. table 1

<td> 1</td><td> 2</td>

<td>sorbic aldehyde (2,4-hexadienal)</td><td> 86</td>

<td>terpinen-4-ol</td><td> 616</td>

<td>toluic aldehyde</td><td> 240</td>

<td>γ-undecalactone</td><td> 28</td>

<td>undecanal</td><td> 34</td>

<td>1-undecanol</td><td> 14</td>

<td>vanillin</td><td> 1216</td>

The preservative system preferably comprises 1 to 100 ppm of at least one essential oil. More preferably the preservative system comprises 1 to 50 ppm of at least one essential oil, even more preferably 1 to 32.5 ppm.

Some of the aforementioned essential oils have been found to be particularly advantageous in respect of their influence on the flavor profile of tea based beverages containing these oils. They are listed in Table 2 below. In each case, their minimum inhibitory concentrations and their preferred concentrations are given.

Table 2

Particularly preferred essential oils

<td>Relationship</td><td>MIC (ppm)</td><td>Favorable concentration (ppm)</td>

<td>lemon aldehyde</td><td> 228</td><td> 1-30</td>

<td>citral dimethyl acetal</td><td> 198</td><td> 1-30</td>

<td>cumic alcohol</td><td> 450</td><td> 1-40</td>

<td>T rans, trans-2,4-decadienal</td><td> 8</td><td> 1-20</td>

<td>3,7-dimethyl-1-octanol</td><td> 15,8</td><td> 1-20</td>

<td>ethyl pyruvate</td><td> 1392</td><td> 1-40</td>

<td>myrtenol</td><td> 137</td><td> 1-20</td>

<td>Piperonyl acetate</td><td> 242</td><td> 1-20</td>

Tea extract

The tea extract can be obtained by any suitable method. Preferably the tea leaves are extracted with hot water for between 20 minutes and 5 hours. The extract can be dried to a powder, reconstituted to an acidic beverage, or concentrated to a syrup from which a tea based beverage can be made.

Tea is known to have some antibacterial and antiviral properties by itself. However, a concentration of about 3% has to be exceeded to find the onset of suppression of yeast and mold growth. At lower concentrations than those stated, which are typical for tea-based beverages, tea acts as a nutrient that increases the possibility of deficiencies due to the presence of microorganisms. The beverage should therefore contain from 0.01 to 3% tea solids, with an amount of about 0.14% being particularly preferred.

Other factors

The water quality can seriously compromise the shelf life of the drink. This is an important factor in the preparation of a beverage, particularly a cold fill tea based beverage. For this purpose it will often be important to minimize the yeast content of the water used at all stages of production. Prior art methods include chlorination / dechlorination and irradiation with UV rays.

The ambient temperature stable beverages produced by the process of the invention may be still or carbonated with carbon dioxide. The mere introduction of carbon dioxide appears

It provides a preservative effect, so the formulation of the carbon dioxide product need not be the same as that of a still product.

Tea-based beverages usually contain sugar or some other sweetener to counterbalance the sometimes tart taste of the tea. Most of the microorganisms that can grow in tea-based beverages feed on sugar, nitrogen source, oxygen, zinc, magnesium, potassium, phosphate, and vitamins. It is therefore preferable to limit the sugar content to 8 to 10 degrees brix, however, up to 60 degrees brix may be used when the product is a tea blend.

The oxygen content can be minimized by pre-pasteurization or heat treatment, or by nitrogen purging. The mineral content of a tea based beverage can be minimized by using EDTA, citrate or a water softener. For example, microorganisms can grow in tea if the concentration of magnesium ions exceeds 0.2 ppm and they only need trace amounts of zinc.

If desired, the preservative system may also include ascorbic acid, a well-known food preservative that is most commonly known as vitamin C.

The present invention also relates to a process for producing an ambient stable tea-based, cold-fill, beverage. The process comprises adding cinnamic acid, dimethyl bicarbonate, and at least one essential oil to the tea extract.

Cinnamic acid is freely soluble in essential oils, benzene, ether, acetone, glacial acetic acid, and carbon disulfide. However, the acid is not readily soluble in tea, and one should not want to contaminate the tea based beverage with any of the above-mentioned chemicals. Since the preservative system of the present invention contains one or more essential oils, it may be necessary to include a solubility enhancing step prior to adding the cinnamic acid to the tea solution. This can be achieved by spray-drying cinnamic acid on a powdered carrier (which may optionally be sugar-based) and adding the powder to tea, converting the acid into a salt thereof, or dissolving the cinnamic acid in a small amount of an organic solvent such as ethanol or propylene glycol. The essential oil can be spray dried in the same way.

The invention will now be described in the following examples with reference to the accompanying drawings.

Example 1

Experiments with ready-to-drink tea (RDT)

Figure 1 shows the results of a Saccharomyces cerevisiae X2180-1B yeast growth control experiment in a matrix of tubes with Ready-to-Drink tea, 0.14% tea. The matrix of 30 ml tubes each containing 10 ml RTD tea, pH 3.4, containing 1-175 ppm cinnamic acid. Tubes were inoculated with 10⁴ cells / ml of yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate, DMDC, was added in concentrations ranging from 1 to 0 ppm. Tubes were then incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Figure 2 shows the combined effect of citral dimethyl acetal, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of Ready to Drink tea containing 0.14% tea. The matrix of 30 ml tubes each containing 10 ml RTD tea, pH 3.4, all contained 100 ppm citral acetal and 1-175 ppm cinnamic acid. Tubes were inoculated with 10⁴cells / ml of yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate, DMDC, was added to the samples at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Comparison of this Figure with Figure 1 shows significantly less growth in some tubes supporting yeast growth in the presence of the essential oil component, citral dimethyl acetal.

Figure 3 shows the combined effect of cumic alcohol, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of Ready-To-Drink tea, 0.14% tea. The matrix of 30 ml tubes each containing 10 ml RTD tea, pH 3.4, all contained 100 ppm cumic alcohol and 1-175 ppm cinnamic acid. Tubes were inoculated with 10⁴ cells / ml of yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate was added at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Comparison of this Figure with Figure 1 shows significantly less growth in some tubes supporting yeast growth in the presence of the essential oil component cumic alcohol.

Figure 4 shows the combined effect of citral, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of Ready to Drink tea containing 0.14% tea. The matrix of 30 ml tubes each containing 10 ml RTD tea, pH 3.4, all contained 100 ppm citral and 1-175 ppm cinnamic acid. Tubes were inoculated with 10⁴ cells / ml of yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate was added at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Comparison of this Figure with Figure 1 shows significantly less growth in some tubes supporting yeast growth in the presence of the essential oil component, citral.

Figure 5 shows the combined effect of 3,7-dimethyloctanol, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of Ready-To-Drink tea, 0.14% tea. The matrix of 30 ml tubes each containing 10 ml RTD tea, pH 3.4, all contained 50 ppm 3,7-dimethyl octanol and 1-175 ppm cinnamic acid. Tubes were inoculated with 10⁴ cells / ml of yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate was added at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Comparison of this Figure with Figure 1 shows significantly less growth in some tubes supporting yeast growth in the presence of the essential oil component, 3,7-dimethyl octanol.

Figure 6 shows the combined effect of myrtenol, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of Ready to Drink tea tubes containing 0.14% tea. The matrix of 30 ml tubes each containing 10 ml RTD tea, pH 3.4, all contained 100 ppm myrtenol and 1-175 ppm cinnamic acid. Tubes were inoculated with 10⁴ cells / ml of yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate was added at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Comparison of this Figure with Figure 1 shows significantly less growth in some tubes supporting yeast growth in the presence of the essential oil component myrtenol.

Figure 7 shows the combined effect of piperonyl acetate, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of Ready to Drink tea containing 0.14% tea. The matrix of 30 ml tubes each containing 10 ml RTD tea, pH 3.4, all contained 100 ppm piperonyl acetate and 1-175 ppm cinnamic acid. Tubes were inoculated with 10⁴ cells / ml yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate was added at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Comparison of this Figure with Figure 1 shows significantly less growth in some tubes supporting yeast growth in the presence of the essential oil component, piperonyl acetate.

PL 203 188 B1

Figure 8 shows the combined effect of trans, trans-2,4-decadienal, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of Ready to Drink tea containing 0.14% tea. The matrix of 30 ml tubes each containing 10 ml RTD tea, all containing 15ppm trans, trans-2,4-decadienal and 1-175ppm cinnamic acid. Tubes were inoculated with 10⁴ cells / ml of yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate was added at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Comparison of this Figure with Figure 1 shows significantly less growth in some tubes supporting yeast growth in the presence of the essential oil component, trans, trans-2,4-decadienal.

Figure 9 shows the combined effect of δ-decanolactone, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of Ready to Drink tea containing 0.14% tea. The matrix of 30 ml tubes each containing 10 ml RTD tea, all contained 100 ppm δ-decanolactone and 1-175 ppm cinnamic acid. Tubes were inoculated with 10⁴ cells / ml of yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate was added at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Comparison of this Figure with Figure 1 shows significantly less growth in some tubes supporting yeast growth in the presence of the essential oil component, δ-decanolactone.

Figure 10 shows the combined effect of citral dimethyl acetal, cumic alcohol, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of Ready to Drink tea tubes containing 0.14% tea. The matrix of 30 ml tubes each containing 10 ml RTD tea, all contained 25 ppm citral dimethyl acetal, 35 ppm cumic alcohol and 1-175 ppm cinnamic acid. Tubes were inoculated with 10⁴ cells / ml of yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate was added at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Comparison of this Figure with Figure 1 shows significantly less growth in some tubes supporting yeast growth in the presence of essential oil components, citral dimethyl acetal and cumic alcohol.

Example 2

Experiences with a synthetic cold soft drink

Figure 11 shows the results of a Saccharomyces cerevisiae X2180-1B yeast growth control experiment in a matrix tube with a cold soft drink containing 0% tea. The synthetic non-alcoholic cold drink contained 8 wt% glucose, 3 g / L citric acid, 1 g / L potassium orthophosphate, 0.1 g / L magnesium chloride, and 0.1 g / L yeast extract. The matrix of 30 ml tubes each containing 10 ml soft drink, pH 3.4, containing cinnamic acid in the range 1-175 ppm. Tubes were inoculated with 10⁴ cells / ml of yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate, DMDC, was added at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Figure 12 shows the combined effect of citral dimethyl acetal, cinnamic acid, and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of synthetic soft drink, 0% tea. The synthetic soft drink contained 8% by weight of glucose, 3 g / l of citric acid, 1 g / l of potassium orthophosphate, 0.1 g / l of magnesium chloride and 0.1 g / l of yeast extract. The matrix of 30 ml tubes each containing 10 ml soft drink pH 3.4, all containing 100 ppm citral dimethyl acetal and 1-175 ppm cinnamic acid. Tubes were inoculated with 10⁴ cells / ml Saccharomyces yeast

Cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate, DMDC, was added at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Comparison of this Figure with Figure 11 shows significantly less growth in some tubes supporting yeast growth in the presence of the essential oil component, citral dimethyl acetal.

Figure 13 shows the combined effect of cumic alcohol, cinnamic acid, and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of synthetic soft drink, 0% tea. The synthetic soft drink contained 8% by weight of glucose, 3 g / l of citric acid, 1 g / l of potassium orthophosphate, 0.1 g / l of magnesium chloride and 0.1 g / l of yeast extract. The matrix of 30 ml tubes each containing 10 ml soft drink pH 3.4, all containing 100 ppm cumic alcohol and 1-175 ppm cinnamic acid. Tubes were inoculated with 10⁴ cells / ml of yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate, DMDC, was added at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Comparison of this Figure with Figure 11 shows significantly less growth in some tubes supporting yeast growth in the presence of the essential oil component cumic alcohol.

Figure 14 shows the combined effect of citral, cinnamic acid, and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of synthetic soft drink, 0% tea. The synthetic soft drink contained 8% by weight of glucose, 3 g / l of citric acid, 1 g / l of potassium orthophosphate, 0.1 g / l of magnesium chloride and 0.1 g / l of yeast extract. The matrix of 30 ml tubes each containing 10 ml soft drink pH 3.4, all containing 100 ppm citral and 1-175 ppm cinnamic acid. Tubes were inoculated with 10⁴ cells / ml of yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate, DMDC, was added at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Comparison of this Figure with Figure 11 shows significantly less growth in some tubes supporting yeast growth in the presence of the essential oil component, citral.

Figure 15 shows the combined effect of 3,7-dimethyloctanol, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of test tubes of a synthetic soft drink, 0% tea. The synthetic soft drink contained 8% by weight of glucose, 3 g / l of citric acid, 1 g / l of potassium orthophosphate, 0.1 g / l of magnesium chloride and 0.1 g / l of yeast extract. The matrix of 30 ml tubes each containing 10 ml soft drink pH 3.4, all containing 50 ppm 3,7-dimethyl octanol and 1-175 ppm cinnamic acid. Tubes were inoculated with 10⁴ cells / ml of yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate, DMDC, was added at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Comparison of this Figure with Figure 11 shows significantly less growth in some tubes supporting yeast growth in the presence of the essential oil component, 3,7-dimethyl octanol.

Figure 16 shows the combined effect of myrtenol, cinnamic acid, and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of synthetic soft drink, 0% tea. The synthetic soft drink contained 8% by weight of glucose, 3 g / l of citric acid, 1 g / l of potassium orthophosphate, 0.1 g / l of magnesium chloride and 0.1 g / l of yeast extract. The matrix of 30 ml tubes each containing 10 ml soft drink pH 3.4, all containing 100 ppm myrtenol.

And cinnamic acid in the range 1 to 175 ppm. Tubes were inoculated with 10⁴ cells / ml of yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate, DMDC, was added at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Comparison of this Figure with Figure 11 shows significantly less growth in some tubes supporting yeast growth in the presence of the essential oil component myrtenol.

Figure 17 shows the combined effect of piperonyl acetate, cinnamic acid, and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of synthetic soft drink, 0% tea. The synthetic soft drink contained 8% by weight of glucose, 3 g / l of citric acid, 1 g / l of potassium orthophosphate, 0.1 g / l of magnesium chloride and 0.1 g / l of yeast extract. The matrix of 30 ml tubes each containing 10 ml soft drink pH 3.4, all containing 100 ppm piperonyl acetate and 1-175 ppm cinnamic acid. Tubes were inoculated with 10⁴ cells / ml of yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate, DMDC, was added at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Comparison of this Figure with Figure 11 shows significantly less growth in some tubes supporting yeast growth in the presence of the essential oil component, piperonyl acetate.

Figure 18 shows the combined effect of trans, trans-2,4-decadienal, cinnamic acid, and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of synthetic soft drink, 0% tea. The synthetic soft drink contained 8% by weight of glucose, 3 g / l of citric acid, 1 g / l of potassium orthophosphate, 0.1 g / l of magnesium chloride and 0.1 g / l of yeast extract. The matrix of 30 ml tubes each containing 10 ml soft drink pH 3.4, all containing 15 ppm trans, trans-2,4-decadienal and 1-175 ppm cinnamic acid. Tubes were inoculated with 10⁴ cells / ml of yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate, DMDC, was added at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Comparison of this Figure with Figure 11 shows significantly less growth in some tubes supporting yeast growth in the presence of the essential oil component, trans, trans-2,4-decadienal.

Figure 19 shows the combined effect of δ-decanolactone, cinnamic acid and DMDC on the growth of yeast Saccharomyces cerevisiae X2180-1B in a matrix of test tubes of synthetic soft drink, 0% tea. The synthetic soft drink contained 8% by weight of glucose, 3 g / l of citric acid, 1 g / l of potassium orthophosphate, 0.1 g / l of magnesium chloride and 0.1 g / l of yeast extract. The matrix of 30 ml tubes each containing 10 ml of a pH 3.4 synthetic soft drink, all containing 100 ppm δ-decanolactone and 1-175 ppm cinnamic acid. Tubes were inoculated with 10⁴ cells / ml of yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate, DMDC, was added at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Comparison of this Figure with Figure 11 shows significantly less growth in some tubes supporting yeast growth in the presence of the essential oil component, 8-decanolactone.

Figure 20 shows the combined effect of citral dimethyl acetal, cumic alcohol, cinnamic acid and DMDC on the growth of the yeast Saccharomyces cerevisiae X2180-1B in a matrix of tubes of synthetic soft drink, 0% tea. The synthetic soft drink contained 8% by weight of glucose, 3 g / l of citric acid, 1 g / l of potassium orthophosphate, 0.1 g / l of magnesium chloride and 0.1 g / l of yeast extract. The matrix of 30 ml tubes each containing 10 ml of synthetic soft cold drink

PH 3.4, all containing citral dimethyl acetal in an amount of 25 ppm, cumic alcohol in an amount of 35 ppm, and cinnamic acid in the range 1-175 ppm. Tubes were inoculated with 10⁴ cells / ml of yeast Saccharomyces cerevisiae X2180-1B. Immediately after inoculation, dimethyl dicarbonate, DMDC, was added at concentrations ranging from 1 to 250 ppm. Tubes were incubated for 14 days at 25 ° C to allow surviving yeasts to grow out.

After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Comparison of this Figure with Figure 11 shows significantly less growth in some tubes supporting yeast growth in the presence of the essential oil components, citral dimethyl acetal and cumic alcohol.

Example 3

Effective concentrations of essential oils

Figure 21 shows the effective concentrations of the trans essential oil, trans-2,4-decadienal. The yeast Saccharomyces cerevisiae X2180-1B was cultivated in 30 ml bottles containing RTD tea containing 0.14% tea containing 0, 15 ppm or 30 ppm cinnamic acid. Rows of tubes also contained trans, trans-2,4-decadienal at concentrations ranging from 0-16 ppm. After inoculating the tubes with 10⁴ Tubes were incubated for 14 days at 25 ° C for yeast cells to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.

Figure 22 shows the effective concentrations of the essential oil component, citral. The yeast Saccharomyces cerevisiae X2180-1B was grown in 30 ml bottles containing RTD tea containing 0.14% tea and 0.15 ppm or 30 ppm cinnamic acid. Rows of tubes also contained citral in concentrations ranging between 0-120 ppm. After inoculating the tubes with 10⁴ Tubes were incubated for 14 days at 25 ° C for yeast cells to allow surviving yeasts to grow out. After 14 days, the yeast population was measured by measuring the optical density at 600 nm in a sample diluted 11 times and the blank value was subtracted.