KR102302385B1 - A natural coloring agent composition - Google Patents

Abstract

The present application relates to a natural coloring agent composition comprising a fermented plant, the composition comprising nitrite having the concentration of 700 ppm or more and free sugar having the concentration of 1,000 ppm or less, wherein the free sugar contains lactose. The natural coloring agent composition can be prepared such that nitrite is included in a high concentration from natural raw materials without inoculation of reducing bacteria, and can exhibit excellent coloring effects when applied to meat products, instead of synthetic nitrite, to meat products.

Description

A natural coloring agent composition

This application relates to a natural coloring agent composition.

Meat products such as ham and sausage processed with meat as raw materials are manufactured to exhibit their own bright red or red color, and nitrite is added as a colorant for the purpose of developing the color of the meat product. The nitrite reacts with the myoglobin contained in the meat to give the meat a bright red color, and also plays a role in increasing the shelf life of food by inhibiting C. botulinum, which generates toxins, and preventing the growth of microorganisms.

However, in the case of synthetic nitrite, which is often added as a coloring agent, due to the fact that it is a chemically synthetic additive, consumers are increasingly avoiding salted meat products with synthetic nitrite added. Attempts are continuing. As a method for producing natural nitrite, a method for obtaining nitrite by inoculating a plant or extract thereof rich in nitrate with a reducing bacterium having an activity capable of reducing nitrate to nitrite has been proposed.

Korean Patent Publication No. 2018-0031438 discloses a method for producing natural nitrite by inoculating a spinach extract with Lactobacillus parminisis and performing fermentation, "Study on optimization of spinach-derived natural nitrite production" (Kim Tae-Kyung et al. ., KOREAN J. FOOD SCI. TECHNOL . Vol. 49, No. 4, pp. 459~461 (2017)) also disclosed a method of inoculating Staphylococcus carnosus or various Lactobacillus strains to produce natural nitrite using spinach extract. Therefore, inoculation of reducing bacteria is essentially used to produce natural nitrite. However, there is a problem that a process for producing a natural plant extract that meets a sufficient amount of nitrite content or a reduction technology is not secured in Korea.

Accordingly, in the present application, including a high content of natural nitrite, a natural coloring agent composition that contains a smaller amount of nitrite than a conventional natural coloring agent composition and has sufficient color development effect as well as suitable properties when added to meat products and processed devised.

An object of the present application is to provide a natural coloring agent composition containing a high concentration of nitrite, which is processed so that the meat product can exhibit its own meat color.

Also, an object of the present application is to provide a composition for the production of meat products.

In addition, the present application aims to provide a meat product.

Another object of the present application is to provide a method for developing meat color.

An aspect of the present application is a natural colorant composition comprising a fermented plant, wherein the composition contains nitrite at a concentration of 700 ppm or more, and free sugar at a concentration of 1,000 ppm or less, wherein the free sugar contains lactose It provides a natural coloring agent composition.

Another aspect of the present application provides a composition for preparing a meat product including the natural colorant composition, wherein the natural colorant composition is included in an amount of 0.5% by weight or less.

Another aspect of the present application provides a meat product including the natural colorant composition and meat, wherein the residual nitrite ions (NO ₂ ⁻ ) in the meat product is 70 ppm or less.

Another aspect of the present application provides a method for color-developing meat, comprising mixing the natural color-developer composition with a raw meat material.

The natural coloring agent composition of the present application includes a plant fermented product, contains nitrite at a concentration of 700 ppm or more, and contains free sugar at a concentration of 1,000 ppm or less.

The nitrite (nitrite) is a nitrite ion (NO ₂ ⁻ ) and a salt combined, specifically sodium nitrite (NaNO ₂ ) It may be.

Since the natural coloring agent composition contains nitrite, it can be applied to food, such as meat-containing food, and used for the purpose of making the characteristic bright red and red colors of meat products appear.

The nitrite may be produced by nitrate reductase in the plant. Unlike the prior art of producing nitrite by reducing bacteria by inoculating a plant containing nitrate with an external reducing bacteria, the nitrite contained in the natural coloring agent composition of the present application is present in a plant or extract of a plant containing nitrate. It can be prepared without inoculation of reducing bacteria by activating nitrate reductase. Therefore, the process of separately culturing the reducing bacteria or the process of purchasing it can be omitted, and there is an advantage in that nitrite can be prepared in a more economical and easy way. The natural coloring agent composition of the present application has an excellent coloring effect when applied to meat products, and the cost, equipment, labor, etc. required for manufacturing the same can be reduced. In addition, since it does not undergo the inoculation process of reducing bacteria, there is no risk of problems such as generation of unintended substances that may be caused by the reducing bacteria, and the process of removing the reducing bacteria or by-products produced therefrom is not required. There is this.

The natural coloring agent composition of the present application contains the nitrite at a concentration of 700 ppm or more. Specifically, the nitrite is 700 ppm or more, 750 ppm or more, 800 ppm, 1,000 ppm or more, 2,000 ppm or more, 2,500 ppm or more, 3,000 ppm or more, 5,000 ppm or more, 8,000 ppm or more, 10,000 ppm or more, 20,000 ppm or more, 30,000 ppm or more, 40,000 ppm or more, 50,000 ppm or more, 60,000 ppm or more, 70,000 ppm or more, 80,000 ppm or more, 90,000 ppm or more, 100,000 ppm or more, 110,000 ppm or more, 120,000 ppm or more, 130,000 ppm or more, 150,000 ppm or more, 180,000 ppm or more Or it may be included in a concentration of 200,000 ppm or more.

As the nitrite is included in the concentration in the above range in the natural coloring composition, the composition of the present application may cause the red color inherent in meat products to appear when applied to meat products, and a similar level of coloring effect to the case of using synthetic nitrites In addition, compared to the case of using a natural nitrite using reducing bacteria, the natural color developer composition of the present invention can exhibit an excellent or similar level of color development effect even when a small amount is used.

The natural coloring agent composition of the present application contains free sugars at a concentration of 1,000 ppm or less, and the free sugars include lactose.

Specifically, the free sugar may be included in a concentration of 1,000 ppm or less, 900 ppm or less, 800 ppm or less, 700 ppm or less, 500 ppm or less, 300 ppm or less, 200 ppm or less, 100 ppm or less, or 50 ppm or less. Among the free sugars, reducing sugars may cause browning in the process of sterilizing them at high temperature after being applied to the product, which may negatively affect the appearance of the product. , when applied to meat products, it can reduce browning even when exposed to high-temperature sterilization conditions, and this can be expected to have a synergistic effect in terms of color and appearance improvement of the product along with the color development effect by nitrite.

More specifically, the natural coloring agent composition of the present application may contain the fructose at a concentration of 1,000 ppm or less, or may not contain glucose and sucrose.

The natural coloring agent composition of the present application may further include 8 or more amino acids selected from the group consisting of aspartic acid, glutamic acid, glycine, alanine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine and arginine at a concentration of 500 ppm or more. can Specifically, it may include 8 or more, 9 or more, 10 or more. In addition, specifically, the amino acid is 500 ppm or more, 600 ppm or more, 800 ppm or more, 1,000 ppm or more, 1,500 ppm or more, 2,000 ppm or more, 2,500 ppm or more, 3,000 ppm or more, 5,000 ppm or more, 7,000 ppm or more, 10,000 ppm or more , 12,000 ppm or more, 15,000 ppm or more, 17,000 ppm or more, 20,000 ppm or more, or may be included in a concentration of 25,000 ppm or more. The nitrite-containing composition prepared by inoculating the reducing bacteria may contain an amino acid in an amount less than the above range, and more specifically, amino acids such as glycine, methionine, isoleucine, leucine, tyrosine, lysine, histidine, and arginine. may not be included. As the composition of the present application contains the various types of amino acids as described above at a relatively high concentration, a positive effect may be exhibited in terms of nutrition and taste. For example, amino acids such as aspartic acid, glutamic acid, and glycine are known to produce umami, so that when the composition of the present application is applied to a meat product, it can have an advantageous effect in terms of flavor. In particular, the color developer composition of the present application may be applied to meat products and used for the purpose of inducing color development thereof. A desirable synergistic effect may be exhibited together with the effect.

The natural coloring agent composition of the present application is selected from the group consisting of hypoxanthine, guanosine, xanthine, inosine, xanthosine and IMP.2Na·7.5H ₂ O It may further include at least one nucleic acid-based compound. The natural coloring agent composition may not contain adenine and/or AMP.2Na.

The natural coloring agent composition may further include acetic acid at a concentration of 1,000 ppm or more. Specifically, the acetic acid has a concentration of 1,000 ppm or more, 2,000 ppm or more, 3,000 ppm or more, 5,000 ppm or more, 7,000 ppm or more, 10,000 ppm or more, 12,000 ppm or more, 20,000 ppm or more, 30,000 ppm or more, 40,000 ppm or more, or 45,000 ppm or more. may be included as In addition, the composition may include succinic acid. The nitrite-containing composition prepared by inoculating the reducing bacteria contains acetic acid at 300 ppm or less or contains only a small amount of succinic acid. It is useful as a natural coloring agent composition rather than a coloring agent composition.

The natural coloring agent composition may include γ-aminobutyric acid (GABA) at a concentration of 25 mg/L or more. Specifically, the GABA is 25 mg/L or more, 50 mg/L or more, 100 mg/L or more, 200 mg/L or more, 500 mg/L or more, 900 mg/L or more, 1,000 mg/L or more, 1,500 mg It may be a concentration of at least /L, at least 1,700 mg/L, at least 2,000 mg/L, or at least 2,100 mg/L.

The natural coloring agent composition according to the present invention can be prepared by the following method.

The natural coloring agent composition, preparing an extract from a plant containing nitrate; activating nitrate reductase in the plant extract; and inactivating the nitrate reductase. The extraction is a process of exporting the components contained in the plant to the outside, and the plant may be juiced or crushed. The juice may be obtained from a plant using a conventional juicer, green juicer, or the like. The crushing may be to obtain a liquid and a plant body in the plant by crushing or milling the plant. The plant extract may include nitrate and nitrate reductase in the plant. When preparing the extract through the extraction method as described above, there is an advantage that the activity of the nitrate reductase contained in the extract can be maintained without being lost or inhibited. For example, there is an effect of preventing the inhibition of the activity of nitrate reductase that may be caused by heat, solvent, or an enzyme input from the outside. Activating the nitrate reductase may include maintaining a pH range through a buffer system. Specifically, maintaining the pH range may include maintaining the pH within the range of pH 6 to pH 9, specifically pH 6.5 to pH 9, pH 6.5 to 8.5, pH 6.5 to pH 8, pH 6.5 to pH It may be maintained in the range of 7.5 or pH 6.5 to pH 7. When it undergoes a metabolic process, the pH may be reduced by by-products produced accordingly, which may reduce the activity of nitrate reductase. Therefore, when the pH is maintained in the above range, the nitrate reductase activity is not reduced and the nitrite conversion rate can be maintained high.

The natural coloring agent composition of the present application may not contain reducing bacteria having nitrate reducing activity. The reducing bacteria are, Staphylococcus genus, Micrococcus genus, Lactobacillus genus, Enterococcus genus, Lactococcus genus, Streptococcus It may be a microorganism of the genus Streptococcus genus, Pediococcus genus or Leuconostoc genus, more specifically Staphylococcus carnosus ) or Staphylococcus vitul Linus ( Staphylococcus vitulinus ) may be.

The plant fermented product may be obtained by proceeding an enzymatic reaction contained in a plant or an extract thereof.

In the present application, the term “fermentation” refers to a series of metabolic processes in which changes in organic compounds by enzyme action occur.

The plant may be at least one selected from the group consisting of spinach, lettuce, Chinese cabbage, lettuce, cabbage, radish, sesame leaf, chicory, radish leaf, radish, mugwort, kale, gat, leek, water parsley and red beet, but is limited thereto No, any plant containing nitrates can be used. The plant may contain nitrate at a concentration of 3,000 ppm or more, for example, 3,100 ppm or more, 3,200 ppm or more, 3,300 ppm or more, 3,500 ppm or more, 4,000 ppm or more, 4,500 ppm or more, 5,000 ppm or more, 3,000 ppm or more of nitrate. It may be included in a concentration of 10,000 ppm, 3,200 ppm to 800 ppm, 3,500 ppm to 7,000 ppm, 4,000 ppm to 6,000 ppm, but is not limited thereto. can be used

The natural color developer composition of the present application may be in a powder form, and in this case, the nitrite may be included in a concentration of 100,000 ppm or more. The powder may be prepared by freeze-drying or spray-drying. When the composition of the present application is in powder form, nitrite may be included in a higher concentration, and there is an advantage that it is easy to apply to meat products. In addition, the composition in powder form has a more advantageous advantage in the storage and distribution process.

The composition for preparing a meat product of the present application includes the above-mentioned natural color-imparting agent composition. The composition for preparing a meat product is for manufacturing a meat product, and may include pork, pork fat, purified water, and the like, and the components and content may be different depending on the meat product. The natural coloring agent composition may be included in an amount of 5% by weight or less in the composition for preparing a meat product. Specifically, the natural coloring agent composition is 4% by weight, 3% by weight, 2% by weight, 1% by weight, 0.5% by weight or less, 0.3% by weight or less, 0.2% by weight or less, 0.1% by weight or less, 0.08% by weight or less, 0.05 It may be included in an amount of less than or equal to 0.03 wt%, or less than or equal to 0.01 wt%. The content may be changed depending on the content of nitrite in the natural color developer, however, since the natural color developer composition according to the present invention contains a high concentration of sodium nitrite, the amount of the natural color developer composition may be reduced, and the natural color developer composition As described above, when applied to meat products, it has an excellent color development effect that allows the characteristic bright red or red color of meat products to appear. Accordingly, the natural coloring agent composition may be usefully used for producing meat products by coloring meat.

The meat product of the present application includes the natural colorant composition and meat, and the residual nitrite ions (NO ₂ ⁻ ) in the meat product may be 70 ppm or less.

As described above for the natural coloring agent composition, when applied to meat products, it has an excellent color development effect that allows the characteristic bright red or red color of meat products to appear. In addition, it is manufactured using natural raw materials such as plants, and may be used in place of synthetic nitrite. Therefore, as the meat product of the present application contains the natural coloring agent composition, the meat can exhibit a desired bright red or red color, thereby meeting consumer preferences and having an appearance that can arouse appetite. there is

The residual nitrite ions are specifically 70 ppm or less, 65 ppm or less, 60 ppm or less, 55 ppm or less, 50 ppm or less, 40 ppm or less, 30 ppm or less, 20 ppm or less, 15 ppm or less, 10 ppm or less, 5 ppm or less , 2 ppm or less or 1 ppm or less. Since nitrite ions may have a negative effect on health, when the amount of residual nitrite ions is within the above range, there is an advantage in that it is possible to minimize adverse health effects while providing sufficient color effect in meat products.

The method for color-developing meat of the present application includes mixing the natural color-developer composition with a raw meat material.

As described above for the natural coloring agent composition, when applied to meat products, it has an excellent color development effect that allows the characteristic bright red or red color of meat products to appear. Therefore, by mixing the natural color-developer composition with the raw meat material, the meat can be colored to have a desired level of meat color.

Since the natural colorant composition of the present application contains plant-derived natural nitrite, it can color meat by replacing synthetic nitrite and applying it to meat products. The natural coloring agent composition can induce color development of meat products at a level similar to that of meat products using synthetic nitrite, and can significantly improve the appearance quality of meat products by preventing browning caused by reducing sugars due to its low free sugar content. there is an effect In addition, the effect of improving the taste quality of meat products due to the high content of amino acids contained in the natural coloring agent composition of the present application may appear, and an excellent synergistic effect can be expected when considering that the coloring agent composition is applied to food.

However, the effects of the present application are not limited to the above-mentioned effects, and other effects not mentioned will be clearly understood by those skilled in the art from the following description.

1 is a model meat product prepared by mixing a colorant composition prepared by fermenting spinach without inoculation of reducing bacteria with meat (Preparation Examples 1 and 2), and a model meat product to which synthetic nitrite is added (Comparative Preparation Examples 2 and 3) ), and a photograph taken by observing the appearance of a model meat product without nitrite (Comparative Preparation Example 1).

Hereinafter, the present application will be described in detail by way of Examples.

However, the following examples specifically illustrate the present application, and the content of the present application is not limited by the following examples.

[Example 1]

In the present application, by activating nitrate reductase contained in plants, a new natural nitrite-containing fermentation broth containing natural nitrite in a high content was prepared.

Preparation of spinach fermented product

Spinach extracts were obtained by washing domestic spinach (3,000 ppm or more of nitrate) in running water to remove soil and foreign matter, and then squeezing it using a conventional green juicer.

Spinach extract was fermented at a temperature of 20 ° C to 40 ° C in a 2 liter fermenter (Marado-PDA, BIOCNS Co., Korea), and the initial pH was in the range of 8.5 to 9.0 and 0.03 M Na ₂ CO ₃ -NaHCO ₃ buffer solution was used, and fermentation was performed under 200 rpm impeller conditions, and changes in pH and nitrate and nitrite concentrations with time were measured (Table 1).

extract 0 hours 1 hours 2 hours 3 hours 4 hours 5 hours 6 hours 6.5 hours 7 hours 7.5 hours 8 hours 8.5 hours pH 6.09 8.99 8.77 8.65 8.04 7.34 7.19 7.02 6.83 6.74 6.70 6.60 6,54 nitrate
(ppm) 3,273 3,259 3,012 3,454 3,346 2,545 1,761 1,014 438 51 0 0 0 nitrite
(ppm) 0 0 0 0 297 933 1,556 2,082 2,602 2,874 2,948 2,897 2,899 Conversion rate (%) 0% 0% 0% 0% 9% 29% 48% 64% 79% 88% 90% 89% 89%

As a result, as the reaction was carried out, the pH gradually decreased, but the buffering effect of the buffer solution was sufficiently exhibited, and the conversion rate increased to about 90% when about 7 hours had elapsed from the reaction proceeding, resulting in a very good conversion effect. It was confirmed that the

[1-2] Drying of fermented products

Spinach Fermentation Concentrate Preparation

An inactivation process was performed to remove the activity of nitrate converting enzyme from the spinach fermented product prepared through the process of Example 1-1. After inactivating the enzyme by applying a high temperature of 60℃ or higher to the spinach fermented product, 5% diatomaceous earth is added using a filter press (JUNGDO 1000, JUNGDO Co., Korea) and filtered through a 15 cc filter cloth, followed by primary filtration. , the residual diatomaceous earth and fine substances were further removed by secondary filtration using a 5 μm MF filter. Thereafter, using a UHT sterilizer, sterilization was performed at a temperature of 95-100° C. for 1 minute and 30 seconds. Even after performing the deactivation, filtration, and sterilization processes as described above, the occurrence of nitrite loss did not appear significantly. In addition, a concentrate of the nitrite-containing composition of the present application was prepared by performing concentration using a centrifugal thin film concentrator in a method with the least loss of nitrite (Table 2).

nitrite (ppm) Concentration (27.5 Brix) 34,728

Spinach Fermented Powder Manufacturing

A powder of spinach extract was prepared by spray-drying and freeze-drying by different spraying methods for the concentrate, and the concentration of nitrite contained therein was measured and compared. In the case of spray drying, maltodextrin was mixed with the concentrate to adjust the solid content to 35-38 Brix level. The ratio of spinach concentrate and maltodextrin was 85:15, 88:12, and 90:10, respectively, and spray drying was performed (In let temperature: 180 °C, Out let temperature: 90 °C, sample injection rate: 10 ml/ min). In the case of freeze-drying, the concentrate was pre-frozen in a deep freezer at -70° C. without adding an excipient, and then a freeze dryer was used. As the freeze-drying conditions, the vacuum degree was 20 kPa, and the temperature inside the chamber was slowly raised from -40°C to 30°C, and drying was performed for 72 hours. The concentration of nitrite contained in the spray-dried powder (SD powder) and the freeze-dried powder (FD powder) of the three ratios as described above was measured and shown in Table 3 below.

Furtherance nitrite (ppm) SD powder (85:15) 67,806 SD powder (88:12) 72,659 SD powder (90:10) 80,780 FD Powder (100) 104,917

[Example 2]

Production of meat products using nitrite-containing composition and confirmation of color development effect

[2-1] Manufacture of model meat products

Using the spinach fermented powder prepared in Example 1, mixed with pork hind legs and fat, a model meat product was prepared at the mixing ratio shown in Table 4 below. In Preparation Example 1 and Preparation Example 2, the spinach fermented powder of Example 1 was added at concentrations of 20 ppm and 100 ppm, respectively, Comparative Preparation Example 1 did not add nitrite (negative control), Comparative Preparation Example 2 and Comparative Preparation Example 2 Preparation Example 3 was prepared by adding 20 ppm and 100 ppm of synthetic nitrite, respectively (positive control). In the case of the fermented spinach powder, 0.03% and 0.149% were added to the meat products of Preparation Examples 1 and 2, respectively, in terms of 100,000 ppm.

Raw materials Comparative Preparation Example 1 Comparative Preparation Example 2
(20ppm) Comparative Preparation Example 3
(100ppm) Preparation Example 1
(20ppm) Preparation 2
(100ppm) pork 60.00 60.00 60.00 60.00 60.00 pork fat 20.00 20.00 20.00 20.00 20.00 Purified water 16.70 16.70 16.70 16.60 16.50 isolate soy protein 2.00 2.00 2.00 2.00 2.00 refined salt 1.00 1.00 1.00 1.00 1.00 phosphate 0.30 0.30 0.30 0.30 0.30 synthetic nitrite 0.002 0.01 Spinach Fermented Powder 0.030 0.149 Sum 100 100 100 100 100

[2-2] Color measurement of nitrite-added meat products

The appearance of the model meat products of Preparation Examples 1 and 2 and Comparative Preparation Examples 1 to 3 was observed, and chromaticity was measured. The meat products were pulverized with a mixer, put in a Petri dish, and repeated three times to measure Hunter L, a, and b values. For the surface color, a Chromameter (CR-400, Minolta Co., Japan) calibrated with a standard white plate (L=97.40, a=-0.49, b=1.96) was used. Based on the measured L, a, and b values, color difference (ΔE) values were calculated and analyzed for comparison (Table 5). *ΔE is the color difference from Comparative Preparation Example 1 (negative control), **ΔE is the color difference from Comparative Preparation Example (positive control) of the same nitrite concentration.

treatment area L a b *ΔE **ΔE Comparative Preparation Example 1 68.43 4.36 10.04 Comparative Preparation Example 2 (20ppm) 67.46 6.36 8.49 2.71 Preparation Example 1 (20ppm) 67.85 7.69 8.26 3.81 1.40 Comparative Preparation Example 3 (100ppm) 68.34 7.84 8.25 3.91 Preparation 2 (100ppm) 67.69 6.82 8.18 3.17 1.21

As a result of observing the appearance of the model meat product, in Comparative Preparation Example 1, which is a negative control to which nitrite was not added, almost no red color was observed, whereas in Preparation Examples 1, 2, and Comparative Preparation Examples 2 and 3, Comparative Preparation Example 1 It was confirmed that the color was red even when viewed with the naked eye (FIG. 1). As a result of chromaticity measurement, the difference in brightness between the meat products of Preparation Examples 1 and 2 and Comparative Preparation Examples was not large. 2 and 3 showed a similar pattern. In the case of the color difference ΔE value, there was a large difference from Comparative Preparation Example 1, which is a negative control, but there was no significant difference in the color difference value between the positive controls and Preparation Examples 1 and 2.

From the above results, it was confirmed that even when fermented spinach powder prepared without reducing bacteria was added to meat products and used, a color development effect similar to that of meat products containing synthetic nitrites was obtained.

[2-3] Measurement of residual nitrite ions

The amounts of residual nitrite ions in the model meat products of Preparation Examples 1 and 2 and Comparative Preparation Examples 1 to 3 were measured using the diazotization method. 0.5 N sodium hydroxide, 12% zinc sulfate, pH 9.0 ammonium acetate and distilled water were added to 10 g of each sample, and homogenized, heated and filtered to prepare a test solution. To the prepared test solution, sulfanilamide solution, N-(1-nphthyl)ethylenediamine solution, and distilled water were added and reacted for 20 minutes. The absorbance was measured at a wavelength of 540 nm using a spectrophotometer, and the content (μg) of nitrite ions contained in 20 ml of the test solution was calculated by substituting it into the calibration curve. Based on this, the concentration of nitrite ions was calculated and shown in Table 6 below. It is expected that the fermented spinach powder can be used in a minimum amount so that residual nitrite can be minimized while sufficiently exhibiting the color effect.

Comparative Preparation Example 1 Comparative Preparation Example 2
(20ppm) Comparative Preparation Example 3
(100ppm) Preparation Example 1
(20ppm) Preparation 2
(100ppm) nitrite ion
(ppm) 0 4.0 35.0 8.0 27.0

[Example 3]

Component analysis of nitrite-containing composition according to the present application

It was confirmed whether any components other than nitrite were included in the spinach fermented product prepared in Example 1 above. A sample prepared by extracting spinach in Example 1-1 and fermenting it through pH adjustment (hereinafter, “fermented spinach”), a sample that has undergone inactivation and concentration as in Example 1-2 (hereinafter, “fermented spinach”) Concentrate'), and a powder sample prepared through the freeze-drying process as in Example 1-2 (hereinafter, 'fermented spinach powder') was used, and as a control, nitrite reducing bacteria were inoculated and prepared by the conventional technique One plant fermented product (hereinafter, 'reducing bacteria fermented product') was used. The conventional fermented plant was prepared by mixing purified water with dried celery powder, and then inoculating the reducing bacteria ( Staphylococcus camosus ) to perform fermentation.

[3-1] Amino Acid Analysis

The types and contents of amino acids contained in the spinach fermented product of the present application, the spinach fermented concentrate, the spinach fermented powder and the fermented reducing bacteria prepared through the prior art were measured (Table 7).

Specifically, 9.9 ml of distilled water was added to 0.1 ml of each sample solution, thoroughly mixed, and then centrifuged (10,000 rpm, 10 minutes, 4° C.) and the supernatant was filtered with a 0.25 μm syringe filter. Amino acids in the filtrate were analyzed using an amino acid analysis equipment (High Speed Amino Acid Analyzer, L-8900, Hitachi Co., Japan) under the following analysis conditions. As a column, a 2622SC-PH ion exchange column (4.6x60 mm, Hitachi, Co., Japan) was used. The mobile phase is in gradient mode, Pump 1 uses sodium acetate buffer (MCI buffers PH1, PH4, RG) at a column temperature of 57 °C, a flow rate of 0.4 ㎖/min, and Pump 2 uses a ninhydrin solution (R1, R2) with a flow rate of 0.35 ㎖/min. was done with 10 μl was injected as an injection volume, and the detector used a dual channel with Channel 1: UV-570 nm and Channel 2: UV-440 nm.

(ppm) Spinach Fermented Spinach Fermented Concentrate Spinach Fermented Powder Reducing bacteria fermented product Aspartic acid 27.03 68.85 402.99 31.70 Threonine 38.17 ND ND 47.54 Serine 39.52 Trace 393.96 75.88 Glutamic acid 179.20 893.71 3,893.06 145.56 Glycine 30.93 159.99 492.11 ND Cysteine ND ND ND ND Alanine 62.52 535.07 1,693.99 18.85 Valine 109.11 891.48 2,710.17 32.81 Methionine 28.99 Trace 374.73 ND Isoleucine 47.83 593.67 1,322.64 ND Leucine 100.64 1,243.50 2,487.55 ND Tyrosine 50.69 Trace 1,366.18 ND Phenylalanine 134.44 2,362.49 3,318.69 74.82 Lysine 59.82 370.57 1,077.16 ND Histidine ND ND ND ND Arginine 12.73 ND ND ND

As a result, it was found that a total of 14 amino acids were generated and included in the samples such as spinach fermented product prepared in Example 1 of the present application. It was converted to a different type of compound and showed some differences in spinach fermented concentrate and spinach fermented powder. Threonine was present in the fermented product of reducing bacteria, which is a control prepared by inoculation of reducing bacteria, but threonine was not present in samples such as the spinach fermented product prepared in Example 1. In addition, glycine, methionine, isoleucine, leucine, tyrosine, lysine, and arginine were not detected in the fermented reducing bacteria, but all of the above amino acids were detected in the fermented spinach of the present application, and arginine was also found in the spinach fermented concentrate and spinach fermented powder. All amino acids except for were detected. In addition to the glycine in the spinach fermented product sample, it was confirmed that a large amount of amino acids such as aspartic acid and glutamic acid were also included. Glycine, aspartic acid, and glutamic acid are known as amino acids that give a umami taste. The nitrite-containing composition of the present application containing these amino acids When silver is added to meat, it can have a positive effect in terms of taste as well as its color development effect.

[3-2] Analysis of nucleic acid-based compounds

The types and contents of nucleic acid-based compounds contained in the spinach fermented product of the present application, the spinach fermented concentrate, the spinach fermented powder and the fermented reducing bacteria prepared through the prior art were measured (Table 8).

Specifically, 200 μl of each filtered liquid sample solution was centrifuged and injected into high-performance liquid chromatography (HPLC, Waters, Milford, MA, USA / Pump: Waters 510, Injector: Waters 712 WISP) to inject a nucleic acid-based compound was analyzed. A UV detector (254 nm, Waters, Milford, MA, USA) and a μ-Bondapackcolumn (3.9×300 mm) were used, and the analysis temperature of the column was 30°C. As a mobile phase solution, a 1% triethylamine solution adjusted to pH 6.5 using phosphoric acid was used, and the rate was 1 ml/min.

(ppm) Adenine Hypoxanthine AMP.2Na Guanosine Xanthine Inosine Xanthosine IMP.2Na·7.5H ₂ O Spinach Fermented ND trace ND trace 35.54 0.47 0.53 7.67 Spinach Fermented Concentrate ND 377.0 ND 45.0 577.0 115.0 ND ND Spinach Fermented Powder ND 984.0 ND 85.0 1,387.0 176.0 ND ND Reducing bacteria fermented product 2.26 ND 11.8 ND ND ND ND ND

As a result, adenine and AMP.2Na were detected in the fermented reducing bacteria prepared by inoculating the reducing bacteria, but not in the spinach fermented product or its concentrate or fermented powder. In addition, reducing bacteria are fermented hypoxanthine, guanosine, xanthine, inosine, cytosine and glass IMP.2Na · 7.5H ₂ O, such as a nucleic acid-based compounds are not detected in the water is generated from spinach fermented product or a concentrate thereof, the powder exists was confirmed to be In the case of some compounds, they were included only in the spinach ferment and were removed in the process of concentration and drying, but in the case of hypoxanthine, guanosine, xanthine, and inosine, it was confirmed that the concentration was increased.

[3-3] Organic Acid Analysis

The types and contents of organic acids contained in the spinach fermented product of the present application, the spinach fermented concentrate, the spinach fermented powder and the fermented reducing bacteria prepared through the prior art were measured (Table 9).

Specifically, 9.9 ml of distilled water was added to 0.1 ml of each sample solution, thoroughly mixed, and then centrifuged (10,000 rpm, 10 minutes, 4° C.) and the supernatant was filtered with a 0.25 μm syringe filter. Thermo Scientific Dionex ICS-3000 System was used to measure the type and content of organic acids contained in each sample solution, and analysis was performed under the following conditions. As a column, an aminex HPX-87H ion exclusion column (7.8×300 mm; Bio-Rad, Hercules, CA, USA) was used. The mobile phase was flowed at a temperature of 33 °C with 0.005N sulfuric acid and acetonitrile (95:5, v/v) solution at a flow rate of 0.6 ml/min in gradient mode, and 25 μl was injected. The detector was a Conductivity Detector.

Citric acid malic acid succinic acid lactic acid Acetic acid Spinach Fermented ND ND 125.7 ND 1,027.1 Spinach Fermented Concentrate trace trace 723.1 trace 12,402.9 Spinach Fermented Powder 2,360.4 1,966.9 5,349.2 1,203.4 45,393.7 Reducing bacteria fermented product 87.0 1,274.2 29.2 149.3 243.7

As a result, adenine and AMP.2Na were detected in the fermented reducing bacteria prepared by inoculating the reducing bacteria, but not in the spinach fermented product or its concentrate or fermented powder. In addition, reducing bacteria are fermented hypoxanthine, guanosine, xanthine, inosine, cytosine and glass IMP.2Na · 7.5H ₂ O, such as a nucleic acid-based compounds are not detected in the water is generated from spinach fermented product or a concentrate thereof, the powder exists was confirmed to be In the case of some compounds, they were included only in the spinach ferment and were removed in the process of concentration and drying, but in the case of hypoxanthine, guanosine, xanthine, and inosine, it was confirmed that the concentration was increased.

[3-4] Free sugar analysis

The types and contents of free sugars contained in the spinach fermented product of the present application, the spinach fermented concentrate, the spinach fermented powder, and the fermented reducing bacteria prepared through the prior art were measured (Table 10).

Specifically, 9.9 ml of distilled water was added to 0.1 ml of each sample solution, thoroughly mixed, and then centrifuged (10,000 rpm, 10 minutes, 4° C.) and the supernatant was filtered with a 0.25 μm syringe filter. Dionex ICS-5000 system was used to measure the type and content of free sugar contained in each sample solution, and the following analysis was performed. As columns, a CarboPac PA1 guard column (Dionex, 50 × 4 mm) and a CarboPac PA1 analytical column (Dionex, 250 × 4 mm) were used while maintaining at 30 °C. 400 mM NaOH was used for the mobile phase A solution, and pure water was used for the mobile phase B solution. In gradient mode, 0 min, 95% B, 5 min, 0% B, 30 min, 10% B, 30.1 min, 95% B, 40 min 95% B, 1.0 ml/min flow rate and 5 μl was injected. . ECD was used as the detector.

Trehalose Fructose Glucose Sucrose Lactose Spinach Fermented 12.45 27.63 ND ND trace Spinach Fermented Concentrate ND ND ND ND 230.5 Spinach Fermented Powder ND ND ND ND 793.6 Reducing bacteria fermented product ND 1,370.9 1,868.9 2,756.4 ND

As a result, it was found that trehalose and lactose, which were not detected in the fermented by reducing bacteria, were present in the fermented spinach. In particular, lactose was also included in the fermented spinach concentrate and fermented powder that had undergone filtration and drying processes. In addition, as a result of fermentation using reducing bacteria, glucose and sucrose were present in very high contents, but were not detected at all in the spinach fermented product.

In the case of a fermented product prepared by inoculating reducing bacteria, as described above, a large amount of reducing sugar is included, and when applied to a product sterilized at a high temperature, browning may be induced and the appearance may be negatively affected. On the other hand, in the case of the nitrite-containing composition of the present application, in addition to the excellent color development effect by nitrite, since a small amount of reducing sugar is included, it has the effect of minimizing browning. may appear

[3-5] GABA (γ-aminobutyric acid) analysis

The content of GABA (γ-aminobutyric acid) contained in the spinach fermented product of the present application, the spinach fermented concentrate, the spinach fermented powder and the fermented reducing bacteria prepared through the prior art was measured (Table 11).

Specifically, for derivatization for analysis first, Water's AccQ Fluor™ reagent kit was used. A reaction reagent was prepared by dissolving 1 ml of acetonitrile (Vial 2B) in 6-aminoquinolyl-n-hydroxysuccinimidyl carbamate (Vial 2A). 70 μl of borate buffer solution (Vial 1) was added to 10 μl of standard solution and extract, vortexed, and left at room temperature for 1 minute. 20 μl of the reaction reagent was added, vortexed, and then reacted for 10 minutes in a water bath at 55 °C. After completion of the reaction, it was cooled to room temperature and analyzed using HPLC (Waters 2690 system). A Mightysil RP-18 GP column (4.6 × 250 mm, 5 μm, Kanto Chemical, Tokyo, Japan) was used as a column, and as the mobile phase, 200 ml of AccQ-Tag Eluent A concentrate was diluted in 2 ℓ of HPLC water, and the mobile phase was B used AccQ-Tag Eluent B. The solvent composition maintained the initial A:B at 90:10 by gradient elution, 70:30 (0.6 ml/min) until 30 min, 0:100 (0.8 ml/min) until 31 min, 0: 100 (0.8 mL/min), 90:10 (0.6 mL/min) up to 39 min, and 90:10 (0.6 mL/min) up to 50 min. The sample injection amount was 10 μl, and the detector was Ex. 250 nm, Em. Analyzed at 395 nm.

Spinach Fermented Spinach Fermented Concentrate Spinach Fermented Powder Reducing bacteria fermented product GABA (mg/L) 54.0 940.0 2,150.0 10.0

As a result, compared to the fermented product using reducing bacteria, GABA was found to be present at a concentration five times higher in the spinach fermented product, and its fermented concentrate and fermented powder contained much higher amounts of GABA. has been confirmed to have been

In the above, representative embodiments of the present application have been exemplarily described, but the scope of the present application is not limited to the specific embodiments as described above, and those of ordinary skill in the art are within the scope described in the claims of the present application. can be changed appropriately.

Claims (14)

A natural coloring agent composition comprising a plant fermented product, comprising:
The composition contains nitrite at a concentration of 700 ppm or more, and contains free sugar at a concentration of 1,000 ppm or less,
The free sugar contains lactose,
The nitrite is produced by the nitrate reductase (nitrate reductase) in the plant, a natural coloring agent composition.
The method according to claim 1,
The plant is at least one selected from the group consisting of spinach, lettuce, Chinese cabbage, lettuce, cabbage, radish, sesame leaf, chicory, radish leaf, radish, mugwort, kale, gat, leek, water parsley and red beet.
3. The method according to claim 2,
The plant will contain nitrate in a concentration of 3,000 ppm or more, a natural colorant composition.
delete The method according to claim 1,
The composition further comprises at least 8 amino acids selected from the group consisting of aspartic acid, glutamic acid, glycine, alanine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine and arginine at a concentration of 500 ppm or more. A colorant composition.
The method according to claim 1,
The composition comprises at least one nucleic acid system selected from the group consisting of hypoxanthine, guanosine, xanthine, inosine, xanthosine and IMP.2Na·7.5H _{2 O} Which further comprises a compound, a natural coloring agent composition.
The method according to claim 1,
The composition further comprises acetic acid at a concentration of 1,000 ppm or more, a natural color developer composition.
The method according to claim 1,
The composition contains GABA (γ-aminobutyric acid) at a concentration of 25 mg/L or more, a natural color developer composition.
The method according to claim 1,
The composition does not contain reducing bacteria having nitrate reducing activity, a natural color developer composition.
10. The method according to any one of claims 1 to 3, 5 to 9,
The nitrite is contained in a concentration of 5,000 ppm or more, a natural color developer composition.
10. The method according to any one of claims 1 to 3, 5 to 9,
The natural color-imparting composition is in the form of a powder, and it contains nitrite at a concentration of 100,000 ppm or more.
A composition for producing a meat product comprising the natural coloring agent composition of claim 1, comprising:
The composition for producing a meat product, wherein the natural colorant composition is included in an amount of 5% by weight or less.
A meat product comprising the natural colorant composition of claim 1 and meat, the meat product comprising:
Residual nitrite ions (NO ₂ ^- ) in the meat product is 70 ppm or less, meat product.
A method for color-developing meat, comprising mixing the natural color-developer composition of claim 1 with raw meat.

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