WO2009091092A1

WO2009091092A1 - Canned kimchi with improved storability and enhanced functionality

Info

Publication number: WO2009091092A1
Application number: PCT/KR2008/000329
Authority: WO
Inventors: Kun Young Park; Sook Hee Rhee; Hye Ji Kang
Original assignee: Pusan National University Industry-University Cooperation Foundation
Priority date: 2008-01-18
Filing date: 2008-01-18
Publication date: 2009-07-23
Also published as: JP2010510813A; JP5017378B2

Abstract

Canned kimchi is provided. The canned kimchi comprises at least one main kimchi material selected from the group consisting of Chinese cabbage, leaf mustard, joung radishes, radishes, green onions, sesame leaves, Korean lettuces and Korean leeks, 0.01 to 1.0 part by weight of at least one additive selected from chitosan, vitamin C and grapefruit seed extract, based on 100 parts by weight of the main kimchi material, and 25 to 55 parts by weight of a filling solution containing fermentation water and at least one salted-fermented seafood selected from salted- fermented anchovy sauce and salted-fermented shrimp, based on 100 parts by weight of the main kimchi material. The number of Leuconostoc sp. in the canned kimchi is in the range of 1.0 x 102 CFU/ml to 5.0 x 104 CFU/ml. Deaeration and pasteurization are performed to control the number of lactic acid bacteria in the canned kimchi of the present invention within a specified range, to reduce a loss of texture of the kimchi, and to suppress the evolution of gas, thus achieving improved stability of the kimchi. In addition, the antimutagenic and anticarcinogenic activities of the canned kimchi according to the present invention can be effectively enhanced.

Description

CANNED KIMCHI WITH IMPROVED STORABILITY AND ENHANCED FUNCTIONALITY

Technical Field

[1] The present invention relates to canned kimchi with improved storability and enhanced functionality. More specifically, the present invention relates to canned kimchi in which the number of lactic acid bacteria is controlled within a specified numerical range to achieve improved storability and enhanced functionality while maintaining the inherent texture of fermented kimchi.

[2]

Background Art

[3] With the recent increase in the number of women who participate in social activities, improvements in income level and changes in dietary life, there has been a growing demand for instant food that is less time consuming in cooking and is convenient to eat. Further, there is an increasing tendency to buy and eat commercial kimchi products rather than to make kimchi. As kimchi gains wider international recognition and popularity, kimchi is exported to foreign countries as well as overseas Koreans. Thus, the need for commercial kimchi products is steadily increasing. The use of ready-made kimchi products advantageously saves the time required to cook kimchi food. As the national income of Korea steadily rises, Korean people spend more time to participate in leisure activities and enjoy opportunities for dining out. Under these circumstances, there is a need for high-quality kimchi products that are convenient to consume.

[4] However, kimchi continues to be fermented by a variety of fermentation-related microorganisms and enzymes during storage. Continued fermentation of kimchi brings about various problems, such as excessive sourness, tissue softening and generation of an unpleasant smell, eventually resulting in a deterioration in quality and a loss of storability thus making the kimchi non-edible. The period of circulation of kimchi is very short, for example, less than 7 days at room temperature and 2-4 weeks even under refrigeration conditions. After such a short period of circulation, gas is evolved to swell the package, which causes damage and separation of the kimchi from the packaging container to deteriorate the marketability of the product.

[5] Thus, it is necessary to develop a method by which the shelf life of kimchi can be extended in order to enhance the commercial value of kimchi as a product. A variety of approaches associated with the use of radiation and the addition of preservatives have been proposed to improve the storability of kimchi, but consumers tend to shun away from kimchi that has undergone the chemical treatments. In view of this, attempts have been made concerning the use of additives, for example, the addition of a buffer to suppress a fluctuation in pH, the addition of Na-acetate and Na-malate capable of buffering against acid flavor and the addition of a phosphate/Na-citrate mixed salt. Much research has been conducted to retard the fermentation rate and prolong the edible period of kimchi by the addition of preservatives, such as sorbic acid, polybutyl benzoate, and sodium dehydroacetate, and by the addition of a citric acid/citrate mixture as a pH-adjusting agent. It was also reported that the removal of fermentable sugar from kimchi showed positive effects on the improvement of the storability of the kimchi. In addition, a great deal of research has been focused on retort pouch storage, heat sterilization, addition of preservatives, addition of supplementary materials, addition of chitosan, canning processes, etc. In particular, research is actively underway to add naturally occurring preservatives to inhibit kimchi from being ripened.

[6] Many methods have been suggested to solve the problems (e.g., poor storability and short period of circulation) associated with the commercialization of kimchi. A representative method for improving the marketability of commercialized kimchi is the use of metal cans for long-term packaging of kimchi. Metal packaging containers are superior in terms of three main functions of food packaging containers, i.e. long-term preservation, convenience and marketability, compared to any other packaging containers, and are easy to handle and carry. Problems encountered with the preparation and use of canned foods includes food spoilage due to poor sealing and incomplete sterilization and dissolution of heavy metals (e.g., tin) from containers. Further, organic acids of kimchi may induce corrosion of metal cans and cause problems in terms of stability during storage.

[7] Meanwhile, heavy metals contained in processed food may be derived from raw materials of the processed food or may be eluted from packaging containers in the course of the manufacture, processing and distribution of the processed food. Heavy metals accumulated in the body may be responsible for various chronic poisonings. However, according to a study on pH changes with varying storage periods and levels of heavy metals (e.g., Pb and Sn) with the lapse of time in agricultural canned products having relatively low pH values, the contents of Pb and Sn as heavy metals in the canned products increased with increasing storage period, a relatively large amount of Pb was dissolved in canned products having relatively low pH values, and the level of Sn in canned kirrchi products having pH of 4 or above was lower than other samples. Further, there are some reports that the lead and tin contents of canned products at low pH values were much lower than the standard values even after storage for 12 months.

[8] Conventional commercially available canned kirrchi products adopt heat sterilization to achieve improved storability. However, in view of the fact that kirrchi contains live lactic acid bacteria, heat sterilization causes a loss of taste and flavor inherent to kirrchi, generates a heating smell and softens the tissue of kirrchi.

[9] In view of the foregoing problems, the present inventors have tried to effectively control the number of lactic acid bacteria in kirrchi, and as a result, have succeeded in developing canned kirrchi with improved storability, good kirrchi flavor and enhanced functionality and stability resulting from the addition of additives.

[10]

Disclosure of Invention Technical Problem

[11] It is an object of the present invention to provide canned kirrchi in which the number of Leuconostoc sp. is controlled within a specified numerical range by pasteurization to achieve good flavor, improved storability and enhanced functionality while maintaining the inherent texture of fermented kirrchi.

[12]

Technical Solution

[13] In order to accomplish the above object of the present invention, there is provided canned kirrchi comprising at least one main kirrchi material selected from the group consisting of Chinese cabbage, leaf mustard, joung radishes, radishes, green onions, sesame leaves, Korean lettuces and Korean leeks, QOl to 1.0 part by weight of at least one additive selected from chitosan, vitamin C and grapefruit seed extract, based on 100 parts by weight of the main kirrchi material, and 25 to 55 parts by weight of a filling solution containing fermentation water and at least one salted-fermented seafood selected from salted-fermented anchovy sauce and salted-fermented shrimp, based on 100 parts by weight of the main kirrchi material wherein the number of Leuconostoc sp. in the canned kirrchi is in the range of 1.0 x 10 ² CFU/ml to 5.0 x 10⁴ CFU/ml.

[14] The number of Lactobacillus sp. in the canned kirrchi is preferably less than 1.0 x 10 i CFU/ml. [15] The salted-fermented seafood is preferably present in an amount of 4 to 8 parts by weight, based on 100 parts by weight of the main kirrchi material.

[16] The fermentation water is preferably prepared by adding seasoning including 1.4 to

2.8 parts by weight of garlic, 06 to 1.2 parts by weight of ginger, 1.0 to 2.0 parts by weight of sugar and 13 to 26 parts by weight of shredded radishes to salt water and fermenting the seasoning to a pH of 3.8 to 4.1.

[17]

Advantageous Effects

[18] The canned kirrchi of the present invention is produced using pasteurization to achieve improved storability, high sensory quality and enhanced functionality. According to a method for producing the canned kirrchi of the present invention, a mixture of salted Chinese cabbage and seasoning is fermented to obtain white kirrchi to shorten the fermentation time and make the flavor better. Red pepper powder is added to the fermented white kirrchi and is then contained in a can to prevent the color of the final kirrchi from being deteriorated due to the low chromatidty. A filling solution, which is prepared by adding salted-fermented anchovy sauce and salted- fermented shrimp to fermentation water remaining after removal of the fermented Chinese cabbage, is added to the can to make the flavor of the final kirrchi better without affecting the number of bacteria in the kirrchi. The fermented Chinese cabbage is washed by a combination of sodium hypochlorite treatment and ultrasonic washing to efficiently remove bacteria and reduce a loss of texture arising from the sodium hypochlorite treatment. Deaeration and pasteurization are performed after filling into a can to decrease the number of bacteria and suppress the evolution of gas, thus achieving improved stability of the kirrchi. Chitosan and vitamin C are subsequently added to effectively enhance the antimutagenic and anticarcinogenic activities of the final canned kirrchi.

[19]

Brief Description of the Drawings

[20] FIG. 1 shows the results of sensory tests and the number of lactic acid bacteria after red pepper powder was added in the course of the production of canned kirrchi according to the present invention.

[21] FIG. 2 shows the results of sensory tests and the number of lactic acid bacteria after salted-fermented seafood was added to fermentation water in the course of the production of canned kirrchi according to the present invention. [22] FIG. 3 shows the results of sensory tests and the number of lactic acid bacteria after chitosan was added in the course of the production of canned kimchi according to the present invention.

[23] FIG. 4 shows the results of sensory tests and the number of lactic acid bacteria when packing rates were varied in the course of the production of canned kimchi according to the present invention.

[24] FIG. 5 shows the number of lactic acid bacteria and physicochemical properties when deaeration was performed after filling into cans in the course of the production of canned kimchi according to the present invention.

[25] FIG .6 shows the number of lactic acid bacteria and physicochemical properties when pasteurization was performed after filling into cans in the course of the production of canned kimchi according to the present invention.

[26] FIG. 7 shows variations in the number of bacteria in canned kimchi products of the present invention during long-term storage at 2O⁰C and 35⁰C.

[27] FIG. 8 shows variations in the body weight and dietary intake of rats after canned kimchi of the present invention was added to diets of the rats.

[28] FIG. 9 shows the ability of canned kimchi according to the present invention to inhibit the formation of micronuclei using mouse peripheral reticulocytes.

[29]

Best Mode for Carrying Out the Invention

[30] The present invention will now be described in greater detail with reference to the accompanying drawings.

[31] The canned kimchi of the present invention uses at least one main kimchi material selected from the group consisting of Chinese cabbage, leaf mustard, young radishes, radishes, green onions, sesame leaves, Korean lettuces and Korean leeks. There is no particular restriction on the kind of the canned kimchi according to the present invention. Examples of such kimchi products include, but are not limited to, Chinese cabbage kimchi, leaf mustard kimchi, cubed white radish kimchi, white kimchi, young radish kimchi, Korean lettuce kimchi, Korean leek kimchi, sesame leaf kimchi and green onions kimchi.

[32] The canned kimchi of the present invention comprises a filling solution containing salted-fermented seafood and fermentation water. The filling solution is present in an amount of 25 to 55 parts by weight, based on 100 parts by weight of the main kimchi material.

[33] The salted-fermented seafood is preferably selected from salted-fermented anchovy sauce, salted-fermented shrimp and mixtures thereof and is used to enhance the sensory quality of the final canned kimchi. The filling solution is prepared by boiling a blend of the salted- fermented seafood and the fermentation water in a standard ratio. The salted- fermented seafood is preferably present in an amount of 4 to 8 parts by weight, based on 100 parts by weight of the main kimchi material.

[34] The fermentation water is prepared by salting seasoning including 1.4 to 2.8 parts by weight of garlic, 06 to 1.2 parts by weight of ginger, 1.0 to 2.0 parts by weight of sugar and 13 to 26 parts by weight of chopped radishes in salt water and fermenting the seasoning to a pH of 3.8 to 4.1. If the contents of the seasoning materials are out of the respective ranges, the sensory quality of the canned kimchi according to the present invention is undesirably deteriorated.

[35] The canned kimchi of the present invention may be Chinese cabbage kimchi, joung radish kimchi, cubed white radish kimchi, leek kimchi, Korean lettuce kimchi, sesame leaf kimchi or Korean leek kimchi. The kind and content of the seasoning may be varied depending on the kind of the kimchi.

[36] The number of Leuconostoc sp. in the canned kimchi of the present invention is limited within the range of 1.0 x 10² CFU/ml to 5.0 x 10⁴ CFU/ml. When the number of Leuconostoc sp. in the canned kimchi is below 1.0 x 10 ² CFU/ml, the flavor of kimchi cannot be maintained. Meanwhile, when the number of Leuconostoc sp. in the canned kimchi is above 5.0 x 10 ⁴ CFU/ml, the evolution of gas in the can is promoted, resulting in a deterioration in the storage stability of the canned kimchi.

[37] The number of Lactobacillus sp. in the canned kimchi of the present invention is preferably less than 1.0 x 10¹ CFU/ml. When the number of Lactobacillus sp. in the canned kimchi is more than 1.0 x 10 ¹ CFU/ml, the kimchi tastes sour and may be fermented and ripened within the can.

[38] The canned kimchi of the present invention can be produced in accordance with the following sequential steps: Salting of Chinese cabbage → Mixing/fermentation of the salted Chinese cabbage with seasoning → Washing → Addition of additives → Filling → Pasteurization.

[39] The salting of Chinese cabbage is carried out by cutting Chinese cabbage into appropriate sizes, salting the Chinese cabbage pieces in salt water, washing the salted cabbage pieces with flowing tap water several times, and draining the washed cabbage pieces.

[40] A fermentation solution is prepared and mixing/fermentation is carried out. The fermentation solution is prepared by mixing wheat flour with water, boiling the mixture to make paste, adding the paste to salt water containing sun-dried salt, adding the salted Chinese cabbage and seasoning composed of garlic, ginger, sugar and chopped radishes to the salt water, and adjusting the final salinity using sun-dried salt. The fermentation solution is added to the salted Chinese cabbage to make white kimchi, and then the white kimchi is fermented at 2O⁰C until the pH reaches 3.8 to 4.1. According to prior art methods, a kimchi material and mixed seasoning are independently fermented, mixed together and contained in a can to produce canned kimchi. However, the kimchi fails to have the inherent ripened flavor. In contrast, according to the present invention, since the main kimchi material and the seasoning are fermented together to make white kimchi, the time required for the fermentation is shortened and the flavor is effectively enhanced.

[41] Subsequently, the fermented white kimchi is washed with tap water and cut into appropriate sizes. Then, the fermented Chinese cabbage pieces are introduced into an aqueous solution of sodium hypochlorite, washed by sonication in a sterilization and/or ultrasonic washer, and dehydrated/dried. A combination of sodium hypochlorite treatment and ultrasonic washing is preferably employed to wash the fermented Chinese cabbage pieces, and as a result, the frequency and time of the hypochlorous add treatment can be shortened to efficiently remove bacteria present in the fermented white kimchi.

[42] Thereafter, the solid, i.e. a mixture of the Chinese cabbage and the seasoning, is contained in a can, and then a filling solution (fermentation water + salted-fermented seafood) is added thereto. The filling solution is added for the purpose of maintaining the flavor of kimchi and facilitating the sterilization of the solid. The addition of the filling solution facilitates the heat transfer during subsequent deaeration and pasteurization to assist in raising the internal temperature of the can to the sterilization temperature, thus allowing the solid to be uniformly sterilized. Although the inherent flavor of the Chinese cabbage is important, it is necessary to compensate the loss of the flavor of the Chinese cabbage by washing. This compensation is effected by the filling solution to enhance the flavor of the final kimchi.

[43] The fermented Chinese cabbage having undergone the sterilization treatments (i.e. sodium hypochlorous add treatment and ultrasonic washing), red pepper powder, and at least one additive selected from chitosan, vitamin C and grapefruit seed extract are mixed with the fermented chopped radishes to prepare kimchi.

[44] The red pepper powder is sterilized with ethanol before use. Alternatively, red pepper powder produced at Youngyang, Korea, which contains a small number of bacteria, may be used without sterilization. The canned kimchi of the present invention is characterized in that the chromatidty of the canned kimchi be increased without any noticeable increase in the number of bacteria by adding the red pepper powder before filling into a can rather than by adding the red pepper powder to make a mixed seasoning.

[45] The grapefruit seed extract is added to kill lactic acid bacteria. No Lactobacillus sp. is observed, which indicates that the addition of the grapefruit seed extract is effective in ensuring sufficient storability. The chitosan and the vitamin C independently do not contribute to a drastic reduction in the number of bacteria, but they can be combined with another additive to exert a synergistic effect. Of course, it is important to control the initial number of bacteria in the Chinese cabbage. More importantly, the functions of the additives and the sterilization treatments are combined to further enhance the sterilization effects.

[46] The selected additive is preferably used in an amount of QOl to 1.0 part by weight, based on 100 parts by weight of the main kimchi material. When the additive is used in an amount of less than QOl parts by weight, the effect of adding the additive is negligible. Meanwhile, when the additive is used in an amount of more than 1.0 part by weight, the number of lactic acid bacteria is not effectively controlled.

[47] The kimchi is contained in a can, deaerated and pasteurized at 65⁰C for 30 minutes to control the number of lactic acid bacteria to a specified numerical range. The deaeration and the pasteurization are performed to inhibit the formation of gas. The chitosan and the vitamin C can be added to enhance the antimutagenic and anticar- cinogenic activities of the final canned kimchi.

[48] The canned kimchi of the present invention suppresses the growth of Lactobacillus sp. and the proliferation of other bacteria so that the number of the bacteria is maintained at a steady state within a particular range to effectively prolong the shelf life of the canned kimchi without gas evolution.

[49]

Mode for the Invention

[50] EXAMPLES

[51] Example 1

[52] 1. SaltingandfermentationofChinesecabbage

[53] Chinese cabbage ( Brassicacampestris L. ) and radishes (Raphanussativus L.) were purchased from the Bujun market, Busan, Korea under the trademarks Garak Shin #1 and Chungun, respectively. Red pepper (Capsicumannum L.) powder was purchased from a factory, where clean red pepper powder was processed, belonging to the Youngyang Agricultural Cooperative, Korea. Garlic (Allium sativumL.) and ginger ( ZingiberofficinaleRosc.) were purchased from the Bujun market. Clean salted- fermented anchovy sauce (Daesang Corp., Korea), salted-fermented shrimp (Hasunjung General Food Co., Ltd., Korea) and sun-dried salt (Wooil Co., Ltd., Korea) were used as salted-fermented seafood products.

[54] The Chinese cabbage was cut into four segments, salted in 10% salt water for 12 hours, washed with flowing tap water three times, and drained for 3 hours. Wheat flour and water were mixed together in a ratio of 2:8 and boiled to make paste. 2.5% by weight of the paste was added to 2.5% salt water containing the sun-dried salt, and then seasoning composed of 1.4 parts by weight of garlic, Q6 parts by weight of ginger, 1.0 part by weight of sugar and 13 parts by weight of chopped radishes with respect to 100 parts by weight of the salted Chinese cabbage was added to the salt water to prepare a fermentation solution. Sun-dried salt was used to adjust the final salinity of the fermentation solution to 2.5%. The fermentation solution was added to the salted Chinese cabbage in a ratio of 1:1.1 and fermented at 2O⁰C to a pH of 3.8-3.9 to make white kimchi.

[55]

[56]

[57] 2. Controlofthenumberoflacticacidbacteriaincannedkimchi

[58] The number of bacteria in final canned kimchi was controlled within a specified numerical range in accordance with the following steps. Specifically, the steps were divided into sub-steps by washing and after washing, and a detailed explanation thereof will be provided below.

[59]

[60] A. Controlofthenumberoflactic acid bacteria bvwashing

[61] The fermented Chinese cabbage was washed once with tap water and cut into pieces having a size of 3 an x 3 cm. The fermented Chinese cabbage pieces were introduced into a 400 ppm aqueous solution of sodium hypochlorite (YAKURI PURE CHEMICAL CO., LTD./TYOTO JAPAN), sterilized for 5 minutes or subjected to ultrasonic washing in an ultrasonic washer (GM saver Co. Ltd., Korea), where ultrasonic waves of about 27,000 Hz were generated, for up to 5 minutes, and dehydrated/dried for 30 minutes.

[62] The sodium hypochlorous acid treatment was conducted only once to control the initial number of bacteria in the Chinese cabbage and the ultrasonic washer was operated to examine the effects of the ultrasonic washing on the reduction of the number of bacteria.

[63] The pH was measured using a pH meter (Istek model 735-P, Korea) at room temperature, and the acidity was measured by using AOAC method. According to the AOAC method, 20 ml of the sample was diluted 20-fold with distilled water and 10 ml of the dilute solution was taken for the acidity test. 1 ml of Q 1% phenolphthalein as an indicator was added to the dilution solution, followed by titration with Ql N NaOH until the indicator turned pink ('end point') The titration value was expressed in terms of lactic acid content (%)

[64] mLof O. INNaOH x Normality of NaOH x 0.09 Lactic acid = x l OO

Weight (g) of Sample [65] Table 1 [Table 1]

[66] Effects of ultrasonic washing on the total number of bacteria (x 10⁶CFU/ml) in fermented Chinese cabbage [67] As can be seen from Table 1, the total number of bacteria and the number of yeast in the groups treated with sodium hypochlorous acid (5 min.) and sonication was decreased by a factor of 10¹ when compared to those in the salted Chinese cabbage. The total number of bacteria and the number of yeast increased with increasing ultrasonic washing time, indicating that the ultrasonic washing had no effect. In the groups treated with sonication and sodium hypochlorous acid, the number of Leuconostoc bacteria was lowered by a factor of 10 ¹ but no washing effect could not be observed in Lactobacillus sp.

[68] Based on the fact that the washing effect was varied depending on the washing time, the ultrasonic washing effects were evaluated by dividing the washing time. Further, the initial number of bacteria was varied to examine the removal effects of bacteria because there were variations in the number of bacteria according to the degree of fermentation. After Chinese cabbage was cut into pieces having a size of about 3 cm x4 an and introduced into ultrasonic washers at pH values of 4.1 and 3.8, the total number of bacteria, the number of yeast and the number of lactic acid bacteria were measured at intervals of 30 seconds.

[69] Table 2 [Table 2]

[70] Effects of ultrasonic washing on the total number of bacteria (x 10⁶ CFU/ml) in fermented Chinese cabbage (pH 4.1) [71] Table 3 [Table 3]

[72] Effects of ultrasonic washing on the total number of bacteria (x 10⁶ CFU/ml) in fermented Chinese cabbage (pH 3.8) [73] The results of Table 2 show that the initial numbers of all bacteria, yeast and lactic acid bacteria were on the order of 10 ⁷ CFU/ml and decreased by a factor of 10¹ after the ultrasonic treatment for 30 seconds. The level of 10 ⁶ CFU/ml was maintained without any great change until the ultrasonic washing was conducted for 5 minutes. 5 minutes after the ultrasonic washing, the number of bacteria showed a tendency to increase.

[74] The results of Table 3 show that the total number of bacteria, the number of yeast and the number of lactic acid bacteria in the fermented Chinese cabbage (pH 3.8) were about 10¹ times higher than those in the fermented Chinese cabbage (pH 4.1) 30 seconds after the sonication, the numbers were decreased by a factor of 10 ^!. One minute after the sonication, the total number of bacteria and the number of yeast were further decreased by a factor of 10 ¹ to reach the order of 10⁶ CFU/ml.

[75] That is, the ultrasonic washing led to a decrease in the total number of bacteria and the number of yeast by a factor of 10 ^!. The fact that any bacteria washing effect was observed with the lapse of time is believed to be because the bacteria were washed out from the Chinese cabbage rather than killed in the Chinese cabbage.

[76]

[77] B. Controlofthenumberofbacteriaafterwashing

[78] When a can was filled with the washed fermented Chinese cabbage, effects of supplementary materials and filling fermentation water were examined on the number of bacteria and the enhancement of flavor.

[79] Based on 1 kg of the fermented Chinese cabbage having undergone the sterilization treatments by the treatment with sodium hypochlorous acid and the ultrasonic washing, 64g of the red pepper powder, 5g of chitosan, Ig of vitamin C, 1,500 ppm of grapefruit seed extract (GFSE, 1.5g/10g of distilled water, model DF- 100, FA bank Co., Ltd.) and the shredded radishes fermented in the previous step were mixed together to prepare kimchi. The red pepper powder was sterilized with 70% ethanol before use. Alternatively, red pepper powder produced in Youngyang, Korea, which contains a small number of bacteria, was used without sterilization.

[80] Fermentation water remaining after fermentation of the Chinese cabbage, salted- fermented shrimp and salted-fermented anchovy sauce were mixed in a ratio of 8:Q5:Q5, boiled, and cooled to prepare filling fermentation water.

[81]

[82] ( 1 ) Subsequent addition of red pepper powder

[83] Since supplementary materials were added during fermentation, the influence of different seasoning compositions on the enhancement of flavor and the number of lactic acid bacteria were examined.

[84] Seasoning materials were mixed in a standard ratio and dried to prepare mixed seasoning ('a control group') For comparison, red pepper powder only was used without the addition of any supplementary material, garlic and ginger powders were added with red pepper powder, and red pepper powder was sterilized with 70% ethanol.

[85] The fermented mixed seasoning was dried using a hot-air dryer (J-300M, JBICO

Co., Ltd.), pulverized using a nine-stage mixer (GP-2003/2005, Greenpia Co., Ltd.), and diluted 10-fold with distilled water. The chromatidty of the diluted solution was measured using a Minolta Chroma Meter (CT-310, Japan) based on the following criteria: L (Lightness), a (+: Red, -: Green), b (+: Yellow, -: Blue)

[86] After the red pepper powder was sterilized by dipping it in 70% ethanol for 30 seconds, 1 minute and 2 minutes, the sterilized red pepper powder was measured for the total number of bacteria and chromatidty. There was no marked change in the red pepper powder sterilized with 70% ethanol within 30 seconds, but the number of bacteria tended to increase with time. It is believed that no sterilization effect occurred because colorants escaped from the red pepper powder with time.

[87] To evaluate the sensory effects, the red pepper powder sterilized with 70% ethanol was used to produce canned kimchi, red pepper powder without undergoing sterilization with 70% ethanol was used to produce canned kimchi, a mixture of garlic and ginger powders and red pepper powder without undergoing sterilization with 70% ethanol was used to produce canned kimchi, and a conventional canned kimchi using mixed dry seasoning were prepared (a control group)

[88] Referring to the sensory test results shown in FIG. 1, the control group using the mixed seasoning had low points in terms of appearance and high points in terms of moldy smell, bitter flavor, etc., the group using red pepper powder only had low points in terms of the same items, the group using the mixture of red pepper powder, garlic and ginger obtained similar results to the group using mixed seasoning except appearance. Based on the overall acceptability, the group using red pepper powder only gained the highest score and the group using mixed seasoning gained the lowest score.

[89] Lactobacillus bacteria were not observed and there were no drastic variations in the number of Leuconostoc sp. in the control group and the other groups, demonstrating that all seasoning compositions did not greatly affect the numbers of bacteria.

[90]

[91] (2) Addition of salted-fermented seafood

[92] After salted-fermented anchovy sauce, salted-fermented shrimp and a mixture of salted-fermented anchovy sauce and salted-fermented shrimp were used as filling solution materials to produce canned kimchi products, the sensory tests of the canned kimchi products were conducted. Snce different salted-fermented seafood materials are used in different regions, people feel various flavors of the salted-fermented seafood materials. The sensory tests were conducted using salted-fermented seafood materials produced in the Seoul-Gyeonggi and Busan-Gyeongnam provinces, Korea.

[93] As shown in FIG. 2, all groups in the Seoul-Gyeonggi province showed similar results in terms of appearance, bitter flavor, metal flavor, sour flavor, etc. Based on the overall acceptability, the group using salted-fermented anchovy sauce gained the lowest score and the group using the mixture of salted-fermented anchovy sauce and salted-fermented shrimp gained the highest score.

[94] In the Busan-Gyeongnam province, the group using the mixture of salted- fermented anchovy sauce and salted-fermented shrimp gained slightly low scores in terms of sour flavor and bitter flavor when compared to the other groups. Further, there were no marked differences in terms of sour and hot flavors in terms of appearance and texture between the group using the mixture of salted-fermented anchovy sauce and salted- fermented shrimp and the other groups. Based on the overall acceptability, the group using salted-fermented anchovy sauce gained the highest score, which is similar to that of the group using the mixture of salted-fermented anchovy sauce and salted- fermented shrimp.

[95] Lactobacillus bacteria were not observed and there was only a slight difference between the number of Leuconostoc sp. and the total number of bacteria in all canned kimchi products, demonstrating that the kind of the salted-fermented seafood materials did not greatly affect the storability of the canned kimchi products. Therefore, it was determined that the use of flavor-creating and nutritionally significant salted-fermented seafood materials is effective in flavor enhancement and the mixture of salted- fermented anchovy sauce and salted-fermented shrimp can be selectively used to produce desirable canned kimchi.

[96]

[97] (3) Addition of chitosan

[98] Chitosan was added to improve the storability of final canned kimchi. The effects of chitosan on the number of bacteria, texture and sensory quality were examined.

[99] After chitosan was added in amounts of Q5%, 1%, 1.5% and 2% by weight with respect to the weight of the Chinese cabbage, the texture of canned kimchi products was evaluated (FIG. 3) As a result, the groups using Q5 and 1% by weight of chitosan showed improved texture, whereas there was only a slight difference in texture between the groups using 1.5 and 2% by weight of chitosan and the control group.

[100] The sensory tests of the groups using 05, 1% and 2% by weight of chitosan were conducted. As a result, the group using 2% by weight of chitosan gained high scores in terms of moldy smell, bitter flavor, etc. but gained the lowest score in terms of appearance. The group using Q5% by weight of chitosan gained low scores in terms of moldy smell and bitter flavor and gained the best score in terms of appearance. All groups did not show much difference in terms of texture. The overall acceptability was better in this order: the group using 05 % by weight of chitosan > the group using 1% by weight of chitosan > the group using 2% by weight of chitosan. Therefore, it is understood that the group using Q5% by weight of chitosan showed the best sensory quality.

[101] As shown in FIG. 3, there was no noticeable difference between the group using chitosan and chitosan oligosaccharide and the group using no chitosan and chitosan oligosaccharide at the initial stage and even after 8 weeks of storage.

[102] These observations are believed to be because the canned kimchi was produced after bacteria were controlled and chitosan did not greatly affect the storability of the canned kimchi. However, the addition of chitosan is more effective from the viewpoint of improvement in the sensory quality and texture of the canned kimchi.

[103]

[104] (4) Variations according to packing rates

[105] Fermentation water plays a role in enhancing the flavor of Chinese cabbage and assists in raising the internal temperature of canned kimchi during sterilization to more efficiently sterilize the canned kimchi. The effects of the fermentation water were evaluated in canned kimchi by varying the ratio between the solid kimchi material and the fermentation water.

[106] The content ratio between the solid kimchi material and the fermentation water in ordinary canned kimchi whose production conditions were established was 128:32 (g/g) 12Og of the solid was blended with 4Og of the fermentation water, and then the blend was contained in a can to produce canned kimchi. Also, 11Og of the solid was blended with 50g of the fermentation water, and then the blend was contained in a can to produce canned kimchi. Variations in the number of bacteria in the canned kimchi products were observed.

[107] As shown in FIG. 4, a decrease in the content of the solid and an increase in the amount of the fermentation water tended to somewhat decrease the number of Leu- conostoc sp. and the total number of bacteria. It appears that the amount of the fermentation water was increased during pasteurization to increase the temperature of the canned kimchi products, thus shortening the time required to reach the sterilization temperature to allow the sterilization to continue for a long time, thereby achieving improved sterilization effects.

[108] In connection with the internal temperature of the canned kimchi products, the variations in packing rate led to a decrease in the number of bacteria to some extent, but caused poor sensory quality. In other words, the maintenance of the fermentation water in an optimal ratio is more effective in improving the sensory quality than the addition of a large amount of the fermentation water.

[109]

[110] (5) Control of the number of bacteria by deaeration

[111] After the kimchi was contained in cans, steam at about 97⁰C was passed through the cans to remove air from the kimchi and increase the temperature of the cans (deaeration) Then, the cans were seamed and pasteurized to evaluate the sterilization effects and observe whether gas was evolved.

[112] After the deaeration, the number of Leuconostoc sp. and the total number of bacteria were lowered from 10⁷ cells/ml to 10⁴ cells/ml. The subsequent pasteurization decreased the total number of bacteria and the number of Leuconostoc sp. from 10⁴ cells/ml to 10² cells/ml. Referring to FIG. 5, Lactobacillus sp. bacteria were completely killed by deaeration. Gas was formed in the can having undergone no deaeration and pasteurization, and as a result, the can was swollen with the gas. Fermentation occurred in the canned kimchi that was allowed to stand at room temperature for three days without undergoing deaeration and pasteurization due to the presence of air. Three days after the production, the kimchi was at the stage of maturity. These results suggest that the deaeration made the pasteurization efficient to decrease the number of bacteria, thus greatly assisting in ensuring the storability of the canned kimchi.

[113]

[114] (6) Control of the number of bacteria by pasteurization, etc.

[115] The canned kimchi was subjected to pasteurization at 65⁰C for 30 minutes. Bacteria were inoculated into Chinese cabbage juice media. The media were treated with grapefruit seed extract and chitosan to evaluate the influence of the supplementary materials on the pasteurization.

[116] The Chinese cabbage juice media were prepared by the following procedure. First, the outer leaves of Chinese cabbage were removed. The remaining Chinese cabbage was pressed using a juicer to obtain a juice. The juice was filtered through a cotton cloth, followed by centrifugation at 3,000 x g for 20 minutes. Salt was added to the centrifuged juice until its content become 2.5% and passed through a membrane filter (Q45 /M) The supplementary materials were added to the media in amounts necessary for the respective steps, and then bacteria were inoculated thereinto. The bacteria were plate-cultivated, and then their number was counted.

[117] The bacteria were divided into two groups by the number (10⁴ CFU/ml and 10⁸ CFU/ ml) to evaluate whether the initial number of bacteria was influenced by the ster- ilization effects. Specifically, the following experimental groups were prepared: groups treated with grapefruit seed extract (1,500 ppm), groups treated with chitosan (Q5%), groups having undergone pasteurization (65⁰C, 30min), groups treated with both grapefruit seed extract and chitosan, groups treated with grapefruit seed extract and having undergone pasteurization, groups treated with chitosan and having undergone pasteurization, and groups treated with both grapefruit seed extract and chitosan and having undergone pasteurization. The obtained results are shown in FIG. 6.

[118] Large sterilization effects were attained in most of the groups whose initial number of bacteria was 1 x 10⁴ CFU/ml. Lactobacillus and Leuconostoc bacteria were completely killed in the group treated with grapefruit seed extract only. Lactobacillus and lO'-lO² CFU/ml of Leuconostoc bacteria were removed from the groups where pasteurization was performed. Lactobacillus and Leuconostoc bacteria were completely killed in the groups treated with grapefruit seed extract.

[119] In the meanwhile, different profiles were found in the groups whose initial number of bacteria was 1 x 10⁸ CFU/ml. Lactobacillus bacteria were completely killed and 10⁴-10 5 CFU/ml of Leuconostoc bacteria were removed in all groups except the group treated with chitosan and having undergone pasteurization. Lactobacillus bacteria were not found and 10⁵ CFU/ml of Leuconostoc bacteria were killed in the group treated with both grapefruit seed extract and chitosan and having undergone pasteurization.

[120] The bacteria were completely killed in the groups treated with grapefruit seed extract, regardless of whether the initial number of bacteria was small or large. Particularly, the fact that Lactobacillus bacteria were not found suggests that the grapefruit seed extract was effective in ensuring the storability of kimchi. The use of chitosan did not directly contribute to a marked decrease in the number of bacteria, but chitosan is believed to show sterilization effects when being combined with another additive. Of course, it is important to control the initial number of bacteria in the Chinese cabbage. More importantly, a combination of the functions of the additives actually used for the production of canned kimchi and the sterilization treatments has been found to produce synergistic effects. The best results were attained in the groups treated with grapefruit seed extract and chitosan and having undergone pasteurization.

[121]

[122] ComparativeExample 1

[123] The number of lactic acid bacteria in commercially available canned kimchi (Wang Kimchi, SAMJIN G.F. Co., Ltd., Korea) was measured. The canned kimchi was composed of 75% of Chinese cabbage, 3.15% of purified salt, 2.65% of leeks, 2.61% of red pepper powder, 1.15% of white sugar, Q33% of garlic, Q17% of ginger, Ql% of L-glutamine sodium and 14.84% of purified water. The pH and the acidity (lactic acid) of the canned kimchi were found to be 4.2 and Q81, respectively. Neither Leu- conostoc bacteria nor Lactobacillus bacteria were observed. This is because the fermented kimchi was sterilized at a high temperature of 95⁰C for 30 minutes to kill the lactic acid bacteria. As a result, the inherent texture of fermented kimchi could not be kept.

[124]

[125] ComparativeExample 2

[126] The number of lactic acid bacteria in commercially available canned kimchi

(Penguin Canned Kimchi, Gumhan Co., Ltd., Korea) was measured. The canned kimchi was composed of 84.21% of Chinese cabbage cultivated in Korea, red pepper powder, radishes, garlic and ginger. The pH and the acidity (lactic acid) of the canned kimchi were found to be 3.7 and 1.04, respectively. Neither Leuconostoc bacteria nor Lactobacillus bacteria were observed. This is because the lactic acid bacteria necessary for fermentation were killed by high-temperature sterilization. As a result, the inherent texture of fermented kimchi could not be kept.

[127]

[128] Testresults 1: Storabilityofcannedkimchi

[129] The number of Leuconostoc bacteria in the Chinese cabbage canned kimchi of

Example 1 was controlled within the range of 1.0 x 10² CFU/ml to 5.0 x 10⁴ CFU/ml. In addition, the canned kimchi of Example 1 was controlled such that Lactobacillus bacteria were not present therein. The Chinese cabbage canned kimchi products of Example 1 were stored at 2O⁰C and 35⁰C for a long period of time to examine variations in the number of bacteria therein. As shown in FIG. 7, the number of Leuconostoc bacteria was maintained constant (1.0 x 10² CFU/ml), Lactobacillus bacteria were not observed and there was no marked variation in the total number of bacteria during storage at room temperature for six months. No marked variation in the number of bacteria was also shown at 35⁰C, which is an optimal temperature for the growth of lactic acid bacteria. Consequently, it has proven that the storability of the canned kimchi of Example 1 was not damaged even after long-term storage.

[130]

[131] Testresults 2 : Functionality of cannedkimchi

[132] Chitosan and vitamin C, which are ingredients with high anticarcinogenic and anti- oxidative activity, were added to the canned kimchi products of Example 1 to enhance the health functionality of the canned kimchi products.

[133]

[134] Antimutagenic activity

[135] Kimchi containing no chitosan and vitamin C was introduced into cans ('control group') Canned kimchi containing chitosan only, canned products containing vitamin C only and canned kimchi containing both chitosan and vitamin C were extracted with methanol. The functionality of the canned kimchi products was evaluated. In an Ames test using Salmonellatyphimurium TAlOO, the antimutagenic activities of the canned kimchi products against aflatoxin B _x (AFB₁, Sgma), an indirect- acting mutagen, were evaluated.

[136] First, the canned kimchi products were ljophilized and powdered. To each of the powders was added methanol in an amount 20 times greater than the powder. The mixture was stirred for 12 hours. The stirring was repeated twice. The resulting mixture was filtered and concentrated using a rotary evaporator (EYELA, Tokyo Rikakikai Co., Japan) to obtain a methanol extract. These extracts were diluted with DMSO. The dilute solutions were used to treat cells.

[137] N-methyl-N'-nitro-N-nitrosoguanidine (MNNG, Aldrich Chemical Corp., U.S.A.), a direct-acting mutagen, was dissolved in distilled water. Salmonella typhimurium TAlOO, which is a histidine-requiring strain of Salmonella typhimurium LT-2, was furnished from Dr. Ames B.N, University of California in Los Angeles (UCLA), U.S.A. and used for the following experiments.

[13S] Table 4

[Table 4]

[139] Antimutagenic activities of methanol extracts of canned kimchi products against

AFBi-induced mutation [140] Table 5

[Table 5]

[141] Antimutagenic activities of methanol extracts of canned kimchi products against MMNG-induced mutation. [142] As is apparent from Table 4, as for the low-concentration methanol extracts (1.25 mg/plate), the kimchi containing no additive showed an inhibition rate of 16% before filling into cans and an inhibition rate of 4% after filling into cans and sterilization; the group containing chitosan only showed an inhibition rate of Wc; the group containing vitamin C only showed an inhibition rate of 2%; and the group containing both chitosan and vitamin C showed an inhibition rate of 23%. As for the high- concentration methanol extracts (2.5 mg/plate), the antimutagenic activities of the kimchi containing no additive were about 37% before filling into cans and 22% after filling into cans and sterilization; and the antimutagenic activity of the group containing both chitosan and vitamin C was 46%.

[143] The antimutagenic activities of the methanol extracts of canned kimchi products against MMNG, a direct-acting mutant, were evaluated. As for the low-concentration methanol extracts (1.25 mg/plate), the antimutagenic activities of the kimchi containing no additive were 14% before filling into cans and 3% after filling into cans and sterilization; the antimutagenic activity of the group containing chitosan only was 12%; the antimutagenic activity of the group containing vitamin C only was 11%; and the antimutagenic activity of the group containing both chitosan and vitamin C was 16%. As for the high-concentration methanol extracts (2.5 mg/plate), the antimutagenic activities of the kimchi containing no additive were 33% before filling into cans and 23% after filling into cans; the antimutagenic activity of the group containing chitosan only was 33%; the antimutagenic activity of the group containing vitamin C only was 32%, which was similar to that of the group containing chitosan only; and the antimutagenic activity of the group containing both chitosan and vitamin C was about 46%. The results showed similar profiles to those obtained when AFB₁ was used as a mutant (Table 5)

[144] In conclusion, the higher the concentration of the methanol extracts, the better the inhibitory effects. The antimutagenic activities of the canned kimchi products having undergone all treatments were lower than those of the kimchi products before filling into cans. The addition of chitosan or vitamin C contributed to the improvement of antimutagenic activity to some extent. The addition of both chitosan and vitamin C led to the highest antimutagenic activity.

[145]

[146] Anticarcino genie activity

[147] The inhibitory effects of the canned kimchi products on the growth of HT-29 human colon adenocarcinoma cells and AGS human gastric adenocarcinoma cells were observed to evaluate the anticarcinogenic activity in vitro of the canned kimchi products.

[148] AGS human gastric adenocarcinoma cells furnished from the Korean cell line bank (Seoul National University College of Medicine) were cultured in RPMI 1640 supplemented with penicillin/streptomydn and 10% fetal bovine serum (FBS) in a CO ₂ incubator at 37⁰C with 5% relative humidity.

[149] For MTT assay, the cells (i.e. 100 μJt) were seeded at a density of 1 x 10⁴ cells/ml per well of a 96- well plate and 80 jΛ of a culture medium was added thereto. The samples were diluted with PBS to have different concentrations and 20 jΛ was added to each well. 20 jΛ of PBS was added to one column ('100% survival group') The plate was incubated in a CO₂ incubator at 37⁰C with 5% relative humidity for 72 hours, and the absorbance was measured at 540 nm to calculate the number of viable cells.

[150] The average value of one column from the respective wells was obtained and expressed as a percent of the average value of the control group (i.e. 100% survival group) This percent corresponds to the viability of the test group, as calculated by the following equation.

[151]

ViaUhty CA) = ^0D°f^ S™P _xl00

OD of control group

[152] Table 6 [Table 6]

[153] Anticarcinogenic activities of methanol extracts of canned kimchi products on human gastric adenocarcinoma cell lines and human colon adenocarcinoma cell lines [154] As is evident from Table 6, when the samples were used in an amount of 1.0 mg/ml in the AGS human gastric adenocarcinoma cells, the anticarcinogenic activities of the group containing no additive were 6% before filling into cans and 5% after filling into cans and having undergone pasteurization, both of which did not show much difference. In contrast, the anticardnogenic activities of the group containing chitosan only and the group containing vitamin C were 13% and 12%, respectively, which were slightly higher than those of the group containing no additive. The anticardnogenic activity of the group containing both additives was highest (24%) Smilar results were obtained when the samples were used in an amount of 2.0 mg/ml. Spedfically, the anticardnogenic activities of the group containing no additive were relatively low, i.e. 12% before filling into cans and 10% after filling into cans and having undergone pasteurization. In contrast, the anticardnogenic activities of the group containing chitosan only and the group containing vitamin C were 36% and 31%, respectively. The anticardnogenic activity of the group containing both additives was highest (48%) From these results, it can be concluded that the addition of chitosan and/or vitamin C showed inhibitory effects on the growth of cancer cells.

[155] When the samples were used in an amount of 1.0 mg/ml in theHT-29human colon adenocardnoma cells, the anticardnogenic activities of the group containing no additive were 5% before filling into cans and 4% after filling into cans and having undergone pasteurization, both of which did not show much difference. In contrast, the anticardnogenic activities of the group containing chitosan only and the group containing vitamin C were 7% and 6%, and the anticardnogenic activity of the group containing both additives was highest (13%) Smilar results were obtained when the samples were used in an amount of 2.0 mg/ml. Spedfically, the anticardnogenic activities of the group containing chitosan only and the group containing vitamin C were 12% and 8%, respectively. The anticardnogenic activity of the group containing both additives was highest (26%) From these results, it can be concluded that the addition of chitosan and/or vitamin C showed inhibitory effects on the growth of cancer cells.

[156]

[157] Testresults 3: Stability (toxicity) ofcannedkimchi

[158] To examine the safety of the canned kimchi products against heavy metals and reaction products that might be formed by the reactions of internal coating materials and the kimchi when the kimchi was stored in cans, a determination was made as to whether toxic materials were contained in the canned kimchi products. In the following test examples, epoxyphenol cans and laminated cans were used.

[159] [iω] TestExample 1: Invitrotests

[161] 1. Toxicity test using Salmonellatyphimurium TAlOO [162] The canned kimchi of Example 1 was stored for 4 months and tested for toxicity of the can materials. 2 ml of top agar was poured into a sterilized cap test tube and 100 jΛ of the strain and 100 jΛ of the diluted sample were added thereto. The mixture was mildly vortexed, poured onto a nutrient agar plate, solidified and cultured at 37⁰C for 24 or 48 hours to determine whether toxicity was observed.

[163] Table 7 [Table 7]

[164] Inhibitory effects of canned kimchi products on the growth of Salmonella typhimurium TAlOO [165] The results of Table 7 show that no toxicity was observed in the common Chinese cabbage kimchi and the Chinese cabbage kimchi in the laminated can, but the kimchi in the epoxyphenol can was very weakly toxic (< 20%)

[166] [167] 2. Cytotoxicity [168] The cytotoxicity of the canned kimchi samples was reviewed. Specifically, the toxicity of methanol extracts of the canned kimchi samples to 3T3-L1 cells as normal cells was evaluated by varying the coating materials of cans.

[169] Table 8 [Table 8]

[170] Viability of 3T3-L1 cells in culture media containing methanol extracts of canned kimchi samples

[171] As can be seen from Table 8, the normal cells showed survival rates as high as 95% at concentrations of 1.0-4.0 mg/ml in all groups. The toxicity of the methanol extracts to the normal cells was increased at concentrations higher than 4.0 mg/ml. The reason is due to the high concentrations of the methanol extracts rather than due to the different coating materials of the cans. Consequently, no toxicity to the normal cells resulting from the coating materials of the cans was found.

[172]

[173] TestExample 2: Invivotests

[174] 1. Toxicity in rats

[175] Male Sprague-Dawley (SD) rats (Korea Research Institute of Chemical Technology, Daejeon, Korea) weighing about 80g were acclimatized to the laboratory environment for six weeks. The animals were maintained at 22 ± I⁰C, 55 ± 5% relative humidity on a 12 h light-dark c}cle and supplied with a laboratory standard diet and water ad libitum prior to starting the experiment.

[176] The samples added to experimental diets were ljophilized and powdered. The ingredients of the powders were analyzed in accordance with AOAC standard test methods. Specifically, the moisture content was quantitatively analyzed by a heating/ drying method (105⁰C) under ambient pressure, the crude protein content was determined by the micro- Kj eldahl method, the crude fat content was determined by the soxhlet extraction method, and the crude ash content was determined by dry ashing (55O⁰C) Taking into consideration the contents of the proteins, fats, carbohydrates and fibers, a normal diet was prepared based on AIN-93M. The AIN-93 diet was used as a control group. Kimchi (Chonggafood, Korea) and two canned products of the kimchi using different coating materials were ljophilized and powdered. The powders were used in the same amount (5%) to prepare diets having the same energy level in consideration with the other general ingredients. As the other ingredients of the experimental diets, casein was purchased from PC&S (U.SA.); methionine, L-cystine, choline bitartrate, cellulose, corn starch, corn oil, soybean oil and tert - butylhydroquinone (TBHQ) were purchased from Sgma Chemical Corp. (U.SA.); dextrinized-corn starch mineral mixture and vitamin mixture were purchased from Dylnc (U.SA.); and sucrose were purchased from a chemical distributor in Korea.

[177] Table 9 [Table 9]

[178] Composition (%) of experimental diets [179] i) Variations in body weight and dietary intake [180] There was no large difference in increment of body weight between the normal group supplied with the AIN-93 diet only and the groups supplied with the kimchi samples. The difference in body weight between the normal group and the groups supplied with various kimchi samples was 5- 1Og four weeks after the experiments. Thereafter, the difference in body weight was less than 1Og in all groups. The average dietary intake was 25-3Og and increased gradually as the body weight of the rats increased with time (FIG. 11) This tendency was similar in all groups.

[181] [182] ii) Variations in weight of organs [183] The weights of the organs of the rats were measured to evaluate the influence of the dietary intake on the organs.

[184] Table 10 [Table 10]

[185] Influence of the canned kimchi products on the organs [186] Table 10 shows that the weights of the livers, which are responsible for detoxi- fication, of the control (kirrchi A), the epoxyphenol canned kirrchi (CK-E) and the laminated canned kirrchi groups (CK-L) were somewhat increased when compared to those of the livers of the normal group. All groups except the normal group showed similar increments in the weight of the livers. The weights of the kidneys of the groups supplied with the kirrchi, particularly the group supplied with the epoxyphenol canned kirrchi, were larger than those of the kidneys of the normal group. The weights of the spleens, which are involved in immune system responses, of the groups supplied with the kirrchi, particularly the group supplied with the epoxyphenol canned kirrchi, were larger than those of the spleens of the normal group without any significant difference. The weights of the reproductive organs of the groups supplied with the kirrchi were larger than those of the reproductive organs of the normal group without any significant difference.

[187]

[188] iii) Liver toxicity in serum

[189] The activity of serum aminotransferase was determined using a kit (Asan Pharmaceutical Co., Ltd., Korea) made according to Reitman and Frankel method in accordance with the following procedure. Alanine transaminase (containing 1,780 mg of DL-alanine and 29.2 mg of α-ketoglutaric acid per 100 ml) and 1.0 ml of a matrix were introduced into the kit. The mixture was pre-incubated at 37⁰C for 5 minutes, and then 02 ml of serum was added thereto. Alanine transaminase was reacted for 30 minutes and aspartate transaminase was reacted for 60 minutes in the pre-incubated mixture. 1.0 ml of a chromogenic reagent (containing 19.8 mg of 2,4-dinitrophenylhydrazine per 100 ml) and 1.0 ml of 04 N NaOH were sequentially added, mixed together and allowed to stand at room temperature for 10 minutes. The absorbance was measured at a wavelength of 505 nm. The activity was expressed in Karmen unit per ml of serum based on the standard calibration curve.

[190] Table 11

[Table 11]

[191] Effects of canned kimchi diets on serum aminotransferase in rats after six weeks [192] As can be seen from Table 11, the AST (Karmen unit/ml of serum) of the normal group supplied with the AIN-93 diet only was 134.8, which was similar to the AST values of the group (111.1) supplied with the commercial kimchi, the group (113.1) supplied with the epoxyphenol canned kimchi and the group (116.3) supplied with the laminated canned kimchi. The ALT (Karmen unit/ml of serum) of the normal group supplied with the AIN-93 diet only was 409, which was similar to the ALT values of the group (36.1) supplied with the commercial kimchi, the group (33.5) supplied with the epoxyphenol canned kimchi and the group (32.8) supplied with the laminated canned kimchi.

[193] [194] iv) Kidney toxicity test [195] The level of blood urea nitrogen (BUN) is increased by various factors, for example, tissue collapse by high protein intake and fasting, elimination disorders, kidney disorders and uremia. The level of blood urea nitrogen was measured using a kit (Asan Pharmaceutical Co., Ltd., Korea) made by urease testing to examine kidney toxicity. Serum was added to a urease buffer containing Ql ml of urease and 20 ml of a buffer and allowed to react at 37⁰C for 15 minutes. To the reaction mixture were added salicylic acid and alkalic hypochlorous acid. The resulting mixture was allowed to react at 37⁰C for 5 minutes to prepare indophenol. The color of the indophenol was compared with the color of a urea nitrogen reference solution (60mg/ 100ml urea-N) (Colorimetric determination) [196] The creatinine content was measured by the Jaffe modified kinetic method.

According to the kinetic method, sodium lauryl sulfate and borax are used to make proteins and carbides as complexes dark, and creatinine only is decolored with an acidic reagent to compare the colors. 3 ml of a picric acid reagent was added to serum and allowed to stand in a water bath at 37⁰C for 20 minutes. The absorbance of the serum mixture was measured. Then, two drops of an acidic reagent were added to the serum mixture and allowed to stand in a water bath at 37⁰C for 5 minutes. The absorbance of the mixture was measured. The creatinine content was determined by calculating the difference between the two absorbance values and comparing the difference with the standard value (creatinine reference solution)

[197] Table 12 [Table 12]

[198] Effects of canned kimchi diets on the level of blood urea nitrogen and creatinine content in rats after six weeks

[199] Table 12 shows that the BNU value of the control group supplied with the AIN-93 diet was 13.3 mg/dl and the BNU values of the group supplied with the commercial kimchi, the group supplied with the epoxyphenol canned kimchi and the group supplied with the laminated canned kimchi were 12.1 mg/dl, 11.7 mg/dl and 11.7 mg/ dl, respectively, which showed no significant difference with the control group. Further, the BNU values of the group supplied with the epoxyphenol canned kimchi and the group supplied with the laminated canned kimchi were similar to the BNU value of the group supplied with the commercial kimchi.

[200] The creatinine content of the control supplied with the AIN-93 diet only was 1.1 mg/ dl, and the creatinine contents of the group supplied with the commercial kimchi, the group supplied with the epoxyphenol canned kimchi and the group supplied with the laminated canned kimchi were 1.0 mg/dl, 1.2 mg/dl and 1.2 mg/dl, respectively, which fall within the normal range.

[201] [202] 2. Formation of micronuclei using mouse peripheral reticulocytes [206] Each of the canned kimchi samples, which were produced by filling kimchi into an epoxyphenol can and a laminated can, was administered orally once to ICR mice (Korea Research Institute of Chemical Technology, Daejeon, Korea) Sx hours after the administration, mitomycin C, a substance inducing the formation of micronuclei, was injected into the ICR mice to form micronuclei in the ICR mice. 48 hours after the injection, blood samples were drawn from the tail veins of the mice and the density of the micronuclei formed was measured.

[204] The ICR mice was male, aged 6-7 weeks, and weighed about 35g. The animals were supplied with a standard diet. Free ai-cess to food and water was available for this time period. The animals were maintained at 22 ± I⁰C, 55 ± 5% relative humidity on a 12 h light-dark cycle.

[205] The mitomycin C (MMC, Ql mg/ml) as a positive control group was purchased from Sgma, U. SA. and dissolved in physiological saline. The mitomycin C solution was intraperitoneally administered in an amount of Ql ml/10g-body weight to the animals. Samples were prepared using sterilized distilled water and administered orally to the mice. The amounts of the samples administered were 500 mg and 1,000 mg per 1 kg of body weight.

[206] A slide glass was pre-heated at 7O⁰C, and 10 μJl of a solution (1 mg/ml) of acridine orange in distilled water was dropped onto the center of the slide glass. The acridine orange solution was homogeneously spread on the slid glass using a glass rod to construct an acridine orange-coated slide. The acridine orange-coated slide was sealed and stored until use.

[207] 48 hours after the MMC administration, blood samples (5 μJt) were drawn from the tails of the mice, dropped onto the acridine orange-coated slide, covered with a cover glass, and allowed to stand at 4⁰C for 2 hours to sufficiently react the cells with the acridine orange. The slide was observed under a fluorescence microscope (Olympus, model U-ULH, Japan) at a magnification of 400 x. 2,000 reticulocytes were counted from type I to type III. The number of reticulocytes carrying micronuclei was counted to determine the density of the micronuclei formed.

[208] As shown in FIG. 9, the ability of the group supplied with the epoxyphenol canned kimchi and the group supplied with the laminated canned kimchi to inhibit the formation of micronuclei was better than that of the control group and was similar to that of the group supplied with the kimchi.

[209]

Industrial Applicability

[210] The canned kimchi of the present invention exhibits greatly improved storability and is advantageous in terms of sensory characteristics, functionality and stability. Therefore, the canned kirrchi of the present invention is very useful in the food industry.

Claims

[1] Canned kimchi comprising at least one main kimchi material selected from the group consisting of Chinese cabbage, leaf mustard, joung radishes, radishes, green onions, sesame leaves, Korean lettuces and Korean leeks, 001 to 1.0 part by weight of at least one additive selected from chitosan, vitamin C and grapefruit seed extract, based on 100 parts by weight of the main kimchi material, and 25 to 55 parts by weight of a filling solution containing fermentation water and at least one salted-fermented seafood selected from salted- fermented anchovy sauce and salted-fermented shrimp, based on 100 parts by weight of the main kimchi material wherein the number of Leuconostoc sp. in the canned kirrchi is in the range of 1.0 x 10 ² CFU/ml to 5.0 x 10⁴ CFU/ml.

[2] The canned kimchi according to claim 1, wherein the number of Lactobacillus sp. in the canned kimchi is less than 1.0 x 10 ¹ CFU/ml.

[3] The canned kimchi according to claim 1, wherein the salted- fermented seafood is present in an amount of 4 to 8 parts by weight, based on 100 parts by weight of the main kimchi material.

[4] The canned kimchi according to claim 1, wherein the fermentation water is prepared by adding seasoning including 1.4 to 2.8 parts by weight of garlic, 06 to 1.2 parts by weight of ginger, 1.0 to 2.0 parts by weight of sugar and 13 to 26 parts by weight of chopped radishes to salt water and fermenting the seasoning to a pH of 3.8 to 4.1.