초록 ▼

1. 1차년도 (1997년) □ 어종별 정밀 선도판 정법의 탐색 및 비파괴 측정법 탐색 ○ 기존 신선도 측정방법 검토 - 화학적 방법(VBN, TMA, K값 등) - 물리적, 미생물학적 방법(총균수 등) ○ 새로운 선도 측정방법 검토 - ATPase 활성 - 단백질 분해양상 - Diamine류 검출 - DNA, RNA 분해양상 ○ 비파괴에 의한 선도측정법 검토 - Light Technique - 화상분석 등2. 2차년도 (1998년) □ 어종별 정밀 선도 측정법의 개발 및 비파괴 선도

1. 1차년도 (1997년) □ 어종별 정밀 선도판 정법의 탐색 및 비파괴 측정법 탐색 ○ 기존 신선도 측정방법 검토 - 화학적 방법(VBN, TMA, K값 등) - 물리적, 미생물학적 방법(총균수 등) ○ 새로운 선도 측정방법 검토 - ATPase 활성 - 단백질 분해양상 - Diamine류 검출 - DNA, RNA 분해양상 ○ 비파괴에 의한 선도측정법 검토 - Light Technique - 화상분석 등2. 2차년도 (1998년) □ 어종별 정밀 선도 측정법의 개발 및 비파괴 선도측정 시스템 개발 ○ 적정 비파괴 선도측정 시스템 선정 및 최적화 연구 - Image analysis의 적용 가능성 정밀검토 - Light technique을 이용한 비파괴 측정법 개발 ○ 새로운 선도 측정방법 확립 및 최적화 연구 - Ca-ATPase 활성이 간이 측정 장치 및 대중성 어종별 data base 개발 ○ 선도 신속측정 시스템 개발 - Enzyme assay를 이용한 선도 신속 측정법 개발3. 3차년도(1999년) □ 객관적 선도 및 비파괴 방법에 의한 정밀 선도 측정방법 확립 ○ 비파괴 방식에 의한 선도 측정기술의 실용화를 위한 보완 연구 ○ 선도 간이측정장치의 성능개선 연구 ○ 선도 신속측정시스템 실용화 연구 - 현장 유통 시료의 전처리방법 개선 및 어종별 자료축적 연구 ○ 개발기술의 산업화 현장 적용연구

Abstract ▼

I. Title Studies on the Development of Freshness Evaluation Technology in Fishery Products II. Objective and Significance By agreement of GATT/BOP on October in 1989, all fishery products had been imported freely into Korea. And fishing areas were reduced increasingly because of ocean pollution

I. Title Studies on the Development of Freshness Evaluation Technology in Fishery Products II. Objective and Significance By agreement of GATT/BOP on October in 1989, all fishery products had been imported freely into Korea. And fishing areas were reduced increasingly because of ocean pollution accompanied with great governmental industrialization policy, land reclamation. Therefore, it was seriously expected that large amounts of fishery products were imported into Korea. Actually, yearly importing amount of fresh and frozen fishery products have been increased since 1997. The systems for import inhibition of those products was still not ready domestically. For inspection of those products organoleptic test which known to be very subjective evaluation method were regulated in fishery product inspection law. Therefore, it was needed that technical barrier for import inhibition of fishery products were made promptly. In this study, it was carried out to develop the objective technology for freshness evaluation of fishery products including non-destructive metohds such as use of NIR spectroscopy, image analysis system, aroma scan system. III. Results and discussins 1. Invetigation of establshed methods for freshness evaluation 1) Chemical methods VBN content and K-value of alaska pollack, mackerel, squid and shortneck clam during storage at 5℃ was investigated. Though same stage of freshness, VBN content was different by fish species. therefore, it was thought that indiscriminate application of established specification of VBN content to all species of fishery products was not realistic. By the way, K-value of alaska pollack and mackerel was comparatively agree with established judging standard of freshness 2) Physical methods Water holding capacity of alaska pollack, mackerel, squid and shortneck clam during storage at 5℃ was decreased with elapse of storage time. But, differences among the freshness stage such as fresh, moderately putrid and putrid were too small to utilize as a quality index. 3) Microbial methods As a results of determining total viable count of mackerel at 30℃, Microbial content was not exceed $10^3$ in the putrid samples. By the established standard, it was noted that $10^6$ of microbes meant in initial stage of putrifaction. 2. Studies on the new methods for freshness evaluation 1) Study on the measuring Ca-ATPase activity To develop the new technology for freshness evaluation, relationship between Mf Ca-ATPase activity and quality attributes including soluble protein, water holding capacity and breaking strenth of alaska pollack, sardine and squid at different storage temperature was investigated. Significant correlationship(r=0.96) was shown, suggesting that Ca-ATPase activity was usable index for frshness evaluation. 2) SDS-PAGE patterns SDS-PAGE patterns of alaska pollack, sardine and squid at different storage temperature was observed. Protein subunit of samples stored below -10℃ was not changed significantly but the biological aspects and quality attributes of them were seriously changed. 3) Changes of DNA & RNA pattern DNA and RNA content of mackerel stored at 30℃ was examined. During the storage, partial fragmentation of DNA and RNA was observed, but their total content was not changed. Reaction products were observed in genomic DNA of alaska pollack. But to use quality index further study about reaction condition of PCR, sequence and length of primer and slection of amplifying position were requried. 4) Changes of polyamines As a result of determining putrescine, cadaverine and spermine with different stage of frshness, putrescine and cadaverine content in fresh and moderately putrid samples was very low, but high in putrid sample. Spermine was decreased with elapse of storage time. 3. Investigation of non-destructive freshness evaluation systems 1) Applicatin of aroma scan system Aroma scan system was applied to alaska pollack and mackerel with different stage of freshness such as fresh, moderately putrid and putrid. Significant differences in each group was not observed, guggesting that aroma scan system was not fitted to freshness evaluation system. 2) Application of image analysis system The changes of skin, eyes and gill in mackerel with different stage of freshness were investigated by image analysis system. Although the canges of skin and eyes was observed, but quantification of that was very difficultwhile the change of color in gill was recognized significantly 3) Application of near-infrared(NIR) spectroscophy Because NIR spectrum of mackerel with different temperature showed different patterns of energy absortion it was very important to keep temperature constantly in measuring NIR spectrum. K-value was lowest in the hindhead part of mackerel. 4. Development of objective technolgy for freshness evaluation 1) Development of a simple Ca-ATPase activity measuring system Small sized simple system for measuring Ca-ATPase activity was developed to determine freshness in the field. The size of this system was 230(W) ×330(L) ×100(H)mm and attached specially holographic gratings, and CCD detecter. 2) Development of reagents for measuring Ca-ATPase activity For measuring Ca-ATPase activity using simple system, reaction reagent(buffer A) which was stable in room temperature and coloring reagent(buffer B) were developed 3) Development of new technology for measuring Ca-ATPase activity A simple and accurate technology for measuring Ca-ATPase activity using fish muscle was developed. Operating procedure was composed of weighing muscle, dilution, reaction with with buffer A, inactivation, filtration, colorization and activity measurement. 10 samples could be analyzed in 1 hour by this simle method 4) Examination of conditions for utilizing new technology To utilize the new simple method stability of buffers, proper concentrations of muscle Ca-ATPase activity and stability at various temperature were investigated. Buffer A was stable for 48 hours at room temperature and proper concentration of muscle Ca-ATPase was shown as a dilution ratio of 40 ∼ 100. As muscle Ca-ATPase activity was dependent on temperature, it was thought that developed technology could be utilized for freshness evaluation of fishery products. 5) Relationship between Mf Ca-ATPase activity and muscle Ca-ATPase activity As a result of investigating the relationship between Mf Ca-ATPase activity and muscle Ca-ATPase activity. above 0.96 of correlationship was found, suggesting measuring the muscle Ca-ATPase activity could be utilized as a objective index of freshness. 6) Freshness evaluation of imported fishes by muscle Ca-ATPase activity measuring method As a result of freshness evaluation of imported frozen alaska pollack and mackerel which were distributed in the local market, freshness of them shown to be seriously low. 7) Improvement of muscle Ca-ATPase activity measuring system Simple Ca-ATPase activity measuring system was reduced to the size of 200(W) ×250(L) ×85(H)mm and optical density of reaction solution could be checked as it was standing in tube. As electronic control system was attached inside, one-touch measurement could be carried out. 5. Development of non-destructive freshness evaluation technology 1) Non-deatructive technology using NIR NIR spectra of mackerel during storage at 5℃ was obtained and differenciated 2ndly. Calibration equation of K-value vs NIR spectrum wea shown to be r= 0.914 and standard error 2.547 at term 8. High correlation coefficient of 0.959 was exhibited when plotted to actual K-value between calculated K-value by the calibration equation. As a result of evaluating mackerel in the market by calibration equation 0.940 of correlation coefficient was found. Therefore, it was thought that NIR technolgy was applicable to non-destructive freshness evaluation of mackerel. 6. Development of rapid freshness measuring technology Relatively high correlationship(r=0.92) was recognized between real microbes y aerobic plate count and relative light unit(RLU) by ATP bioluminescence assay in mackerel skin. And microbials attached to mackerel skin were also measurable by the correlationship equation. Only thirty minutes were needed to determine the microbes in the mackerel skin by ATP bioluminescence assay. 7. Review on the use of biosensor for freshness evaluation Many biosensors such as enzyme biosensor for quantification of hypoxanthine and inosine, amperometric biosensor for quantification of histamine, putricine and cadaverine, and biosensors for octopine, ornithine and arginine in invertebrates were developed to determine the freshness of fishery products.

목차 Contents

• 요 약 문...1
  Summary...9
  제 1 장 서 론...25
  제 2 장 재료 및 방법...29
  제 1 절 실험재료...29
  1. 원료수산물...29
  제 2 절 실험방법...29
  제 2 절 실험방법...29
  1. Ca-ATPase활성의 측정...29
  2. 보수력의 측정...29
  3. Diamine류의 정량...30
  4. SDS-PAGE에 의한 근원섬유 단백질의 subunit 분석...30
  5. Genome DNA의 정제...30
  6. RAPD(Random Amplified polymorpic DNA) 분석...30
  7. Aroma Scan System을 이용한 선도변화 측정...31
  8. 화상분석(Image Analysis system)...31
  9. NIR을 이용한 신선도 측정...31
  10. ATP bioluminescence assay...31
  11. ATP calibration curves...32
  12. APC(aerobic plate count)...32
  13. Ca-ATPase activity 간이측정장치...32
  14. K-value의 측정...33
  15. Polyamine류 측정...33
  16. Soluble protein의 측정...34
  17. Isolated protein 함량...34
  제 3 장 결과 및 고찰...35
  제 3 장 결과 및 고찰...35
  제 3 장 결과 및 고찰...35
  제 1 절 기존의 신선도 측정방법 검토...35
  1. 화학적 선도 측정법...35
  2. 물리적 선도 측정법...38
  3. 미생물학적 선도 측정법...41
  제 2 절 새로운 선도측정방법 검토...42
  제 2 절 새로운 선도측정방법 검토...42
  1. Mf Ca-ATPase 활성 측정법 검토...42
  2. 선도저하에 따른 단백질 분해양상 측정...72
  3. DNA, RNA변화 양상 측정...77
  4. 선도저하에 따른 Diamine류 검출...84
  제 3 절 비파괴 방식에 의한 선도 측정법 탐색...86
  제 3 절 비파괴 방식에 의한 선도 측정법 탐색...86
  1. Aroma Scan System에 의한 선도측정 가능성 검토...86
  2. Image analysis System에 의한 선도 측정 가능성 검토...90
  3. NIR에 의한 선도측정법의 검토...95
  제 4 절 객관적 신선도 측정기술 개발...98
  제 4 절 객관적 신선도 측정기술 개발...98
  1. Ca-ATPase 활성의 간이 측정장치 개발...98
  2. Ca-ATPase 활성측정용 반응시약 및 발색시약 개발...98
  3. Ca-ATPase 활성의 간이 측정법 개발...98
  4. 개량된 Ca-ATPase 활성 측정법의 검토...101
  5. Mf Ca-ATPase 활성과 Ca-ATPase 활성의 간이 측정법과의 관계...103
  6. 근육 Ca-ATPase활성법에 의한 수입수산물의 신선도 평가...110
  7. 근육 Ca-ATPase 활성 간이 측정장치의 개선...112
  제 5 절 비파괴 방식에 의한 신선도 측정기술 개발...114
  제 5 절 비파괴 방식에 의한 신선도 측정기술 개발...114
  1. L