JP4556497B2 - Non-destructive evaluation method of freshness of fresh seafood, animal meat or poultry meat, estimation method and kit of remaining days of fresh fish - Google Patents
Non-destructive evaluation method of freshness of fresh seafood, animal meat or poultry meat, estimation method and kit of remaining days of fresh fish Download PDFInfo
- Publication number
- JP4556497B2 JP4556497B2 JP2004165384A JP2004165384A JP4556497B2 JP 4556497 B2 JP4556497 B2 JP 4556497B2 JP 2004165384 A JP2004165384 A JP 2004165384A JP 2004165384 A JP2004165384 A JP 2004165384A JP 4556497 B2 JP4556497 B2 JP 4556497B2
- Authority
- JP
- Japan
- Prior art keywords
- fresh
- freshness
- color
- coloring
- value
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- 235000014102 seafood Nutrition 0.000 title claims description 60
- 238000000034 method Methods 0.000 title claims description 51
- 241000251468 Actinopterygii Species 0.000 title claims description 45
- 235000013372 meat Nutrition 0.000 title claims description 21
- 235000013594 poultry meat Nutrition 0.000 title claims description 20
- 241001465754 Metazoa Species 0.000 title claims description 19
- 238000011156 evaluation Methods 0.000 title claims description 8
- 238000004040 coloring Methods 0.000 claims description 53
- 108010093894 Xanthine oxidase Proteins 0.000 claims description 51
- 102100033220 Xanthine oxidase Human genes 0.000 claims description 51
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical compound O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 claims description 40
- LRFVTYWOQMYALW-UHFFFAOYSA-N 9H-xanthine Chemical compound O=C1NC(=O)NC2=C1NC=N2 LRFVTYWOQMYALW-UHFFFAOYSA-N 0.000 claims description 33
- 238000006243 chemical reaction Methods 0.000 claims description 22
- 125000003831 tetrazolyl group Chemical group 0.000 claims description 22
- 238000003860 storage Methods 0.000 claims description 21
- UGQMRVRMYYASKQ-UHFFFAOYSA-N Hypoxanthine nucleoside Natural products OC1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 UGQMRVRMYYASKQ-UHFFFAOYSA-N 0.000 claims description 20
- 229940075420 xanthine Drugs 0.000 claims description 17
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 claims description 16
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 claims description 16
- 229940116269 uric acid Drugs 0.000 claims description 16
- 235000013305 food Nutrition 0.000 claims description 13
- 239000002798 polar solvent Substances 0.000 claims description 13
- 239000003086 colorant Substances 0.000 claims description 12
- 238000011161 development Methods 0.000 claims description 11
- 239000000203 mixture Substances 0.000 claims description 11
- 230000001066 destructive effect Effects 0.000 claims description 10
- 230000000007 visual effect Effects 0.000 claims description 9
- 230000008014 freezing Effects 0.000 claims description 8
- 238000007710 freezing Methods 0.000 claims description 8
- 238000005259 measurement Methods 0.000 claims description 8
- 238000011088 calibration curve Methods 0.000 claims description 7
- 230000008878 coupling Effects 0.000 claims description 7
- 238000010168 coupling process Methods 0.000 claims description 7
- 238000005859 coupling reaction Methods 0.000 claims description 7
- 235000015278 beef Nutrition 0.000 claims description 2
- 244000144977 poultry Species 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims 2
- 235000019688 fish Nutrition 0.000 description 36
- 235000015170 shellfish Nutrition 0.000 description 22
- 238000002835 absorbance Methods 0.000 description 14
- 102000004190 Enzymes Human genes 0.000 description 13
- 108090000790 Enzymes Proteins 0.000 description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 12
- 239000002904 solvent Substances 0.000 description 12
- ZKHQWZAMYRWXGA-UHFFFAOYSA-N Adenosine triphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)C(O)C1O ZKHQWZAMYRWXGA-UHFFFAOYSA-N 0.000 description 11
- 239000000243 solution Substances 0.000 description 10
- 239000000758 substrate Substances 0.000 description 10
- GRSZFWQUAKGDAV-UHFFFAOYSA-N Inosinic acid Natural products OC1C(O)C(COP(O)(O)=O)OC1N1C(NC=NC2=O)=C2N=C1 GRSZFWQUAKGDAV-UHFFFAOYSA-N 0.000 description 9
- 230000018109 developmental process Effects 0.000 description 9
- 235000013902 inosinic acid Nutrition 0.000 description 9
- VMGAPWLDMVPYIA-HIDZBRGKSA-N n'-amino-n-iminomethanimidamide Chemical compound N\N=C\N=N VMGAPWLDMVPYIA-HIDZBRGKSA-N 0.000 description 9
- 230000008569 process Effects 0.000 description 9
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 9
- 239000000523 sample Substances 0.000 description 8
- 238000000354 decomposition reaction Methods 0.000 description 7
- 238000009826 distribution Methods 0.000 description 7
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 6
- NTYJJOPFIAHURM-UHFFFAOYSA-N Histamine Chemical compound NCCC1=CN=CN1 NTYJJOPFIAHURM-UHFFFAOYSA-N 0.000 description 6
- 239000011521 glass Substances 0.000 description 6
- 238000010521 absorption reaction Methods 0.000 description 5
- UDMBCSSLTHHNCD-KQYNXXCUSA-N adenosine 5'-monophosphate Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(O)=O)[C@@H](O)[C@H]1O UDMBCSSLTHHNCD-KQYNXXCUSA-N 0.000 description 5
- 210000003205 muscle Anatomy 0.000 description 5
- XTWYTFMLZFPYCI-KQYNXXCUSA-N 5'-adenylphosphoric acid Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP(O)(O)=O)[C@@H](O)[C@H]1O XTWYTFMLZFPYCI-KQYNXXCUSA-N 0.000 description 4
- XTWYTFMLZFPYCI-UHFFFAOYSA-N Adenosine diphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(O)=O)C(O)C1O XTWYTFMLZFPYCI-UHFFFAOYSA-N 0.000 description 4
- 241001553014 Myrsine salicina Species 0.000 description 4
- 229950006790 adenosine phosphate Drugs 0.000 description 4
- 238000006911 enzymatic reaction Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 150000001298 alcohols Chemical class 0.000 description 3
- 230000002528 anti-freeze Effects 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
- 229960001340 histamine Drugs 0.000 description 3
- 238000007726 management method Methods 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- -1 methanol and ethanol Chemical class 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 241000555825 Clupeidae Species 0.000 description 2
- UDMBCSSLTHHNCD-UHFFFAOYSA-N Coenzym Q(11) Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(O)=O)C(O)C1O UDMBCSSLTHHNCD-UHFFFAOYSA-N 0.000 description 2
- 241001149724 Cololabis adocetus Species 0.000 description 2
- 241000287828 Gallus gallus Species 0.000 description 2
- 102000004316 Oxidoreductases Human genes 0.000 description 2
- 108090000854 Oxidoreductases Proteins 0.000 description 2
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- WERYXYBDKMZEQL-UHFFFAOYSA-N butane-1,4-diol Chemical compound OCCCCO WERYXYBDKMZEQL-UHFFFAOYSA-N 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 235000005911 diet Nutrition 0.000 description 2
- 230000037213 diet Effects 0.000 description 2
- 238000011059 hazard and critical control points analysis Methods 0.000 description 2
- 230000001771 impaired effect Effects 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 238000003908 quality control method Methods 0.000 description 2
- 235000019512 sardine Nutrition 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- AUHDWARTFSKSAC-HEIFUQTGSA-N (2S,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)-2-(6-oxo-1H-purin-9-yl)oxolane-2-carboxylic acid Chemical compound [C@]1([C@H](O)[C@H](O)[C@@H](CO)O1)(N1C=NC=2C(O)=NC=NC12)C(=O)O AUHDWARTFSKSAC-HEIFUQTGSA-N 0.000 description 1
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 1
- CVOFKRWYWCSDMA-UHFFFAOYSA-N 2-chloro-n-(2,6-diethylphenyl)-n-(methoxymethyl)acetamide;2,6-dinitro-n,n-dipropyl-4-(trifluoromethyl)aniline Chemical compound CCC1=CC=CC(CC)=C1N(COC)C(=O)CCl.CCCN(CCC)C1=C([N+]([O-])=O)C=C(C(F)(F)F)C=C1[N+]([O-])=O CVOFKRWYWCSDMA-UHFFFAOYSA-N 0.000 description 1
- ZKHQWZAMYRWXGA-KQYNXXCUSA-J ATP(4-) Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)[C@H]1O ZKHQWZAMYRWXGA-KQYNXXCUSA-J 0.000 description 1
- ZKHQWZAMYRWXGA-KQYNXXCUSA-N Adenosine triphosphate Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)[C@H]1O ZKHQWZAMYRWXGA-KQYNXXCUSA-N 0.000 description 1
- 241000962514 Alosa chrysochloris Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 241000624562 Cololabis saira Species 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 239000007995 HEPES buffer Substances 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- UGQMRVRMYYASKQ-KQYNXXCUSA-N Inosine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C2=NC=NC(O)=C2N=C1 UGQMRVRMYYASKQ-KQYNXXCUSA-N 0.000 description 1
- 229930010555 Inosine Natural products 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- KDCGOANMDULRCW-UHFFFAOYSA-N Purine Natural products N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 description 1
- 241000736084 Scomber japonicus Species 0.000 description 1
- 241000282898 Sus scrofa Species 0.000 description 1
- 241000234314 Zingiber Species 0.000 description 1
- 235000006886 Zingiber officinale Nutrition 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 230000002745 absorbent Effects 0.000 description 1
- 229920000122 acrylonitrile butadiene styrene Polymers 0.000 description 1
- LNQVTSROQXJCDD-UHFFFAOYSA-N adenosine monophosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(CO)C(OP(O)(O)=O)C1O LNQVTSROQXJCDD-UHFFFAOYSA-N 0.000 description 1
- 239000003708 ampul Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 238000009529 body temperature measurement Methods 0.000 description 1
- 230000006652 catabolic pathway Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 238000009614 chemical analysis method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007515 enzymatic degradation Effects 0.000 description 1
- CCIVGXIOQKPBKL-UHFFFAOYSA-M ethanesulfonate Chemical compound CCS([O-])(=O)=O CCIVGXIOQKPBKL-UHFFFAOYSA-M 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000008397 ginger Nutrition 0.000 description 1
- 238000005469 granulation Methods 0.000 description 1
- 230000003179 granulation Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 229960003786 inosine Drugs 0.000 description 1
- 229940028843 inosinic acid Drugs 0.000 description 1
- 239000004245 inosinic acid Substances 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 125000004433 nitrogen atom Chemical group N* 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000010979 pH adjustment Methods 0.000 description 1
- 229920000058 polyacrylate Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 229920000915 polyvinyl chloride Polymers 0.000 description 1
- 239000004800 polyvinyl chloride Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000000275 quality assurance Methods 0.000 description 1
- 238000013441 quality evaluation Methods 0.000 description 1
- 150000003242 quaternary ammonium salts Chemical group 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000007226 seed germination Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000005846 sugar alcohols Polymers 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 235000019640 taste Nutrition 0.000 description 1
- 235000019583 umami taste Nutrition 0.000 description 1
- 208000019206 urinary tract infection Diseases 0.000 description 1
- 238000011179 visual inspection Methods 0.000 description 1
Images
Landscapes
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Description
本発明は、生鮮魚介類、獣肉または家禽肉の鮮度の非破壊的評価法、生鮮魚類の生可食残存日数の推定方法およびそのためのキットに関するものである。 The present invention relates to a non-destructive evaluation method for freshness of fresh seafood, animal meat or poultry meat, a method for estimating the number of days remaining for fresh edible food, and a kit therefor.
食品の安全性確保や品質の保証に消費者の関心が高まっている。生産者等には原料の生産・流通に関して安全性を確保し、併せてその情報を開示していくトレーサビリティシステムの構築が求められている。食肉や農産物に関しては牛肉トレーサビリティ法の制定、SEICAの普及、ICタグ等の試みが実施されているが、生鮮魚介類に関してはトレーサビリティシステムは端を発したばかりと言える。 Consumers are increasingly interested in ensuring food safety and quality assurance. Producers, etc. are required to establish a traceability system that ensures safety in the production and distribution of raw materials and also discloses the information. The beef traceability law has been enacted for meat and agricultural products, SEICA has been widely used, and IC tags have been tried. However, for fresh seafood, the traceability system has just started.
これまで提案されたトレーサビリティシステムは消費者の安全・安心というよりは、むしろ生産者側や流通業者の生産物に対する管理に重点が置かれている。魚介類を生で食する我が国では、トレーサビリティに関する消費者のニーズとして、生産履歴(どこで漁獲・養殖されたか、生簀の情報、薬剤使用の有無など)や流通履歴(加工場所、流通及び配送過程など)よりは、あと何日生で食べられるか?一番おいしい時期はいつか?に大きな関心があるように考えられる。 The traceability system proposed so far focuses on the management of producers and distributors' products rather than the safety and security of consumers. In Japan, where fish and shellfish are eaten raw, the needs of consumers regarding traceability include production history (where fished and farmed, ginger information, presence or absence of drug use, etc.) and distribution history (processing location, distribution and delivery process, etc.) ) How many days can you eat more than? When is the most delicious time? There seems to be a great interest in.
生鮮魚介類、獣肉または家禽肉(以下、「生鮮魚介類等」と略称することがある)の品質保持技術は現在のところ、冷蔵・冷凍技術をおいて他にない。従って、温度管理をいかに厳格にするかによって最高の品質管理が可能になるものと考えられる。 At present, there is no other technology for maintaining the quality of fresh seafood, animal meat or poultry meat (hereinafter sometimes abbreviated as “fresh seafood and the like”), including refrigeration and freezing techniques. Therefore, it is considered that the highest quality control is possible depending on how strict the temperature control is.
しかし、生鮮魚介類等が流通過程の途中で品質が劣化するような温度にさらされた場合であっても、生鮮魚介類等の保蔵中の任意の時点での品質状態を目視で即座に判断することは難しい。 However, even when fresh fish and shellfish are exposed to temperatures that degrade their quality during the distribution process, the quality status at any point during storage of fresh fish and shellfish is immediately judged visually. Difficult to do.
生鮮魚介類の鮮度が今どのような状態にあるのかを見極める計測法や装置は、種々開発実用化されている(特許文献1〜5)。しかしながら従来技術では、検体として、魚肉等を採取し試料溶液等を調製する必要があった。これらの従来技術は、破壊検査法であり、生鮮魚介類の商品価値は当然損なわれる。
Various measuring methods and devices for determining the state of freshness of fresh seafood are being developed and put to practical use (
一方、生鮮魚介類の品質管理や安全管理にHACCPを導入することは世界の趨勢である。HACCPの基準に基づけば生鮮魚介類の品質は生成ヒスタミン量によって判断される。ヒスタミンはバクテリアの生産する脱炭酸酵素によって遊離ヒスチジンから生成される。従って、魚介類を生で食べるレベルの品質評価法としてヒスタミン法はふさわしくないことは明らかである。 On the other hand, the introduction of HACCP for quality control and safety management of fresh seafood is a global trend. Based on HACCP standards, the quality of fresh seafood is judged by the amount of histamine produced. Histamine is produced from free histidine by decarboxylase produced by bacteria. Therefore, it is clear that the histamine method is not suitable as a quality evaluation method for the level of eating seafood.
また、我が国では生鮮魚介類は生食されることが多く、生で食べられる期間はあと何日かという生可食残存日数も併せて推定できればよいが、このような生可食残存日数を非破壊的に推定する方法は知られていなかった。
本発明が解決しようとする手段は、生鮮魚介類等の鮮度を非破壊で評価する方法、生鮮魚類の生可食残存日数の推定方法およびそのためのキットを提供することである。 The means to be solved by the present invention is to provide a method for non-destructively evaluating the freshness of fresh fish and shellfish, a method for estimating the number of days remaining for fresh edible food, and a kit therefor.
(1) ヒポキサンチンがキサンチンオキシダーゼによってキサンチンと尿酸に分解する反応に共役して発色する発色剤、ヒポキサンチン、キサンチンオキシダーゼおよび不凍性の極性溶媒を含む発色剤組成物を透明容器に封入して発色媒体とし、該発色媒体を生鮮魚介類、獣肉または家禽肉と一緒に保蔵し、任意の時点で発色媒体の発色強度を測定し、測定した発色強度の強弱をもとに生鮮魚介類、獣肉または家禽肉の鮮度を推定することを特徴とする生鮮魚介類、獣肉または家禽肉の鮮度の非破壊的評価方法。
(2) 前記発色剤がテトラゾリウム塩であることを特徴とする(1)に記載の生鮮魚介類、獣肉または家禽肉の鮮度の非破壊的評価方法。
(3) 前記鮮度の推定を、予め発色強度とK値の検量線を作成し、測定した発色強度からK値を求めることを特徴とする(1)または(2)に記載の生鮮魚介類、獣肉または家禽肉の鮮度の非破壊的評価方法。
(4) 前記発色強度の測定および鮮度の推定を、色見本を用いて目視法で行うことを特徴とする(1)ないし(3)のいずれか1項に記載の生鮮魚介類、獣肉または家禽肉の鮮度の非破壊的評価方法。
(5) ヒポキサンチンがキサンチンオキシダーゼによってキサンチンと尿酸に分解する反応に共役して発色する発色剤、ヒポキサンチン、キサンチンオキシダーゼおよび不凍性の極性溶媒を含む発色剤組成物を透明容器に封入して発色媒体とし、該発色媒体を生鮮魚類が保蔵された保蔵容器内に配置し、任意の時点で発色媒体の発色強度を測定し、測定した発色強度の強弱をもとに生鮮魚類のK値を推定し、式(1)
RDVt=[Tt(K2−K’)/(K2−K0)](1)
(ただし式(1)中、RDVt:t℃貯蔵における生可食残存日数
Tt:t℃保存における生食としての許容限界日数
K2:生食として食せる許容限界のK値
K0:非常に新鮮な生鮮魚類のK値
K’:測定した時点のK値の推定値を示す。)
で表される式により生可食残存日数を算定することを特徴とする生鮮魚類の生可食残存日数の推定方法。
(6) 前記発色剤がテトラゾリウム塩であることを特徴とする(5)に記載の生鮮魚類生可食残存日数の推定方法。
(7) 前記発色強度の測定および鮮度の推定を、色見本を用いて目視法で行うことを特徴とする(5)または(6)に記載の生鮮魚類の生可食残存日数の推定方法。
(8) ヒポキサンチンがキサンチンオキシダーゼによってキサンチンと尿酸に分解する反応に共役して発色する発色剤、ヒポキサンチン、キサンチンオキシダーゼおよび不凍性の極性溶媒を含む発色剤組成物を透明容器に封入したことを特徴とする生鮮魚介類、獣肉または家禽肉の鮮度推定用のキット。
(9) ヒポキサンチンがキサンチンオキシダーゼによってキサンチンと尿酸に分解する反応に共役して発色する発色剤、ヒポキサンチン、キサンチンオキシダーゼおよび不凍性の極性溶媒を含む発色剤組成物を透明容器に封入したことを特徴とする生鮮魚類生可食残存日数推定用のキット。
(10) 前記発色剤がテトラゾリウム塩であることを特徴とする(8)または(9)に記載のキット。
(1) A coloring agent composition containing hypoxanthine, xanthine oxidase, and an antifreezing polar solvent , which is colored by coupling to a reaction in which hypoxanthine is decomposed into xanthine and uric acid by xanthine oxidase, is sealed in a transparent container. The coloring medium is stored together with fresh seafood, animal meat or poultry meat, the coloring intensity of the coloring medium is measured at an arbitrary time, and fresh seafood, animal meat is measured based on the strength of the measured coloring intensity. Alternatively, a non-destructive evaluation method for freshness of fresh seafood, animal meat or poultry meat, characterized by estimating freshness of poultry meat.
(2) The nondestructive evaluation method for freshness of fresh seafood, animal meat or poultry meat according to (1), wherein the color former is a tetrazolium salt.
( 3 ) The fresh fish and shellfish according to (1) or (2) , wherein the freshness is estimated by preparing a calibration curve of the color intensity and the K value in advance, and obtaining the K value from the measured color intensity. A non-destructive method of assessing the freshness of animal or poultry meat.
( 4 ) The fresh fishery products, meat or poultry according to any one of (1) to ( 3 ), wherein the color intensity is measured and the freshness is estimated by a visual method using a color sample. A non-destructive method for assessing the freshness of meat.
( 5 ) A coloring agent composition containing hypoxanthine, xanthine oxidase and a non-freezing polar solvent is colored in a transparent container, which is coupled with a reaction in which hypoxanthine is decomposed into xanthine and uric acid by xanthine oxidase. The coloring medium is placed in a storage container in which fresh fish is stored, and the coloring intensity of the coloring medium is measured at an arbitrary point in time, and the K value of fresh fish is determined based on the strength of the measured coloring intensity. Estimate and formula (1)
RDV t = [T t (K 2 −K ′) / (K 2 −K 0 )] (1)
(In the formula (1), RDV t : remaining days of raw edible in storage at t ° C.
T t : Tolerable limit number of days as raw food when stored at t ° C
K 2 : K value of allowable limit that can be eaten as raw food
K 0 : K value of very fresh fresh fish
K ′: Indicates an estimated value of the K value at the time of measurement. )
A method for estimating the number of days remaining in raw edible for fresh fish, characterized in that the number of days remaining in raw edible is calculated by an expression represented by:
(6) the method of estimating the fresh fish raw edible residual days according to the color former is characterized in that it is a tetrazolium salt (5).
( 7 ) The method for estimating the number of days remaining for fresh fish edible, according to (5) or (6) , wherein the color intensity is measured and the freshness is estimated by a visual method using a color sample.
( 8 ) A coloring agent composition containing hypoxanthine, xanthine oxidase, and an antifreezing polar solvent that is colored by coupling to a reaction in which hypoxanthine is decomposed into xanthine and uric acid by xanthine oxidase is sealed in a transparent container. A kit for estimating the freshness of fresh seafood, animal meat or poultry meat.
( 9 ) A coloring agent composition containing hypoxanthine, xanthine oxidase, and an antifreezing polar solvent that is colored by coupling to the reaction of hypoxanthine decomposed into xanthine and uric acid by xanthine oxidase is sealed in a transparent container. A kit for estimating the number of days remaining for raw fish edible.
( 10 ) The kit according to ( 8 ) or ( 9 ), wherein the color former is a tetrazolium salt.
請求項1ないし5に記載された本発明方法により、生鮮魚介類等の鮮度を非破壊的に評価できるので、生鮮魚介類等の商品価値を損ねることがなく、従来の目視法に比べて生鮮魚介類等の鮮度をより客観的に評価をすることができる。特に請求項5に記載された発明は、目視法により生鮮魚介類等の鮮度を簡便に推定することができる。
According to the method of the present invention described in
請求項6ないし9に記載された本発明方法により、簡便な方法で、生鮮魚類の生可食残存日数を推定することができる。特に請求項9に記載された発明は、目視法により生鮮魚類の生可食残存日数を簡便に推定することができる。 According to the method of the present invention described in claims 6 to 9, the number of days of fresh edible life of fresh fish can be estimated by a simple method. In particular, the invention described in claim 9 can easily estimate the number of days remaining for fresh edible fish by visual inspection.
請求項10ないし13に記載された本発明のキットにより、生鮮魚介類等の鮮度を非破壊的に評価し、また生鮮魚類生可食残存日数を推定することができる。
According to the kit of the present invention described in
生鮮魚介類等の鮮度指標K値は生鮮魚介類等の筋肉中に含まれるアデノシン三リン酸(以下、「ATP」と略称する)の分解過程から関数として導き出される。ATPはアデノシン二リン酸(以下、「ADP」と略称する)を経てアデノシン一リン酸(アデニル酸、以下、「AMP」と略称する)へと脱リン酸される。続いて、塩基部分の構造の一部が変化してイノシン酸(以下、「IMP」と略称する)となる。IMPはカツオ節の旨み成分としてよく知られている。ATPの分解に際してここまでの過程は比較的早い。IMPの蓄積は旨み成分の蓄積であり、ここまでの分解は生鮮魚介類等の美味しさにとって重要な過程である。IMP は脱リン酸されてイノシン(以下、「HxR」と略称する)となり、さらに酵素反応を受けてヒポキサンチン(以下、「Hx」と略称する)となる。Hxはさらに酵素分解を受けてキサンチン・尿酸へと分解していく。この一連の過程の中で、IMPの分解速度が比較的小さいため、
[(HxR+Hx)/(ATP+ADP+AMP+IMP+HxR+Hx)×100(%)] (2)
を便宜的にK値と定義すると、K値は死後の時間経過と温度の関数となり、K値の増加速度が死後の生鮮魚介類等に起きる諸変化と概略平行する。
The freshness index K value of fresh seafood and the like is derived as a function from the decomposition process of adenosine triphosphate (hereinafter abbreviated as “ATP”) contained in muscle of fresh seafood and the like. ATP is dephosphorylated to adenosine monophosphate (adenylic acid, hereinafter abbreviated as “AMP”) via adenosine diphosphate (hereinafter abbreviated as “ADP”). Subsequently, part of the structure of the base moiety changes to become inosinic acid (hereinafter abbreviated as “IMP”). IMP is well known as a flavor component of skipjack. The process up to this point is relatively fast when ATP is decomposed. The accumulation of IMP is the accumulation of umami components, and the decomposition so far is an important process for the taste of fresh seafood and the like. IMP is dephosphorylated to inosine (hereinafter abbreviated as “HxR”), and further subjected to an enzymatic reaction to hypoxanthine (hereinafter abbreviated as “Hx”). Hx is further decomposed into xanthine and uric acid by enzymatic degradation. In this series of processes, because the degradation rate of IMP is relatively small,
[(HxR + Hx) / (ATP + ADP + AMP + IMP + HxR + Hx) × 100 (%)] (2)
Is defined as a K value for convenience, the K value is a function of time and temperature after death, and the rate of increase in K value is approximately parallel to various changes occurring in fresh fish and shellfish after death.
生鮮魚介類等の鮮度指標K値に関する上記式(2)は、以下の式(3)のように簡略化することも可能である。 The above equation (2) relating to the freshness index K value of fresh fish and shellfish can be simplified as the following equation (3).
[(HxR+Hx)/(IMP+ HxR+Hx)×100(%)] (3)
式(3)はATP、ADP、AMPを省略したもので、これはATPからIMPまでの分解が極めて短時間に進むことから提唱されたもので、K値とほぼ同じ値を示す。
[(HxR + Hx) / (IMP + HxR + Hx) × 100 (%)] (3)
Formula (3) omits ATP, ADP, and AMP, which is proposed because the decomposition from ATP to IMP proceeds in a very short time, and shows almost the same value as the K value.
一般に、生鮮魚介類ではK値20%以下なら生食可能であるが、それ以上だと加熱調理する必要がある。そして、60%を超えると腐敗が始まって食用に不可になるといわれる。一方、鶏肉では、K値が20%以下なら極めて鮮度が高く、30%以下なら良好、50%以上では問題があるといわれている。 Generally, fresh fish and shellfish can be eaten raw if the K value is 20% or less, but if it is more than that, it is necessary to cook by heating. And when it exceeds 60%, it is said that rot begins and it becomes inedible. On the other hand, chicken is said to be extremely fresh when the K value is 20% or less, good when 30% or less, and problematic when 50% or more.
本発明者らは生鮮魚介類等の代謝過程であるATPの分解過程でHxからキサンチンおよび尿酸が生成する酵素反応に着目し、この酸化反応と共役して発色する発色剤(ホルマザン色素)の発色強度を計測することにより、任意の時点の生鮮魚介類等の鮮度を非破壊的に評価する方法を見出したものである。 The present inventors paid attention to the enzyme reaction that xanthine and uric acid are produced from Hx during the decomposition process of ATP, which is a metabolic process of fresh fish and shellfish, and the coloring of a color former (formazan dye) that develops color coupled with this oxidation reaction. The present inventors have found a method for nondestructively evaluating the freshness of fresh seafood and the like at an arbitrary time by measuring the strength.
すなわち、生鮮魚介類等の鮮度を非破壊的に評価する本発明は、ヒポキサンチンがキサンチンオキシダーゼによってキサンチンと尿酸に分解する反応に共役して発色する発色剤、ヒポキサンチン、キサンチンオキシダーゼおよび溶媒を含む発色剤組成物を透明容器に封入して発色媒体とし、該発色媒体を生鮮魚介類、獣肉または家禽肉と一緒に保蔵し、任意の時点で発色媒体の発色強度を測定し、測定した発色強度の強弱をもとに生鮮魚介類、獣肉または家禽肉の鮮度を推定することを特徴とするとするものである。 That is, the present invention for non-destructively evaluating the freshness of fresh seafood, etc. includes a color former, hypoxanthine, xanthine oxidase and a solvent that develops color by coupling to the reaction of hypoxanthine decomposed into xanthine and uric acid by xanthine oxidase. The coloring agent composition is sealed in a transparent container to form a coloring medium, and the coloring medium is stored together with fresh seafood, animal meat or poultry meat, and the coloring intensity of the coloring medium is measured at any point in time, and the measured coloring intensity The freshness of fresh seafood, animal meat or poultry meat is estimated based on the strength of the food.
本発明における、生鮮魚介類等とは、種々の魚や貝などの魚介類、牛、豚、羊等の獣肉、鶏肉等の家禽肉を意味する。 In the present invention, fresh seafood and the like mean seafood such as various fish and shellfish, animal meat such as cow, pig and sheep, and poultry meat such as chicken.
本発明における発色剤とは、ATPの分解過程でHxからキサンチン・尿酸を生成させるキサンチンオキシダーゼ(EC1.1.3.22、以下、「XOD」と略称する)反応に共役して発色しホルマザン色素を生成する発色剤であれば特に限定されないが、表1に示したテトラゾリウム塩を挙げることができる。なお、表1に示したテトラゾリウム塩が反応して生成するホルマザン色素の発色の吸収極大の波長を合わせて示す。 The color former in the present invention is a formazan dye produced by coupling with a xanthine oxidase (EC 1.1.3.22, hereinafter abbreviated as “XOD”) reaction that produces xanthine / uric acid from Hx during the ATP decomposition process. Although it will not specifically limit if it is a color former to perform, The tetrazolium salt shown in Table 1 can be mentioned. In addition, the wavelength of the absorption maximum of color development of the formazan dye produced by the reaction of the tetrazolium salt shown in Table 1 is also shown.
表1に示した波長は吸収極大の例であり、測定条件によっては若干シフトする場合があるが、この波長を中心として短波長側あるいは長波長側20〜30nm付近の波長を採用することができる。発色強度の測定は、色見本による目視法、光電比色計または分光光度計の計器測定のいずれでもよい。 The wavelength shown in Table 1 is an example of an absorption maximum, and may be slightly shifted depending on the measurement conditions. However, a wavelength near the short wavelength side or the long wavelength side of 20 to 30 nm can be adopted with this wavelength as the center. . The color development intensity may be measured by a visual method using a color sample, an instrument measurement using a photoelectric colorimeter or a spectrophotometer.
テトラゾリウム塩は窒素原子を4個有する五員環化合物の第4級アンモニウム塩で、酸化還元酵素の反応に共役して、それぞれ色調の異なるホルマザン色素を生成する。これらテトラゾリウム塩酸化還元酵素・コルチコイド・糖類・アスコルビン酸のような還元性物質の検出や、種子の発芽能力の試験、生細胞の計数・生体染色・***や慢性肉芽種の臨床検査などに使用される他、動物実験法代替試薬として実験動物に代わりin vitroでの培養細胞の毒性を調べることによって客観的に毒性を測定する試薬としても用いられる。 The tetrazolium salt is a quaternary ammonium salt of a five-membered ring compound having four nitrogen atoms, and forms a formazan dye having a different color tone, coupled to the reaction of the oxidoreductase. For detection of reducing substances such as tetrazolium hydrochloride reductase, corticoids, sugars, ascorbic acid, seed germination ability, live cell counting, vital staining, urinary tract infection, and clinical examination of chronic granulation species In addition to being used as a substitute for animal experiments, it is also used as a reagent for objectively measuring toxicity by examining the toxicity of cultured cells in vitro instead of experimental animals.
すなわち、生鮮魚介類等の代謝過程であるATPの分解過程でHxからキサンチンおよび尿酸が生成するXOD反応に共役して発色するホルマザン色の発色強度は対象生鮮魚介類等のHxの濃度を示す一方で、過去の温度履歴X時間を表す。K値および温度履歴が分かることで今の鮮度がどの状態にあるかの詳細が分かる。 That is, the formazan color intensity that develops in conjunction with the XOD reaction produced by xanthine and uric acid from Hx during the decomposition process of ATP, which is a metabolic process of fresh seafood, etc., indicates the concentration of Hx in the target fresh seafood, etc. Represents the past temperature history X time. By knowing the K value and the temperature history, the details of the current freshness state can be understood.
XODの反応過程を水が溶媒の例で下記に示した。 The reaction process of XOD is shown below with water as an example of a solvent.
ヒポキサンチン+O2+H2O→キサンチン+2H++O2 −
キサンチン+O2+H2O→尿酸+2H++O2 −
O2 −はスーパーオキシドアニオンである。このスーパーオキシドアニオンがテトラゾリウム塩と反応して発色する(図1参照)。
Hypoxanthine + O 2 + H 2 O → xanthine + 2H + + O 2 −
Xanthine + O 2 + H 2 O → uric acid + 2H ++ O 2 −
O 2 − is a superoxide anion. The superoxide anion reacts with the tetrazolium salt to develop color (see FIG. 1).
テトラゾリウム塩のピークの波長の吸光度、すなわち発色強度は、XODの反応で生成したキサンチンおよび尿酸量を反映するとともに、酵素XODにより分解されたHxの量に相当することから、予めランバート−ベールの法則に基づき、濃度既知のHxを用いて作成しておいた検量線(吸光度と濃度との関係式)から、発色媒体中で分解されたHxの量の推定が可能である。発色強度が強いほど、基質のHxが分解されていることを示し、すなわち生鮮魚介類等の鮮度が悪いことを示すので、発色強度を測定することにより鮮度を容易に推定することができる。 The absorbance at the peak wavelength of the tetrazolium salt, that is, the color development intensity reflects the amount of xanthine and uric acid produced by the reaction of XOD and corresponds to the amount of Hx decomposed by the enzyme XOD. Based on the above, it is possible to estimate the amount of Hx decomposed in the coloring medium from a calibration curve (relational expression between absorbance and concentration) prepared using Hx having a known concentration. The stronger the coloring intensity, the more the substrate Hx is decomposed, that is, the freshness of fresh seafood and the like is poor. Therefore, the freshness can be easily estimated by measuring the coloring intensity.
本発明の方法で検出された酵素XODにより分解されたHxの量は、生鮮魚介類等の生体内で起きている任意の時点の酵素XODの反応にほかならないから、その生鮮魚介類等の生体内で生成されたHxの量にほぼ等しい。従って、生鮮魚介類等の生体内で起きている酵素XODのHxの分解程度から、すなわち発色強度から鮮度指標K値を推定する関数をつくり検量線を作成しておけば、本発明の方法による発色強度から、任意の時点の生鮮魚介類等のK値を推定することができる。 The amount of Hx decomposed by the enzyme XOD detected by the method of the present invention is nothing but the reaction of the enzyme XOD occurring at any time in the living body such as fresh fish and shellfish. It is approximately equal to the amount of Hx produced in the body. Therefore, if a calibration curve is prepared by creating a function for estimating the freshness index K value from the degree of Hx decomposition of the enzyme XOD occurring in the living body such as fresh fish and shellfish, that is, according to the method of the present invention. The K value of fresh fish and shellfish at any time can be estimated from the color intensity.
本発明の発色媒体には、Hx、XOD、発色剤を溶解させるために溶媒が必要である。溶媒は、Hx、XOD、発色剤を溶解するものであれば特に限定されないが、極性溶媒を用いることができる。本発明の発色媒体は、冷蔵あるいは冷凍環境下で使用するため、氷結しない冷蔵条件下では水を溶媒としてもよいが、氷結するような冷蔵あるいは冷凍環境下では、水は凍るため溶媒としては不適当である。氷点下でも凍らない不凍性の極性溶媒としては、例えばメタノール、エタノールなどの一価アルコール、エチレングリコール(EG)、1,4-ブタンジオールなどの二価アルコール、グリセロールなどの三価アルコール、ポリエチレングリコール(PEG)、グルシトールなどの多価アルコールを例示することができる。 The coloring medium of the present invention requires a solvent for dissolving Hx, XOD, and the coloring agent. The solvent is not particularly limited as long as it dissolves Hx, XOD, and a color former, and a polar solvent can be used. Since the coloring medium of the present invention is used in a refrigerated or frozen environment, water may be used as a solvent under refrigerated conditions that do not freeze, but in a refrigerated or frozen environment that freezes, water is frozen and is not suitable as a solvent. Is appropriate. Examples of non-freezing polar solvents that do not freeze below freezing include monohydric alcohols such as methanol and ethanol, dihydric alcohols such as ethylene glycol (EG) and 1,4-butanediol, trihydric alcohols such as glycerol, and polyethylene glycol. Examples thereof include polyhydric alcohols such as (PEG) and glucitol.
しかし、上述の採用可能な溶媒を用いても、本発明の方法では最良の結果が導き出されるとは限らない。これらの溶媒を採用するに当たっては、それぞれ最適濃度があることを考慮しなければならない。 However, the use of the above employable solvents does not always lead to the best results with the method of the present invention. In employing these solvents, it must be taken into account that each has an optimum concentration.
本発明の発色媒体において、Hx、XOD、発色剤および溶媒を含む発色剤組成物を封入する透明容器は、Hxからキサンチン・尿酸を生成させるXOD反応に共役して発色するホルマザン色素の強度を計測できるものであれば、材質および形状は特に限定されない。 In the coloring medium of the present invention, a transparent container enclosing a coloring agent composition containing Hx, XOD, a coloring agent and a solvent measures the intensity of the formazan dye that develops color in conjunction with the XOD reaction that generates xanthine / uric acid from Hx. If possible, the material and shape are not particularly limited.
透明容器の材質としては、ポリアクリレート、ABS樹脂、ポリ塩化ビニール等の高分子化合物あるいはガラス等を挙げることができる。形状は、試験管状、アンプル状、袋状いずれでもよいが、発色剤の発色強度を分光光度計で計測する場合は、内径10mm、容量7〜8mlの試験管状のものが使いやすい。 Examples of the material of the transparent container include polymer compounds such as polyacrylate, ABS resin, and polyvinyl chloride, and glass. The shape may be any of a test tube shape, an ampoule shape, and a bag shape, but when measuring the color intensity of the color former with a spectrophotometer, a test tube shape having an inner diameter of 10 mm and a capacity of 7 to 8 ml is easy to use.
しかし、生鮮魚介類等のK値を計測する従来の方法では、生鮮魚介類の筋肉から測定用試料を調製しなければならず、破壊法であり、生鮮魚介類の商品価値を損なう欠点があった。また従来法は化学分析法であり、実験室的で熟練を要し、簡便な現場向き計測法ではなかった。 However, the conventional method for measuring the K value of fresh seafood, etc. requires preparation of a measurement sample from the muscle of fresh seafood, which is a destructive method and has the disadvantage of impairing the commercial value of fresh seafood. It was. In addition, the conventional method is a chemical analysis method, requires laboratory skills, and is not a simple on-site measurement method.
XODの基質はプリン化合物分解経路で生じるHxであり、生鮮魚介類等の筋肉中ではATPの分解途中で生じるHxであるが、本発明では生鮮魚介類等の筋肉中のHxである必要はない。すなわち、Hxそのものを基質として用いることによっても、温度履歴から鮮度を推定することが可能であることを見出した。本発明の方法では、生鮮魚介類等の筋肉を採取する必要がないため、非破壊法であり、生鮮魚介類等の商品価値を損ねることがない。従って、発色媒体は、鮮度を測定しようとする生鮮魚介類等と一緒に保蔵すればよい。魚や貝等の生鮮魚介類の場合は、トロ箱等の保蔵容器に発色媒体を装入すればよい。 The substrate of XOD is Hx generated in the purine compound degradation pathway, and it is Hx generated in the course of ATP degradation in muscle such as fresh seafood, but in the present invention, it is not necessary to be Hx in muscle such as fresh seafood. . That is, it has been found that the freshness can be estimated from the temperature history by using Hx itself as a substrate. In the method of the present invention, since it is not necessary to collect muscles such as fresh seafood, it is a non-destructive method and does not impair the commercial value of fresh seafood. Therefore, the coloring medium may be stored together with fresh seafood or the like whose freshness is to be measured. In the case of fresh seafood such as fish and shellfish, a coloring medium may be placed in a storage container such as a toro box.
XOD反応は一般酵素反応と同じように温度依存性である。予め、Hx、XOD、発色剤および溶媒を適当な透明容器に封入した発色媒体を用意し、この発色媒体を水揚げ後の生鮮魚介類の入ったトロ箱等貯蔵容器に装入しておけば、その発色媒体が置かれた温度環境に応じた発色強度を示す。すなわち、ある任意の時点の発色強度は鮮度指標K値を示すと共に、生鮮魚介類等の置かれた環境の温度履歴にも相当する。 The XOD reaction is temperature dependent like the general enzyme reaction. Prepare a coloring medium in which Hx, XOD, a coloring agent and a solvent are sealed in a suitable transparent container in advance, and charge this coloring medium into a storage container such as a toro box containing fresh seafood after landing. The color intensity corresponding to the temperature environment where the color medium is placed is shown. That is, the color intensity at a certain arbitrary time point indicates the freshness index K value, and also corresponds to the temperature history of the environment where the fresh seafood is placed.
生鮮魚介類が適正に冷蔵・冷凍保存されていれば、すなわち、温度が氷点下程度に保たれていれば、XODの反応は進まないから発色しないが、氷点下よりも高い温度に置かれた場合は、XOD反応が進むから発色する。この発色媒体の発色強度を計測することによって、今置かれた生鮮魚介類等の鮮度の状態(K値)を把握することが可能になる。 If fresh seafood is properly refrigerated and frozen, that is, if the temperature is kept below the freezing point, the reaction of XOD will not proceed and the color will not develop, but if it is placed at a temperature higher than the freezing point Color develops as the XOD reaction proceeds. By measuring the coloring intensity of the coloring medium, it is possible to grasp the freshness state (K value) of the fresh fish and shellfish placed now.
従って、予め発色媒体の発色強度とK値の関係を求めて検量線を作成しておけば、発色媒体の発色強度からK値を求めることができる。 Accordingly, if a calibration curve is prepared by previously obtaining the relationship between the color intensity of the color medium and the K value, the K value can be obtained from the color intensity of the color medium.
生鮮魚介類の鮮度低下の様式を予め計測し、一定時間毎の発色強度をデータベース化しておけば、その時の発色強度から、その時の鮮度の状態を把握することが可能となるだけでなく、以下の生可食残存日数の式により生可食残存日数を推定することも可能となる。 If you measure the freshness decline of fresh fish and shellfish in advance and create a database of color intensity at regular intervals, you can not only grasp the state of freshness at that time, but also the following. It is also possible to estimate the remaining days of raw edible by the formula of the remaining days of raw edible.
図2は、貯蔵温度とK値上昇率の関係を示すグラフであり、このグラフより1日当たりのK値上昇率を求めると、例えば、5℃貯蔵では1日当たりのK値は生可食限界値に達するまで12.5%ずつ増大していくことになる。図2はこの状態を示したもので、−10℃以下の貯蔵であればK値の増加は穏やかであるが、−10℃を超えるとK値は急激に増大している。 Figure 2 is a graph showing the relationship between storage temperature and K values increase rate, when determining the daily K values increase rate from the graph, for example, 1 day K value Namaka diet limits at 5 ° C. storage It will increase by 12.5% until the value is reached. FIG. 2 shows this state. The K value increases moderately when stored at −10 ° C. or lower, but the K value increases rapidly above −10 ° C.
貯蔵中のK値変化が一次反応であると仮定して、貯蔵中におけるK値変化を図3に示した。ここで、K 2 は生可食限界のK値、K 0 は限りなく新鮮な状態のK値、すなわち漁獲あるいは水揚げ直後の生鮮魚介類のK値を示す。
貯蔵管理の徹底を図ることにより、任意の時点におけるK値は積算温度の影響を表していると考えることができる。それ故に、任意の時点のK値(K)は、
K=ΣEt×Xt=(K2−K0)Σ(Xt/Tt) (4)
と表すことができる。ここでEtはt℃におけるK値上昇率で、(K2−K0)/Ttで表すことができる。Ttはt℃で保存された生鮮魚介類の生可食(刺身)としての許容日数を、Xtはt℃に保存された日数を表す。
Assuming that the change in K value during storage is a primary reaction, the change in K value during storage is shown in FIG. Here, K 2 indicates a K value of Namaka diet limits, K 0 is K value of fresh as possible, i.e. the K value of the catch or immediately after landing fresh seafood.
By carrying out thorough storage management, it can be considered that the K value at an arbitrary time point represents the influence of the integrated temperature. Therefore, the K value (K) at any time is
K = ΣE t × X t = (K 2 -K 0) Σ (X t / T t) (4)
It can be expressed as. Here, Et is the K value increase rate at t ° C., and can be expressed as (K 2 −K 0 ) / T t . Tt represents the allowable number of days as fresh edible (sashimi) of fresh seafood stored at t ° C, and Xt represents the number of days stored at t ° C.
上記式(4)のKが求まれば、任意の時点以降のt℃貯蔵における生可食残存日数(RDVt)は式(1)で表すことができる。 If K of the equation (4) is obtained, the raw edible residual number of days at t ° C. storage after any time (RDV t) can be expressed by Equation (1).
RDVt=[Tt(K2−K’)/(K2−K0)] (1)
なお、式(1)は、
RDVt=Tt[K2/(K2−K0)−Σ(Xt/Tt)] (5)
と書き直すこともできる。
RDV t = [T t (K 2 −K ′) / (K 2 −K 0 )] (1)
In addition, Formula (1) is
RDV t = T t [K 2 / (K 2 −K 0 ) −Σ (X t / T t )] (5)
It can also be rewritten.
図3にRDVtの関係式を示した。例えば、−20℃に貯蔵した場合に、刺身として食べることが可能な残存日数(RDV−20)を式(1)あるいは式(5)により求めることができる。 FIG. 3 shows a relational expression of RDV t . For example, it can be determined when stored in -20 ° C., the possible residual number of days to eat as sashimi (RDV -20) Equation (1) or Equation (5).
RDV−20=85(20−K’)/(20−3)=100−5K’
となり、請求項1ないし6に記載した本発明により生鮮魚類任意の時点のK値を求めれば、−20℃に貯蔵した時の生鮮魚類の刺身として食べられることが可能な残存日数を式(1)により求めることができる。
RDV- 20 = 85 (20-K ') / (20-3) = 100-5K'
When the K value of fresh fish at any time is determined according to the present invention as set forth in
さらに、保存温度毎に検量線を作成し、生鮮魚介類の種別や鮮度変化の特徴や様式をデータベース化することで、個々の種類だけでなく、多種の生鮮魚類にも対応が可能となる。鮮度の推定を簡便に行うには例えば、図4に示したように発色強度と生可食残存日数や鮮度値K値を印刷した色見本を発色媒体に貼付しておけば、発色媒体の発色強度を目視で測定し、生可食残存日数や鮮度を簡便に推定することができる。 Furthermore, by creating a calibration curve for each storage temperature and creating a database of types of fresh fish and shellfish and characteristics and styles of changes in freshness, it is possible to support not only individual types but also various types of fresh fish. In order to easily estimate the freshness, for example, as shown in FIG. 4 , if a color sample printed with the color intensity, the number of days remaining edible, and the freshness value K is pasted on the color medium, the color of the color medium can be estimated. The strength can be measured visually to easily estimate the number of days remaining in raw edible food and the freshness.
最近、トレーサビリティの一手段として電子荷札(ICタグ)や温度記録子(データロガ)の利用が始まりつつある。電子荷札は信号を記録する部分と発信する部分からなり、生産者や生産状況の記録、流通の記録等の情報が得られる。温度記録子は一定時間毎の温度を一定回数分記録する機能をもつ。 Recently, the use of electronic tags (IC tags) and temperature recorders (data loggers) has started as a means of traceability. The electronic tag includes a signal recording part and a transmission part, and information such as a producer, production status record, and distribution record can be obtained. The temperature recorder has a function of recording the temperature every certain time for a certain number of times.
前者は任意の時点の情報をインプットすることは可能であっても、任意の時点の対象物の状態を測定するものではない。後者は任意の時点における対象物の置かれた環境を計測するものであるが、任意の時点の対象物の状態は計測できない。両者とも、その情報を読み取ったり、解析するためには、コンピュータ(以下、「PC」と略称する)などに取り込む必要があり、PCやスキャナーなどの解析装置、読み取り装置が必要である。両者とも電子媒体であるが、非対象物の状態を計測するものではない。 Although the former can input information at an arbitrary time, it does not measure the state of an object at an arbitrary time. The latter measures the environment where the object is placed at an arbitrary time, but cannot measure the state of the object at an arbitrary time. In both cases, in order to read or analyze the information, it is necessary to import the information into a computer (hereinafter abbreviated as “PC”), and an analysis device such as a PC or a scanner, or a reading device is required. Both are electronic media, but do not measure the state of non-objects.
一方、本発明の方法はある意味で温度記録計と類似する点を持つ。温度記録計は一定時間毎の温度を一定回数記録するが、PCに取り込んで処理することによって、積算温度や平均温度、温度の変化等の解析が可能である。これらの内、積算温度は時間の関数であり、被対象物の温度履歴の総和に等しい。一定に温度環境に被対象物が置かれた場合、特に本発明の被対象物のように温度依存性のある酵素反応を指標とするものでは、冷蔵・冷凍環境下と、常温環境下とでは同じ積算温度であっても後者の反応時間は早いものとなる。すなわち、被対象物の置かれた温度環境に依存する。このことは、被対象物がどのような温度環境に置かれていたかの温度履歴を示す。 On the other hand, the method of the present invention has a point similar to the temperature recorder in a sense. The temperature recorder temperature a predetermined number of times recording every predetermined time interval, by treating capture on your PC, accumulated temperature and average temperature, it is possible to analyze the changes in temperature. Of these, the integrated temperature is a function of time and is equal to the sum of the temperature history of the object. When an object is placed in a constant temperature environment, especially in an object that uses temperature-dependent enzyme reaction as an index, such as the object of the present invention, in a refrigerated / frozen environment and a room temperature environment. Even at the same integrated temperature, the latter reaction time is fast. That is, it depends on the temperature environment where the object is placed. This indicates a temperature history of what temperature environment the object is placed in.
本発明の方法は、被対象物(生鮮魚介類等)の任意の時点の状態(鮮度)を計測するものであり、その状態からどのような温度環境下に置かれていたかの追跡が可能であり、その時点での鮮度の状態の把握が可能であり、さらに任意の時点での被対象物の状態から将来の被対象物の状態変化を推定することができる。そして、本発明の方法は非電子的(非電磁波情報媒体)である。 The method of the present invention measures the state (freshness) of an object (fresh seafood, etc.) at an arbitrary point in time, and it is possible to trace under what temperature environment the state was placed. It is possible to grasp the state of freshness at that time, and it is possible to estimate the future state change of the object from the state of the object at an arbitrary time. The method of the present invention is non-electronic (non-electromagnetic information medium).
なお、Hx、XOD、発色剤および溶媒を含む発色剤組成物を封入した透明容器は、生鮮魚介類等の鮮度推定用または生鮮魚介類生可食残存日数推定用のキットとして最適である。 A transparent container enclosing a color former composition containing Hx, XOD, a color former and a solvent is most suitable as a kit for estimating freshness of fresh seafood or the like, or estimating the number of days remaining for fresh seafood.
以下に、本発明の実施形態を更に詳細に説明する。 Hereinafter, embodiments of the present invention will be described in more detail.
本発明の生鮮魚介類の生可食残存日数推定方法は、K値を指標にした生可食残存日数をXODによるHxの酸化反応に共役して発色するテトラゾリウム塩発色強度から推定する原理に基づくものである。 The method for estimating the number of days remaining for fresh edible fish and shellfish according to the present invention is based on the principle of estimating the number of days left for raw edible using the K value as an index from the color intensity of tetrazolium salt that develops color coupled to the oxidation reaction of Hx by XOD. Is.
本発明を実施する一形態としては、極性溶媒に基質Hxとテトラゾリウム塩を加え、例えば、テトラゾリウム塩としてMTTを用いた場合はTris−塩酸緩衝液でpH7.8に調製した溶液をガラス管等の透明容器に注入し、所定単位のXODを加えて密栓し発色媒体を調製し、この発色媒体を貯蔵される生鮮魚介類等と一緒にセットする。保蔵容器の置かれた環境が常温で温度履歴が増加する場合は発色し、氷蔵された場合は発色しない(図5)。 As one mode for carrying out the present invention, a substrate Hx and a tetrazolium salt are added to a polar solvent. For example, when MTT is used as a tetrazolium salt, a solution prepared to pH 7.8 with a Tris-hydrochloric acid buffer is used. Pour into a transparent container, add a predetermined unit of XOD, seal tightly to prepare a coloring medium, and set this coloring medium together with fresh fish and shellfish to be stored. When the storage container is placed in an environment where the temperature history increases at room temperature, the color develops, and when stored in ice, the color does not develop (FIG. 5 ).
使用するXODの最適pHは弱塩基性領域にあるため、使用するテトラゾリウム塩や極性溶媒の種類によっては緩衝液を用いてpHを一定に保つ必要がある。 Since the optimum pH of the XOD to be used is in a weakly basic region, it is necessary to keep the pH constant by using a buffer depending on the type of tetrazolium salt or polar solvent to be used.
pH調整はTris-塩酸の他に、HEPES(2-[4-(2-ヒドロキシル)-1-ピペラジニル]エタンスルホン酸)などの使用が可能であるが、使用するテトラゾリウム塩の種類によって使い分ける必要がある。また、テトラゾリウム塩の種類によっても条件が異なる。 In addition to Tris-hydrochloric acid, HEPES (2- [4- (2-hydroxyl) -1-piperazinyl] ethanesulfonic acid) can be used for pH adjustment. However, it is necessary to use it properly depending on the type of tetrazolium salt used. is there. Moreover, conditions differ also with the kind of tetrazolium salt.
具体的には、極性溶媒の一定量に基質(Hx)とテトラゾリウム塩の一定量を作成し、このpHを調整した後、内径10mmのガラス管等の透明容器に収めた発色液を準備する。次に、一定の酵素単位(unit)になるように溶解したXODの酵素液を用意する。酵素液は一定量をバイアル瓶に溶解しておき、必要に応じて、その一部を注射筒などで取り出し、発色液と混合して透明容器を密封して発色媒体とする。あるいは、酵素の必要単位をガラス管等の透明容器に取り、必要時に溶媒に溶解して酵素液とし、これを発色液と混合して透明容器を密封し、発色媒体としてもよい。 In concrete terms, to create a certain amount of a certain amount of substrate (Hx) and a tetrazolium salt of a polar solvent, after adjusting the pH, to prepare a coloring solution of matches in a transparent container of the glass tube or the like of internal diameter 10mm . Next, an enzyme solution of XOD dissolved so as to be a constant enzyme unit (unit) is prepared. A predetermined amount of the enzyme solution is dissolved in a vial, and if necessary, a part of the enzyme solution is taken out with a syringe or the like, mixed with the coloring solution, and the transparent container is sealed to obtain a coloring medium. Alternatively, a necessary unit of the enzyme may be taken in a transparent container such as a glass tube, dissolved in a solvent when necessary to obtain an enzyme solution, and this may be mixed with a coloring solution to seal the transparent container, thereby forming a coloring medium.
発色媒体を生鮮魚介類等を収蔵あるいは貯蔵する容器に入れて、経時的に発色する強度を光電比色計で測定し、その吸光度から、その時点の鮮度の度合い、すなわちK値を推定する。予め作成しておいた色見本と目視で比べることで発色強度を測り、鮮度を推定しても構わない。また、実験室的に分光光度計を用いてもよい(図6)。 The coloring medium is put in a container for storing or storing fresh seafood, etc., the intensity of color development with time is measured with a photoelectric colorimeter, and the degree of freshness at that time, that is, the K value is estimated from the absorbance. The color intensity may be measured by visual comparison with a color sample prepared in advance, and the freshness may be estimated. A spectrophotometer may be used in the laboratory (FIG. 6 ).
発色媒体の透明容器としては、内径10mm、容量7〜8mlのガラス管が使いやすい。発色強度は予め作成した色見本と比較する目視法、あるいはランバート−ベールの法則(Lambert-Beer Low)に基づいた光電比色系や分光光度計で定量されるが、後者の場合、光路長はガラス管の内径そのものであり、10mmであることが望ましい。 As a transparent container for the coloring medium, a glass tube having an inner diameter of 10 mm and a capacity of 7 to 8 ml is easy to use. The intensity of color development is quantified with a visual color method compared with a color sample prepared in advance or with a photoelectric colorimetric system or spectrophotometer based on the Lambert-Beer Law (Lambert-Beer Low). The inner diameter of the glass tube itself is preferably 10 mm.
図4に本発明の発色媒体を用いて生可食残存日数を推定する一実施形態を示す。図4に示した発色媒体は、試験管状の内径10mm、容量7〜8mlのガラス管に上述の発色剤組成物を注入し、密栓したものである。発色媒体は、温度履歴の増加によって、発色剤が発色するので、例えば図示したように発色媒体に刺身としての残存賞味期限を色見本で印刷したものを添付しておけば、発色媒体の発色強度を目視で観察することにより、発色媒体と一緒に保蔵した生鮮魚介類の生可食残存日数を簡便に推定することができる。 FIG. 4 shows an embodiment for estimating the number of days remaining raw edible using the coloring medium of the present invention. The color forming medium shown in FIG. 4 is obtained by injecting the above color former composition into a test tube having an inner diameter of 10 mm and a capacity of 7 to 8 ml and sealing it. Since the color former develops color with increasing temperature history, the color development strength of the color development medium can be obtained by attaching a color sample of the remaining shelf life as sashimi to the color development medium as shown in the figure. By visually observing this, it is possible to easily estimate the number of days remaining for fresh edible fresh fish and shellfish stored together with the coloring medium.
上述したランバート−ベールの法則とは、光が均質な物質層を透過するときの光の吸収に関する法則で、吸収層の厚さ依存性と溶液の濃度依存性を合わせて吸収の強さを示し、下記の式で表される。 The Lambert-Beer law described above is a law related to light absorption when light passes through a homogeneous material layer, and indicates the intensity of absorption by combining the thickness dependency of the absorption layer and the concentration dependency of the solution. Is represented by the following formula.
I=I0×e−κcl (6) I0:入射光の強度
I=I0×10−εcl (7) I:透過光の強度
l:吸収層の厚さ(光路長)
c:吸収物質の濃度
κ:モル吸収係数(定数)
ε:モル吸光係数(定数)、cm-1・mol-1・l
A=log10(I0/I) (8) A:吸光度
従って、吸光度が解れば物質の濃度が解る。
I = I 0 × e −κcl (6) I 0 : Intensity of incident light I = I 0 × 10 −εcl (7) I: Intensity of transmitted light
l: Absorbing layer thickness (optical path length)
c: Absorbent concentration
κ: molar absorption coefficient (constant)
ε: molar extinction coefficient (constant), cm −1 · mol −1 · l
A = log 10 (I 0 / I) (8) A: Absorbance Accordingly, if the absorbance is known, the concentration of the substance is known.
酵素濃度・基質濃度・発色剤濃度の検討
基質Hxにテトラゾリウム塩MTTの存在下でそれぞれの濃度の酵素XODを作用させ、生成したホルマザン色素の発色強度を吸光度565ナノメートルで測定した。Hxの濃度を0.5グラム毎リットルとし、XODの濃度を0.1・0.3・0.5・1.0酵素単位とし、エタノール(以下、「EtOH」と略称する)80パーセント不凍液中で室温(25.5℃)で反応させた(図7)。
Examination of Enzyme Concentration / Substrate Concentration / Coloring Agent Concentration Enzyme XOD of each concentration was allowed to act on substrate Hx in the presence of tetrazolium salt MTT, and the color intensity of the produced formazan dye was measured at an absorbance of 565 nanometers. The concentration of Hx is 0.5 gram per liter, the concentration of XOD is 0.1, 0.3, 0.5 and 1.0 enzyme units, and ethanol (hereinafter abbreviated as “EtOH”) in 80 percent antifreeze. At room temperature (25.5 ° C.) (FIG. 7 ).
テトラゾリウム塩MTT、酵素XODの存在下でそれぞれの濃度の基質Hxを添加し、生成したホルマザン色素の発色強度を吸光度565ナノメートルで測定した。MTTの濃度は0.5グラム毎リットル、XODの濃度は0.3酵素単位とし、HxRの濃度を0.1、0.25、0.4、0.55グラム毎リットルとし、EtOH80パーセント不凍液中で室温(25.5℃)で反応させた(図8)。 Substrate Hx of each concentration was added in the presence of tetrazolium salt MTT and enzyme XOD, and the color intensity of the produced formazan dye was measured at an absorbance of 565 nanometers. The MTT concentration is 0.5 gram per liter, the XOD concentration is 0.3 enzyme unit, the HxR concentration is 0.1, 0.25, 0.4, 0.55 gram per liter, in EtOH 80 percent antifreeze. At room temperature (25.5 ° C.) (FIG. 8 ).
基質Hx、酵素XODの存在下でそれぞれの濃度のテトラゾリウム塩MTTを添加し、生成したホルマザン色素の発色強度を吸光度565ナノメートルで測定した。Hxの濃度は0.4グラム毎リットル、XODの濃度は0.3酵素単位とし、MTTの濃度を0.1、0.25、0.4、0.55グラム毎リットルとし、EtOH80パーセント不凍液中で室温(25.5℃)で反応させた(図9)。 The tetrazolium salt MTT at each concentration was added in the presence of the substrate Hx and the enzyme XOD, and the color intensity of the produced formazan dye was measured at an absorbance of 565 nanometers. Hx concentration is 0.4 gram per liter, XOD concentration is 0.3 enzyme unit, MTT concentration is 0.1, 0.25, 0.4, 0.55 gram per liter, in EtOH 80 percent antifreeze At room temperature (25.5 ° C.) ( FIG. 9 ).
その結果、XOD:0.3酵素単位毎4ミリリットル、Hx:1.6ミリグラム毎4ミリリットル、MTT:1.6ミリグラム毎4ミリリットルが最適濃度であった。 As a result, XOD: 0.3 enzyme unit was 4 milliliters per unit, Hx: 1.6 milligrams per 4 milliliters, and MTT: 1.6 milligrams per 4 milliliters were optimum concentrations.
そこでXOD:0.3酵素単位毎4ミリリットル、Hx:1.6ミリグラム毎4ミリリットル、MTT:1.6ミリグラム毎4ミリリットルの濃度のEtOH80パーセント溶液からなる発色剤組成物を調整し、内径10mmのガラス管に7ml注入したのち密栓し発色媒体を調製し、鮮度測定用キットを作成した。 Therefore, a color former composition comprising an 80% EtOH solution having a concentration of XOD: 0.3 enzyme unit per 4 ml, Hx: 1.6 milligram per 4 ml, MTT: 1.6 milligram per 4 ml, and having an inner diameter of 10 mm. After injecting 7 ml into a glass tube, the tube was tightly sealed to prepare a coloring medium, and a kit for measuring freshness was prepared.
K値と発色強度との関係
実施例1で作成したキットをサンマ・マサバ・イワシの小型魚種とともに5℃および0℃環境下に置いた。発色媒体の発色強度の経時変化を測定するとともに、サンマ・マサバ・イワシの5℃および0℃環境下、HPLCでK値を測定し、検量線を作成した(図10,図11)。
Relationship between K value and coloring intensity The kit prepared in Example 1 was placed in a 5 ° C. and 0 ° C. environment together with a small fish species of saury, chub mackerel and sardines. The color development intensity of the chromogenic medium was measured over time, and the K value was measured by HPLC under the conditions of 5 ° C. and 0 ° C. of saury, masaba and sardines to prepare a calibration curve (FIGS. 10 and 11 ).
サンマ・マサバ・イワシのK値変化のパターンはほぼ同じで、これらと発色媒体との間には良好な相関関係(0℃:R=0.95、5℃:R=0.95)があった。 Pacific saury pattern K value variation of Mackerel-sardine similar, good correlation between these and the coloring medium body (0 ℃: R = 0.95,5 ℃ : R = 0.95) is there were.
水揚げ時に実施例1で作成した鮮度測定用キットを用意し、このキットをトロ箱等収蔵容器中の魚介と一緒に置き、漁港から東京まで輸送した。その際、温度計測用のセンサーも同時にトロ箱等収蔵容器内の数カ所に設置した。試験開始時は10℃あったが、まもなく0℃にまで下降した。10時間後東京へ到着したが、その時の温度上昇はほとんどなかった。キットの発色媒体の発色は無色のままであった。トロ箱等収蔵容器をそのまま常温に置き、なおも観察を続けた。18時間後にわずかに着色したので吸光度を測定し、0.40の吸光度を得た。この時の温度は1℃であった。この吸光度から鮮度値K値は15.2%と推測され、生食可と判定された(Y=7.95X+12)。24時間後にはトロ箱等収蔵容器内の氷も溶け、発色媒体の吸光度は1.19、K値は19.9%で食限界と判定された(Y=14.5X+2.6)。この時の温度は5℃であった。30時間を超える頃には常温となり、腐敗が始まり悪臭が漂っていた。
A freshness measurement kit prepared in Example 1 was prepared at the time of landing, and this kit was placed together with the seafood in a storage container such as a toro box and transported from the fishing port to Tokyo. At that time, sensors for temperature measurement were also installed at several locations in the storage container such as a trolley box. Although it was 10 degreeC at the time of a test start, it fell to 0 degreeC soon. I arrived in
請求項1〜5に記載された本発明方法は、生鮮魚介類等の鮮度を非破壊的に推測することができるので、生鮮魚介類等の流通、販売において有用である。 Since the freshness of fresh fishery products etc. can be estimated nondestructively, the method of this invention described in Claims 1-5 is useful in distribution and sale of fresh fishery products.
請求項6〜9に記載された本発明方法は、生鮮魚介類の生可食残存日数を非破壊的に推測することができるので、生鮮魚介類の流通、販売において有用である。 The method of the present invention described in claims 6 to 9 is useful in the distribution and sale of fresh seafood since it can non-destructively estimate the number of days remaining for fresh seafood.
請求項10〜13に記載されたキットは、生鮮魚介類等の鮮度推定用もしくは生鮮魚介類生可食残存日数推定用のキットとして有用である。
The kit described in
Claims (10)
RDVt=[Tt(K2−K’)/(K2−K0)](1)
(ただし式(1)中、RDVt:t℃貯蔵における生可食残存日数
Tt:t℃保存における生食としての許容限界日数
K2:生食として食せる許容限界のK値
K0:非常に新鮮な生鮮魚類のK値
K’:測定した時点のK値の推定値を示す。)
で表される式により生可食残存日数を算定することを特徴とする生鮮魚類の生可食残存日数の推定方法。 A coloring agent composition containing hypoxanthine, xanthine oxidase and a non-freezing polar solvent that is colored by coupling to the reaction of hypoxanthine decomposed into xanthine and uric acid by xanthine oxidase is sealed in a transparent container to form a coloring medium. The coloring medium is placed in a storage container in which fresh fish is stored, the coloring intensity of the coloring medium is measured at an arbitrary time, and the K value of the fresh fish is estimated based on the strength of the measured coloring intensity. Formula (1)
RDV t = [T t (K 2 −K ′) / (K 2 −K 0 )] (1)
(In the formula (1), RDV t : remaining days of raw edible in storage at t ° C.
T t : Tolerable limit number of days as raw food when stored at t ° C
K 2 : K value of allowable limit that can be eaten as raw food
K 0 : K value of very fresh fresh fish
K ′: Indicates an estimated value of the K value at the time of measurement. )
A method for estimating the number of days remaining in raw edible for fresh fish, characterized in that the number of days remaining in raw edible is calculated by an expression represented by:
The kit according to claim 8 or 9 , wherein the color former is a tetrazolium salt.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2004165384A JP4556497B2 (en) | 2004-06-03 | 2004-06-03 | Non-destructive evaluation method of freshness of fresh seafood, animal meat or poultry meat, estimation method and kit of remaining days of fresh fish |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2004165384A JP4556497B2 (en) | 2004-06-03 | 2004-06-03 | Non-destructive evaluation method of freshness of fresh seafood, animal meat or poultry meat, estimation method and kit of remaining days of fresh fish |
Publications (3)
Publication Number | Publication Date |
---|---|
JP2005345263A JP2005345263A (en) | 2005-12-15 |
JP2005345263A5 JP2005345263A5 (en) | 2007-07-05 |
JP4556497B2 true JP4556497B2 (en) | 2010-10-06 |
Family
ID=35497790
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2004165384A Expired - Lifetime JP4556497B2 (en) | 2004-06-03 | 2004-06-03 | Non-destructive evaluation method of freshness of fresh seafood, animal meat or poultry meat, estimation method and kit of remaining days of fresh fish |
Country Status (1)
Country | Link |
---|---|
JP (1) | JP4556497B2 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR101899426B1 (en) * | 2017-05-25 | 2018-09-17 | (주)큐브바이오 | Enzyme Composition for analysing purine metabolite concentration such as hypoxanthine and xanthine for tumor diagnosis |
Families Citing this family (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2009079966A (en) * | 2007-09-26 | 2009-04-16 | Shizuoka Prefecture | Method of determining maturity of meat |
US20110262943A1 (en) * | 2008-10-17 | 2011-10-27 | Tokyo University Of Marine Science And Technology | Reagent kit for measuring freshness |
RU2492470C1 (en) * | 2011-12-27 | 2013-09-10 | Государственное научное учреждение Всероссийский научно-исследовательский институт мясной промышленности им. В.М. Горбатова Российской академии сельскохозяйственных наук | Method for determination of species identity, freshness and thermal condition of meat raw material |
KR101396268B1 (en) * | 2013-06-28 | 2014-05-16 | 이종구 | Method of manufacture compound |
CN103675220B (en) * | 2013-12-09 | 2015-05-13 | 浙江工商大学 | Method utilizing two texture indexes to jointly determine tuna flesh freshness |
CN103776828B (en) * | 2014-02-14 | 2016-04-13 | 武汉工程大学 | A kind of color change label monitoring the fresh activity of food and preparation method thereof |
JP6534555B2 (en) * | 2014-09-26 | 2019-06-26 | パナソニック株式会社 | Storage state estimation device, storage state estimation method and storage state information output device |
JP2018203439A (en) * | 2017-06-01 | 2018-12-27 | 国立大学法人 鹿児島大学 | Method for managing fresh products and device for managing fresh products |
JP2021076536A (en) * | 2019-11-12 | 2021-05-20 | 東芝テック株式会社 | Freshness measuring system |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH02257872A (en) * | 1989-03-30 | 1990-10-18 | Kanzaki Paper Mfg Co Ltd | Indicator for storage state |
JPH08338837A (en) * | 1995-06-14 | 1996-12-24 | Seiichi Kitamura | Method and material for checking freshness of food |
WO2003096309A1 (en) * | 2002-05-10 | 2003-11-20 | National Food Research Institute | Method of judging storage condition of food and beverage, medicine, etc. and indicator therefor |
JP2004028735A (en) * | 2002-06-25 | 2004-01-29 | Macnica Inc | Method for preparing workpiece and workpiece |
-
2004
- 2004-06-03 JP JP2004165384A patent/JP4556497B2/en not_active Expired - Lifetime
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH02257872A (en) * | 1989-03-30 | 1990-10-18 | Kanzaki Paper Mfg Co Ltd | Indicator for storage state |
JPH08338837A (en) * | 1995-06-14 | 1996-12-24 | Seiichi Kitamura | Method and material for checking freshness of food |
WO2003096309A1 (en) * | 2002-05-10 | 2003-11-20 | National Food Research Institute | Method of judging storage condition of food and beverage, medicine, etc. and indicator therefor |
JP2004028735A (en) * | 2002-06-25 | 2004-01-29 | Macnica Inc | Method for preparing workpiece and workpiece |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR101899426B1 (en) * | 2017-05-25 | 2018-09-17 | (주)큐브바이오 | Enzyme Composition for analysing purine metabolite concentration such as hypoxanthine and xanthine for tumor diagnosis |
WO2018216864A1 (en) * | 2017-05-25 | 2018-11-29 | (주)큐브바이오 | Enzyme composition for analyzing concentration of purine metabolites hypoxanthine and xanthine for tumor diagnosis |
CN108929895A (en) * | 2017-05-25 | 2018-12-04 | 高普生物有限公司 | Hypoxanthine and xanthine concentration analysis enzymatic compositions as the purine metabolism object for diagnosing tumor |
Also Published As
Publication number | Publication date |
---|---|
JP2005345263A (en) | 2005-12-15 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Hong et al. | The importance of ATP-related compounds for the freshness and flavor of post-mortem fish and shellfish muscle: A review | |
Howgate | A review of the kinetics of degradation of inosine monophosphate in some species of fish during chilled storage | |
JP4556497B2 (en) | Non-destructive evaluation method of freshness of fresh seafood, animal meat or poultry meat, estimation method and kit of remaining days of fresh fish | |
Dalgaard | Freshness, Quality and Safety in Seafoods F-FE 380A/00 [May 2000] | |
Lorenzon et al. | Stress effect of two different transport systems on the physiological profiles of the crab Cancer pagurus | |
Marsh et al. | A method for accurate measurements of the respiration rates of marine invertebrate embryos and larvae | |
Kohlbach et al. | Dependency of Antarctic zooplankton species on ice algae‐produced carbon suggests a sea ice‐driven pelagic ecosystem during winter | |
Heinemann et al. | Conditions of Mytilus edulis extracellular body fluids and shell composition in a pH‐treatment experiment: acid‐base status, trace elements and δ11B | |
Reppas et al. | Practical urinalysis in the cat: 1: Urine macroscopic examination ‘tips and traps’ | |
Ghaffari et al. | Thermotolerance divergence revealed by the physiological and molecular responses in two oyster subspecies of Crassostrea gigas in China | |
Heising et al. | Non-destructive sensing of the freshness of packed cod fish using conductivity and pH electrodes | |
Heising et al. | Mathematical models for the trimethylamine (TMA) formation on packed cod fish fillets at different temperatures | |
Mohamat-Yusuff et al. | Imposex in Thais gradata as a biomarker for TBT contamination on the southern coast of Peninsular Malaysia | |
Teuber et al. | Respiration rates of tropical Atlantic copepods in relation to the oxygen minimum zone | |
Rahman et al. | Expeditious prediction of post-mortem changes in frozen fish meat using three-dimensional fluorescence fingerprints | |
Liao et al. | Freshness related fluorescent compound changes in Japanese dace fish (Tribolodon hakonensis) eye fluid during storage | |
Escribano et al. | Influence of food quantity and temperature on development and growth of the marine copepod Calanus chilensis from northern Chile | |
Ding et al. | Rapid and nondestructive evaluation of fish freshness by near infrared reflectance spectroscopy combined with chemometrics analysis | |
Anusankari et al. | Optical determination of carbon dioxide and oxygen by a fluorescent membrane to evaluate the freshness of meat products | |
Cameron et al. | Effects of temperature and ocean acidification on the extrapallial fluid pH, calcification rate, and condition factor of the king scallop Pecten maximus | |
Thomas et al. | The use of novel optode sensor technologies for monitoring dissolved carbon dioxide and ammonia concentrations under live haul conditions | |
Daryanani et al. | Statolith chemistry: a new tool to understand the ecology and provenance of octopus | |
Fan et al. | Estimating freshness of ice storage rainbow trout using bioelectrical impedance analysis | |
Simon et al. | Predicting transport survival of brindle and red rock lobsters Jasus edwardsii using haemolymph biochemistry and behaviour traits | |
JP2005345263A5 (en) |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20070517 |
|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20070517 |
|
RD02 | Notification of acceptance of power of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7422 Effective date: 20080430 |
|
A711 | Notification of change in applicant |
Free format text: JAPANESE INTERMEDIATE CODE: A711 Effective date: 20080423 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20100216 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20100415 |
|
TRDD | Decision of grant or rejection written | ||
A711 | Notification of change in applicant |
Free format text: JAPANESE INTERMEDIATE CODE: A711 Effective date: 20100628 |
|
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20100629 |
|
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20100628 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20100712 |
|
R150 | Certificate of patent or registration of utility model |
Free format text: JAPANESE INTERMEDIATE CODE: R150 Ref document number: 4556497 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20130730 Year of fee payment: 3 |
|
S111 | Request for change of ownership or part of ownership |
Free format text: JAPANESE INTERMEDIATE CODE: R313117 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20130730 Year of fee payment: 3 |
|
R360 | Written notification for declining of transfer of rights |
Free format text: JAPANESE INTERMEDIATE CODE: R360 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20130730 Year of fee payment: 3 |
|
R370 | Written measure of declining of transfer procedure |
Free format text: JAPANESE INTERMEDIATE CODE: R370 |
|
S111 | Request for change of ownership or part of ownership |
Free format text: JAPANESE INTERMEDIATE CODE: R313113 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20130730 Year of fee payment: 3 |
|
R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |