Survey of Histamine Content in Seafood on the Market

June 2005 127 16 12 16, Survey of Histamine Content in Seafood on the Market Kimiko K6C,Hirofumi UH=>N6B6, Tetsuya S=>C9D and Kazuo S6>ID (Tokyo Metropolitan Institute of Public Health: 3 24 1, Hyakunin-cho, Shinjuku-ku, Tokyo 169 0073, Japan; Corresponding author) Food poisoning involving histamine has occurred almost every year for 20 years in Tokyo, and is usually due to ingestion of fish with lean meat, such as sardine, mackerel, horse mackerel and so on. Therefore, we were investigated the levels of histamine and 4 non-volatile amines (tyramine, putrescine, cadaverine, spermidine) in 637 samples on the market. The water activity of samples in which histamine was detected at 5 mg/100 g and over was examined. Histamine, tyramine, putrescine, cadaverine and spermidine were detected in 66, 43, 26, 64 and 5 samples, and the detection ranges were 5 340, 5 51, 5 42, 5 180 and 5 8 mg/100 g, respectively. Most of the samples in which histamine was detected were semi-dried round and split sardine. Water activity of 24 samples of semi-dried round and split sardine in which histamine was detected was in the range of 0.68 0.96. (Received December 16, 2004) Key words: histamine; non-volatile amine; survey; fish; big-eye sardine; water activity; seafood 1) 16) 20 17) 19) 13), 16), 20) 24) 169 0073 3 24 1 1. 11 11 15 8 637 1) 299 109 63 35 35 32 15 8 6 6 5 4 3 3 3 3 2 1 1 1 1 1 1 23 637 2) 73 7 337 5 4

128 10 5 78 4 11 5 7 7 15 3 66 16 2. 1) (99 ) 100 mg 0.5 mol/l 100 ml 1,000 mg/ml 100 mg/ml 0.5 mol/l 250 mg/ml 50 mg/ml 10 mg/ml 10 mg/ml 10 mg/ml 0.5 mol/l 2) 1,6-10 mg/ml 0.5 mol/ L 3. 1) (HPLC) LC-10 ATVP RF-10AXL, CTO-10 Vol. 46, No. 3 AVP SIL-10ADVP 2) AW-2 Hygroskop DT Rotronic 3) MOV-212 (U) 4. 17) (TLC) HPLC Scheme 1 5mg/100 g (TLC) Kieselgel 60 Merck 28 (9 : 1) I) 0.01 / 365 nm II) 0.1 / HPLC Inertsil ODS-80A (5 mm, 250 mm 4.6 mm i.d.) 40 (62 : 38) 1.5 L/min 325 nm 525 nm) 5. Sample 10 g add H 2 O15mL homogenize add 10 trichloroacetic acid solution 20 ml make up to 50 ml with water keep at room temp. for 10 min filter (No. 5A paper) Filtrate Test solution for TLC Sample solution Mixed standard solution (Qualitative analysis) 0.5 ml 0.2 1.0 ml make up to 1 ml with water add internal standard 0.5 ml add Na 2 CO 3 0.2 g add 1 dansyl chloride/acetone solution 1 ml stand at 45 in a waterbath (or at room temp. overnight) add 10 proline solution 0.5 ml stand for 10 min add toluene 5.0 ml shake Toluene layer evaporate Residue make up to 1.0 ml with acetonitrile Test solution for HPLC (Quantitative analysis) Scheme 1. Analytical procedure for non-volatile amines

June 2005 129 Table 1. Detection and Contents of Non-volatile Amines in Commercial Fish No. Fish Number of samples Detected (content: mg/100 g) Histamine Tyramine Putrescine Cadaverine Spermidine 1 Sardine (Iwashi-rui) 299 54 (6 340) 38 (5 51) 23 (5 42) 60 (5 180) 5 (5 8) 2 Skipjack and frigate mackerel (Katsuo-rui) 32 6 (5 82) 5 (6 48) 1 (6) 0 0 3 Horse mackerel (Aji-rui) 109 2 (12, 30) 0 0 1 (30) 0 4 Mackerel (Saba-rui) 35 2 (10, 10) 0 0 0 0 5 Pacific saury (Sanma) 35 1 (16) 0 0 1 (9) 0 6 Blue sprat (Kibinago) 4 1 (30) 0 2 (8, 10) 2 (16, 81) 0 7 Tuna (Maguro-rui) 63 0 0 0 0 0 8 Marline and Swordfish (Kajiki-rui) 15 0 0 0 0 0 9 Spanish mackerel (Sawara-rui) 8 0 0 0 0 0 10 Shishamo 6 0 0 0 0 0 11 Yellowtail (Buri-rui) 6 0 0 0 0 0 12 Flying fish (Tobiuo-rui) 5 0 0 0 0 0 13 Japanese sand lance (Ikanago) 3 0 0 0 0 0 14 Pacific herring (Nishin) 3 0 0 0 0 0 15 Atka mackerel (Hokke) 3 0 0 0 0 0 16 Acropoma japonicum (Hotarujako) 3 0 0 0 0 0 17 Gizzard shad (Konoshiro) 2 0 0 0 0 0 18 Butterfish (Ibodai) 1 0 0 0 0 0 19 Barracuda (Kamasu-rui) 1 0 0 0 0 0 20 Big-eyed herring (Sappa) 1 0 0 0 0 0 21 Dolphinfish (Shiira) 1 0 0 0 0 0 22 Salmon (Sake-rui) 1 0 0 0 0 0 23 Opah (Mandai) 1 0 0 0 0 0 Total 637 66 (5 340) 43 (5 51) 26 (5 42) 64 (5 180) 5 (5 8) Table 2. Detection and Contents of Non-volatile Amines in Processed Fish Products Form Number of samples Detected (content: mg/100 g) Histamine Tyramine Putrescine Cadaverine Spermidine Raw (Fresh) 73 1 (16) 0 0 1 (9) 0 Seasoned raw (Ajitsuke nama) 7 0 0 0 0 0 Semi-dried, round and split (Himono) 337 52 (6 340) 35 (5 51) 25 (5 42) 60 (7 180) 5 (5 8) Seasoned and dried (Ajitsuke himono) 5 0 0 0 0 0 Brine-soaked and dried (Kusaya) 4 0 0 0 0 0 Boiled and dried whole (Niboshi) 10 0 0 0 0 0 Boiled and dried whitebait (Shirasuboshi) 5 0 0 0 0 0 Paste products (Neriseihin) 78 5 (7 34) 1 (23) 0 3 (5 70) 0 Pickled (Suzuke) 4 0 0 0 0 0 Seasoned and boiled (Tsukudani) 11 1 (8) 1 (5) 0 0 0 Boiled meat (Namari) 5 0 0 0 0 0 Dried strip (Katsuobushi) 7 0 0 0 0 0 Stock (Dashi) 7 0 0 0 0 0 Seasoned powder for sprinkling (Furikake) 15 6 (5 82) 6 (5 48) 1 (6) 0 0 Canned (Kanzume) 66 0 0 0 0 0 Salted viscera preserves (Shiokara) 3 1 (12) 0 0 0 0 Total 637 66 (5 340) 43 (5 51) 26 (5 42) 64 (5 180) 5 (5 8) 6. 5g 105 4 1. 637 4 Table 1 Table 2 66 5 340 mg/100 g 69 mg/100 g 10

130 5 51 mg/100 g 17 mg/100 g 43 7 5 42 mg/ 100 g 11 mg/100 g 26 4 5 180 mg / 100 g 47 mg/100 g 64 10 5 8 mg/100 g 6mg/100 g 5 1 1) 299 54 18 6 340 mg/100 g 61 mg/100 g 32 6 19 5 82 mg/100g 22 mg/100 g 109 2 2 12 30 mg/100 g 21 mg/100 g 35 2 6 10 mg/100 g 35 1 3 16 mg/100 g 4 1 25 30 mg/100 g 82 2) 337 52 15 6 340 mg/100 g 82 mg/100 g 15 6 40 5 82 mg / 100 g 22 mg/100 g 78 5 6 7 34 mg/100 g 19 mg/100 g 73 1 1 16 mg/100 g 11 1 9 8mg/100 g 3 1 33 12 mg/100 g Vol. 46, No. 3 1 20 1/3 17), 24) 100 mg/100 g Table 3 1 2 42 g 1 3 139 mg 24) 100 mg 1 139 mg 1 1 61 mg 2 1 20 30 mg 3 5 3) Table 3. Water Activity and Water Content of Big-eye Sardine in which Histamine was Detected (Urume-iwashi, 100 mg/100 g and over) No. Sample Histamine concentration (mg/100 g) Weight (g) Histamine (mg)/fish Water activity Water content ( ) 1 Big-eye sardine 340 18 61 0.82 32 2 Big-eye sardine 330 42 139 0.95 59 3 Big-eye sardine 280 9 25 0.75 26 4 Big-eye sardine 220 13 29 0.95 53 5 Big-eye sardine 200 8 16 30 6 Big-eye sardine 200 10 20 0.90 45 7 Big-eye sardine 190 12 23 0.73 27 8 Big-eye sardine 170 2 3 16 9 Big-eye sardine 170 18 31 0.96 62 10 Big-eye sardine 170 20 34 0.95 55 11 Big-eye sardine 150 9 14 29 12 Big-eye sardine 130 15 20 55 13 Big-eye sardine 100 11 11 0.71 21 Not tested, Salted and dried whole

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