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Extractive Components of the Squid Ink

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Extractive Components of the Squid Ink Fisheries Science 63(6), 939-944 (1997) Extractive Components of the Squid Ink Takaaki Shirai,*1 Naoki Kikuchi,*1 Shingo Matsuo,*1 Hiroshi Inada,*2 Takeshi Suzuki,*1 and Toshiyuki Hirano*1,3 *1Department of Food Science and Technolog y, Tokyo University of Fisheries, Konan, Minato, Tokyo 108, Japan *2Department of Marine Science and Technology , Tokyo University of Fisheries, Konan, Minato, Tokyo 108, Japan *3School of Marine Science and Technology , Tokai University, Orido, Shimizu, Shizuoka 424, Japan (Received September 30, 1996) An extract was prepared with trichloroacetic acid from the inks of neon flying, boreal clubhook, and boreo pacific gonate squids and free amino acids, trimethylamine oxide, 5'-ATP and related com pounds, betaines, and other extractive components were analyzed. The squid inks were rich in taurine and hydroxyproline. Higher amounts of aspartic acid, glutamic acid, alanine, leucine, and arginine were detected in the boreal clubhook and boreo pacific gonate squid inks. Each free amino acid of the neon flying squid ink was lower than those of the boreal clubhook and boreo pacific gonate squid inks. A small amount of trimethylamine oxide and large amounts of homarine and glycinebetaine were detect ed in three squid inks. Ethanol extract was prepared to determine the extraction capability. There were some composition similarities between ethanol and trichloroacetic acid extracts. Ethanol was inferior in extraction of inor ganic ions, even in the case of the hot-ethanol extraction. The hot-water extract showed that amounts of glycine, ƒ¿-aminobutyric acid, Cl-, and PO43- were higher than those of the trichloroacetic acid ex tract of the neon flying squid ink. Key words: squid ink, extractive component, taurine, extraction method Squidis one of the major marine food resources and pos from June though July 1995 and they were immediately siblythe most important of those that are not yet fully ex frozen at -35•Ž and transported to our laboratory in the ploited,and occur in all oceans and seas. The squid is frozen condition under -18•Ž. These squids were thawed knownto synthesize melanin from tyrosine via 3,4-di at 5•Ž for 2-3 h. Five ink-sacs were removed from five hydroxyphenylalaninein the ink1) and its ink has been half-frozen individuals of neon flying squids and used as traditionally utilized as food products commonly in samples. Ink-sacs were removed from several bodies of Japan.2)On the other hand, squid ink is reported to have boreal clubhook squid and were combined and divided an anti-bacterial activity.3) Recently, the antitumor pep into two samples. Each ink-sac of boreo pacific gonate tideglyconfraction was isolated and purified from squid squid was removed from several squid bodies and were inkby Takaya et al. 4)Squid ink has been utilized as a food combined and used as one sample. materialand is attracting attention in the seafood market in Japan, however, little is known about the food chemis Proximate Composition tryof the extractive components. The squid ink sample was dried at 108•Ž and its The neon flying squid Ommastreohes bartrami is one of moisture was measured. Crude protein was determined by the major commercial squids in Japan and boreo club the Kjeldahl method. Lipid was extracted with the chlo hook Onychoteuthis borealijaponica and boreo pacific roform/methanol mixture of Bligh and Dyer7) and dried in gonate Gonatopsis borealis squids are also caught as an air-oven. Ash was measured in an oven at 550•Ž. bycatch of the squid jigging fishery for neon flying squid in the Pacific Ocean.5,6) The inks of these three squids were Preparation of Extracts therefore used as samples in this study and extracts were Three grams of the neon flying squid ink and 1 g each of prepared with ethanol and trichloroacetic acid and the ex the boreal clubhook and boreo pacific gonate squid inks tractive components were estimated. were respectively extracted with 50 ml of 5% trichloroace tic acid in a glass pestle homogenizer cooled with ice. The Materials and Methods homogenate was centrifuged at 8,000 rpm for 15 min at 4•Ž. The residue was washed with 20 ml of 5% trichloro Squid Ink acetic acid and centrifuged twice more. The supernatants Neon flying squid, boreal clubhook squid, and boreo were combined and trichloroacetic acid was removed with pacificgonate squid were jigged in the North Pacific Ocean diethyl ether. Then the lower aqueous layer was evaporat- 940 Shirai et al. ed and diluted to 50 ml with water. This extract was desig Table 1. Proximate composition of the squid inks nated as the trichloroacetic acid extract. (%) Ethanol extract was prepared as follows. The squid ink sample (1-5 g) was homogenized with 50 ml of 80% ethanol in a glass pestle homogenizer cooled with ice and centrifuged at 8,000 rpm for 15 min at 4•Ž. The residue was extracted with 80% ethanol and centrifuged twice more. The supernatants were combined and evaporated to remove ethanol. Then the aqueous solution was defatted with diethyl ether. The lower layer was evaporated and Five individuals of the neon flying squid ink and two individuals each of the boreal clubhook and boreo pacific gonate squid inks were analyzed and values are diluted to 50 ml with water. This extract was named as expressed as means with S.D. and ranges. Carbohydrate was calculated temporarily the ethanol extract. by the difference [100% -(moisture + crude protein + lipid + ash)]. Since squid ink has been traditionally and often utilized with a small amount of liquors in food products and some times cooked, hot-ethanol extraction was also employed. Extractive Components The squid ink sample was homogenized with 80% ethanol Acid, alcohol, heating, etc. are generally known to in a glass pestle homogenizer and then heated under a affect the degree of protein denaturation and extraction reflex condenser in a boiling-water bath for 15 min. The capability physically and chemically.9) Little information homogenate was centrifuged and the residue was washed about the extractive components of squid ink has been with 80% ethanol. The residue was washed with 80% reported yet, so the trichloroacetic acid, ethanol, hot ethanol and centrifuged twice more. The supernatants ethanol, and hot-water extractions were adopted in this were combined and evaporated to remove ethanol. The study to analyze the effects of these deprotein treatments aqueous solution was defatted with diethyl ether and the on squid ink. lower layer was again evaporated. The extract was diluted Table 2 shows the composition of the trichloroacetic to 50 ml with water. This extract was named as the acid extractable components of the three squid inks. Five hot-ethanol extract. individuals of the neon flying squid ink, two samples of Hot-water extraction was also adopted with 3 g each the boreal clubhook squid ink, and one sample of the from five individuals of the neon flying squid ink in the boreo pacific gonate squid ink were analyzed and shown in same manner as the above hot-ethanol extraction substitut the table. Values of the inks of neon flying and boreal club ing water for ethanol. This extract was designated as the hook squids are expressed as means with S.D. and hot-water extract. differences from the means, respectively. Among the free amino acids of the trichloroacetic acid Analysis of Extractive Components extracts shown in Table 2, the inks of neon flying, boreal Free amino acids, 5•Œ-adenosine triphosphate (ATP) and clubhook, and boreo pacific gonate squids were rich in tau related compounds, trimethylamine and its oxide, and rine and hydroxyproline. Higher amounts of aspartic acid, cyclic and ƒÖ-betaines were measured with the previously glutamic acid, alanine, leucine, and arginine were detected reported methods.8) Na+, K+, Cl-, and PO43- were also de in the boreal clubhook and boreo pacific gonate squid termined by the reported methods.8) inks. Each free amino acid of the neon flying squid ink was Extractive nitrogen of the above extracts was deter lower than those of the boreal clubhook and boreo pacific mined by the Kjeldahl method. gonate squid inks. A portion of the extract was dried at 110•Ž and the Trimethylamine oxide was known to be detected in residue was designated as a dry matter. higher amounts in squid muscles,10-13)however, small amounts of less than 40 mg/ 100 g were detected in the Results and Discussion three squid inks in this study. Each amount of glycinebetaine of the neon flying squid Proximate Composition ink among five individuals fluctuated. Large amounts of Five samples of the neon flying squid ink and two sam homarine and glycinebetaine were detected in the squid ples each of the boreal clubhook and boreo pacific gonate inks and these values were of similar levels to or higher squid inks were analyzed and values are expressed as than those of the squid mantle muscles amounting to 60 means with S.D. and differences from the means, respec 300 mg/ 100 g for homarine and 300-930 mg/ 100 g for tively. Table 1 shows moisture, crude protein, lipid, and glycinebetaine, respectively.10-12) ash of the squid inks. Moisture of the boreal clubhook The squid inks seemed to be rich in Cl-. A large amount squid ink was higher than those of the neon flying squid of PO43- was detected in the boreo pacific gonate squid ink and boreo pacific gonate squid inks. Crude protein of the and this value was almost four times higher than that of boreo pacific gonate squid was higher than those of the the mantle muscle of boreo pacific gonate squid.10) The ink boreal clubhook and neon flying squid inks.
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