Analisis komposisi kimia pada ikan

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Analisis proksimat komposisi lemak pada ikan dapat dilakukan menggunakan metode-metode standar analisis Food Savety Regulation of Japan (Yamaji dan Sekiguchi, 1991). Kandungan air diukur melalui proses pengeringan menggunakan oven pada suhu 105 derajat Celcius. Kandungan lemak mentah dihitung menggunakan metode Soxhlet dengan bantuan diethyl ether untuk ekstraksi. Kandungan protein mentah dihitung menggunakan metode Kjedahl dan dengan mengalikan kandungan protein dengan konstanta 6,25. Kandungan abu dihitung melalui proses pemanasan pada suhu 550 derajat Celcius.

Untuk analisis kelas lemak, lemak mentah diamati dengan kromatografi kolom gel silika dan di-elusi dengan kloroform untuk lemak netral, dilanjutkan perlakuan menggunakan methanol untuk lemak kompleks. Campuran larutan eluent ini kemudian dipisahkan dengan alat evaporator berputar. Lemak yang dihasilkan kemudian dianalisis dengan kromatografi lapisan tipis melalui sistem solvent: n-heksana-ether-asam asetat (80:30:1, vol/vol/vol). Kelas-kelas lemak yang terpisah diidentifikasi berdasarkan referensi standar otentiknya.

Nilai karbonil (CV) dihitung dengan metode standar (Toyota, 1991). Senyawa reaktif thiobarturat (TBA) dihitung dengan metode distilasi uap yang telah dimodifikasi (Shibata dan Kinumaki, 1979).

Ikan yang digunakan dalam proses analisis komposisi lemak adalah spesies Y. lipolytica NRBC-10073. Ikan sampel dimasukkan ke dalam wadah steril yang mengandung unsur lemak. Untuk mengamati proses terjadinya pengurangan lemak pada sampel melalui fermentasi, wadah kemudian diinkubasi pada suhu 20 derajat Celcius selama 72 jam, dan ditambah 10 volume diethyl ether untuk ekstraksi lemak mentah. Lemak mentah yang didapatkan kemudian dipisahkan dari lemak netral dan lemak kompleks menggunakan kromatografi kolom. Eksperimen ini dilakukan selama ini tiga kali pengulangan. Untuk mengamati perubahan oksidasi lemak dalam sampel dengan fermentasi, dan wadah diinkubasi pada suhu 20 derajat Celcius, sampel pada waktu inkubasi 0,24 dan 48 jam. Semua sampel ikan pada wadah kemudian disimpak dalam almari pendingin hingga proses analisis selesai. Nilai karbonil (CV) dan senyawa reaktif tiobarbiturat (TBA) ikan dihitung dengan metode tersebut di atas. Eksperimen ini dilakukan secara kuaterner.

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