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ADOCX/557213CA497959FC0B9306AC) Please download to view (https://docslide.com.br/download/link/determination-of-vitamin-adocx) Determination of Vitamin A Setelah ekstraksi, penentuan dilakukan pada pelarut ekstraksi cair. kolorimetri Penentuan Melalui metode ini, karotenoid dan vitamin A telah ditetapkan. Penentuan karotenoid. Karotenoid ditentukan pada 450 nm. Setelah menguap fase eter dari larutan yang diekstraksi, ekstraksi dilarutkan kembali dengan 1 ml heksana. Tentukan O.D.
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fase ini pada 450 nm. Penentuan vitamin A. Tahap heksana diperoleh sebelumnya diambil lagi dan terkonsentrasi dalam ruang hampa. diLarutkan kembali ekstrak dalam sebuah kloroform. Kemudian, dengan volume kloroform, tambahkan empat volume pereaksi asam trifluoroasetat dibuat dengan mencampur 1 v asam trifluoroasetat dengan tiga volume kloroform. Kemudian, amati DO pada 620 nm Determination of Vitamins D Penentuan D2 Vitamin, D3, dan Metabolit mereka Jika sampel mengandung semua vitamian metabolit D, maka Anda dapat melakukan kromatografi cair dalam kondisi berikut: Kolom: kolom analisis asam lemak Pelarut: MeCN (asetonitril), 55% Campuran air / asam asetat (4 ml per liter asetat) 45% Laju alir: 1 ml / menit Panjang gelombang: 265 nm Pelarut suhu: 25 Suhu oven: 40 Determination of Vitamin E Sejumlah metode dapat digunakan untuk menentukan vitamin ini. 1. kolorimetri Penentuan Setelah ekstraksi dan evaporasi, kembali melarutkan residu menggunakan nheptana. Tambahkan 1 ml larutan dipyridil, kemudian menentukan absorbansi pada 460
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nm. Metode berasal dari yang satu ini telah direkomendasikan untuk digunakan dengan besi klorida dengan pembacaan pada 510 nm. 2. Penentuan dengan Kromatografi Cair Menggunakan ekstrak dibuat seperti yang dijelaskan sebelumnya, dilanjutkan dengan penentuan HPLC dengan ketentuan sebagai berikut: Kolom: LiChrosorb RP 8,25 cm x, 4,6 mm, 5 pM Pelarut: metanol / air (92:8) Laju alir: 1,5 ml / menit Panjang gelombang: 288 nm Pelarut suhu: 25 Suhu oven: 40 * Untuk beberapa makanan, ketiga vitamin (A, D, E) dapat ditentukan, atau hanya vitamin A dan E secara bersamaan. Extraction of Hydrosoluble Vitamins 1. Prinsip umum adalah bahwa sampel harus dihancurkan sehalus mungkin. 2. Ekstraksi enzimatik dilakukan dengan enzim amilolitik atau proteolitik Alur: menghomogenkan sampel setelah menghancurkannya halus dan cepat (jika perlu) Timbang sampel g 5-10 ke dalam labu 250 ml dan tambahkan 65 ml HCl 0,1 M Panas pada 100 derajat C selama 30 menit dalam air mandi Dinginkan dan menyesuaikan diri dengan pH4.5 dengan 2,5 M NaOAc Tambahkan 50mg -amilase dan 50 mg takadiastase, diinkubasi pada 37 derajat Celsius pada oven selama semalam Tuang kuantitatif ke
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dalam labu ukur 100 ml dan mengatur volume dengan air Saring supernatan lebih lanjut dan lakukan untuk filtrat jika diperlukan Ascorbic Acid Extraction 1. Dalam breaker berat menjadi sekitar 0,1 mg kuantitas produk tertentu sebagai fungsi dari isi C diasumsikan vitamin zat makanan sampel. 2. Tuang ke dalam labu 50 ml volumetrik menggunakan asam metafosfat 0,4%. 3. Bawa volume sampai 50 ml dengan solusi ini. *. Untuk analisis cairan, pippet sampel langsung ke labu dan mengatur volume sampai 50 ml dengan solusi metafosfat 0,4% asam. 4. Saring larutan a membran selulosa asetat (0,2 mg), kemudian lulus substansi disaring melalui cartridge SEP-PAK C18 (yang disediakan oleh Waters, sebuah divisi dari Millipore). Hilangkan 2 ml pertama, kemudian mengumpulkan 5 ml untuk analisis oleh RP-HPLC. (Catatan: Harus menunjukkan bahwa beberapa penulis dapat mengusulkan penentuan asam askorbat dan dehydro-L-sacorbic yang teknik ekstraksi adalah berbeda dari yang sebelumnya) Determination of Water Soluble Vitamins 1. Penentuan Vitamin B1 Thiamine 1,1 Fluorimetric Penentuan Setelah aksi ferricyanide kalium di hadapan kalium, menentukan fluoresensi
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menggunakan filter utama 360-365 nm dan 460-480 nm filter sekunder. 1,2 mikrobiologi Penentuan Sejumlah laktobasilus dapat digunakan, seperti Lactobacillus fermentum dan Lactobacillus viridiceus ATCC 1270 C, tergantung pada metode
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kimia penentuan. Perhitungan statistik aktivitas didasarkan pada metode enam poin. 1,3 Penentuan Kromatografi Cair
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kromatografi kondisi Kolom: C 18, 25 cm x 4,6 mm, 5µm Pelarut: metanol 0,005 M natrium asetat disesuaikan menjadi 4,5
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pH (30-70) Laju alir: 1 ml / menit Deteksi: eksitasi: 366 nm emisi: 435 nm
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