Download - Fortif Bioavai Nutr 2014
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PENGUMUMAN
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PEMBAGIAN KELOMPOK FORTIFIKASI
No. klp Nama No. klp Nama
1. Purbo Carito 5. Larasati Novitasari
Bonardo T. Manurung Kadek Intan Putri P.
Yanni Hadiwibowo Izzata Ishmah
2. Mustiko Warih 6. Natalia Fransisca P.
Zakia Dwi Utami K. Mutmainah R. Indriati
Agata Pinastika Kena Veto Anggitaningrum
3. Gugum Kaelani S 7. Mentari Sekar Putri
Maurits Karindahang Asti Rahma Puspita
Yudha Atnawilastra Indri Astuti
4. Winta Hayu Pramesti 8. Alif Zulfahmi (10007)
Andi Suares Sirait Monica (10092)
Kevin Tanumihardja Mentari (09668)
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PEMBAGIAN KELOMPOK FORTIFIKASI
No. klp Nama No. klp Nama
9. Rati Satiti 13. Yulius Damara D.P.
Ratih Kumala Dewi Laras Sulastri Suher
Diah Fitalina Syani Alfian Mey Pramita D.
10. Puspita Kumala Sari 14. Kamilia Indraputri S.
Fanny Arivia Davanti Istna Nafi Azzahrani
Gina Noviani Karlinda Siti Hanum Indah L.
11. Anindhita Dewanti N. 15. Chrysanta Aurelia
Fariz Nurpita Aziz Margareta Parahita W.
Putu Ayu Kartika U. Henri Suhardja
12. Kartika Wulan Sari 16. Rr. Ardinta Fajrini S.
Fahmi Irfan Nadia Citta Paramita
Monica Yudha C.I. Natasha Rona P.I.
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Keterangan :Setiap kelompok diharuskan menyusun
satu paper tentang fortifikasi dalam baha-sa Indonesia, sengan sumber minimal satu makalah ilmiah berbahasa Inggris dari jurnal ilmiah internasional (misal J. Food Science, J. Food Chemistry)
Makalah yang digunakan sebagai sumber utama harus dilampirkan dalam bentuk hard copy pada paper yang dikumpulkan ke dosen pengampu (bu Sri Naruki)
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Guna keperluan presentasi, kelompok mahasiswa juga diminta membuat ‘power point’ papernya masing-masing
Presentasi paper oleh mahasiswa dilaksa-nakan pada tanggal 1 dan 8 Oktober 2014
Nama mahasiswa yang diketik tebal dan berwarna merah bertugas sebagai ketua (penanggung jawab) kelompok
Mahasiswa yang namanya tidak tercantum dalam tabel, akan disertakan pada ‘kelompok formulasi’ (dosen pengampu: Prof. Dr. Mary Astuti, MS)
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LANJUTAN KULIAH MINGGU LALU (10 September 2014)
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I
IRON DEFICIENCY ELIMINATION ACTION
(IDEA)
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DEFISIENSI ZAT BESI > 1 miliar orang (66 – 88% dari populasi
dunia) menderita efisiensi Fe. Sekitar 90% dari jumlah tersebut berada di negara yang sedang berkembang
PENANGGULANGAN
1. Penerapan diet yang terencana dengan baik, sehingga dapat memenuhi kebutuhan Fe tubuh
2. Pemberian tablet suplemen zat besi
3. Fortifikasi zat besi
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1. PENERAPAN DIET YANG TERENCANA DENGAN BAIK
Merupakan cara terbaikSulit diterapkan di negara yang sedang
berkembang, seperti Indonesia ada kecenderungan pola makan yang kaya akan bahan nabati & miskin bahan hewani (Tab. 1)
2. PEMBERIAN TABLET SUPLEMEN ZAT BESITidak dianjurkan karena ada resiko over dosisSebaiknya dilakukan di bawah pengawasan
dokterKadang timbul efek samping : mual, sembelit,
muntah
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Tabel 1. Kontribusi protein (%) dari kelompok bahan makanan menurut Kota / Desa
Kelompok bahan makanan
Kota Desa
Padi-padian 41,1 47,8
Daging / ayam 3,9 2,7
Telur 4,5 2,7
Ikan 15,7 13,4
Susu 0,7 0,5
Kacang-kacangan 28,3 27,4
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Kelebihan asupan zat besiAsupan 200 mg/hari dalam waktu lama
timbul akumulasi Fe di jaringan lunak, seperti hati dan limfa, serta persendian
Gejala : lemah, penurunan berat badan, sakit pada persendian, pembengkakan hati, bahkan sirosis hati
Kelebihan Fe dapat pula terjadi pada individu yang sering menerima transfusi darah. Misal : penderita talasemia
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3. FORTIFIKASI Fe (vs IDEA)Fortifikasi Fe merupakan cara yang
menjanjikan dalam memerangi defisiensi Fe
ILSI-CHP (International Life Sciences Institute – Center for Health Promotion) : pada tahun 1997 mendirikan Project IDEA
FOKUS : membantu mengurangi beban global defisiensi Fe melalui FORTIFIKASI MAKANAN
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MENGAPA FORTIFIKASI MAKANAN ?
1. Essential part of any strategy to reduce iron deficiency
2. Secara sosial mudah diterima karena tidak mengubah food habit
3. Secara legal, penerapannya dapat diwajibkan oleh pemerintah
4. Relatif mudah dimonitor5. Cost-effective
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TUJUAN PROJECT IDEA1. Menjalin kerjasama dengan sektor publik &
privat untuk memerangi defisiensi Fe2. Melakukan identifikasi vehicle/carrier &
fortifikan Fe yang sesuai di masing-masing negara
3. Mengamankan bantuan unt program khusus4. Melakukan evaluasi pengaruh fortifikasi Fe
terhadap status Fe populasi target5. Mendorong program-program pendidikan
untuk meningkatkan kepedulian masyarakat6. Mengembangkan sistem monitoring dan
pengawasan
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AKTIVITAS DI CHINA
Studi penambahan NaFeEDTA ke kecap kedelai (dikonsumsi oleh >70% popu-lasi) defisiensi Fe menurun
Tahun 2004 : program fortifikasi Fe pada kecap kedelai dimulai secara nasional
Harapan : lebih dari 100 juta rakyat akan menuai keuntungan dari program tersebut
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KEUNGGULAN KECAP (IKAN & KEDE-LAI) SEBAGAI VEHICLE :
1. Berupa cairan memungkinkan penyebar-an fortifikan menjadi lebih homogen
2. Merupakan produk fermentasi bahan berprotein tinggi sehingga kaya akan asam amino Lys, His, dan Cys, yang diketahui dapat memacu absorpsi Fe
3. Warna : coklat gelap dan citarasa = kuat dapat menutupi perubahan warna & menyamarkan perubahan citarasa akibat fortifikasi
4. Jumlah konsumsi = terbatas resiko overdosis Fe dapat dihindari
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AKTIVITAS DI VIETNAM Bekerjasama dengan The National
Institute of Nutrition (vehicle = saus ikan)
MENGAPA VEHICLE NYA SAUS IKAN ?
1. Dikonsumsi oleh > 80% populasi (10 – 15 mL/kapita/hari)
2. Terdapat jaringan pabrik saus ikan di bawah supervisi The Ministry of Fishery memudahkan pengaturan proses fortifikasi
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MACAM AKTIVITAS PROJECT IDEA DI VIETNAM :
1. Field survey2. Bioavailability study3. Efficacy study4. Effectiveness study5. National lauch of fortified fish sauce
which is composed of : (a) Production & distribution, (b) Quality assurance, (c) Social marketing & education, and (d) Monitoring & surveillance (penga-wasan)
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1. FIELD SURVEYHasil survey membuktikan bahwa
fortifikasi dapat dilakukan melalui sedikit modifikasi pada proses, yaitu sebelum tahap pembotolan
Biaya : terbukti affordable (dapat ditanggung / cukup murah)
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2. EFFICACY STUDYPada tahun 2000 dilakukan efficacy
study selama 6 bulan Subyek : 152 perempuan pekerja pabrik,
17 – 49 tahun, Hb > 8 tapi < 12 g/dLHasil : konsumsi fortified fish sauce
(1mg Fe sebagai NaFeEDTA/mL saus ikan) sebanyak 10 Ml/kapita/hari dapat menu-runkan prevalensi anemia
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3. EFFECTIVENESS STUDY Program berskala besar dengan 14.000
populasi di pedesaan, dilakukan sejak tahun 2001
Area dibagi menjadi dua kelompok : 1 (= kontrol) dan 2 (=mengkonsumsi fortified fish sauce, 7,5 mg Fe/kpt/hari)
Subyek perempuan, 16 – 49 tahunHasil: prevalensi anemia (Hb < 12 g/dL)
dan defisiensi Fe (feritin serum < 12 ng/mL) MENURUN
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5a. Production & distributionFortifikasi diterapkan pada saus ikan
dengan mutu low dan mediumTahun ke-1 : dua pabrik besar di
Vietnam Utara dan Tengah memperkenalkan fortified fish sauce
Program diperluas ke 30 pabrik yang memproduksi 80% saus ikan di Vietnam
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5b. Quality assuranceProgram jaminan mutu, baik di
pabrik maupun di pasar, dilaksanakan dibawah Ministry of Health
Untuk memperbaiki manajemen mutu, diperkenalkan pula GMP dan HACCP
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5c. Social marketing & education Dilakukan untuk mendidik
masyarakat tentang :
1) Pentingnya Fe bagi kesehatan
2) Makanan yang baik sebagai sumber Fe
3) Pentingnya fortified fish sauce bagi masyarakat
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5d. Monitoring & surveillanceDilakukan untuk menjamin terlaksa-
nanya fortifikasi dengan baikPengawasan dikembangkan untuk
mengevaluasi IDA (iron deficiency anemia) dan perbaikan konsumsi makanan di Vietnam
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AKTIVITAS DI FILIPINA
Bekerjasama dengan FNRI (The Food and Nutrition Research Institute) tentang the stability and acceptability of several alternatives on iron fortification of rice, the national staple.
Hasil : metoda yang menjanjikan adlh
a. Ekstrusi beras dengan fero sulfat
b. Ekstrusi beras dengan micronized ferric phosphate (mfpp)
c. Coating beras dengan mfpp
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Bekerjasama dengan FNRI (The Food and Nutrition Research Institute) tentang the stability and acceptability of several alternatives on iron fortification of rice, the national staple.
Hasil : metoda yang menjanjikan adalah
a. Ekstrusi beras dengan fero sulfat
b. Ekstrusi beras dengan micronized ferric phosphate (mfpp)
c. Coating beras dengan mfpp
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Dilakukan pula efficacy study pada Juli 2004 selama 6 bulan
Subyek : 240 anak SD, anemia, 6-8 tahun, dibagi dalam 4 kelompok
Diet : beras biasa (untuk kelompok kontrol) + 3 fortified rice untuk 3 kelompok uji (10 mg Fe/anak/hari)
Hasil : setelah 3 dan 6 bulan, terdapat penurunan jumlah penderi-ta iron deficiency anemia
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AKTIVITAS DI INDONESIAProject IDEA bekerjasama dengan
UNICEF menyelenggarakan beberapa workshops untuk mendorong fortifikasi Fe di Indonesia
Depkes melalui SK Menkes No.632/ 1998 : terigu yang diproduksi dan dijual di Indone-sia harus difortifikasi dengan Fe, Zn, tiamin, riboflavin, dan asam folat
Kadar Fe terigu = 60 ppm
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BIOAVAILABILITY OF NUTRIENTS
BIOAVAILABILITAS ZAT GIZI
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PENGANTAR
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BIOAVAILABILITAS (Brody, 1994) : the percentage of the nutrient in the food that is absorbed and used for physiological purpose (such as red blood cell formation for iron)
BIOAVAILABILITAS : the extent to which nutrients added to food are absorbed and reach the plasma (Bauernfeind & Lachance, 1991)
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MENGAPA BIOAVAILABILITAS = PENTING ??
Bioavailabilitas yang tinggi : MERUPAKAN SALAH SATU SYARAT FORTIFIKAN
Syarat fortifikan :
1. Bioavailabilitas : tinggi
2. Reaktivitas fortifikan dapat mengaki-batkan diskolorisasi maupun penyim-pangan bau, rasa, dan flavor
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2. Stabilitas selama penyiapan & penyim-panan makanan
3. Kompatibilitas dengan nutrien lain
4. Harga
5. Kehalalan
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FAKTOR YANG BERPENGARUH TERHA-DAP BIOAVAILABILITAS ZAT GIZI (Walker & Roll, 1992) 1. Agronomic and environmental
factors : Pupuk membuat produksi kebun
meningkat komposisi produk bisa berubah (misal : kadar senyawa fitat)
Hujan asam : mempengaruhi kelarutan mineral
Penelitian tentang hal ini : masih sedikit
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2. Food processing :a. Pemanasan (suhu + waktu) mengaki-
batkan : Kehilangan vitamin CReaksi Maillard menghasilkan senya-
wa kimia yang dapat membentuk hydrolysis-resistant metal complexes dapat menurunkan bioavailabilitas mineral
b. Fermentasi : bisa menaikkan bioavai-labilitas beberapa zat gizi
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3. Interaksi mineral dengan kompo-nen lain dalam bahan pangan
Komponen oksalat, tanin membentuk stable insoluble mineral complexes inhibitor absorpsi mineral
An increase in dietary fibre cause negative mineral balance for one or more minerals in man and other animals
Fitat (myo-inositol hexaphosphate) binds minerals in the order Zn > Fe > Ca > Mg > Cu menurunkan bioavailabilitas
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Konsumsi obat ada beberapa obat yang mempengaruhi bioavailabilitas zat gizi (contoh lihat Tabel 1)
Tabel 2 tentang factors influencing the bioavailability of vitamins added to food
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Table 1. Some drugs group and drugs that may cause nutrient depletion and nutrient
deficienciesDrug group Drugs Deficiency
Antacids Na-bikarbonat, Al-hidroksida
Folat, fosfat, kalsium, copper
Antibiotic TetrasiklinGentamicin
KalsiumPotasium, Mg
Antibacterial Boric acid Riboflavin
Anti-inflammatory
AspirinPrednison
Vit C, folat, FeKalsium
Anti cancer Methotrixate Folat, kalsium
Diuretic Forusemide K, Ca, Mg
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Table 2. Factors influencing the bioavailability of vitamins added to food
Vitamin Factor Effect
ß-Carotene
FatLecithinProtein
Pectin + methil celulloseTocopherol
EnhancesEnhancesConvertion to vit A : increasesGreater than 10% : decreasesIncreases
Thiamin Sulfites Decreases
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Table 2. Factors influencing the bioavailability of vitamins added to food (lanjutan)
Vitamin Factor Effect
Retinyl ester
Fat, proteinPUFA
IncreasesDecreases
Riboflavin
Glucose polymer, Na-alginate, hemi-cellulose
Increases
Folic acid FiberVitamin C
No effectPromotes
Vitamin B6
Hemicellulose Minor reduction
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FACTORS INFLUENCING DIETARY IRON ABSORPTIONIron status (body’s need for iron)Form of ironComposition of the mealBulk in dietSize of doseBalance between plus- and minus-
factors in meal (see the next page)
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Iron status
Form of iron
Absorption of reduced ferrous (Fe2+) is greater than the oxidized ferric (Fe3+)
Composition of the meal
Iron status Iron absorption, %
Normal hemoglobin level 2 – 10
Low hemoglobin evel Up to 35
Kind of the meal Iron absorption, %
Vegetables food (rice, wheat, corn, spinach)
+ 5
Meat + 20
Fish + 10
Vegetables + meat + 10 – 15
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Bulk in diet
High fiber or cellulose in diet resulted in poor absorption of iron
Size of dose
Plus- and minus- factors in meal- Meat, fish, ascorbic acid : positive correlation
- Phytate, Ca, polyphenols : negative correlation
Intake of iron Absorption Absorption
0.25 mg/kg BW 32 % 0.08 mg/kg BW
4 mg/kg BW 4.1 % 0.164 mg/kg BW
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Balance beetwen plus- and minus- factors in meals a. Plus-factors :- Meat, chicken, fish, sea food- Ascorbic acid (fruits, juices,
potatoes, vegetable)- Some fermented food
(sauerkraut, fermented soy-sauce, etc)
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b. Minus-factors :- Phytates and inositol hexaphosphates
(bran products, oat products, high extraction flour, nuts, etc)
- Calcium in different form (milk, cheese, etc)
- Iron-binding phenolic compounds (tea, coffe, cacao, certain spices, vegetables, etc)
- Other factors : soy protein, manganese
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METODA PENENTUAN BIOAVAILABILITAS
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Bioavailabilitas dapat ditentukan secara
IN VIVO memakai makhluk hidup atau hewan percobaan / hewan coba / hewan laboratorium. Misal : tikus, babi, manusia
Metoda IN VITRO hasil : availability (bukan bioavailability) zat gizi
Metoda IN SITU
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METODA IN SITU : memakai organ hewan hidup (organ dalam kondisi hidup dan masih menempel di hewan yang bersangkutan) hewan masih hidup atau dibuat pingsan
Misal :
- Memakai tikus yang dibuat pingsan
- Memakai sapi hidup yang telah mengalami
surgical interventionAda yang tetap menganggap in vivo
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PENELITIAN BIOAVAILABILITAS YANG VALID :
Sedapat mungkin memakai manusia sebagai subyek.
Subyek memiliki kesamaan umur, jenis kelamin, status fisiologis & kesehatan
Nutrient-enriched food yang diberikan untuk diuji harus diolah dan disajikan dengan cara seperti yang biasa dilakukan / dikonsumsi
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PEMAKAIAN HEWAN LABORATORIUM/ HEWAN
PERCOBAAN Dapat dilakukan asalkan keterbatasan mereka dimengerti
Contoh : kera, babi, anjing, kelinci, tikus, ayam jago
Misal pada tikus : 1. Tikus dapat menghasilkan vitamin C
secara endogenous tikus tak dapat dipakai untuk penelitian biaoavailabilitas vitamin C
2. Tikus tidak dapat disuruh mengasup seju-mlah tertentu makanan (kecuali melalui force feeding)
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1. HEWAN BESAR SEBAGAI MODEL DALAM PENELITIAN HUMAN NUTRITION Pendapat : supaya valid model harus
manusia TETAPI : a. Mahal (10 x babi; 100 x tikus)b. Mencari kelompok subyek yang
homogen : sulitc. Kondisi lingkungan, sosial dan
fisiologikal : dapat berubah sebelum penelitian selesai
d. Pada penelitian tertentu : kadang ada dampak negatif penggunaan subyek manusia = tidak boleh
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Oleh karena itu : sering digunakan hewan sebagai subyek penelitian
Yang harus diperhatikan :Perbedaan antara hewan dan manusia
antara lain : anatomi (GI tract & liver), fisiologi (pertumbuhan dan metabolisme), respons imunologikal, kebutuhan akan zat gizi.
BABI : sangat mirip (anatomi, fisiologi) dengan manusia (lihat Tabel 3)
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Table 3. Similarities between pigs and man
Dental characteristicsRenal morphology & physiologyEye structure & visual acuitySkin morphology & phisiologyCardiovascular anatomy & phisiologyDigestive anatomy & phisiologyProportions of fat-free body at common stages of life
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Keunggulan lain dari babi :Patuh; ukuran cukup besar sehingga :- memudahkan dalam sampling size- memungkinkan surgical interventionSiklus reproduksi : pendek (115 hari)Mau memakan banyak jenis makananMudah diperoleh secara komersial CONTOH : penelitian tentang
aterosklrosis hasil sangat mirip dengan manusia
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2. HEWAN KECIL SEBAGAI MODEL DALAM PENELITIAN HUMAN NUTRITION
Animal experiments have played a vital role in human biological research (including human nutrition)
Rodents : small, have a short generation time and large litter size reduces cost of experiment
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METODA ANALISIS: BIOAVAILABILITAS VITAMIN (pada makanan konvensional)
Penelitian dengan subyek manusia dapat dilakukan dengan 3 metoda :
1. Change in the PLASMA LEVEL after ingestion of the food
2. Measurement of URINARY LOSS after ingestion of the food with prior tissue saturation
3. RADIOTRACER METHOD for vitamin absorption from GI tract
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1. CHANGE IN PLASMA LEVEL OF VITAMIN
Compare folate absorption from different food products rat and chick bioassay has also been used
Examine the effect of drug such as folate absorption
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2. PENGUKURAN URINARY EXCRETION OF VITAMIN
Pernah dilakukan untuk mengevaluasi absorpsi riboflavin dari beberapa produk makanan, khususnya tentang pengaruh food additive pada absorpsi riboflavin
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3. RADIOLABELLINGDapat memakai hewan percobaanUntuk menentukan bioavailabilitas
vitamin B6 dari produk makanan hewani
Pernah diterapkan dalam studi tentang efek thermal processing pada bioavailabilitas vitamin
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Tabel 4. Metoda penentuan bioavailabilitas vitamin yang ditambahkan ke makanan
Vitamin Makanan Metoda
Asam folat
Bread, other cereal
Frozen food
Folate excretion
Chick bioassay
B6 Enriched cornmeal
Rat bioassay, labelling
Riboflavin Breakfast cereal Urinary excretion
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BIOAVALABILITY OF VITAMIN ADDED TO INFANT FORMULA
It was commonly examine by comparing BLOOD LEVEL of vitamins of concern in breast- and formula-fed infants
Contoh : studi tentang bioavailabilitas vitamin K diukur kadar vitamin K serum dan feses
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METODA ANALISIS : BIOAVAILABILITAS MINERAL (pada makanan konvensional)
ANTARA LAIN :1. Metoda Radiolabelling : paling akurat,
baik pada penelitian dengan hewan coba maupun manusia
2. Balance Method (Neraca Mineral)3. Depletion-Repletion Method (for Fe)4. Metoda Digesti dan Dialisis In Vitro5. In Vivo Rat Intestinal Perfusion
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6. Pengujian Absorpsi Mineral Secara In Situ
7. Penentuan Absorpsi Mineral dengan Metoda Usus Terbalik (Inverted Sac)
8. Change in Serum Level
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Tabel 4. Metoda penentuan bioavailabilitas mineral yang ditambahkan ke bahan makanan
Mineral Metoda
Kalsium Fecal calcium
Calcium balance
Radioisotopic methode
Besi Labelling In vitro dialysis
Hb repletion
Seng Metabolic balance
Intestinal perfusion
Isotropic tracer
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1. Metoda radiolabelling (radioisotopic methode)
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Example : Effect of ascorbic acid intake on nonheme-iron absorption from a complete diet (Cook and Reddy, 2001)
There were perfomed 4 separate measurement of iron absorption in 10 women and 2 men (20-38 y, iron status were normal – serum ferritin > 12 µg/L)
Iron absorption tests were performed in each subject by using radioactive iron label
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Sumber Fe : 59FeCl3Macam diet : a. SS (self selection) diet + 59FeCl3 b. HC (high-vitamin C) diet + 59FeCl3 c. LC (low-vitamin C) diet + 59FeCl3Study design : 5 d 5 d 5 d 5d hamburger a b c
59Fe analyse
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Standard meal : hamburger Radioactivity was measured in duplicate
10-mL blood samplesPercentage absorption was calculated
based on the total blood volume estimated from the height and weight of each subject
The red cell incorporation of absorbed radioactivity was assumed to be 80% in all subject
The volume of blood is about one-eleventh (7-9%) of the body weight man (70 kg) = 5L and woman = 4,7L
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Daily nutrient intake during the 3 labeling period
Nutrient SS diet HC diet LC diet
Energy, CHO, P, L See text See text See text
Iron (mg)- Nonheme- Heme- TOTAL
11.40.7
12.1
16.61.0
17.6
12.80.8
13.6
Vitamin C (mg) 90 247 51
Others
- Animal tissue (g)- Tea (ml)- Fiber (g)
10910313.4
8491
16.0
10310112.5
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The absorption data in human subjects
Subject
Iron absorption (% of dose)
Standard meal
(hamburger)
SS diet HC diet LC diet
MaleFemale
4.812.1521.516.182.9013.96
etc
1.181.8624.272.162.425.20etc
3.951.7717.153.163.367.21etc
3.181.629.188.784.354.01etc
Mean 8.00 4.57 7.69 5.69
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2. Metoda Neraca Mineral (Balance Method)
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Tikus jantan yang telah melewati masa adaptasi 4 hari, dipelihara di kandang metabolit selama 7 hari
Tikus diberi pakan dan minum secara ad libitum
Asupan mineral: bisa melalui asupan pakan atau force feeding
Setiap hari, sisa pakan ditimbang (untuk menge-tahui asupan mineral bila asupan mineral bukan melalui force feeding, melainkan melalui asupan pakan), sedangkan eksresi urin dan feses dikumpul-kan dan disimpan di almari es
Pada akhir pemeliharaan, dilakukan analisa mineral terhadap feses, urin, dan pakan
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INGAT : Untuk penelitian tentang neraca
mineral Fe : dipakai tikus yang sebelumnya dibuat anemia
Cara pembuatan anemia : tikus diberi pakan standar bebas Fe (sekitar 14 hari) sampai dicapai kadar Hb 6,0 g/dl
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PERHITUNGAN (misal untuk mineral Zn)A = absorpsi Zn = asupan Zn – Zn fesesApparent digestibility = % A/I
= (absorpsi Fe : asupan Fe) x 100R = Retensi = retained Zn = (absorpsi Zn – Zn urin)% R/A = (retensi Zn : absorpsi Zn) x 100% R/I = (retensi Zn : asupan Zn) x 100
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Example : Zinc balance in rats : influence of the
heat treatment of a casein-glucose-fructose mixture (Navarro, et al, 2000)
------------------------------------------------------------Weanling Wistar rats, weighing 40 + 0.5g
were used for balance assays (8 animals/group)
Animals were allowed free access to food and water
Each animal was placed in an individual cage in an environmentally controlled room kept at 20-22oC, with light/dark cycle and 55-70% humidity
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The test involved a preliminary 4-day adaptation period during which solid intake and body weight changes were monitored, followed by a second period lasting 7 days in which, moreover, feces and urine were collected to perform the Zn balance. Urine and feces were collected as a 1-week pool for each animal during this period
The feces were lyophilized, weighed, and homogenized
Urine were collected in 0.5% HCl solution, filtered, and diluted with deionized distilate water.
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DIET PREPARATIONComposition : 37.5% starch, 29.83%
sugar, 8% olive oil, 5% cellulose, 3.81% mineral mixture and 0.16% vitamin mixture.
Sample C and HC (16.90%) were used as sources of protein and partial sources of carbohydrate to prepare the C-D and H-CD diets, respectively.
C = raw casein-glucose-fructose, and HC = heated (150oC for 1 hour) casein-glucose-fructose.
The nutrient composition : see Table 1
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Table 1. Nutrien content of experimental diets containing unheated casein-glucose-fructose (C-D) and heated casein-glucose-fructose (HC-D)
Diet
C-D HC-DMoisture (g/100g)Protein (g/100g)Fat (g/100g)Ashes (g/100Ca (mg/g)Mg (mg/g)Fe (g/g)Cu (g/g)Zn (g/g)
3.9510.558.043.305.930.46
56.208.05
35.08
4.3610.494.833.465.790.46
55.257.57
32.64
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To control for possible contamination during the collection of feces and urine, blank capsules were manipulated in the same way as those used for the animals
The following indices were calculated from the data obtained for Zn intake and fecal and urinary excretion :
1. Absorbed Zn = (ingested Zn – fecal Zn)
2. Retained Zn = (absorbed Zn – urinary Zn)
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3. Apparent digestibility = % A/I
= (absorbed Zn/ingested Zn) x 100
4. Proportion of ingested Zn retained in the body = % R/I
= (retained Zn/ingested Zn) x 100
5. Proportion of absorbed Zn retained in the body = % R/A
= (retained Zn/absorbed Zn) x 100
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Figure. % A/I, %R/A, and % R/I in rats fed different diet. C-D, diet containing raw casein-glucose-fructose; HC-D, diet containing heated casein-glucose-fructose
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ConclusionThe heated casein-glucose-fructose
increased the urinary Zn excretion and lowered Zn absorption and retention.
The effect of browning products generated during food processing should be taken into account, especially in diets containing marginally adequate levels of Zn, to prevent possible deficiency.
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3. Penentuan Bioavailabilitas Fe dengan Metoda Deplesi-
Replesi
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Tikus jantan sapihan (umur 21 hari) dibuat anemia (Hb < 6 g/dl) periode deplesi
Tikus kemudian dipelihara dalam kandang individual; diberi diet + Fe (0, 6, 12, dan 24 mg/kg diet) selama 14 hari periode replesi
Sebagai standar : dipakai fero sulfatPada awal dan akhir periode replesi,
dilakukan penentuan Hb darah dan penimbangan BB
Bioavailabilitas Fe dinyatakan sebagai NILAI HRE
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PERHITUNGAN NILAI HREHRE = hemoglobin regeneration
efficiency% HRE = [(mg Hb akhir – mg Hb awal) x
100] : mg asupan FeAsumsi : a) volume darah tikus (dalam satuan ml)
= 6,7% dari BB (dalam satuan g) b) kadar Fe hemoglobin (dalam satuan
g/L) = 0,335%Jadi berat Fe Hemoglobin (mg) = [berat badan (g) x Hb (g/L) x
6,7 x 0,335] : 10.000 (Sumber : Hernandez, et al., 2003)
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Example : Iron bioavailability and utilization in rats are lower from lime-treated corn flour than from wheat flour when they are fortified with different sources of iron (Hernandez et al., 2003)Diet were based on wheat and lime-treated corn
flour, with and without fortification with iron (30 mg iron/kg flour) i.e. reduced iron (ri), ferrous sulfate (fs), ferrous fumarate (ff) and ferric citrate (fc).
Diets were prepared with 95% lime-treated corn flour and 82% wheat flour, with protein content of 8 and 9%, respectively
Male anf female Sprague-Dawley rats, 21-23 d old, were fed the casein diet (9% protein and 1.4mg iron/100g) for 10 d (depletion period)
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After depletion phase, the rats were weighed and a blood sample was obtained from the tail vein to measure initial Hb.
The anemic rats were assigned to 10 groups (n = 6)
Each group was fed one of 10 experimental diet during the repletion period (14 d).
The control diet consisted of unfortified wheat or lime-treated corn flour
The composition of 10 experimental diets : see Table 1.
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Table 1. Composition of experimental dietsSample Mois-
tureAsh Protein Fat Crude
fiberCarbo -hydrate
Phy-tates
g/100g
LCF
LCF+ri
LCF+fs
LCF+ff
LCF+fc
8.7
9.9
8.5
8.5
8.4
1.3
1.3
1.2
1.3
1.3
8.6
7.6
8.3
8.2
7.8
4.3
4.2
4.2
4.3
4.2
1.5
1.3
1.8
1.7
1.6
75.6
75.7
76.0
76.0
76.7
0.503
0.500
0.494
0.492
0.514
WF
WF+ri
WF+fs
WF+ff
WF+fc
10.1
10.1
8.5
8.6
8.4
0.6
0.6
0.7
0.7
0.6
10.2
10.3
11.0
11.1
10.6
1.5
1.4
1.6
1.5
1.6
0.5
0.4
0.5
0.5
0.4
77.1
77.2
77.7
77.6
78.4
0.125
0.110
0.105
0.123
0.107
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*Note : Fe balance = g Fe intake – g Fe feces
Diet
Fe in diet
Fe intake
Fe excreted
Fe balance*
Fe ab-sorption
mg/100g mg %
LCF
LCF+ri
LCF+fs
LCF+ff
LCF+fc
2.52
5.39
5.62
5.62
5.93
2.68
6.07
6.68
8.05
7.58
1.90
4.35
3.90
4.77
4.64
0.78
1.72
2.78
3.28
2.94
28.9
27.9
41.5
40.6
39.0
WF
WF+ri
WF+fs
WF+ff
WF+fc
2.23
5.42
5.63
5.90
4.12
2.50
5.43
6.23
6.90
5.98
1.09
2.84
3.42
3.46
3.55
1.41
2.59
2.81
3.44
2.43
56.9
47.8
45.6
49.7
40.6
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*Note: mg Fe in Hb = (body weight x Hb x6.7x 0.335)/10.000
Diet
Weight Hemoglobin Fe in Hb*
Initial Final Initial Final Initial Finalg g/L mg
LCF
LCF+ri
LCF+fs
LCF+ff
LCF+fc
60
56
60
65
55
69
72
76
82
75
60
57
56
56
56
110
108
125
135
142
0.87
0.73
0.76
0.83
0.68
1.78
1.74
2.15
2.51
2.38
WF
WF+ri
WF+fs
WF+ff
WF+fc
60
72
62
80
80
74
82
75
91
97
57
57
57
60
60
109
123
156
140
145
0.77
0.94
0.80
1.09
1.06
1.85
2.26
2.61
2.90
3.16
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Note : % HRE = [mg Fe hemoglobin (final) – mg Fe hemoglobin (initial) x 100]/ mg Fe consumed
Diet
Gain HRE
Weight Hb Iron Hb
g g/L mg %
LCF
LCF+ri
LCF+fs
LCF+ff
LCF+fc
9
16
16
17
20
50
51
69
79
86
0.91
1.01
1.39
1.68
1.70
33.8
17.2
21.0
20.8
22.5
WF
WF+ri
WF+fs
WF+ff
WF+fc
14
10
12
12
16
52
66
99
80
85
1.08
1.32
1.81
1.81
2.10
41.4
22.8
29.5
26.2
34.7
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ConclusionIron bioavailability were higher from fortified and
unfortified wheat flour than from the lime-treated corn flour
Fortification of lime-treated corn flour with reduced iron has no effect on iron bioavailability, probably due to the high phytate content
Othe iron compounds must be selected to fortify lime-treated corn flour when intended for public nutrition programs.
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4. Penentuan Availabilitas Fe dengan Metoda Digesti &
Dialisis Secara In Vitro
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Example : Proteolytic digestion of meat is not necessary for iron solubilization (Carpenter dan Mahoney, 1989)
Digesti pertama : memakai PEPSIN, pada kondisi ASAM (mencerminkan digesti lambung)
Digesti kedua : memakai PANKREATIN, pada kondisi pH NETRAL (mencerminkan kondisi usus)
Kantong dialisis : MW cut off 6.000 - 8.000
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PRINSIP ANALISAMeal sampel + HCl sampai pH = 2,0 + 0,05Dilakukan penambahan pepsin (5 mL, 1 mg
pepsin /mL) dan FeCl3 (0,5 mL; 1000 mg FeCl3 /L)
Selanjutnya dilakukan inkubasi 37oC, 3 jam, dalam shaking waterbath
Hasil digesti pepsin (digested treatment) ditambah Tris 0,1M sampai pH 7,0
Pankreatin (5 mL, 10mg/mL) ditambahkan ke dalam digested treatment.
Kantong dialisis (MW cut off : 6.000 – 8.000) yang berisi 5 mL Tris 0,1M pH 7,0 dimasuk-kan ke digested treatment tersebut
Dilakukan inkubasi (shaking wb, 37oC, 3 jam)
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Lanjutan . . .Pada akhir inkubasi, kantong dialisis
diambil dan dilakukan analisa kadar Fe terhadap cairan didalam kantong dialisis
Fe dalam kantong = Fe yang dapat melewati membran dialisis = soluble iron = dialyzable iron dianggap = AVAILABLE IRON
Fe total dalam kantong
% Dialyzable Fe = --------------------------------------- x 100
Fe total dalam sample meal
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Diagram analisisMeal sample + HCl Pengaturan pH = 2,0 + 0,05
Penambahan pepsin dan sumber Fe yang diuji
Inkubasi 37oC, 3 jam, dalam shaking waterbath
Penambahan Tris sampai pH 7,0
Kantong dialisis Pemasukan kantong dialisis
(berisi 5 mL Tris 0,1M pH 7,0) ke cairan hasil digesti
Penambahan pankreatin ke cairan hasil digesti
Inkubasi 37oC, 3jam, dalam shaking waterbath
Analisis kadar Fe cairan di dalam kantong dialisis
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5. In Vivo Rat Intestinal Perfusion sama dengan in situ (?)
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IN VIVO RAT INTESTINAL PERFUSION
Sprague-Dawley rats, 250 to 350 g, were anesthetized with isoflurane
The abdomen opened, the ligament Treitz (duodenal-jejunal junction) identified
Polyethylene catheter (2.5 mm outer diame-ter x 1 mm inner diameter) were intalled with the proximale infusion catheter placed 5 cm beyond the ligament Treitz
The collection catheter positioned 20 cm distally
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Animal core temperature was monitored to be at 37oC by rectal thermometer
Intraintestinal perfusion were perfused at 0.4 mL/min. The residence time of the perfusate in the intestinal lumen was approximately 10 min.
The perfusion effluent was collected, imme-diately frozen, and subsequently analyzed.
(Courtesy : Yustin, et al., 2004)
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6. Pengujian Absorpsi Mineral Secara In Situ
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PENGUJIAN ABSORPSI IODIUM SECARA IN SITU (Mary Astuti, 1988 )
Tikus dipuasakan selama 24 jam, diberi minum ad libitum
Dilakukan anestesi dg sodium pento-barbital (dosis 40 mg/kg BB tikus)
Tikus diletakkan di meja bedah dan dilaku-kan pembedahan abdomen secara longi-tudinal
Usus halus dikeluarkan, bagian atas diikat dg benang sutera
Kira-kira 2-3 cm duodenum bagian ats diberi lubang dan dimasuki kanula plastik, kemudian diikat
P ada bagian pangkal ileum diikat dan diberi lubang serta dipasang kanula gelas, kemudian diikat
Kanula atas dihubungkan dg slang plasting pada buret yang berisi NaCl 0,9%
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Usus dicuci dengan 50 mL NaCl 0,9% (0,5
mL/menit) sampai seluruh kotoran keluarSetelah usus bersih, dilakukan perfusi dengan
memakai 50 mL larutan yang diuji (0,5 mL/ menit), yang berupa larutan NaI atau KIO3
Cairan yang keluar dari kanula bawah ditampung dalam bekerglass
Dilakukan analisis kadar I yang masuk maupun yang keluar
Kadar I yang diserap = {I yang masuk – I yang keluar}
Jumlah I yang diserap Persen absorpsi = x 100 Jumlah I yang masuk
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7. Penentuan Absorpsi Mineral dengan Metoda Usus Terbalik
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ANALISA ABSORPSI KALSIUM DENGAN METODA USUS TERBALIK (Rahmawati, 2004)
Tikus jantan (200-350g; dipuasakan 24 jam) dianestesi
Diambil usus bagian duodenum (10 cm) dan dicuci dengan NaCl 0,9%
Usus dibalik, ujungnya diikat dengan benang, dan diisi dengan NaCl 0,9% (1 mL/5 cm usus) = larutan serosa, dan diikat
Dipersiapkan larutan mukosa yang diuji (yogurt) dalam larutan NaCl 0,9%
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ANALISA ABSORPSI KALSIUM DENGAN METODA USUS TERBALIK (lanjutan)
Usus dimasukkan ke larutan mukosaInkubasi dalam shaking waterbath, 37oC,
90 menit, dengan aliran O2 (100 gelem-bung/menit)
Pada akhir inkubasi, dilakukan analisa kadar Ca pada larutan serosa
% Absorpsi = (kadar Ca serosa : kadar Ca mukosa awal) x 100%
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CALCIUM BIOAVAILABILITY IN PIG FED A DRY MILK-BASED DIET
Subyek : babi jantan, usia 2 bln; 16,4 + 0,3 kgAda 2 kelompok babi @ 9 ekor : klp milk
(milk-Ca diet) dan klp CCM (calcium citrate-malate high available Ca source) diet, lama diet : 10 minggu
Balance Ca dan P : dari 10 hari penelitian, 2 minggu sebelum penyembelihan
At slaugter : tibia (T) and two main metatar-sals (MT) (finger III and IV) were excised and broken to measure bending moment (at fracture point). Fresh bone ash content, density, length, diameter
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Result : Bone parameters
Fresh weight (g)
(g)
-----------------------
T MT
Ash weight
(g)
-----------------
T MT
Diameter
(mm)
-----------------
T MT
MILK 131 21.3 36.6 5.6 10.3 13.9
CCM 127 20.5 35.7 5.4
9.9 13.7
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