LAPORAN PRAKTIKUM BIOKIMIA ASAM AMINO DAN PROTEIN
Disusun Oleh : Kelompok
: 4 dan 9
Nama
:
1. Abidatussoleha
(201610410311043)
2. Fahrozal
(201610410311088)
3. Dyah Ayu Pipin L
(201610410311128) (201610410311128)
4. Andri Saputra Dunggio
(201610410311158) (201610410311158)
5. Dwi Trisnawati
(201610410311168) (201610410311168)
6. Dinda Dwi Yulyandari
(201610410311187) (201610410311187)
7. Dinda Muji Nurhandini
(201610410311171) (201610410311171)
8. Nur Sheilla Alda
(201610410311193) (201610410311193)
9. Nur ‘Ilmi Hikmatul M M
(201610410311196)
Kelas
: Farmasi D
PROGRAM STUDI FARMASI FAKULTAS ILMU KESEHATAN UNIVERSITAS MUHAMMADIYAH MALANG 2018
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KATA PENGANTAR
Segala Puji bagi Allah SWT karena atas petunjuk dan hidayah-Nya serta dorongan dari semua pihak sehingga kami dapat menyelesaikan laporan praktikum ini dengan baik dan seksama. Laporan praktikum mengenai
“Asam Amino dan Protein”
ini disusun dengan
sistematis untuk memenuhi salah satu tugas praktikum mata kuliah biokimia, program studi farmasi, Fakultas Ilmu Kesehatan, Universitas Muhammadiyah Malang. Dengan selesainya laporan resmi praktikum ini, maka tidak lupa kami mengucapkan banyak terima kasih kepada semua pihak yang terlibat dalam penyusunan laporan ini, khususnya kepada : 1. Dosen pembimbing kami 2. Dan asisten laboratorium “Biokimia Farmasi” serta teman-teman teman-teman yang saling membantu dalam menyelesaikan laporan resmi praktikum ini. Kami menyadari bahwa laporan praktikum ini masih jauh dari sempurna dan tidak luput dari kekurangan-kekurangan, baik dari segi materi maupun teknis penulisan. Oleh karena itu saran dan kritik yang membangun dari rekan-rekan pembaca sangat dibutuhkan untuk penyempurnaanya. Semoga laporan praktikum ini dapat memberikan manfaat untuk rekan-rekan yang membaca.
Malang,17 Mei 2018
Penulis
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DAFTAR ISI
KATA PENGANTAR ......................................... ............................................................... ............................................. .......................................... ...................ii DAFTAR ISI ............................................ ................................................................... ............................................. ............................................. ............................ ..... iii A. Tujuan Praktikum...................... Praktikum ............................................ ............................................. ............................................. ......................................... ................... 1 B. Dasar Teori ............................... ...................................................... .............................................. .............................................. ......................................... .................. 1 Sumber Asam Amino ................................ ...................................................... ............................................ ............................................. ........................... .... 1 Asam Amino dan Karakteristiknya .......................... ................................................ ............................................. .................................. ........... 2 Klasifikasi Protein Berdasarkan Daya Larut dan Komposisi Kimianya Kimian ya .......................... .......................... 3 Golongan-golongan Golongan-golongan Asam Amino ............................................................ ................................................................................... ....................... 3 Sifat-sifat Asam Amino ........................................... ................................................................. ............................................ .................................. ............ 3 Asam Amino yang diperlukan Tubuh ............................................ ................................................................... .................................. ........... 4 C. Alat dan Bahan ............................................. ................................................................... ............................................ ............................................. ....................... 6 D. Prosedur Kerja ............................................. ................................................................... ............................................ ............................................. ....................... 7 E. Skema Kerja ......................................... ............................................................... ............................................. .............................................. .............................. ....... 9 G. Hasil Praktikum ........................................... ................................................................. ............................................ ........................................... ..................... 17 I. Kesimpulan .................................................. ........................................................................ ............................................ ........................................... ..................... 26 DAFTAR PUSTAKA .......................................... ................................................................. ............................................. ....................................... .................28 Lampiran ............................................ .................................................................. ............................................ ............................................ .................................... .............. 29
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A. Tujuan Praktikum
Percobaan ini dilakukan untuk mengetahui cara mengidentifikasi sifat dan reaksi dari asam amino dan protein B. Dasar Teori
Sumber Asam Amino
Protein adalah sumber asam amino yang mengandung unsur C,H,O dan N yang tidak dimiliki oleh lemak dan karbohidrat. Molekul protein mengandung gula terpor belerang, dan ada jenis protein yang mengandung unsur logam seperti besi dan tembaga te mbaga (Winarno, 1997). Protein merupakan polimer alami yang terdiri atas sejumlah unit asam amino yang berikatan satu dengan lainnya lewat ikatan amida (atau peptida). Jaring laba-laba, bulu hewan, dan otot, putih telur, dan hemoglobin (molekul yang mengangkut oksigen dalam tubuh ke tempat yang memerlukan) ialah beberapa contoh protein (Lehninger, 1990). Protein yang ditemukan kadang-kadang berkonjugasi dengan makromolekul atau mikromolekul seperti lipid, polisakarida dan mungkin fosfat. Protein terkonjugasi yang dikenal antara lain nukleoprotein, fosfoprotein, metaloprotein, lipoprotein, flavoprotein dan glikoprotein. Protein yang diperlukan organisme dapat diklasifikasikan menjadi dua golongan utama, ialah pertama; protein sederhana, yaitu protein yang apabila terhidrolisis hanya menghasilkan asam amino, dan kedua protein terkonjugasi, yaitu protein yang dalam hidrolisis tidak hanya menghasilkan asam amino, tetapi menghasilkan juga komponen organik ataupun komponen anorganik yang disebut “gugus prosthetic” (Sumarno, dkk., 2002). Pada umumnya asam amino diperoleh sebagai hasil hidrolisis protein, baik menggunakan enzim maupun asam. Dengan cara ini diperoleh campuran bermacammacam asam amino dan untuk menentukan jenis asam amino maupun kuantitas masingmasing asam amino perlu diadakan pemisahan antara asam-asam amino tersebut
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Asam Amino dan Karakteristiknya Karakteristiknya
Asam amino sendiri tidak berwarna dan tidak dapat dideteksi secara visual pada kromatografi atau cara analisis lainnya. Dengan mengubahnya menjadi senyawa yang berwarna, kita dapat melihatnya. Reaksi warna yang penting dari asam amino adalah reaksinya dengan ninhydrin karena intensitas warna yang terbentuk pada reaksi ninhidrin ini sebanding dengan konsentrasi asam aminonya maka reaksi ini dapat dipakai untuk analisa kuantitatif. Contohnya: reaksi ninhydrin ini dipakai pada alat analisa otomatik asam amino, suatu alat untuk memisahkan asam amino dengan memakai kolom penukar ion dan ditentukan konsentrasi relatifnya. Pada polimerisasi asam amino, gugus -OH yang merupakan bagian gugus karboksil satu asam amino dan gugus -H yang merupakan bagian gugus amina asam amino lainnya akan terlepas dan membentuk air. Oleh sebab itu, reaksi ini termasuk dalam reaksi dehidrasi. Molekul asam amino yang telah melepaskan molekul air dikatakan disebut dalam bentuk residu asam amino (Anonim, 2011). Asam amino terdiri dari sebuah gugus amino, sebuah gugus karboksil, sebuah atom hidrogen, dan gugus R yang terikat pada sebuah atom C yang dikenal sebagai karbon α, serta gugus R merupakan rantai cabang (Winarno, 2008) . Asam amino dan protein secara umum mempunyai sifat-sifat fisik yang sama.Dari keseluruhan asam amino yang terdapat di alam hanya 20 asam amino yang yangbiasa dijumpai pada protein. Dari struktur umumnya, asam amino mempunyai dua gugus pada tiapmolekulnya, yaitu gugus amino dan gugus karboksil, yang digambarkan sebagai strukturion dipolar. Gugus amino dan gugus karboksil pada asam amino menunjukkan sifat-sifatspesifiknya. Karena asam amino mengandung kedua gugus tersebut, senyawa ini akan memberikan reaksi kimia yang yang mencirikan gugus-gugusnya. Sebagai contoh adalahreaksi asetilasi dan esterifikasi (Girindra, 1993). Asam amino dan protein mempunyai sifat fisik yang sama (memiliki gugus asam dan basa). Kelarutan protein dalam air juga berbeda tergantung ion positif dan ion negatif yang terdapat pada protein. Protein yang dihidrolisa akan terurai menjadi beberapa asam amino. Aktifitas biologis dari protein tergantung dari bentuk tiga dimensi asam amino
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Destruksi atas bentuk tiga dimensi suatu protein disebut denaturasi. Bentuk tiga dimensi tergantung atas ikatan hidrogen, ikatan inter-ionik/jembatan garam, dan ikatan disulfida. Suatu agen zat tertentu yang dapat berinteferensi dengan ikatan hidrogen, ikatan inter-ionik, dan ikatan disulfida dapat mendenaturasi protein. Perubahan pada protein akibat dari denaturasi adalah : berkurangnya daya larut lar ut enzim, hilangnya aktivitas protein, dan berubah/hilangnya berubah/hilangnya antigen. Klasifikasi Protein Berdasarkan Daya Larut dan Komposisi Kimianya
a)
Simple Protein : protein yang hanya mengandung 1-α 1- α asam amino atau derivatnya. derivatnya. Contoh : albumin, globulin, glutein, protamin, albuminoid, histon, dll.
b)
Conjugated Protein : protein yang bergabung dengan zat lain yang bukan protein (gugus prostetik). Contoh : nukleoprotein, glikoprotein, fosfoprotein, lipoprotein, dll.
Golongan-golongan Golongan-golongan Asam Amino
Asam amino dengan rantai samping alifatik
Glisin, alanin, valin, leusin, isoleusin Asam amino dengan rantai samping gugus hidroksil
Serin, treonin, tirosin Asam amino dengan rantai samping mengandung sulfur
Sistein, metionin
Asam amino dengan rantai samping mengandung amida Asam aspartat, asparagine, asam glutamat, glutamine
Asam amino dengan rantai samping mengandung alkali Arginin, lisin, histidin Asam amino dengan rantai samping mengandung cincin aromatis
Fenilalanin, tirosin, triptofan
Asam amino yang tidak terdapat pada protein Β-alanin, taurin, ¥-aminobutirat, ornitin, sitrulin
Sifat-sifat Asam Amino
Kristal putih, larut dalam air, asam/basa kuat
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Mempunyai sifat optis aktif
Bersifat amfoter
Tidak bergerak pada medan listrik dengan pH isoelektrik
Asam Amino yang diperlukan Tubuh
a) Asam amino essensial Asam amino yang mutlak harus ada padamakanan karena tidak dapat disintesa tubuh.
Triptofan, fenilalanin, lisin, treonin, valin, metionin, isoleusin, dll.
b) Asam amino non-essensial Asam amino yang dapat disintesa tubuh, terdapat juga pada makanan sebagai sumber nitrogen.
Pengujian Asam Amino
Ada berbagai cara dalam pengujian terhadap protein yaitu dengan reaksi uji asam amino dan reaksi uji protein. Reaksi uji asam amino sendiri terdiri dari 6 macam uji yaitu: uji millon, uji hopkins cole, uji belerang, uji xantroprotet, dan uji biuret. Sedangkan untuk uji protein, berdasarkan pada pengendapan oleh garam, pengendapan oleh logam dan alkohol. Serta uji koagulasi dan denaturasi protein. a) Tes Uji Biuret Uji ini digunakan untuk menguji adanya ikatan peptida pada suatu asam amino. Uji biuret dikatakan positif (+) bila berwarna ungu/violet (Sumardjo,1998) b) Tes Millon Uji millon umumnya digunakan untuk menunjukkan adanya asam amino tirosin
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triptofan dapat direaksikan dengan pereaksi Hopkins-Cole yang mengandung asam glioksilat. Setelah dicampur dengan pereaksi Hopkins-Cole, asam sulfat dituangkan perlahan-lahan sehingga membentuk lapisan di bawah larutan protein. Beberapa saat kemudian akan terjadi cincin ungu pada batas antara kedua lapisan. Reaksi Hopkins-Cole memberi hasil positif khas untuk gugus indol dalam protein (Poedjadi,1994). d) Tes Ninhidrin Uji ninhidrin bertujuan untuk mengetahui adanya asam amino alfa pada suatu protein. Hasilnya dikatakan positif (+) bila terjadi perubahan warna menjadi ungu. Uji umum untuk protein dan asam amino. Ninhidrin dapat mengubah asam amino menjadi suatu aldehida. Ninhidrin dilakukan dengan menambahkan beberapa tetes larutan ninhidrin yang terlihat tidak warna kedalam sampel, kemudian dipanaskan beberapa menit. Adanya protein ditandai dengan adanya per ubahan warna ungu. (Setiawan,2013) e) Tes Xantoprotein Pada percobaan ini untuk mendeteksi ada tidaknya inti benzena. Tanda (+) menunjukkan adanya inti benzena dalam suatu makanan. Bahan makanan yang mengandung inti benzena setelah diberi larutan asam nitrat pekat dan larutan NaOH akan menghasilkan warna kuning jingga dari warna aslinya.Inti benzena bereaksi dengan larutan Xanthoprotein akan berwarna kuning jingga.Larutan asam nitrat pekat ditambahkan dengan hati-hati ke dalam larutan protein. Setelah dicampur terjadi endapan putih yang dapat berubah menjadi kuning apabila dipanaskan. Reaksi R eaksi yang terjadi adalah nitrasi pada inti benzena yang terdapat pada molekul protein. Jadi reaksi ini positif jika mengandung tirosin, fenil alanin dan triptofan tri ptofan (Poedjadi,1994). f) Tes Logam Berat Uji ini bertujuan untukmembuktikan sifat dari asam amino yaitu akan mengalami
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C. Alat dan Bahan
a) Alat 1. Tabung reaksi 2. Pipet tetes 3. Gelas ukur 4. Hotplate 5. Api bunsen 6. Beaker glass b) Bahan 1. Kasein 0,2% 2. Putih telur 3. Fenol 2% 4. NaOH 10% 5. CuSO4 6. Larutan Hopskin-cole 7. H2SO4 pekat 8. Ninhindrin 0,1% 9. HNO3 pekat 10. Pb acetat 2% 11. HgCl 2% 12. Pereaksi Millon 13. NaOH 13. NaOH
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D. Prosedur Kerja
Tes Biuret
1. Disiapkan 3 tabung reaksi 2. Tabung A berisi 2 ml kasein 0,2%, tabung B berisi 2 ml putih telur dan tabung C berisi 2 ml fenol 2% 3. Masing-masing tabung ditambahkan 1 ml NaOH 10% 4. Masing-masing tabung ditambahkan 3 tetes CuSO 4 5. Diamati perubahan warna (ungu/violet)
Tes Millon
1. Disiapkan 3 tabung reaksi 2. Tabung A berisi 3 ml kasein 0,2%, tabung B berisi 3 ml putih tel ur dan tabung C berisi 3 ml fenol 2% 3. Masing-masing tabung ditambahkan 5 tetes pereaksi Millon 4. Dipanaskan diatas hotplate (tabung dimasukkan ke dalam beaker glass besar yang sudah diisi air) dengan hati-hati. 5. Diamati perubahan warna (merah)
Tes Hopskin-Cole
1. Disiapkan 2 tabung reaksi 2. Tabung A berisi 2 ml kasein 0,2% dan tabung B berisi 2 ml putih telur 3. Masing-masing tabung ditambahkan 1 ml larutan hopskin-cole, lalu dikocok ad
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3. Masing-masing tabung ditambahkan 0,5 ml (8 tetes) tete s) Ninhidrin 0,1% 4. Dipanaskan di atas hotplate (tabung dimasukkan ke dalam beaker glass besa r yang sudah diisi air) selama 10 menit 5. Diamati perubahan warna
Tes Xanthoprotein
1. Disiapkan 3 tabung reaksi 2. Tabung A berisi 2 ml kasein , tabung B berisi 2 ml putih t elur dan tabung C berisi 2 ml fenol 2% 3. Masing-masing tabung ditambahkan 1 ml HNO 3 pekat , lalu ditunggu hingga muncul endapan 4. Dipanaskan di api bunsen, hingga menghasilkan endapan kuning 5. Lalu didinginkan terlebih dahulu dengan air kran (aliran harus kecil) 6. Kemudian masing-masing tabung ditambahkan NaOH 2 ml (32 tetes) 7. Diamati perubahan warna
Tes Logam Berat
1. Disiapkan 4 tabung reaksi 2. Tabung A1 berisi 3 ml putih telur, tabung A2 berisi 3 ml putih telur, tabung B1 berisi 3 ml kasein 0,2% dan tabung B2 B2 berisi 3 ml kasein 0,2%, 3. Tabung A1 A1 dan B1 ditambahkan 5 tetes HgCl 2 2% sedangkan tabung A2 dan B2 ditambahkan 5 tetes Pb asetat 4. Diamati perubahan yang terjadi
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E. Skema Kerja
1. TEST BIURET
a. 2 ml kasein
+
1 ml NaOH 10%
+ 2-3 tetes larutan CuSO4
b.
2 ml putih telur +
1 ml NaOH 10%
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2. TEST MILLON
a. 3 ml kasein 0,2%
+ 5 tetes pereaksi Millon
3 ml putih telur
+ 5 tetes pereaksi Millon
3ml fenol 2%
+ 5 tetes pereaksi Millon
b.
c.
d.
Diamati perubahan warna yang timbul
3. TEST HOPKINS-COLE
a. 2 ml kasein 0,2%
+ 1 ml larutan Hopkins-cole
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c. 2 ml fenol 2%
+ 1 ml larutan Hopkins-cole
+ 10 tetes H2SO4 pekat melalui dinding tabung.
d. Diamati warna yang terbentuk
4. TEST NINHIDRIN
a. 1 ml kasein 0,2%
+ 0,5ml (8 tetes) larutan ninhidrin 0,1%
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5. TEST XANTHOPROTEIN XANTHOPROTEIN
a. 2 ml kasein 0,2%
+ 1 ml larutan HNO3 pekat
b.
2 ml putih telur
+ 1 ml larutan HNO3 pekat
c. 2
2 ml fenol 2%
+ 1 ml larutan HNO3 pekat
d. Perhatikan terbentuknya endapan berwarna putih kasein-putih telur -fenol
kasein-putih telur -fenol
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6. PENGARUH LOGAM BERAT
a. 3 ml kasein 0,2%
b.
3 ml kasein 0,2%
5 tetes HgCl2 2%
5 tetes Pb asetat
c. 3 ml putih telur
5 tetes HgCl2 2%
3 ml putih telur
5 tetes Pb asetat
d.
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F. Bagan Alir
TEST BIURET
Disiapkan 2 ml kasein 0,2%; 2 ml putih telur dan 2 ml fenol 2%
Dimasukkan Dimasukkan ke dalam masing-masing tabung reaksi
DItambahkan 2-3 tetes larutan CuSO4 pada masing-masing tabung
Diamati warna yang timbul pada masingmasing tabung
Ditambahkan 1 ml NaOH 10% pada masing-masing tabung
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TEST HOPKINS-COLE
Disiapkan 2 ml kasein 0,2% dan 2 ml putih telur
Dimasukkan Dimasukkan ke dalam masing-masing tabung reaksi
Ditambahkan 10 tetes larutan H2SO4 pekat melalui dinding tabung
Diamati warna yang timbul pada masingmasing tabung
Ditambahkan 1 ml larutan hopkins-cole pada masing-masing tabung
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TEST XANTHOPROTEIN
Disiapkan masingmasing 2 ml larutan larutan kasein 0,2%, putih telur dan fenol 2%
Dimasukkan pada masing-masing tabung
Ditambahkan 1 ml HNO3 pekat pada masing-masing tabung
Diperhatikan terbentuknya endapan putih
Dipanaskan Dipanaskan dengan dengan api bunsen. bunsen. Endapan Endapan larut larutan berwarna kuning
Didinginka Didinginkan n di bawah keran (aliran kecil)
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G. Hasil Praktikum
Larutan Uji
Pengamatan
Kasein 2% (susu)
- Uji biuret : 2 ml kasein + 1 ml
Hasil
-Uji
biuret
NaOH 10% + 2-3 tetes CuSO4 ->
(mengandung
menghasilkan warna ungu
peptida)
:
+ ikatan
- Uji millon : 3 ml kasein + 5 tetes
-Uji millon : + (terbentuk
pereaksi millon -> menghasilkan
senyawa merkuri dengan
endapan putih keunguan dengan
gugus hidroksifenil yang
larutan bening
berwarna)
- Uji hopkins-cole : 2 ml kasein + 1 ml lar.hopkins cole + 10 tetes
-Uji
hopkins-cole
(terdapat cincin)
:
+
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- Uji xanthoprotein : 2 ml fenol +1
berwarna)
ml HNO3 pekat lalu dipanaskan,
- Uji xanthoprotein : +
muncul endapan, dipanaskan + 3 ml
(mengandung gugus inti
NaOH
benzen)
->
menghasilkan
warna
kuning kecoklatan Putih telur
- Uji biuret : 2 ml putih telur + 1 ml
-Uji
biuret
NaOH 10% + 2 tetes CuSO4 ->
(mengandung
menghasilkan warna ungu
peptida)
- Uji millon : 2 ml putih telur + 5 tetes
pereaksi
millon
->
menghasilkan warna merah
:
+ ikatan
-Uji millon : + (terbentuk senyawa merkuri dengan gugus hidroksifenil yang berwarna)
Uji hopkins-cole : 2 ml putih telur
-Uji
hopkins-cole
:
+
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a. Test Biuret Keterangan :
A = Sampel putih telur B = Sampel kasein 0,2%
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c. Test Hopkins-Cole Keterangan :
A = Sampel putih telur B = Sampel kasein 0,2%
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e. Test Xanthoprotein Keterangan :
A = Sampel kasein 0,2% B = Sampel putih telur
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f. Pengaruh Logam Berat Keterangan :
A = Sampel putih telur B = Sampel kasein 0,2%
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H. Pembahasan Protein merupakan senyawa polimer alami yang tersusun dari berbagai asam amino melalui ikatan peptida. Asam amino merupakan monomer (satuan pembentuk). Protein asam amino adalah satuan senyawa yang mempunyai dua gugus fungsi yaitu gugusan amino dan gugusan karboksil. (Lehninger,1990). Dalam percobaan ini kita menggunakan dua sampel sebagai sumber protein yaitu kasein dan putih telur.
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kemudian ditambahkan dengan 5 tetes pereaksi millon, lalu dipanaskan menghasilkan
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tersebut adalah reaksi nitrasi dimana terjadi subtitusi atom H + dengan NO2 yang akan
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I.
Kesimpulan
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membuktikan bahwa putih telur dan kasein mengalami koagulasi bila direaksikan
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DAFTAR PUSTAKA
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Titles you can't find anywhere else
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Titles you can't find anywhere else
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Titles you can't find anywhere else
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Titles you can't find anywhere else
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.