DAFTAR ISI HALAMAN JUDUL ............................................................................................... i HALAMAN PENGESAHAN PEMBIMBING ...................................................... ii HALAMAN PENGESAHAN PENGUJI .............................................................. iii HALAMAN PERNYATAAN ............................................................................... iv HALAMAN PERSEMBAHAN ..............................................................................v KATA PENGANTAR ........................................................................................... vi DAFTAR ISI........................................................................................................ viii DAFTAR GAMBAR ...............................................................................................x DAFTAR TABEL.................................................................................................. xi DAFTAR LAMPIRAN......................................................................................... xii INTISARI ............................................................................................................ xiii ABSTRACT........................................................................................................... xiv BAB I. PENDAHULUAN .....................................................................................1 A. Latar Belakang Masalah.............................................................................1 B. Perumusan Masalah ...................................................................................2 C. Tujuan Penelitian .......................................................................................2 D. Manfaat Penelitian .....................................................................................2 BAB II. STUDI PUSTAKA ....................................................................................3 A. Tinjauan pustaka ........................................................................................3 1. Kulit .....................................................................................................3 2. Fungsi kulit ..........................................................................................6 3. Khitosan ...............................................................................................8 4. Uji ketoksikan akut dermal ................................................................11 5. Histopatologi Kulit.............................................................................16 B. Landasan Teori.........................................................................................18 C. Hipotesis ..................................................................................................19 BAB III.METODE PENELITIAN ........................................................................20 A. Bahan dan Alat.........................................................................................20 1. Bahan .................................................................................................20 2. Alat.....................................................................................................20
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B. Cara Penelitian .........................................................................................21 1. Identifikasi khitosan...........................................................................21 2. Pembuatan larutan uji ........................................................................21 3. Uji iritasi primer.................................................................................22 4. Histopatologi kulit .............................................................................25 C. Analisis Hasil ...........................................................................................26 1. Kualitatif ............................................................................................26 2. Kuantitatif ..........................................................................................26 3. Analisis statistik .................................................................................27 BAB IV.HASIL DAN PEMBAHASAN ...............................................................28 A. Identifikasi Khitosan................................................................................28 B. Penetapan Dosis, Pelarut dan Formulasi larutan Khitosan ......................32 C. Hasil Uji Iritasi Primer.............................................................................32 1. Hasil pengamatan secara kualitatif berupa eritema dan edema .........37 2. Hasil pengamatan secara kuantitatif berupa indeks iritasi primer .....39 3. Hasil pengamatan berupa perubahan histopatologi ...........................44 BAB V. KESIMPULAN DAN SARAN................................................................50 A. Kesimpulan ..............................................................................................50 B. Saran ........................................................................................................50 DAFTAR PUSTAKA ............................................................................................51 LAMPIRAN...........................................................................................................54
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DAFTAR GAMBAR
Gambar 1. Penampang kulit (Anonim, 2005)........................................................3 Gambar 2. Struktur khitosan ( Morimoto et al., 2002). .........................................9 Gambar 3. Reaksi deasetilasi/transformasi khitin menjadi khitosan (Habibie, 2000). .................................................................................................10 Gambar 4. Skema kerja uji iritasi primer (Lu, 1995)...........................................22 Gambar 5. Skema kerja histopatologi kulit (Bhol et al., 2003). ..........................25 Gambar 6. Spektra absorbansi IR sebagai fungsi panjang gelombang untuk serbuk khitin.......................................................................................29 Gambar 7. Spektra absorbansi IR sebagai fungsi panjang gelombang untuk serbuk khitosan standar......................................................................30 Gambar 8. Spektra absorbansi IR sebagai fungsi panjang gelombang untuk serbuk khitosan uji .............................................................................30 Gambar 9. Skema perlakuan pada punggung kelinci...........................................34 Gambar 10. Foto skor 0 eritema dan 0 edema, yang artinya tanpa eritema dan tanpa edema pada kelompok khitosan 2% tanpa insisi 72 jam ..........37 Gambar 11. Foto skor 1 eritema dan 0 edema, yang artinya eritema sangat sedikit dan tanpa edema pada kelompok khitosan 4% tanpa insisi 72 jam. ..38 Gambar 12. Foto skor 2 eritema dan 0 edema, yang artinya eritema berbatas jelas dan tanpa edema pada kelompok asam laktat 1% insisi 24 jam. .......38 Gambar 13. Nilai Indeks Iritasi Primer untuk Aquades, Khitosan dan Asam Laktat 1% ...........................................................................................43 Gambar 14. Histologi kulit kelinci normal: Epidermis normal, kelenjar rambut menyebar, dermis normal...................................................................46 Gambar 15. Histopatologi kulit kelinci: potongan parasit pada epidermis...........46 Gambar 16. Histopatologi kulit kelinci: penebalan pada epidermis dan proliferasi jaringan ikat longgar ..........................................................................48 Gambar 17. Histopatologi kulit kelinci: infiltrasi sel radang pada dermis ...........46
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DAFTAR TABEL Tabel I.
Skor eritema dan edema (Draize, 1959).............................................13
Tabel II.
Kategori sifat mengiritasi berdasarkan rata-rata gabungan indeks iritasi primer senyawa kimia (Brossia, 1988) ....................................14
Tabel III. Nilai Derajat Deasetilasi untuk Khitin, Khitosan standar, dan Khitosan uji ........................................................................................31 Tabel IV. Alat Uji Iritasi (Lee dan Maibach, 2006)...........................................36 Tabel V.
Data pengamatan iritasi primer secara kuantitatif dalam rata- rata skor eritema dan edema .............................................................................39
Tabel VI. Data pengamatan iritasi primer secara kuantitatif yang berupa indeks iritasi primer.......................................................................................40 Tabel VII. Matriks signifikansi hasil uji analisis Post Hoc Test-Dunnett T3 skor eritema................................................................................................43 Tabel VIII. Hasil pengamatan histopatologi kulit kelinci.....................................45
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DAFTAR LAMPIRAN Lampiran 1. Foto Serbuk Khitosan ……………………………………………….... 54 Lampiran 2. Foto Larutan Khitosan dalam Pelarut Asam Laktat 1% …………….... 55 Lampiran 3 Foto Punggung Kelinci………………………………………………… 56 Lampiran 4 Data pengamatan secara kualitatif berupa eritema dan edema………… 57 Lampiran 5 Data pengamatan iritasi primer secara kuantitatif dalam skor eritema dan edema……………………………………………………………… 58 Lampiran 6 Hasil Pemeriksaan Histopatologi……………………………………….59 Lampiran 7 Perhitungan Derajat Deasetilasi (DD) Khitin, Khitosan standar, dan Khitosan uji……………………………………………………………. 62 Lampiran 8 Perhitungan Indeks Iritasi Primer............................................................ 63 Lampiran 9 Output SPSS Pengaruh Perlakuan Insisi terhadap Skor Eritema............. 65 Lampiran 10 Output SPSS Pengaruh Waktu terhadap Skor Eritema............................ 66 Lampiran 11 Output SPSS Perbandingan Skor Eritema dan Skor Edema antara semua Kelompok Uji…………………………………………………... 67 Lampiran 12 Certificate of Analysis Khitosan Standar.................................................73
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