54 BAB V
KESIMPULAN DAN SARAN
55 DAFTAR PUSTAKA
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64 Lampiran 1. Diagram Alur Penelitian
Preparasi Sampel
Ekstraksi
Optimasi Konsentrasi HAuCl4 dan Komposisi.
Sintesis Nanopartikel Emas
Karakterisasi
Uji Antibakteri
Spektrofotometer UV-Vis
PSA XRD FTIR
Uji Fitokimia
65 Lampiran 2. Bagan Kerja Preparasi Sampel
- Dibersihkan - Dikeringkan
- Dihaluskan dengan mesin penggiling - Ditimbang sebanyak 5 gram
- Direbus dengan akuabides 100 mL hingga suhu 80-90 oC
- Didinginkan
- Disaring dengan menggunakan kertas saring Whatmann 41
Sampel
Ekstrak
66 Lampiran 3. Pembuatan Larutan HAuCl4
- Ditimbang sebanyak 1 gram - Dilarutkan dengan aquaregia
- Dimasukkan ke dalam labu ukur 1000 mL
- Ditambahkan dengan akuades hingga tanda batas - Dihomogenkan
- Dipipet sebanyak 2, 4, 6 dan 8 mL - Dimasukkan ke dalam labu ukur 100 mL
- Ditambahkan dengan akuades hingga tanda batas - Dihomogenkan
Serbuk emas
Larutan HAuCl4 5 mM
Larutan HAuCl4 0,1 mM, 0,2 mM, 0,3 mM, 0,4 mM
67 Lampiran 4. Bagan Kerja Optimasi [HAuCl4]
- Dimasukkan ke dalam erlenmeyer dengan masing-masing konsentrasi 0,1 mM; 0,2 mM; 0,3 mM dan 0,4 mM sebanyak 25 mL
- Ditambahkan 5 mL ekstrak air Pyura sp. ke dalam masing-masing erlenmeyer
- Distirrer selama 7 jam
- Dianalisis dengan menggunakan spektrofotometer UV-Vis
HAuCl4
Data
68 Lampiran 5. Bagan Kerja Optimasi Komposisi
- Dipipet sebanyak 1 mL kedalam Erlenmeyer yang berisi 5 mL HAuCl4 0,3 mM - Distirrer selama
7 jam
- Dipipet sebanyak 1 mL kedalam Erlenmeyer yang berisi 10 mL HAuCl4 0,3 mM - Distirrer selama
7 jam
- Dipipet sebanyak 1 mL kedalam Erlenmeyer yang berisi 15 mL HAuCl4 0,3 mM - Distirrer selama
7 jam
- Dipipet sebanyak 1 mL kedalam Erlenmeyer yang berisi 20 mL HAuCl4 0,3 mM - Distirrer selama
7 jam
AuNP 1:5 AuNP 1:10 AuNP 1:15 AuNP 1:20
- Dianalisis dengan menggunakan
spektrofotometer UV-Vis
Data Ekstrak Pyura sp.
69 Lampiran 6. Bagan Kerja Uji Fitokimia
Catatan : 1. Terbentuknya endapan hijau menandakan adanya tannin
2. Terbentuknya endapan berwarna kuning (penambahan timbal asetat) dan endapan berwarna oranye (penambahan asam sulfat) menandakan adanya kandungan flavonoid
3. Terbentuknya busa yang banyak menandakan adanya saponin
Catatan: 1. Terjadinya perubahan warna dari ungu menjadi biru atau hijau menandakan adanya kandungan steroid
2. Terbentuknya warna merah pada lapisan antarmuka (interface) menujukkan adanya kandungan terpenoid
Uji Terpenoid
- diambil sebanyak 5 mL - ditambahkan 2 mL
CHCl3
- ditambahkan 3 mL H2SO4 secara perlahan hingga terbentuk lapisan
Ekstrak Pyura sp.
- diambil
sebanyak 2 mL - dicampur
dengan 2 mL akuades - ditambahkan
beberapa tetes FeCl3
Hasil
- diambil
sebanyak 2,5 mL - ditambahkan
beberapa tetes akuades
- dikocok dengan kencang
Hasil
Uji Tanin Uji Flavonoid Uji Saponin
- dimasukkan ke dua tabung reaksi
- tabung reaksi pertama ditambahkan beberapa tetes Pb(CH3COO)2
- tabung reaksi kedua
ditambahkan beberapa tetes H2SO4
Hasil
Ekstrak Batang Binahong
- diambil sebanyak 5 mL
- ditambahkan 2 mL anhidrida asetat - ditambahkan 2 mL
H2SO4 secara perlahan
Hasil
Uji Steroid
Hasil
70 Catatan : Terbentuknya endapan berwana oranye pada saat penambahan reagen Dragendorff dan endapan berwarna krem kekuning-kuningan pada saat penambahan reagen Mayer menandakan adanya kandungan alkaloid.
Ekstrak Pyura sp.
- diambil sebanyak 3 mL
- ditambahkan 3 mL HCl 1 % lalu diaduk di atas penangas
- dimasukkan ke dalam dua tabung reaksi berbeda masing-masing sebanyak 1 mL
Uji Alkaloid
- ditambahkan beberapa tetes reagen Dragendorff
- ditambahkan beberapa tetes reagen Mayer
Hasil Hasil
71 Lampiran 7. Bagan Kerja Sintesis Nanopartikel Emas
Larutan Nanopartikel Emas
- Disentrifuse pada kecepatan 14000 rpm selama 15 menit pada suhu 26oC
- Dibiarkan tumbuh selama 8 hari
- Disentrifuse kembali pada 14000 rpm selama 30 menit - Dikeringkan dalam freeze drier pada suhu 60oC selama
24 jam
Nanopartikel Emas Larutan HAuCl4 0,3 mM
- Dipipetkan sebanyak 100 mL ke dalam erlenmeyer
- Distirrer
- Ditambahkann sebanyak 20 mL ekstrak tunikata kedalam erlenmeyer - Distirrer selama 7 jam sambil
dipanaskan
- Diamati perubahan warna yang terjadi
72 Lampiran 8. Bagan Kerja Karakterisasi Nanopartikel
,
Gugus-gugus fungsi yang berperan dalam proses reduksi logam
- Nanopartikel tembaga sebanyak 2 mg dicampur dengan 100 mg KBr kemudian dibuat pelet
- Pelet yang telah dibuat dianalisis dengan FTIR
Analisis FTIR
Koloid Nanopartikel Emas
Spektrofotometri UV-Vis
Konfirmasi terbentuknya nanopartikel serta
Distribusi ukuran nanopartikel - dikarakterisasi
Analisis PSA
- Dipipet sebanyak 4 mL kedalam kuvet
- Dilakukan pengukuran pada panjang gelombang 185 – 700 nm
- Dipipet sebanyak 3 mL kedalam kuvet kuarsa - Dilakukan pengukuran
ukuran partikel
Koloid Nanopartikel Emas
73 Lampiran 9. Bagan Kerja Uji Bioaktivitas Antibakteri
- Ditimbang sebanyak 5 gram dimasukkan kedalam gelas kimia - Dilarutkan dengan 250 mL akuades di dalam labu erlenmeyer
- Dihomogenkan dengan stirrer diatas penangas air hingga mendidih dan diatur pH nya menjadi pH 7.
- Nutrien agar dimasukkan ke dalam tabung reaksi sebanyak ± 5 mL - Disterilkan dalam autoklaf pada suhu 121oC tekanan 2 atm selama 15
menit
- Didinginkan hingga suhu mencapai 40 – 45oC pada kemiringan 30o
- Bakteri Escherichia coli sebanyak 2-3 ose di gores ke dalam medium nutrient agar miring secara aseptis
- Diinkubasi dalam inkubator pada suhu 37oC selama 18-24 jam
- Diswab merata pada permukaan media Mueller Hinton Agar (MHA) yang terdapat pada cawan petri
- Didiamkan selama 5 menit -
- Nanopartikel emas ditimbang sebanyak 7.2 mg kemudian dilarutkan dengan 2 mL akuades steril.
- Kertas cakram dicelupkan kedalam larutan nanopartikel, HAuCl4 0,3 mM, ekstrak tunikata, ampisilin (kontrol positif) dan akuades (kontrol negatif) selama 15 menit.
- Diletakkan diatas media MHA yang telah berisi bakteri - Diinkubasi pada suhu 37oC selama 2×24 jam
- Diamati dan diukur diameter hambat yang terbentuk
Catatan: Diberikan perlakuan yang sama berdasarkan bagan kerja diatas untuk bakteri Staphylococus aureus.
Nutrien Agar
Media Uji Bioaktivitas Bakteri
Daya Hambat Bakteri Nutrien Agar Miring
Bakteri Uji
74 Lampiran 10. Dokumentasi Penelitian
Sampel Pyura sp sebelum dihaluskan
Sampel Pyura sp setelah dihaluskan
Ekstrak Pyura sp. Koloid Nanopartikel Emas
75 Lampiran 11. Hasil Uji Fitokimia
Foto Uji Tanin Foto Uji Flavonoid 1 Foto Uji Flavonoid 2
Foto Uji Saponin Foto Uji Steroid Foto Uji Terpenoid
Foto Uji Alkaloid 1 Foto Uji Alkaloid 2
76 Lampiran 12. Data FTIR Ekstrak H2O Pyura sp
77 Lampiran 13. Data FTIR Nanopartikel Emas
78 Lampiran 14. Data XRD Nanopartikel Emas
79 Lampiran 15. Data PSA Nanopartikel Emas
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