Ekstrak heksan daun kirinyuh memiliki aktivitas antibakteri terhadap bakteri P. acnes dilihat dari hasil persen penghambatan yang didapat. Pada seluruh konsentrasi ekstrak heksan daun kirinyuh terjadi penghambatan pertumbuhan dengan range persen penghambatan yang didapat adalah 13.005% - 50.419%. Hasil MICdidapatkan pada konsentrasi 6.25 mg/ml, yang berarti pada konsentrasi 6.25 mg/ml ekstrak heksan daun kirinyuh sudah dapat menghambat pertumbuhan bakteri P. acnes sebanyak minimal 50% setelah inkubasi selama 72 jam.
Saran bagi penelitian selanjutnya adalah melakukan pengujian kandungan fitokimia ekstrak heksan daun kirinyuh untuk mengetahui senyawa fitokimia yang berperan dalam aktivitas antibakteri. Kemudian, pengujian aktivitas antibakteri lanjutan menggunakan metode time-kill perlu dilakukan untuk mengetahui
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kemampuan aktivitas antibakteri ekstrak heksan daun kirinyuh terhadap bakteri P. acnes yang lebih spesifik yaitu bakteriosidal dan bakteriostatik dan untuk mengetahui waktu optimal yang dibutuhkan ekstrak heksan daun kirinyuh untuk membunuh bakteri P.acnes. Selanjutnya, penelitian menggunakan bakteri P. acnes yang resisten terhadap klindamisin sebaiknya dilakukan, sehingga dapat diamati kemampuan ekstrak heksan daun kirinyuh terhadap bakteri P. acnes resisten.
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30 LAMPIRAN
Lampiran 1. Surat Keterangan Determinasi Tanaman
Lampiran 2. Penetapan Kadar Air Dengan Destilasi Toluen
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Lampiran 3. Sertifikat Propionibacterium acnes ATCC 11827
Lampiran 4. Tanaman Chromolaena odorata (L.) R. M. King & H. Rob
32 Lampiran 5. Penetapan Bobot Tetap
Gambar 3. Hasil Penimbangan Cawan Kosong (a), Hasil Penimbangan Pertama Bobot Tetap (b), dan Hasil Penimbangan Terakhir Bobot Tetap (c)
(a) (b)
33 Lampiran 6. Ekstrak Heksan Daun Kirinyuh
Gambar 4. Ekstrak Heksan Daun Kirinyuh
Lampiran 7. Uji Kelarutan Ekstrak Dalam DMSO
Gambar 5. Kelarutan Ekstrak Dalam DMSO 50% (a) dan Kelarutan Ekstrak Dalam DMSO 100% (b)
34 Lampiran 8. Kultur Bakteri
Gambar 6. Kultur Blood Agar (a), Kultur Tryptone Soy Agar (b), Kultur Brain Heart Infusion (c)
Lampiran 9. Anaerogen, Indikator Anaerob dan Kontainer Airtight
(a) (b)
(c)
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Gambar 7. Anaerogen (a), Cara Pakai Anaerogen (b), Indikator Anaerob (c), Kontainer Airtight (d)
Lampiran 10. Indikator Anaerob Berwarna Putih
Gambar 8. Indikator Anaerob Putih
Lampiran 11. Perlakuan MIC Di 96-well plate
Gambar 9. Konsentrasi Seri EHDK (a), Kontrol Pertumbuhan (b), Kontrol Negatif (c), Kontrol Positif (d), Kontrol Media (e)
(a) (b) (c)
(d) (e)
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Lampiran 12. Optical Density Jam 0 dan 72 pada Perlakuan MIC Tabel V. Optical Density Jam 0 (Sebelum Inkubasi) Kadar (mg/mL) Replikasi 1 Replikasi 2 Replikasi 3 Rata-rata SD EHDK 6.25 0.471 0.571 0.546 0.529 0.052 EHDK 3.125 0.24 0.235 0.236 0.237 0.003 EHDK 1.5625 0.239 0.228 0.237 0.235 0.006 EHDK 0.781 0.22 0.235 0.22 0.225 0.009 EHDK 0.391 0.236 0.235 0.229 0.233 0.004 EHDK 0.195 0.235 0.232 0.23 0.232 0.003 K.Pertumb 0.195 0.197 0.248 0.213 0.030 K.DMSO 1% 0.229 0.246 0.25 0.242 0.011 K.DMSO 2% 0.226 0.247 0.265 0.246 0.020 K.Positif 0.142 0.148 0.196 0.162 0.030 K.Media 0.164 0.197 0.196 0.186 0.019
Keterangan EHDK : Ekstrak Heksan Daun Kirinyuh Tabel VI. Optical Density Jam 72 (Setelah Inkubasi) Kadar (mg/mL) Replikasi 1 Replikasi 2 Replikasi 3 Rata-rata SD EHDK 6.25 0.967 0.984 0.998 0.983 0.016 EHDK 3.125 0.848 0.915 0.835 0.866 0.043 EHDK 1.5625 0.759 0.761 0.771 0.764 0.006 EHDK 0.781 0.762 0.771 0.755 0.763 0.008 EHDK 0.391 0.977 0.971 0.939 0.962 0.020 EHDK 0.195 1.085 1.007 0.993 1.028 0.050 K.Pertumb 1.097 1.105 1.183 1.128 0.048 K.DMSO 1% 0.915 0.999 0.969 0.961 0.043 K.DMSO 2% 0.99 1.052 1.062 1.035 0.039 K.Positif 0.726 0.63 0.681 0.679 0.048 K.Media 0.171 0.201 0.199 0.190 0.017
37 Lampiran 13. Surat Keterangan Analisis Data
38 Lampiran 14. Uji Statistika
(a)
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Gambar 10. Uji Normalitas Data (a), Uji Homogenitas Data dan Uji One Way ANOVA (b), Uji Post-hoc Games Howell (c)
Lampiran 15. Perhitungan a. Perhitungan Kadar Air
Kadar air = Volume air
Berat sampel x 100% Volume air = 0.9 mL Berat Sampe = 10 g Kadar air = 0.9 mL 10 g x 100% = 9%
b. Perhitungan Penetapan Bobot Tetap Bobot porselen = 93.2090 g Bobot porselen + isi = 95.2995 g (1) 0.25% x 95.2995g = 0.2382 g
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95.2995 g ± 0.2382 g = 95.0613 g – 95.5377g
Bobot porselen + isi = 95.2218 g (2) masuk range bobot tetap Berat isi = 95.2218 g – 93.2090 g
= 2.0128 g c. Perhitungan Rendemen
Rendemen = Bobot ekstrak
Bobot simplisia x 100% Bobot ekstrak = 2.0128 g Bobot simplisia = 50.577 g Rendemen = 2.0128 g 50.577 g x 100% = 3.980%
d. Perhitungan Pembuatan Stok 200 mg/ml 200 mg/ml dibuat dalam 2 ml
200 mg/ml x 2 ml = 400 mg 400 mg setara dengan 0.4 g
Ekstrak yang ditimbang sebanyak 0.4 g DMSO 100% di add hingga 2 ml
e. Contoh cara perhitungan persen penghambatan % Penghambatan = (1 − ( ODt72-ODt0
ODgc72-ODgc0)) 𝑥 100% Kons 6.25 mg/ml
Selisih ODt72 – ODt0 = 0.454 Selisih ODgc72 – ODgc0 = 0.915
% Penghambatan = (1 − (0.454
0.915)) 𝑥 100% = 50.419%
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