BAB V KESIMPULAN DAN SARAN
B. Saran
Penelitian ini disarankan dilanjutkan dengan mengambil senyawa yang lebih murni dari buah naga merah (Hylocereus polyrhizus) dengan mengisolasi suatu senyawa fenolik atau senyawa flavonoid. Uji selanjutnya bisa dilakukan dengan pemberian senyawa hasil isolasi pada hewan uji untuk mengetahui efek antioksidan serta uji in vivo untuk mengukur aktivitas fotoprotektif menggunakan senyawa hasil isolasi.
45 DAFTAR PUSTAKA
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Lampiran 1
54
Lampiran 2
Perhitungan nilai rendemen
Hasil fraksinasi dari 5,046 gram ekstrak adalah 2,886 gram atau 57,1938 % dari berat ekstrak.
Oleh karena itu, apabila ekstrak yang digunakan adalah 19,273 gram, maka perkiraan hasil fraksinasi adalah:
19,273 gram 57,1938% = 11,02296 gram
Oleh karena itu, nilai rendemen fraksi kental KBNM-AcOEt terhadap kulit buah naga merah kering (nilai X) adalah:
X =Berat perkiraan hasil fraksinasi
Berat kulit buah naga kering 100% X =!!,"##$% &
'(" & 100%
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Lampiran 3 Perhitungan Rf
Nilai Rf dihitung dengan menggunakan perbandingan sebagai berikut:
Rf = Jarak yang ditempuh solut (cm)
Jarak yang ditempuh fase gerak (cm)
1. Sinar tampak A =7.85 8 = 0,9812 B =6.8 8 = 0.85 2. Sinar UV 254 nm A =7.85 8 = 0,9812 B =6.8 8 = 0.85
3. Sinar UV 366 nm setelah disemprot
A =7.8
8 = 0.975
B =7
56
Lampiran 4
Analisis Total Flavonoid sampel uji
Tabel 13. Uji Flavonoid total standar kuersetin
konsentrasi
(µg/mL) Abs 1 Abs 2 Abs 3
Rerata Absorbansi 400 0.2904 0.2666 0.2633 0.2734 800 0.5042 0.5426 0.5122 0.5196 1200 1.1251 0.9203 1.0753 1.0402 1600 1.3572 1.3900 1.4305 1.3925 2000 1.6538 1.8073 1.7048 1.7219
Gambar 8. Kurva standar kuersetin pada uji total flavonoid
Persamaan regresi kuersetin:
y = 0.0009x - 0.1414
Tabel 14. Uji Flavonoid total Fraksi KBNM-AcOEt
Sampel Replikasi ke- Absorbansi (510 nm)
Fraksi KBNM- AcOEt 1 0.0154 2 0.0151 3 0.0094 Rata- rata 0.0133 SD 0.003380828
Hasil pembacaan Fraksi KBNM- AcOEt
Replikasi 1
y = 0.0009x - 0.1414 0.0154 = 0.0009x - 0.1414 x = 174.2222 µg/mL
57 Replikasi 2 y = 0.0009x - 0.1414 0.0151 = 0.0009x - 0.1414 x = 173.8889 µg/mL Replikasi 3 y = 0.0009x - 0.1414 0.0094 = 0.0009x - 0.1414 x = 167.5556 µg/mL Perhitungan kadar flavonoid
Konsentrasi sampel (ppm) = berat sampel (mg) (volume (L)
Konsentrasi sampel (ppm)= 10.4 mg
0.01 L = 1040 ppm
Total flavonoid Fraksi KBNM- AcOEt (% b/b EQ)
Replikasi 1
Total Flavonoid 1 =kadar flavonoid dalam sampel (ppm)
konsentrasi sampel (ppm) x 100 %
Total Flavonoid 1=174.2222
1040 x 100 %
Kadar Flavonoid =16.7521 % b/b EQ Replikasi 2
Total Flavonoid 2 =kadar flavonoid dalam sampel (ppm)
konsentrasi sampel (ppm) x 100 %
Total Flavonoid 1=173.8889
1040 x 100 %
Kadar Flavonoid =16.7201 % b/b EQ Replikasi 3
Total Flavonoid 3 =kadar flavonoid dalam sampel (ppm)
konsentrasi sampel (ppm) x 100 %
Total Flavonoid 1=167.5556
1040 x 100 %
58
Lampiran 5
Analisis kandungan fenolik total
Tabel 15. Standar asam galat
Konsentrasi (µg/mL) Absorbansi (760 nm) Rerata
Duplo 1 Duplo 2 10 0,119 0,119 0,119 20 0,230 0,231 0,231 30 0,351 0,352 0,352 40 0,432 0,434 0,433 50 0,543 0,544 0,544
Gambar 9. Kurva standar asam galat
Kurva standar asam galat diperoleh persamaan:
Y = 0,010x + 0,020 R² = 0,996
Konsentrasi fenol total larutan Fraksi KBNM-AcOEt
Replikasi 1 y = 0,010x + 0,020 0.174 = 0,010x + 0,020 x = 15.4 µg GAE/mL sampel Replikasi 2 y = 0,010x + 0,020 0.171 = 0,010x + 0,020 x = 15.1 µg GAE/mL sampel Replikasi 3 y = 0,010x + 0,020 0.172 = 0,010x + 0,020
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x = 15.2 µg GAE/mL sampel
Kadar fenol total per berat sampel Fraksi KBNM- AcOEt
Rumus:
Keterangan:
C = Kadar fenol larutan (nilai x)
V = Volume ekstrak yang digunakan (mL) fp = Faktor pengenceran
G = Berat sampel yang digunakan (g)
Replikasi 1
= 15098,039 µg GAE/g sampel
15098,039 µg GAE/g sampel = x mg GAE/100g sampel 15,098039 mg GAE/g sampel = x mg GAE/100g sampel 15,098039 x 100 = x
x = 1509,8039
Replikasi 2
= 14803,922 µg GAE/g sampel
14803,922 µg GAE/g sampel = x mg GAE/100g sampel 14,803922 µg mg GAE/g sampel = x mg GAE/100g sampel 14,803922 x 100 = x
x = 1480,3922
Replikasi 3
= 14901,961 µg GAE/g sampel
14901,961 µg GAE/g sampel = x mg GAE/100g sampel 14,901961 mg GAE/g sampel = x mg GAE/100g sampel 14,901961 x 100 = x
60
Lampiran 6
Uji aktivitas antioksidan sampel uji Sampel kuersetin
Tabel 16. Uji penangkapan radikal bebas DPPH kuersetin replikasi 1
Konsentrasi (µg/mL) Duplo 1 (nm) Duplo 2 (nm) Rata-Rata % Inhibisi 1 0,473 0,472 0,473 31,422 2 0,375 0,371 0,373 45,864 3 0,264 0,262 0,263 61,829 4 0,168 0,169 0,169 75,544 5 0,094 0,093 0,094 86,430 Absorbansi Kontrol 0,689
Tabel 17. Uji penangkapan radikal bebas DPPH kuersetin replikasi 2
Konsentrasi (µg/mL) Duplo 1 (nm) Duplo 2 (nm) Rata-Rata % Inhibisi 1 0,476 0,480 0,478 30,624 2 0,369 0,363 0,366 46,880 3 0,261 0,262 0,262 62,046 4 0,167 0,168 0,168 75,689 5 0,094 0,097 0,096 86,139 Absorbansi Kontrol 0,689
Tabel 18. Uji penangkapan radikal bebas DPPH kuersetin replikasi 3
Konsentrasi (µg/mL) Duplo 1 (nm) Duplo 2 (nm) Rata-Rata % Inhibisi 1 0,473 0,473 0,473 31,350 2 0,371 0,372 0,372 46,081 3 0,264 0,265 0,265 61,611 4 0,169 0,169 0,169 75,472 5 0,091 0,095 0,093 86,502 Absorbansi Kontrol 0,689
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Gambar 10. Grafik uji aktivitas antioksidan kuersetin
Fraksi KBNM-AcOEt
Tabel 19. Uji penangkapan radikal bebas DPPH fraksi KBNM-AcOEt
Kons. ( g/mL) Replikasi 1 Replikasi 2 Rata2 Abs kontrol negatif % inhibisi
Abs 1 Abs 2 Abs 1 Abs 2
100 0.755 0.765 0.746 0.751 0.75425 0.874 13.70137
200 0.671 0.67 0.667 0.668 0.669 0.874 23.45538
300 0.568 0.566 0.661 0.661 0.614 0.874 29.74828
400 0.485 0.484 0.516 0.509 0.4985 0.874 42.96339
500 0.41 0.412 0.459 0.446 0.43175 0.874 50.60069
Gambar 11. Grafik uji aktivitas antioksidan fraksi KBNM- AcOEt
Perhitungan IC50 Kuersetin Persamaan regresi : y = 13.974x + 18.309 Keterangan: y = 50 y = 13.974x + 18.309 50 = 13.974x + 18.309
62 x = 50 − 18.309 13.974 x = 2.267854587 μg/ml IC50 kuersetin adalah 2.26785 μg/mL Fraksi KBNM-AcOEt Persamaan regresi : y = 0.0933x + 4.1018 Keterangan: y = 50 y = 0.0933x + 4.1018 50 = 0.0933x + 4.1018 x = 50 − 4.1018 0.0933 x = 491.9421222 μg/ml
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Lampiran 7 Uji SPF
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Tabel 20. Penetapan SPF Secara In Vitro Fraksi KBNM-AcOEt
konsentrasi ( g/mL) Absorbansi λ max interval 2 nm λ (nm) EE x I (λ) SPF 5 0.001 314 315 0.0839 0.0008 25 0.018 294 295 0.0817 0.0147 50 0.034 292 290 0.015 0.0051 100 0.211 262 - - - Rata- Rata 0.0069 SD 0.0071 Perhitungan SPF Rumus: SPFSpektrofotometrik = CF x Σ290-320 EE (λ) x I (λ) x Abs (λ) Keterangan:
EE (λ) : Spektrum efek erythemal I (λ) : Spektrum intensitas matahari Abs (λ) : Absorbansi
CF : Faktor koreksi (= 10)
Fraksi KBNM-AcOEt konsentrasi 5 g/mL
SPFSpektrofotometrik = 10 x 0.0839 x 0.001
SPFSpektrofotometrik = 0.0008
Fraksi KBNM-AcOEt konsentrasi 25 g/mL
SPFSpektrofotometrik = 10 x 0.0817x 0.018
SPFSpektrofotometrik = 0.0147
Fraksi KBNM-AcOEt konsentrasi 50 g/mL
SPFSpektrofotometrik = 10 x 0.0150 x 0.034
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Lampiran 8
Foto dan dokumentasi
Uji
Gambar 12. Perajangan kulit
buah naga merah
Gambar 14. Proses Evaporasi
Gambar 17. Uji
Kandungan Fenolik Total
Gambar 13. Proses maserasi
Gambar 15. Proses fraksinasi
Gambar 16. Penangkapan