Efek Sitoprotektif Ekstrak Daun Afrika (Vernonia amygdalina Del.) terhadap Sel Vero yang diinduksi Hidrogen Peroksida (H2O2)
Lampiran 1. Hasil Identifikasi Tumbuhan
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Lampiran 2. Persetujuan Etik Penelitian Kesehatan
84
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Lampiran 3. Gambar Tumbuhan Daun Afrika dan Daun Afrika Segar
Tumbuhan daun Afrika
Daun Afrika segar
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Lampiran 4. Gambar Simplisia dan Serbuk Simplisia Daun Afrika
Simplisia daun Afrika
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Serbuk simplisia daun Afrika
Lampiran 5.Hasil Mikroskopik Daun Afrika
1
2
3
4
5
Keterangan:
1. Stomata tipe anomositik
2. Pembuluh kayu (xylem) bentuk spiral
3. Rambut penutup multiseluler
4. Kristal kalsium oksalat bentuk prisma
5. Kristal kalsium oksalat bentuk rosette
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Lampiran 6. Gambar Alat
Laminar Air Flow Class II
ELISA Reader
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Lampiran 7. Bagan Pembuatan Ekstrak Daun Afrika
Serbuk simplisia daun Afrika 500 g
dimasukkan ke dalam wadah kaca berwarna gelap
dimaserasi dengan pelarut n-heksana sebanyak 75
bagian
ditutup wadah kemudian didiamkan selama 5 hari
dipisahkan maserat dan ampas
Maserat I
Ampas
dimaserasi kembali ampas dengan pelarut nheksana sebanyak 25 bagian
dipisahkan kembali maserat dan ampas
Maserat II
Ampas
dikeringkan
Ekstrak kental
dengan cara yang sama
dimaserasi kembali dengan
pelarut etilasetat dan etanol
digabung semua hasil maserat
diuapkan dengan rotary evaporator pada suhu 40-50oC
dipekatkan di atas penangas air
Ekstrak kental
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Lampiran 8. Bagan Alur Penelitian
Serbuk simplisia
(500 g)
dilakukan skrining
fitokimia dan penetapan
kadar air
Diekstraksi dengan cara
maserasi bertingkat
menggunakan pelarut nheksana, etilasetat dan etanol
- Hasil skrining fitokimia
simplisia
- Penetapan kadar air
simplisia
ENDA/EEADA/EEDA
Uji MTT
skrining fitokimia
Viabilitas sel
% jumlah sel hidup
Hasil skrining
fitokimia ekstrak
Ekstrak yang berpotensi
Apoptosis
Ekspresi gen
Sel
apoptosis
Pengikatan antibodi
dengan antigen
% sel yang
apoptosis
Jumlah warna
coklat sedikit
90
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Lampiran 9. Perhitungan Hasil Penetapan Kadar Air Simplisia Daun Afrika
Persen kadar air
-
Volume air (mL )
x 100%
=
Berat sampel (g)
Berat sampel I
= 5, 012 g
Volume penjenuhan toluen
= 1,8 mL
Volume air I
= 2,2 mL
Persen kadar air I
=
- Berat sampel II
2,2−1,8
5,012 g
= 5, 007 g
Volume air I
= 2,2 mL
Volume air II
= 2,6 mL
Persen kadar air II
=
- Berat sampel III
x 100%= 7,98 %
2,6−2,2 mL
5,007 g
x 100%= 7,98 %
= 5, 023 g
Volume air I
= 2,6 mL
Volume air II
= 3,0 mL
Persen kadar air III
=
3,0−2,6 mL
5,023 g
Persen rata-rata kadar air serbuk simplisia
=
x 100%= 7,96%
(7,98+7,98+7,96)%
3
= 7,97%
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Lampiran 10. Perhitungan Konsentrasi Larutan Uji
Dibuat seri konsentrasi ENDA, EEADA, dan EEDA 100 µg/mL, 50 µg/mL, 25
µg/mL, 12,5 µg/mL, 6,25 µg/mL sebagai berikut:
Stok ekstrak = 50 mg dilarutkan dalam 1000 µL
50 mg = 50000 µg
50000
µg
= 50 µg/ µL :
1
mL
Konsentrasi larutan induk
=
- Konsentrasi 50 µg/mL
= dipipet 750 µL dari larutan dengan konsentrasi
1000 µL
1000
= 50000 µg/mL
Dibuat konsentrasi 100 µg/mL
V1 x C1
= V2 x C2
V1 x 50000 µg/mL
= 1500 µL x 100 µg/mL
1500 x 100
V1
=
50000
= 3 µL +1497 µL MK M199
100 µg/mL kemudian ditambahkan dengan 750 µL MK M199.
- Konsentrasi 25 µg/mL
= dipipet 750 µL dari larutan dengan konsentrasi 50
µg/mL kemudian ditambahkan dengan 750 µL MK M199.
- Konsentrasi 12,5 µg/mL
= dipipet 750 µL dari larutan dengan konsentrasi 25
µg/mL kemudian ditambahkan dengan 750 µL MK M199.
- Konsentrasi 6,25 µg/mL
= dipipet 750 µL dari larutan dengan konsentrasi
12,5 µg/mL kemudian ditambahkan dengan 750 µL MK M199.
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Lampiran 11. Perhitungan Pengenceran H2O2
H2O2 30%
BM = 34,0147 g/mol
BJ
= 1,11
Molaritas H2O2
= 30% x
= 30% x
BJ
BM
1,11 g
34,0147 g/mol
= 9,79 M
9790 mM
Untuk membuat 100 mM
= V2 x C2
V1 x C1
V1 x 9790 mM = 1000 µL x 100 mM
1000 x 100
V1
=
9790
= 10, 21 µL + 989,79 µL MK M199
= 10 µL + 990 µL MK M199
Untuk konsentrasi 0,8 mM
V1 x C1
= V2 x C2
V1 x 100 mM = 10000µL x 0,8 mM
10000 x 0,8
V1
=
100
= 80 µL + 9920 µL MK M199
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Lampiran 12. Hasil Analisa Statistik Viabilitas Sel
(a). EEDA
Descriptives
persen viabilitas
95% Confidence Interval for
Mean
N
Mean
Std. Deviation
Std. Error
Lower Bound
Upper Bound
etoh 100
3
63.7198
1.35594
.78285
60.3515
67.0882
etoh 50
3
78.7459
2.50690
1.44736
72.5184
84.9734
etoh 25
3
75.5118
1.79462
1.03613
71.0537
79.9699
etoh 12,5
3
71.6133
.93329
.53884
69.2949
73.9318
etoh 6,25
3
66.7150
1.52534
.88066
62.9258
70.5042
3
100.0000
.00000
.00000
100.0000
100.0000
18
76.0510
12.24538
2.88626
69.9615
82.1405
kontrol sel
Total
Descriptives
persen viabilitas
Minimum
Maximum
etoh 100
62.34
65.05
etoh 50
76.89
81.60
etoh 25
73.61
77.18
etoh 12,5
70.76
72.61
etoh 6,25
65.05
68.05
kontrol sel
100.00
100.00
62.34
100.00
Total
ANOVA
persen viabilitas
Sum of Squares
Between Groups
Within Groups
Total
df
Mean Square
F
2520.058
5
504.012
29.083
12
2.424
2549.141
17
207.961
Sig.
.000
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Post Hoc Tests
Multiple Comparisons
persen viabilitas
Tukey HSD
95% Confidence Interval
(I)
perlakuan
etoh 100
(J)
perlakuan
etoh 50
etoh 25
etoh 25
-15.02607
1.27111
.000
-19.2956
-10.7565
-11.79198
*
1.27111
.000
-16.0615
-7.5224
1.27111
.000
-12.1631
-3.6239
etoh 6,25
-2.99516
1.27111
.245
-7.2647
1.2744
-36.28016
*
1.27111
.000
-40.5497
-32.0106
15.02607
*
1.27111
.000
10.7565
19.2956
etoh 25
3.23409
1.27111
.186
-1.0355
7.5036
etoh 12,5
7.13257
*
1.27111
.001
2.8630
11.4021
etoh 6,25
12.03091
*
1.27111
.000
7.7613
16.3005
kontrol sel
-21.25409
*
1.27111
.000
-25.5237
-16.9845
11.79198
*
1.27111
.000
7.5224
16.0615
-3.23409
1.27111
.186
-7.5036
1.0355
etoh 12,5
3.89849
1.27111
.081
-.3711
8.1680
etoh 6,25
8.79682
*
1.27111
.000
4.5273
13.0664
*
1.27111
.000
-28.7577
-20.2186
7.89350
*
1.27111
.000
3.6239
12.1631
etoh 50
-7.13257
*
1.27111
.001
-11.4021
-2.8630
etoh 25
-3.89849
1.27111
.081
-8.1680
.3711
*
1.27111
.022
.6288
9.1679
*
1.27111
.000
-32.6562
-24.1171
2.99516
1.27111
.245
-1.2744
7.2647
*
1.27111
.000
-16.3005
-7.7613
-4.5273
etoh 100
etoh 100
etoh 100
etoh 6,25
kontrol sel
etoh 100
etoh 50
kontrol sel
Upper Bound
*
kontrol sel
etoh 6,25
Lower Bound
-7.89350
etoh 50
etoh 12,5
Sig.
*
etoh 12,5
kontrol sel
etoh 50
Mean
Difference (I-J) Std. Error
-24.48818
4.89833
-28.38667
-12.03091
etoh 25
-8.79682
*
1.27111
.000
-13.0664
etoh 12,5
-4.89833
*
1.27111
.022
-9.1679
-.6288
kontrol sel
-33.28500
*
1.27111
.000
-37.5546
-29.0154
etoh 100
36.28016
*
1.27111
.000
32.0106
40.5497
etoh 50
21.25409
*
1.27111
.000
16.9845
25.5237
etoh 25
24.48818
*
1.27111
.000
20.2186
28.7577
etoh 12,5
28.38667
*
1.27111
.000
24.1171
32.6562
etoh 6,25
33.28500
*
1.27111
.000
29.0154
37.5546
*. The mean difference is significant at the 0.05 level.
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Homogeneous Subsets
persen viabilitas
a
Tukey HSD
Subset for alpha = 0.05
perlakuan
N
1
2
3
etoh 100
3
63.7198
etoh 6,25
3
66.7150
etoh 12,5
3
71.6133
etoh 25
3
75.5118
etoh 50
3
kontrol sel
3
Sig.
4
75.5118
78.7459
100.0000
.245
.081
.186
1.000
Means for groups in homogeneous subsets are displayed.
a.
Uses Harmonic Mean Sample Size = 3,000.
(b). EEADA
Descriptives
Viabilitas
95% Confidence Interval for
Mean
N
Mean
Std. Deviation
Std. Error
Lower Bound
Upper Bound
etil 100
3
34.8100
.99715
.57570
32.3330
37.2870
etil 50
3
42.3687
.86786
.50106
40.2128
44.5246
etil 25
3
48.6000
1.18655
.68505
45.6524
51.5476
etil 12,5
3
68.9933
.90644
.52333
66.7416
71.2451
etil 6,25
3
65.9033
.37608
.21713
64.9691
66.8376
kontrol sel
3
100.0000
.00000
.00000
100.0000
100.0000
18
60.1126
22.20397
5.23353
49.0708
71.1543
Total
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Descriptives
Viabilitas
Minimum
Maximum
etil 100
33.67
35.52
etil 50
41.37
42.94
etil 25
47.65
49.93
etil 12,5
68.47
70.04
etil 6,25
65.62
66.33
100.00
100.00
33.67
100.00
kontrol sel
Total
Test of Homogeneity of Variances
Viabilitas
Levene Statistic
df1
3.662
df2
5
Sig.
12
.030
ANOVA
Viabilitas
Sum of Squares
Between Groups
Within Groups
Total
Df
Mean Square
F
8373.040
5
1674.608
8.237
12
.686
8381.277
17
2439.666
Sig.
.000
Post Hoc Tests
Multiple Comparisons
Viabilitas
Tukey HSD
95% Confidence Interval
(I)
perlakuan
etil 100
(J)
perlakuan
Std. Error
Sig.
Lower Bound
Upper Bound
*
.67647
.000
-9.8309
-5.2865
-13.79000
*
.67647
.000
-16.0622
-11.5178
-34.18333
*
.67647
.000
-36.4555
-31.9111
etil 6,25
-31.09333
*
.67647
.000
-33.3655
-28.8211
kontrol sel
-65.19000
*
.67647
.000
-67.4622
-62.9178
7.55867
*
.67647
.000
5.2865
9.8309
-6.23133
*
.67647
.000
-8.5035
-3.9591
etil 50
etil 25
etil 12,5
etil 50
Mean
Difference (I-J)
etil 100
etil 25
-7.55867
97
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-26.62467
*
.67647
.000
-28.8969
-24.3525
etil 6,25
-23.53467
*
.67647
.000
-25.8069
-21.2625
kontrol sel
-57.63133
*
.67647
.000
-59.9035
-55.3591
13.79000
*
.67647
.000
11.5178
16.0622
*
etil 12,5
etil 25
etil 100
etil 50
etil 12,5
.67647
.000
3.9591
8.5035
etil 12,5
-20.39333
6.23133
*
.67647
.000
-22.6655
-18.1211
etil 6,25
-17.30333
*
.67647
.000
-19.5755
-15.0311
kontrol sel
-51.40000
*
.67647
.000
-53.6722
-49.1278
34.18333
*
.67647
.000
31.9111
36.4555
26.62467
*
.67647
.000
24.3525
28.8969
20.39333
*
.67647
.000
18.1211
22.6655
*
.67647
.007
.8178
5.3622
-31.00667
*
.67647
.000
-33.2789
-28.7345
etil 100
31.09333
*
.67647
.000
28.8211
33.3655
etil 50
23.53467
*
.67647
.000
21.2625
25.8069
etil 25
17.30333
*
.67647
.000
15.0311
19.5755
etil 12,5
-3.09000
*
.67647
.007
-5.3622
-.8178
-34.09667
*
.67647
.000
-36.3689
-31.8245
65.19000
*
.67647
.000
62.9178
67.4622
etil 50
57.63133
*
.67647
.000
55.3591
59.9035
etil 25
51.40000
*
.67647
.000
49.1278
53.6722
etil 12,5
31.00667
*
.67647
.000
28.7345
33.2789
etil 6,25
34.09667
*
.67647
.000
31.8245
36.3689
etil 100
etil 50
etil 25
etil 6,25
3.09000
kontrol sel
etil 6,25
kontrol sel
kontrol sel
etil 100
*. The mean difference is significant at the 0.05 level.
Homogeneous Subsets
viabilitas
a
Tukey HSD
Subset for alpha = 0.05
perlakuan
N
1
etil 100
3
etil 50
3
etil 25
3
etil 6,25
3
etil 12,5
3
kontrol sel
3
Sig.
2
3
4
5
6
34.8100
42.3687
48.6000
65.9033
68.9933
100.0000
1.000
1.000
1.000
1.000
1.000
1.000
Means for groups in homogeneous subsets are displayed.
a. Uses Harmonic Mean Sample Size = 3,000.
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(c) ENDA
Descriptives
Viabilitas
95% Confidence Interval for
Mean
N
Mean
Std. Deviation
Std. Error
Lower Bound
Upper Bound
nhex 100
3
20.8267
2.01003
1.16049
15.8335
25.8199
nhex 50
3
23.9667
1.24344
.71790
20.8778
27.0555
nhex 25
3
33.3317
2.00415
1.15710
28.3531
38.3102
nhex 12,5
3
42.7033
2.63405
1.52077
36.1600
49.2467
nhex 6,25
3
37.2333
3.02183
1.74465
29.7267
44.7400
kontrol sel
3
100.0000
.00000
.00000
100.0000
100.0000
18
43.0103
27.37600
6.45259
29.3965
56.6240
Total
Descriptives
viabilitas
Minimum
Maximum
nhex 100
18.69
22.68
nhex 50
22.54
24.82
nhex 25
31.24
35.24
nhex 12,5
39.80
44.94
nhex 6,25
34.95
40.66
kontrol sel
100.00
100.00
18.69
100.00
Total
Test of Homogeneity of Variances
Viabilitas
Levene Statistic
2.350
df1
df2
5
Sig.
12
.105
ANOVA
Viabilitas
Sum of Squares
Between Groups
Within Groups
Total
df
Mean Square
12689.229
5
2537.846
51.345
12
4.279
12740.574
17
F
593.125
Sig.
.000
99
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Post Hoc Tests
Multiple Comparisons
viabilitas
Tukey HSD
95% Confidence Interval
(I) perlakuan (J) perlakuan
nhex 100
nhex 50
kontrol sel
Lower Bound
Upper
Bound
1.68894
.468
-8.8130 2.5330
nhex 25
-12.50500
*
1.68894
.000
-18.1780 -6.8320
nhex 12,5
-21.87667
*
1.68894
.000
-27.5497
16.2037
nhex 6,25
-16.40667
*
1.68894
.000
-22.0797
10.7337
kontrol sel
-79.17333
*
1.68894
.000
-84.8463
73.5003
3.14000
1.68894
.468
-2.5330 8.8130
*
nhex 100
1.68894
.001
-15.0380 -3.6920
nhex 12,5
-18.73667
*
1.68894
.000
-24.4097
nhex 6,25
-13.26667
*
1.68894
.000
-18.9397 -7.5937
kontrol sel
-76.03333
*
1.68894
.000
-81.7063
nhex 100
12.50500
*
1.68894
.000
9.36500
*
1.68894
.001
3.6920 15.0380
-9.37167
*
1.68894
.001
-15.0447 -3.6987
nhex 6,25
-3.90167
1.68894
.262
kontrol sel
-66.66833
*
1.68894
.000
nhex 100
21.87667
*
1.68894
.000
16.2037 27.5497
nhex 50
18.73667
*
1.68894
.000
13.0637 24.4097
nhex 25
9.37167
*
1.68894
.001
3.6987 15.0447
nhex 6,25
5.47000
1.68894
.061
kontrol sel
-57.29667
*
1.68894
.000
nhex 100
16.40667
*
1.68894
.000
nhex 50
13.26667
*
1.68894
.000
7.5937 18.9397
nhex 25
3.90167
1.68894
.262
-1.7713 9.5747
nhex 12,5
-5.47000
1.68894
.061
-11.1430
.2030
kontrol sel
-62.76667
*
1.68894
.000
-68.4397
57.0937
nhex 100
79.17333
*
1.68894
.000
73.5003 84.8463
76.03333
*
1.68894
.000
70.3603 81.7063
nhex 25
66.66833
*
1.68894
.000
60.9953 72.3413
nhex 12,5
57.29667
*
1.68894
.000
51.6237 62.9697
62.76667
*
1.68894
.000
57.0937 68.4397
nhex 12,5
nhex 6,25
Sig.
-3.14000
nhex 50
nhex 12,5
Std. Error
nhex 50
nhex 25
nhex 25
Mean Difference
(I-J)
nhex 50
nhex 6,25
-9.36500
13.0637
70.3603
6.8320 18.1780
-9.5747 1.7713
-72.3413
60.9953
-.2030 11.1430
-62.9697
51.6237
10.7337 22.0797
*. The mean difference is significant at the 0.05 level.
100
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Homogeneous Subsets
viabilitas
a
Tukey HSD
Subset for alpha = 0.05
perlakuan
N
1
2
3
nhex 100
3
20.8267
nhex 50
3
23.9667
nhex 25
3
33.3317
nhex 6,25
3
37.2333
nhex 12,5
3
kontrol sel
3
Sig.
4
37.2333
42.7033
100.0000
.468
.262
.061
1.000
Means for groups in homogeneous subsets are displayed.
a. Uses Harmonic Mean Sample Size = 3,000.
101
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83
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Lampiran 2. Persetujuan Etik Penelitian Kesehatan
84
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Lampiran 3. Gambar Tumbuhan Daun Afrika dan Daun Afrika Segar
Tumbuhan daun Afrika
Daun Afrika segar
85
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Lampiran 4. Gambar Simplisia dan Serbuk Simplisia Daun Afrika
Simplisia daun Afrika
86
Universitas Sumatera Utara
Serbuk simplisia daun Afrika
Lampiran 5.Hasil Mikroskopik Daun Afrika
1
2
3
4
5
Keterangan:
1. Stomata tipe anomositik
2. Pembuluh kayu (xylem) bentuk spiral
3. Rambut penutup multiseluler
4. Kristal kalsium oksalat bentuk prisma
5. Kristal kalsium oksalat bentuk rosette
87
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Lampiran 6. Gambar Alat
Laminar Air Flow Class II
ELISA Reader
88
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Lampiran 7. Bagan Pembuatan Ekstrak Daun Afrika
Serbuk simplisia daun Afrika 500 g
dimasukkan ke dalam wadah kaca berwarna gelap
dimaserasi dengan pelarut n-heksana sebanyak 75
bagian
ditutup wadah kemudian didiamkan selama 5 hari
dipisahkan maserat dan ampas
Maserat I
Ampas
dimaserasi kembali ampas dengan pelarut nheksana sebanyak 25 bagian
dipisahkan kembali maserat dan ampas
Maserat II
Ampas
dikeringkan
Ekstrak kental
dengan cara yang sama
dimaserasi kembali dengan
pelarut etilasetat dan etanol
digabung semua hasil maserat
diuapkan dengan rotary evaporator pada suhu 40-50oC
dipekatkan di atas penangas air
Ekstrak kental
89
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Lampiran 8. Bagan Alur Penelitian
Serbuk simplisia
(500 g)
dilakukan skrining
fitokimia dan penetapan
kadar air
Diekstraksi dengan cara
maserasi bertingkat
menggunakan pelarut nheksana, etilasetat dan etanol
- Hasil skrining fitokimia
simplisia
- Penetapan kadar air
simplisia
ENDA/EEADA/EEDA
Uji MTT
skrining fitokimia
Viabilitas sel
% jumlah sel hidup
Hasil skrining
fitokimia ekstrak
Ekstrak yang berpotensi
Apoptosis
Ekspresi gen
Sel
apoptosis
Pengikatan antibodi
dengan antigen
% sel yang
apoptosis
Jumlah warna
coklat sedikit
90
Universitas Sumatera Utara
Lampiran 9. Perhitungan Hasil Penetapan Kadar Air Simplisia Daun Afrika
Persen kadar air
-
Volume air (mL )
x 100%
=
Berat sampel (g)
Berat sampel I
= 5, 012 g
Volume penjenuhan toluen
= 1,8 mL
Volume air I
= 2,2 mL
Persen kadar air I
=
- Berat sampel II
2,2−1,8
5,012 g
= 5, 007 g
Volume air I
= 2,2 mL
Volume air II
= 2,6 mL
Persen kadar air II
=
- Berat sampel III
x 100%= 7,98 %
2,6−2,2 mL
5,007 g
x 100%= 7,98 %
= 5, 023 g
Volume air I
= 2,6 mL
Volume air II
= 3,0 mL
Persen kadar air III
=
3,0−2,6 mL
5,023 g
Persen rata-rata kadar air serbuk simplisia
=
x 100%= 7,96%
(7,98+7,98+7,96)%
3
= 7,97%
91
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Lampiran 10. Perhitungan Konsentrasi Larutan Uji
Dibuat seri konsentrasi ENDA, EEADA, dan EEDA 100 µg/mL, 50 µg/mL, 25
µg/mL, 12,5 µg/mL, 6,25 µg/mL sebagai berikut:
Stok ekstrak = 50 mg dilarutkan dalam 1000 µL
50 mg = 50000 µg
50000
µg
= 50 µg/ µL :
1
mL
Konsentrasi larutan induk
=
- Konsentrasi 50 µg/mL
= dipipet 750 µL dari larutan dengan konsentrasi
1000 µL
1000
= 50000 µg/mL
Dibuat konsentrasi 100 µg/mL
V1 x C1
= V2 x C2
V1 x 50000 µg/mL
= 1500 µL x 100 µg/mL
1500 x 100
V1
=
50000
= 3 µL +1497 µL MK M199
100 µg/mL kemudian ditambahkan dengan 750 µL MK M199.
- Konsentrasi 25 µg/mL
= dipipet 750 µL dari larutan dengan konsentrasi 50
µg/mL kemudian ditambahkan dengan 750 µL MK M199.
- Konsentrasi 12,5 µg/mL
= dipipet 750 µL dari larutan dengan konsentrasi 25
µg/mL kemudian ditambahkan dengan 750 µL MK M199.
- Konsentrasi 6,25 µg/mL
= dipipet 750 µL dari larutan dengan konsentrasi
12,5 µg/mL kemudian ditambahkan dengan 750 µL MK M199.
92
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Lampiran 11. Perhitungan Pengenceran H2O2
H2O2 30%
BM = 34,0147 g/mol
BJ
= 1,11
Molaritas H2O2
= 30% x
= 30% x
BJ
BM
1,11 g
34,0147 g/mol
= 9,79 M
9790 mM
Untuk membuat 100 mM
= V2 x C2
V1 x C1
V1 x 9790 mM = 1000 µL x 100 mM
1000 x 100
V1
=
9790
= 10, 21 µL + 989,79 µL MK M199
= 10 µL + 990 µL MK M199
Untuk konsentrasi 0,8 mM
V1 x C1
= V2 x C2
V1 x 100 mM = 10000µL x 0,8 mM
10000 x 0,8
V1
=
100
= 80 µL + 9920 µL MK M199
93
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Lampiran 12. Hasil Analisa Statistik Viabilitas Sel
(a). EEDA
Descriptives
persen viabilitas
95% Confidence Interval for
Mean
N
Mean
Std. Deviation
Std. Error
Lower Bound
Upper Bound
etoh 100
3
63.7198
1.35594
.78285
60.3515
67.0882
etoh 50
3
78.7459
2.50690
1.44736
72.5184
84.9734
etoh 25
3
75.5118
1.79462
1.03613
71.0537
79.9699
etoh 12,5
3
71.6133
.93329
.53884
69.2949
73.9318
etoh 6,25
3
66.7150
1.52534
.88066
62.9258
70.5042
3
100.0000
.00000
.00000
100.0000
100.0000
18
76.0510
12.24538
2.88626
69.9615
82.1405
kontrol sel
Total
Descriptives
persen viabilitas
Minimum
Maximum
etoh 100
62.34
65.05
etoh 50
76.89
81.60
etoh 25
73.61
77.18
etoh 12,5
70.76
72.61
etoh 6,25
65.05
68.05
kontrol sel
100.00
100.00
62.34
100.00
Total
ANOVA
persen viabilitas
Sum of Squares
Between Groups
Within Groups
Total
df
Mean Square
F
2520.058
5
504.012
29.083
12
2.424
2549.141
17
207.961
Sig.
.000
94
Universitas Sumatera Utara
Post Hoc Tests
Multiple Comparisons
persen viabilitas
Tukey HSD
95% Confidence Interval
(I)
perlakuan
etoh 100
(J)
perlakuan
etoh 50
etoh 25
etoh 25
-15.02607
1.27111
.000
-19.2956
-10.7565
-11.79198
*
1.27111
.000
-16.0615
-7.5224
1.27111
.000
-12.1631
-3.6239
etoh 6,25
-2.99516
1.27111
.245
-7.2647
1.2744
-36.28016
*
1.27111
.000
-40.5497
-32.0106
15.02607
*
1.27111
.000
10.7565
19.2956
etoh 25
3.23409
1.27111
.186
-1.0355
7.5036
etoh 12,5
7.13257
*
1.27111
.001
2.8630
11.4021
etoh 6,25
12.03091
*
1.27111
.000
7.7613
16.3005
kontrol sel
-21.25409
*
1.27111
.000
-25.5237
-16.9845
11.79198
*
1.27111
.000
7.5224
16.0615
-3.23409
1.27111
.186
-7.5036
1.0355
etoh 12,5
3.89849
1.27111
.081
-.3711
8.1680
etoh 6,25
8.79682
*
1.27111
.000
4.5273
13.0664
*
1.27111
.000
-28.7577
-20.2186
7.89350
*
1.27111
.000
3.6239
12.1631
etoh 50
-7.13257
*
1.27111
.001
-11.4021
-2.8630
etoh 25
-3.89849
1.27111
.081
-8.1680
.3711
*
1.27111
.022
.6288
9.1679
*
1.27111
.000
-32.6562
-24.1171
2.99516
1.27111
.245
-1.2744
7.2647
*
1.27111
.000
-16.3005
-7.7613
-4.5273
etoh 100
etoh 100
etoh 100
etoh 6,25
kontrol sel
etoh 100
etoh 50
kontrol sel
Upper Bound
*
kontrol sel
etoh 6,25
Lower Bound
-7.89350
etoh 50
etoh 12,5
Sig.
*
etoh 12,5
kontrol sel
etoh 50
Mean
Difference (I-J) Std. Error
-24.48818
4.89833
-28.38667
-12.03091
etoh 25
-8.79682
*
1.27111
.000
-13.0664
etoh 12,5
-4.89833
*
1.27111
.022
-9.1679
-.6288
kontrol sel
-33.28500
*
1.27111
.000
-37.5546
-29.0154
etoh 100
36.28016
*
1.27111
.000
32.0106
40.5497
etoh 50
21.25409
*
1.27111
.000
16.9845
25.5237
etoh 25
24.48818
*
1.27111
.000
20.2186
28.7577
etoh 12,5
28.38667
*
1.27111
.000
24.1171
32.6562
etoh 6,25
33.28500
*
1.27111
.000
29.0154
37.5546
*. The mean difference is significant at the 0.05 level.
95
Universitas Sumatera Utara
Homogeneous Subsets
persen viabilitas
a
Tukey HSD
Subset for alpha = 0.05
perlakuan
N
1
2
3
etoh 100
3
63.7198
etoh 6,25
3
66.7150
etoh 12,5
3
71.6133
etoh 25
3
75.5118
etoh 50
3
kontrol sel
3
Sig.
4
75.5118
78.7459
100.0000
.245
.081
.186
1.000
Means for groups in homogeneous subsets are displayed.
a.
Uses Harmonic Mean Sample Size = 3,000.
(b). EEADA
Descriptives
Viabilitas
95% Confidence Interval for
Mean
N
Mean
Std. Deviation
Std. Error
Lower Bound
Upper Bound
etil 100
3
34.8100
.99715
.57570
32.3330
37.2870
etil 50
3
42.3687
.86786
.50106
40.2128
44.5246
etil 25
3
48.6000
1.18655
.68505
45.6524
51.5476
etil 12,5
3
68.9933
.90644
.52333
66.7416
71.2451
etil 6,25
3
65.9033
.37608
.21713
64.9691
66.8376
kontrol sel
3
100.0000
.00000
.00000
100.0000
100.0000
18
60.1126
22.20397
5.23353
49.0708
71.1543
Total
96
Universitas Sumatera Utara
Descriptives
Viabilitas
Minimum
Maximum
etil 100
33.67
35.52
etil 50
41.37
42.94
etil 25
47.65
49.93
etil 12,5
68.47
70.04
etil 6,25
65.62
66.33
100.00
100.00
33.67
100.00
kontrol sel
Total
Test of Homogeneity of Variances
Viabilitas
Levene Statistic
df1
3.662
df2
5
Sig.
12
.030
ANOVA
Viabilitas
Sum of Squares
Between Groups
Within Groups
Total
Df
Mean Square
F
8373.040
5
1674.608
8.237
12
.686
8381.277
17
2439.666
Sig.
.000
Post Hoc Tests
Multiple Comparisons
Viabilitas
Tukey HSD
95% Confidence Interval
(I)
perlakuan
etil 100
(J)
perlakuan
Std. Error
Sig.
Lower Bound
Upper Bound
*
.67647
.000
-9.8309
-5.2865
-13.79000
*
.67647
.000
-16.0622
-11.5178
-34.18333
*
.67647
.000
-36.4555
-31.9111
etil 6,25
-31.09333
*
.67647
.000
-33.3655
-28.8211
kontrol sel
-65.19000
*
.67647
.000
-67.4622
-62.9178
7.55867
*
.67647
.000
5.2865
9.8309
-6.23133
*
.67647
.000
-8.5035
-3.9591
etil 50
etil 25
etil 12,5
etil 50
Mean
Difference (I-J)
etil 100
etil 25
-7.55867
97
Universitas Sumatera Utara
-26.62467
*
.67647
.000
-28.8969
-24.3525
etil 6,25
-23.53467
*
.67647
.000
-25.8069
-21.2625
kontrol sel
-57.63133
*
.67647
.000
-59.9035
-55.3591
13.79000
*
.67647
.000
11.5178
16.0622
*
etil 12,5
etil 25
etil 100
etil 50
etil 12,5
.67647
.000
3.9591
8.5035
etil 12,5
-20.39333
6.23133
*
.67647
.000
-22.6655
-18.1211
etil 6,25
-17.30333
*
.67647
.000
-19.5755
-15.0311
kontrol sel
-51.40000
*
.67647
.000
-53.6722
-49.1278
34.18333
*
.67647
.000
31.9111
36.4555
26.62467
*
.67647
.000
24.3525
28.8969
20.39333
*
.67647
.000
18.1211
22.6655
*
.67647
.007
.8178
5.3622
-31.00667
*
.67647
.000
-33.2789
-28.7345
etil 100
31.09333
*
.67647
.000
28.8211
33.3655
etil 50
23.53467
*
.67647
.000
21.2625
25.8069
etil 25
17.30333
*
.67647
.000
15.0311
19.5755
etil 12,5
-3.09000
*
.67647
.007
-5.3622
-.8178
-34.09667
*
.67647
.000
-36.3689
-31.8245
65.19000
*
.67647
.000
62.9178
67.4622
etil 50
57.63133
*
.67647
.000
55.3591
59.9035
etil 25
51.40000
*
.67647
.000
49.1278
53.6722
etil 12,5
31.00667
*
.67647
.000
28.7345
33.2789
etil 6,25
34.09667
*
.67647
.000
31.8245
36.3689
etil 100
etil 50
etil 25
etil 6,25
3.09000
kontrol sel
etil 6,25
kontrol sel
kontrol sel
etil 100
*. The mean difference is significant at the 0.05 level.
Homogeneous Subsets
viabilitas
a
Tukey HSD
Subset for alpha = 0.05
perlakuan
N
1
etil 100
3
etil 50
3
etil 25
3
etil 6,25
3
etil 12,5
3
kontrol sel
3
Sig.
2
3
4
5
6
34.8100
42.3687
48.6000
65.9033
68.9933
100.0000
1.000
1.000
1.000
1.000
1.000
1.000
Means for groups in homogeneous subsets are displayed.
a. Uses Harmonic Mean Sample Size = 3,000.
98
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(c) ENDA
Descriptives
Viabilitas
95% Confidence Interval for
Mean
N
Mean
Std. Deviation
Std. Error
Lower Bound
Upper Bound
nhex 100
3
20.8267
2.01003
1.16049
15.8335
25.8199
nhex 50
3
23.9667
1.24344
.71790
20.8778
27.0555
nhex 25
3
33.3317
2.00415
1.15710
28.3531
38.3102
nhex 12,5
3
42.7033
2.63405
1.52077
36.1600
49.2467
nhex 6,25
3
37.2333
3.02183
1.74465
29.7267
44.7400
kontrol sel
3
100.0000
.00000
.00000
100.0000
100.0000
18
43.0103
27.37600
6.45259
29.3965
56.6240
Total
Descriptives
viabilitas
Minimum
Maximum
nhex 100
18.69
22.68
nhex 50
22.54
24.82
nhex 25
31.24
35.24
nhex 12,5
39.80
44.94
nhex 6,25
34.95
40.66
kontrol sel
100.00
100.00
18.69
100.00
Total
Test of Homogeneity of Variances
Viabilitas
Levene Statistic
2.350
df1
df2
5
Sig.
12
.105
ANOVA
Viabilitas
Sum of Squares
Between Groups
Within Groups
Total
df
Mean Square
12689.229
5
2537.846
51.345
12
4.279
12740.574
17
F
593.125
Sig.
.000
99
Universitas Sumatera Utara
Post Hoc Tests
Multiple Comparisons
viabilitas
Tukey HSD
95% Confidence Interval
(I) perlakuan (J) perlakuan
nhex 100
nhex 50
kontrol sel
Lower Bound
Upper
Bound
1.68894
.468
-8.8130 2.5330
nhex 25
-12.50500
*
1.68894
.000
-18.1780 -6.8320
nhex 12,5
-21.87667
*
1.68894
.000
-27.5497
16.2037
nhex 6,25
-16.40667
*
1.68894
.000
-22.0797
10.7337
kontrol sel
-79.17333
*
1.68894
.000
-84.8463
73.5003
3.14000
1.68894
.468
-2.5330 8.8130
*
nhex 100
1.68894
.001
-15.0380 -3.6920
nhex 12,5
-18.73667
*
1.68894
.000
-24.4097
nhex 6,25
-13.26667
*
1.68894
.000
-18.9397 -7.5937
kontrol sel
-76.03333
*
1.68894
.000
-81.7063
nhex 100
12.50500
*
1.68894
.000
9.36500
*
1.68894
.001
3.6920 15.0380
-9.37167
*
1.68894
.001
-15.0447 -3.6987
nhex 6,25
-3.90167
1.68894
.262
kontrol sel
-66.66833
*
1.68894
.000
nhex 100
21.87667
*
1.68894
.000
16.2037 27.5497
nhex 50
18.73667
*
1.68894
.000
13.0637 24.4097
nhex 25
9.37167
*
1.68894
.001
3.6987 15.0447
nhex 6,25
5.47000
1.68894
.061
kontrol sel
-57.29667
*
1.68894
.000
nhex 100
16.40667
*
1.68894
.000
nhex 50
13.26667
*
1.68894
.000
7.5937 18.9397
nhex 25
3.90167
1.68894
.262
-1.7713 9.5747
nhex 12,5
-5.47000
1.68894
.061
-11.1430
.2030
kontrol sel
-62.76667
*
1.68894
.000
-68.4397
57.0937
nhex 100
79.17333
*
1.68894
.000
73.5003 84.8463
76.03333
*
1.68894
.000
70.3603 81.7063
nhex 25
66.66833
*
1.68894
.000
60.9953 72.3413
nhex 12,5
57.29667
*
1.68894
.000
51.6237 62.9697
62.76667
*
1.68894
.000
57.0937 68.4397
nhex 12,5
nhex 6,25
Sig.
-3.14000
nhex 50
nhex 12,5
Std. Error
nhex 50
nhex 25
nhex 25
Mean Difference
(I-J)
nhex 50
nhex 6,25
-9.36500
13.0637
70.3603
6.8320 18.1780
-9.5747 1.7713
-72.3413
60.9953
-.2030 11.1430
-62.9697
51.6237
10.7337 22.0797
*. The mean difference is significant at the 0.05 level.
100
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Homogeneous Subsets
viabilitas
a
Tukey HSD
Subset for alpha = 0.05
perlakuan
N
1
2
3
nhex 100
3
20.8267
nhex 50
3
23.9667
nhex 25
3
33.3317
nhex 6,25
3
37.2333
nhex 12,5
3
kontrol sel
3
Sig.
4
37.2333
42.7033
100.0000
.468
.262
.061
1.000
Means for groups in homogeneous subsets are displayed.
a. Uses Harmonic Mean Sample Size = 3,000.
101
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