B1J008164 13.
LAMPIRAN
Lampiran 1. Tabel cara kerja Pengukuran Aktivitas Protease Digesti
VOLUME (Ml)
PEREAKSI
Standard
Sampel
Kontrol
Balanko
Tirosin
0.50
-
-
-
Kasein 5%
-
0.50
-
-
Buffer
0.50
0.50
0.50
0.50
Ekstrak
-
0.25
-
-
Akuades
0.25
-
0.75
0.75
TCA
-
-
-
-
standard
enzim
Di Inkubasi selama 60 menit pada suhu 370c
TCA
0.75
0.75
-
0.75
Kasein 1%
-
-
0.50
-
Di masukkan semua tabung kedalama refrigerator selama minimal 1 jam
Sentrifugasi pada kecepatan 6000 rpm selama 15 menit
Ukur absorbansinya pada panjang gelombang 280 nm
Lampiran 2. Tabel Cara Kerja Pengukuran Aktivitas Amilase Digesti
VOLUME (Ml)
PEREAKSI
Standard
Sampel
Kontrol
Balanko
Maltosa
0.75
-
-
-
Amilum 1%
-
0.75
-
-
Ekstrak enzim
-
0.25
-
-
Akuades
0.25
-
0.25
1.0
DNS
-
-
0.50
-
standard
Inkubasi selama 30 menit pada suhu 370c
DNS
0.50
0.50
-
0.50
Amilum 1%
-
-
0.75
-
Di masukkan semua tabung kedalama refrigerator selama 5 menit
Setelah dingin di tambahkan akuades sebanyak 1.5 ml
Ukur absorbansinya pada panjang gelombang 540 nm
Lampiran 3. Tabel cara kerja Penentuan Kadar Protein Supernatan
VOLUME (Ml)
PEREAKSI
Standard
Sampel
Kontrol
Balanko
Kasein
0.1
-
-
-
Ekstrak enzim
-
0.1
-
-
Pereaksi C
5.0
-
-
-
standard
Inkubasi selama 30 menit pada suhu 370c
Pereaksi D
0.5
0.5
-
-
Di kocok segera dan biarkan pada suhu kamar selama paling sedikit 10 menit
Ukur absorbansinyapada panjang gelombang 660 nm
Kadar protein sampel di tentukan dengan kurva standard
Lampiran 4. Analisis Aktivitas Protease pada Hepatopankreas dan Intestine
Univariate Analysis of Variance
Descriptive Statistics
Dependent Variable: Protease mmol/mg/menit (U/menit)
Ukuran
B
S
K
Total
Tempat
H
N
Total
H
N
Total
H
N
Total
H
N
Total
Mean
.58250
.78600
Std. Deviation
.188951
.039758
N
.68425
.77750
.93125
.166765
.078122
.073355
8
4
4
.85438
.60675
.84850
.108054
.090614
.293229
8
4
4
.72763
.65558
.85525
.75542
.238887
.147835
.170907
.186607
8
12
12
24
4
4
Tests of Between-Subjects Effects
Dependent Variable: Protease mmol/mg/menit (U/menit)
Source
Corrected Model
Intercept
Ukuran
Tempat
Ukuran * Tempat
Error
Total
Corrected Total
Type III Sum
of Squares
.372a
13.696
.125
df
5
1
2
.239
.008
.429
14.497
Mean Square
.074
13.696
.063
F
3.123
574.802
2.624
Sig.
.033
.000
.100
.239
.004
.024
10.039
.163
.005
.850
1
2
18
24
.801
23
a. R Squared = .465 (Adjusted R Squared = .316)
Post Hoc Tests
Estimates
Dependent Variable: Protease mmol/mg/menit (U/menit)
Tempat
H
N
Mean
.656
Std. Error
.045
.855
.045
95% Confidence Interval
Lower Bound Upper Bound
.562
.749
.762
.949
Pairwise Comparisons
Dependent Variable: Protease mmol/mg/menit (U/menit)
(I) Tempat
H
N
(J) Tempat
N
H
Mean
Difference
Std. Error
(I-J)
-.200*
.063
.200*
.063
a
Sig.
.005
.005
95% Confidence Interval for
a
Difference
Lower Bound Upper Bound
-.332
-.067
.067
.332
Based on estimated marginal means
*. The mean difference is significant at the .05 level.
a. Adjustment for multiple comparisons: Least Significant Difference (equivalent to no
adjustments).
Univariate Tests
Dependent Variable: Protease mmol/mg/menit (U/menit)
Contrast
Error
Sum of
Squares
.239
df
.429
1
Mean Square
.239
18
.024
F
10.039
Sig.
.005
The F tests the effect of Tempat. This test is based on the linearly independent
pairwise comparisons among the estimated marginal means.
3. Ukuran * Tempat
Dependent Variable: Protease mmol/mg/menit (U/menit)
Ukuran
B
S
K
Tempat
H
N
H
N
H
N
95% Confidence Interval
Lower Bound Upper Bound
.420
.745
Mean
.583
Std. Error
.077
.786
.778
.931
.077
.077
.077
.624
.615
.769
.948
.940
1.093
.607
.848
.077
.077
.445
.686
.769
1.011
Lampiran 5. Hasil Analisis Aktivitas Amilase pada Hepatopankreas dan
Intestine
Univariate Analysis of Variance
Between-Subjects Factors
Ukuran
Tempat
1
2
3
1
2
Value Label
B
S
K
H
N
N
8
8
8
12
12
Descriptive Statistics
Dependent Variable: Amilase mmol/mg/menit (U/menit)
Ukuran
B
S
K
Total
Tempat
H
N
Total
H
N
Total
H
N
Total
H
N
Total
Mean
.62225
Std. Deviation
.231998
N
1.41125
1.01675
.72125
.097288
.452754
.208410
4
8
4
1.14775
.93450
.72125
.174603
.289228
.226366
4
8
4
1.17325
.94725
.68825
.230295
.321034
.207065
4
8
12
1.24408
.96617
.201794
.347242
12
24
4
Tests of Between-Subjects Effects
Dependent Variable: Amilase mmol/mg/menit (U/menit)
Source
Corrected Model
Intercept
Ukuran
Tempat
Ukuran * Tempat
Error
Total
Corrected Total
Type III Sum
of Squares
2.049a
22.403
df
5
1
Mean Square
.410
22.403
F
10.181
556.640
Sig.
.000
.000
.031
1.854
.164
2
1
2
.016
1.854
.082
.390
46.057
2.034
.683
.000
.160
.724
25.177
2.773
18
24
23
.040
a. R Squared = .739 (Adjusted R Squared = .666)
Post Hoc Tests
Estimates
Dependent Variable: Amilase mmol/mg/menit (U/menit)
Tempat
H
N
Mean
.688
Std. Error
.058
1.244
.058
95% Confidence Interval
Lower Bound Upper Bound
.567
.810
1.122
1.366
Pairwise Comparisons
Dependent Variable: Amilase mmol/mg/menit (U/menit)
(I) Tempat
H
N
(J) Tempat
N
H
Mean
Difference
Std. Error
(I-J)
-.556*
.082
.556*
.082
a
Sig.
.000
.000
95% Confidence Interval for
a
Difference
Lower Bound Upper Bound
-.728
-.384
.384
.728
Based on estimated marginal means
*. The mean difference is significant at the .05 level.
a. Adjustment for multiple comparisons: Least Significant Difference (equivalent to no
adjustments).
Univariate Tests
Dependent Variable: Amilase mmol/mg/menit (U/menit)
Contrast
Error
Sum of
Squares
1.854
.724
df
1
18
Mean Square
1.854
.040
F
46.057
Sig.
.000
The F tests the effect of Tempat. This test is based on the linearly independent
pairwise comparisons among the estimated marginal means.
3. Ukuran * Tempat
Dependent Variable: Amilase mmol/mg/menit (U/menit)
Ukuran
B
S
K
Tempat
H
N
H
N
H
N
95% Confidence Interval
Lower Bound Upper Bound
.412
.833
Mean
.622
Std. Error
.100
1.411
.721
1.148
.100
.100
.100
1.201
.511
.937
1.622
.932
1.358
.721
1.173
.100
.100
.511
.963
.932
1.384
Lampiran 6. Kurva Standar Larutan Tirosin
Absorbansi
0,6
Y = a + bx
0,5
0,4
0,3
0,2
0,1
0
0
0,05
0,1
0,25
0,5
1,0
Konsentrasi (mg/ml)
540 nm
Lampiran 7. Kurva Standar Larutan Maltosa
0,01
0,05
0,10
mg/ml
0,25
Lampiran Gambar Penelitian
Gambar 1. Akuarium ikan uji
Gambar 3. Saluran digesti ikan nilem
Gambar 5. Water Heater
Gambar 2. Timbangan Mekanik
Gambar 4. Running Enzim
Gambar 6. Sentrifugase
Gambar 7. Oven
Gambar 8. Timbangan Analitik
Gambar 9. Homogenizer
Gambar 10. Spectrophotometer tipe
U-3900
Lampiran 1. Tabel cara kerja Pengukuran Aktivitas Protease Digesti
VOLUME (Ml)
PEREAKSI
Standard
Sampel
Kontrol
Balanko
Tirosin
0.50
-
-
-
Kasein 5%
-
0.50
-
-
Buffer
0.50
0.50
0.50
0.50
Ekstrak
-
0.25
-
-
Akuades
0.25
-
0.75
0.75
TCA
-
-
-
-
standard
enzim
Di Inkubasi selama 60 menit pada suhu 370c
TCA
0.75
0.75
-
0.75
Kasein 1%
-
-
0.50
-
Di masukkan semua tabung kedalama refrigerator selama minimal 1 jam
Sentrifugasi pada kecepatan 6000 rpm selama 15 menit
Ukur absorbansinya pada panjang gelombang 280 nm
Lampiran 2. Tabel Cara Kerja Pengukuran Aktivitas Amilase Digesti
VOLUME (Ml)
PEREAKSI
Standard
Sampel
Kontrol
Balanko
Maltosa
0.75
-
-
-
Amilum 1%
-
0.75
-
-
Ekstrak enzim
-
0.25
-
-
Akuades
0.25
-
0.25
1.0
DNS
-
-
0.50
-
standard
Inkubasi selama 30 menit pada suhu 370c
DNS
0.50
0.50
-
0.50
Amilum 1%
-
-
0.75
-
Di masukkan semua tabung kedalama refrigerator selama 5 menit
Setelah dingin di tambahkan akuades sebanyak 1.5 ml
Ukur absorbansinya pada panjang gelombang 540 nm
Lampiran 3. Tabel cara kerja Penentuan Kadar Protein Supernatan
VOLUME (Ml)
PEREAKSI
Standard
Sampel
Kontrol
Balanko
Kasein
0.1
-
-
-
Ekstrak enzim
-
0.1
-
-
Pereaksi C
5.0
-
-
-
standard
Inkubasi selama 30 menit pada suhu 370c
Pereaksi D
0.5
0.5
-
-
Di kocok segera dan biarkan pada suhu kamar selama paling sedikit 10 menit
Ukur absorbansinyapada panjang gelombang 660 nm
Kadar protein sampel di tentukan dengan kurva standard
Lampiran 4. Analisis Aktivitas Protease pada Hepatopankreas dan Intestine
Univariate Analysis of Variance
Descriptive Statistics
Dependent Variable: Protease mmol/mg/menit (U/menit)
Ukuran
B
S
K
Total
Tempat
H
N
Total
H
N
Total
H
N
Total
H
N
Total
Mean
.58250
.78600
Std. Deviation
.188951
.039758
N
.68425
.77750
.93125
.166765
.078122
.073355
8
4
4
.85438
.60675
.84850
.108054
.090614
.293229
8
4
4
.72763
.65558
.85525
.75542
.238887
.147835
.170907
.186607
8
12
12
24
4
4
Tests of Between-Subjects Effects
Dependent Variable: Protease mmol/mg/menit (U/menit)
Source
Corrected Model
Intercept
Ukuran
Tempat
Ukuran * Tempat
Error
Total
Corrected Total
Type III Sum
of Squares
.372a
13.696
.125
df
5
1
2
.239
.008
.429
14.497
Mean Square
.074
13.696
.063
F
3.123
574.802
2.624
Sig.
.033
.000
.100
.239
.004
.024
10.039
.163
.005
.850
1
2
18
24
.801
23
a. R Squared = .465 (Adjusted R Squared = .316)
Post Hoc Tests
Estimates
Dependent Variable: Protease mmol/mg/menit (U/menit)
Tempat
H
N
Mean
.656
Std. Error
.045
.855
.045
95% Confidence Interval
Lower Bound Upper Bound
.562
.749
.762
.949
Pairwise Comparisons
Dependent Variable: Protease mmol/mg/menit (U/menit)
(I) Tempat
H
N
(J) Tempat
N
H
Mean
Difference
Std. Error
(I-J)
-.200*
.063
.200*
.063
a
Sig.
.005
.005
95% Confidence Interval for
a
Difference
Lower Bound Upper Bound
-.332
-.067
.067
.332
Based on estimated marginal means
*. The mean difference is significant at the .05 level.
a. Adjustment for multiple comparisons: Least Significant Difference (equivalent to no
adjustments).
Univariate Tests
Dependent Variable: Protease mmol/mg/menit (U/menit)
Contrast
Error
Sum of
Squares
.239
df
.429
1
Mean Square
.239
18
.024
F
10.039
Sig.
.005
The F tests the effect of Tempat. This test is based on the linearly independent
pairwise comparisons among the estimated marginal means.
3. Ukuran * Tempat
Dependent Variable: Protease mmol/mg/menit (U/menit)
Ukuran
B
S
K
Tempat
H
N
H
N
H
N
95% Confidence Interval
Lower Bound Upper Bound
.420
.745
Mean
.583
Std. Error
.077
.786
.778
.931
.077
.077
.077
.624
.615
.769
.948
.940
1.093
.607
.848
.077
.077
.445
.686
.769
1.011
Lampiran 5. Hasil Analisis Aktivitas Amilase pada Hepatopankreas dan
Intestine
Univariate Analysis of Variance
Between-Subjects Factors
Ukuran
Tempat
1
2
3
1
2
Value Label
B
S
K
H
N
N
8
8
8
12
12
Descriptive Statistics
Dependent Variable: Amilase mmol/mg/menit (U/menit)
Ukuran
B
S
K
Total
Tempat
H
N
Total
H
N
Total
H
N
Total
H
N
Total
Mean
.62225
Std. Deviation
.231998
N
1.41125
1.01675
.72125
.097288
.452754
.208410
4
8
4
1.14775
.93450
.72125
.174603
.289228
.226366
4
8
4
1.17325
.94725
.68825
.230295
.321034
.207065
4
8
12
1.24408
.96617
.201794
.347242
12
24
4
Tests of Between-Subjects Effects
Dependent Variable: Amilase mmol/mg/menit (U/menit)
Source
Corrected Model
Intercept
Ukuran
Tempat
Ukuran * Tempat
Error
Total
Corrected Total
Type III Sum
of Squares
2.049a
22.403
df
5
1
Mean Square
.410
22.403
F
10.181
556.640
Sig.
.000
.000
.031
1.854
.164
2
1
2
.016
1.854
.082
.390
46.057
2.034
.683
.000
.160
.724
25.177
2.773
18
24
23
.040
a. R Squared = .739 (Adjusted R Squared = .666)
Post Hoc Tests
Estimates
Dependent Variable: Amilase mmol/mg/menit (U/menit)
Tempat
H
N
Mean
.688
Std. Error
.058
1.244
.058
95% Confidence Interval
Lower Bound Upper Bound
.567
.810
1.122
1.366
Pairwise Comparisons
Dependent Variable: Amilase mmol/mg/menit (U/menit)
(I) Tempat
H
N
(J) Tempat
N
H
Mean
Difference
Std. Error
(I-J)
-.556*
.082
.556*
.082
a
Sig.
.000
.000
95% Confidence Interval for
a
Difference
Lower Bound Upper Bound
-.728
-.384
.384
.728
Based on estimated marginal means
*. The mean difference is significant at the .05 level.
a. Adjustment for multiple comparisons: Least Significant Difference (equivalent to no
adjustments).
Univariate Tests
Dependent Variable: Amilase mmol/mg/menit (U/menit)
Contrast
Error
Sum of
Squares
1.854
.724
df
1
18
Mean Square
1.854
.040
F
46.057
Sig.
.000
The F tests the effect of Tempat. This test is based on the linearly independent
pairwise comparisons among the estimated marginal means.
3. Ukuran * Tempat
Dependent Variable: Amilase mmol/mg/menit (U/menit)
Ukuran
B
S
K
Tempat
H
N
H
N
H
N
95% Confidence Interval
Lower Bound Upper Bound
.412
.833
Mean
.622
Std. Error
.100
1.411
.721
1.148
.100
.100
.100
1.201
.511
.937
1.622
.932
1.358
.721
1.173
.100
.100
.511
.963
.932
1.384
Lampiran 6. Kurva Standar Larutan Tirosin
Absorbansi
0,6
Y = a + bx
0,5
0,4
0,3
0,2
0,1
0
0
0,05
0,1
0,25
0,5
1,0
Konsentrasi (mg/ml)
540 nm
Lampiran 7. Kurva Standar Larutan Maltosa
0,01
0,05
0,10
mg/ml
0,25
Lampiran Gambar Penelitian
Gambar 1. Akuarium ikan uji
Gambar 3. Saluran digesti ikan nilem
Gambar 5. Water Heater
Gambar 2. Timbangan Mekanik
Gambar 4. Running Enzim
Gambar 6. Sentrifugase
Gambar 7. Oven
Gambar 8. Timbangan Analitik
Gambar 9. Homogenizer
Gambar 10. Spectrophotometer tipe
U-3900