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7. LAMPIRAN LAMPIRAN 1. Pembuatan Reagen Bradford dan Larutan Standar Protein 1.1.
Pembuatan Reagen Bradford
Commasive Blue sebanyak 0,01 gram dilarutkan ke dalam 5 ml etanol 95% kemudain ditambah asam fosfor 85% sebanyak 10 ml, kemudian di saring dengan menggunakan kertas saringdan disimpan dalam botol gelap pada suhu rendah. Saat hendak akan digunakan maka harus dilakukan pengenceran sebanyak lima kali. (Bradford, 1976)
1.2.
Pembuatan Larutan Standar Protein
Bovine Serum Albumin (BSA) sebanyak 25 mg dilarutkan ke dalam 10 ml aquadest dan hasil pencampuran dinamakan larutan stock BSA 2500 ppm. Setelah itu diambil 4ml dan diencerkan hingga 100 ml dengan menggunakan aquadest. Setelah itu membuat pengukuran terhadap standar protein terlarut dengan konsentrasi 0,10, 20,30, 40, 50, 60, 70, 80, 90,dan 100. Setelah itu diambil masing – masing 0,1ml larutan standar dari masing – masing konsentrasi kemudian ditambahkan dengan 5ml larutan Bradford dan didiamkan 5 menit,dan dispektrofotometri dengan menggunakan panjang gelombang 595 nm.
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LAMPIRAN 2. Komposisi Larutan Kerja dalam Elektroforesis SDS-Page
Nama Larutan Larutan A (Acrylamide Stock Solution), 100 ml 30% acrylamide, 0,8% bisacrylamide Larutan B (4x Separating Gel Buffer), 100 ml
Komposisi 29,2 gram acrylamide 0,8 gram bis-acrylamide
75 ml 2M Tris-HCl (pH 8,8) --- 1,5 M 4 ml 10% SDS --- 0,4 % 21 ml ddH2O Larutan C 50 ml 1M Tris-HCl (pH 6,8) --- 0,5 M (4x Stacking Gel Buffer), 100 ml 4 ml 10% SDS --- 0,4 % 46 ml ddH2O 10% Ammonium Persulphate 0,5 gram APS (APS) 5 ml DDH2O Buffer Elektroforesis 3 gr Tris --- 25Mm 14,4 gram glycine --- 192Mm 1 gram SDS --- 0,1% ddH2O sampai 1 liter Sampel Buffer 3,028 gram Tris 0,25 ml larutan C 0,4 ml 10% SDS 25 ml gliserol 0,2 mg bromophenol blue 0,031 gram Dithiothreitol (DTT) Sumber : (Bollag et al., 1991 dengan modifikasi)
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LAMPIRAN 3. Kurva Standar BSA (Bovine Serum Albumin) Berat Molekular Weight
3.1. Kurva Standar untuk Mie jahe konsentrasi 0% dan 1% 1.000
0.800 0.600
y = -1.0122x + 1.8568 R² = 0.9635
0.400
Series1 Linear (Series1)
0.200
0.000 0.000 0.500 1.000 1.500 2.000 -0.200
3.2. Kurva Standar untuk Mie Jahe konsentrasi 2%
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LAMPIRAN 4. Analisis SPSS 4.1. Pengujian SPSS untuk Warna Tests of Normality a
perlakuan Mie Kontrol Mie Jahe 1% Mie Jahe 2% Mie Kontrol Mie Jahe 1% Mie Jahe 2% Mie Kontrol Mie Jahe 1% Mie Jahe 2%
L
a
b
Kolmogorov-Smirnov Statistic df Sig. .186 6 .200* .158 6 .200* .333 6 .036 .306 6 .084 .248 6 .200* .173 6 .200* .197 6 .200* .189 6 .200* .183 6 .200*
Statistic .903 .969 .845 .849 .945 .983 .975 .868 .961
*. This is a lower bound of the true significance. a. Lilliefors Significance Correction L a
Duncan
perlakuan Mie Jahe 2% Mie Jahe 1% Mie Kontrol Sig.
N 6 6 6
Subset for alpha = .05 1 2 3 53.3500 66.6883 71.4950 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. a. Uses Harmonic Mean Sample Size = 6.000. a a
Duncan
perlakuan Mie Kontrol Mie Jahe 1% Mie Jahe 2% Sig.
N 6 6 6
Subset for alpha = .05 1 2 3 -1.0500 -.6183 .3700 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. a. Uses Harmonic Mean Sample Size = 6.000. b a
Duncan
perlakuan Mie Jahe 2% Mie Jahe 1% Mie Kontrol Sig.
N 6 6 6
Subset for alpha = .05 1 2 11.6600 15.5400 16.5633 1.000 .307
Means for groups in homogeneous subsets are displayed. a. Uses Harmonic Mean Sample Size = 6.000.
Shapiro-W ilk df 6 6 6 6 6 6 6 6 6
Sig. .392 .882 .143 .154 .696 .964 .922 .217 .830
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4.2. Pengujian SPSS untuk Kelentingan 4.2.1. Tingkat Kelentingan pada Mie Jahe Formulasi A Tests of Normality a
Tensile_Strength
perlakuan Mie Kontrol Mie Jahe 1% Mie Jahe 2%
Kolmogorov-Smirnov Statistic df Sig. .142 6 .200* .209 6 .200* .199 6 .200*
Statistic .985 .906 .954
Shapiro-Wilk df 6 6 6
Sig. .972 .414 .774
Statistic .860 .931 .945
Shapiro-Wilk df 6 6 6
Sig. .187 .591 .697
*. This is a lower bound of the true significance. a. Lilliefors Significance Correction
Tensile_Strength a
Duncan
perlakuan Mie Jahe 2% Mie Jahe 1% Mie Kontrol Sig.
N 6 6 6
Subset for alpha = .05 1 2 3 .01426500 .02583083 .05543833 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. a. Uses Harmonic Mean Sample Size = 6.000.
6.2.2. Tingkat Kelentingan pada Mie Jahe Formulasi B Tests of Normality a
Tensile_Strength
perlakuan Mie Kontrol Mie Jahe 1% Mie Jahe 2%
Kolmogorov-Smirnov Statistic df Sig. .230 6 .200* .180 6 .200* .220 6 .200*
*. This is a lower bound of the true significance. a. Lilliefors Significance Correction
Tensile_Strength a
Duncan
perlakuan Mie Kontrol Mie Jahe 1% Mie Jahe 2% Sig.
N 6 6 6
Subset for alpha = .05 1 2 3 .01475633 .02288317 .03299517 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. a. Uses Harmonic Mean Sample Size = 6.000.
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4.3. Pengujian SPSS untuk Aktivitas Antioksidan 4.3.1. Aktivitas Antioksidan pada Mie Jahe Mentah Tests of Normality a
Kolmogorov-Smirnov Statistic df Sig. .187 6 .200* .266 6 .200* .265 6 .200*
perlakuan Mie Kontrol Mie Jahe 1% Mie Jahe 2%
Inhibisi
Statistic .939 .900 .861
Shapiro-Wilk df 6 6 6
Sig. .647 .373 .192
Shapiro-Wilk df 6 6 6
Sig. .375 .754 .607
*. This is a lower bound of the true significance. a. Lilliefors Significance Correction
Inhibisi a
Duncan
perlakuan Mie Kontrol Mie Jahe 1% Mie Jahe 2% Sig.
N 6 6 6
Subset for alpha = .05 1 2 3 7.6433 27.7850 49.5300 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. a. Uses Harmonic Mean Sample Size = 6.000.
4.3.2. Aktivitas Antioksidan pada Mie Jahe Matang Tests of Normality a
Kolmogorov-Smirnov Statistic df Sig. .241 6 .200* .237 6 .200* .187 6 .200*
perlakuan Mie Kontrol Mie Jahe 1% Mie Jahe 2%
Inhibisi
Statistic .900 .952 .933
*. This is a lower bound of the true significance. a. Lilliefors Significance Correction
Inhibisi a
Duncan
perlakuan Mie Kontrol Mie Jahe 1% Mie Jahe 2% Sig.
N 6 6 6
Subset for alpha = .05 1 2 3 3.2733 10.5083 20.7933 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. a. Uses Harmonic Mean Sample Size = 6.000.
