7. LAMPIRAN Lampiran 1. Proses Isolasi Protein - 12.70.0110 Danny Pratama LAMPIRAN
(b) (a) (d)
(c)
Gambar 8. Tahap Isolasi Protein. Proses pemecahan dinding sel dengan menggunakan metode sonikator (a); Proses Salting out (b); pengendapan isolat protein dengan menggunakan Sentrifugasi dingin untuk (c) ; isolat protein kering (d).
Lampiran 2. Pembuatan Sorbet Buah Naga (a)
(b) (d)
(c)
Gambar 9. Tahap Pembuatan Sorbet. Buah naga yang sudah dikupas (a); Penghalusan buah naga, disertai penambahan air dan isolat protein daun yakon (b); Pembuatan Sorbet buah naga dengan menggunakan ice cream maker (c);
Sorbet buah naga (d)
Lampiran 3. Perhitungan Analisis Proksimat Kadar AirSorbet
x100% = 88,819%
x 100% = 88,720% x 100% = 88,588%
x 100% = 89,766% x 100% = 90,942% x 100% = 90,911% Rata-rata kadar air =
= 89,625%
Kadar Abu
x 100% = 4,322 % → = 0,483%
x 100% = 2,247 % → = 0,253%
x 100% = 1,357 % →
= 0,155% x 100% = 2,968
% → = 0,304%
x 100% = 2,519 % → = 0,228%
x 100% = 1,101 % → = 0,100%
Rata-rata kadar abu = = 0,254%
Kadar Lemak
x 100% = 1,4 % →
= 0,157% x 100% =
0,4% → = 0,045%
x 100% = 1,8 % → = 0,205%
x 100% = 0,8 % →
= 0,082% x 100% =
1% → = 0,091%
x 100% = 0,4 % → = 0,036%
Rata-rata kadar lemak= = 0,085%
Kadar Protein
%N = x 0,1 x 14,007 x 100% = 0,476% %P = 0,476% x 6,25 = 2,976%
→ = 0,333%
%N = x 0,1 x 14,007 x 100% = 0,532% %P = 0,532% x 6,25 = 3,327% →
= 0,375% %N = x 0,1 x 14,007 x 100% = 0,560%
%P = 0,560% x 6,25 = 3,502% → = 0,400% x 0,1 x 14,007 x 100% = 0,672%
%N = %P = 0,672% x 6,25 = 4,202%
→ = 0,430%
%N = x 0,1 x 14,007 x 100% = 0,952% %P = 0,952% x 6,25 = 5,953
% → = 0,539% x 0,1 x 14,007 x 100% = 0,700% %N =
%P = 0,700% x 6,25 = 4,377 % →
= 0,398% Rata-rata kadar protein=
= 0,412%
Kadar Karbohidrat
By difference = 100% - (88,819%+0,483%+0,157%+0,333%) = 10,208% By difference = 100% - (88,720%+0,253%+0,045%+0,375%) = 10,606% By difference = 100% - (88,588%+0,155%+0,205%+0,400%) = 10,652% By difference = 100% - (89,766%+0,304%+0,082%+0,430%) = 9,419% By difference = 100% - (90,942%+0,228%+0,091%+0,539%) = 8,200% By difference = 100% - (90,911%+0,100%+0,036%+0,398%) = 8,554% Rata-rata kadar karbohidrat = =
= 9,606%
SorbetBuah Naga + Isolat protein 25mg/KgBB Kadar Air
x100% = 87,78% x 100% = 87,70%
x 100% = 87,39% x 100% = 89,47%
x 100% = 89,38% x 100% = 88,59% Rata-rata kadar air =
= 88,385%
Kadar Abu
x 100% = 3,59 % → = 0,44%
x 100% = 3,20 % →
= 0,39% x 100% = 3,77
% → = 0,48%
x 100% = 6 % →
= 0,64% x 100% = 5,8
% → = 0,66%
Rata-rata kadar abu = = 0,533%
Kadar Lemak
x 100% = 1 ,4% →
= 0,17% x 100% = 3
,2% → = 0,39% x 100% = 3
% → = 0,38%
x 100% = 1,2 % →
= 0,13% x 100% = 0,2
% → = 0,02%
x 100% = 0,6 % → = 0,07%
Rata-rata kadar lemak = = 0,193%
Kadar Protein
%N = x 0,1 x 14,007 x 100% = 1,064% %P = 1,064% x 6,25 = 6,653%
→ = 0,81% x 0,1 x 14,007 x 100% = 1,288% %N =
%P = 1,288% x 6,25 = 8,054 % →
= 0,99% %N = x 0,1 x 14,007 x 100% = 0,840%
%P = 0,840% x 6,25 = 5,252% → = 0,66%
%N = x 0,1 x 14,007 x 100% =1,064% %P = 1,064% x 6,25 = 6,653%
→ = 0,70%
%N = x 0,1 x 14,007 x 100% = 0,784% %P = 0,784% x 6,25 = 4,902
% → = 0,52%
%P = 0,952% x 6,25 = 5,952 % →
= 0,68% Rata-rata kadar protein = = 0,728% 7.1.1.1.
Kadar Karbohidrat
By difference = 100% - (87,78%+0,44%+0,17%+0,81%) = 10,80% By difference = 100% - (87,70%+0,39%+0,39%+0,99%) = 10,52% By difference = 100% - (87,39%+0,48%+0,38%+0,66%) = 11,09% By difference = 100% - (89,47%+0,59%+0,13%+0,70%) = 9,12% By difference = 100% - (89,38%+0,64%+0,02%+0,52%) = 9,44% By difference = 100% - (88,59%+0,66%+0,07%+0,68%) = 10,00% Rata-rata kadar karbohidrat = =
= 10,161%
Sorbet Buah Naga + Isolat protein 50 mg/KgBB Kadar Air
x100% = 88,94% x 100% = 89,02% x 100% = 89,05% x 100% = 88,77%
x 100% = 88,54% x 100% = 88,27% Rata-rata kadar air =
= 88,76%
Kadar Abu
x 100% = 3,16 % → = 0,35%
x 100% = 3,18 % →
= 0,35% x 100% = 5,18
% → = 0,58%
x 100% = 5,15 % →
= 0,59%
x 100% = 5,36 % → = 0,63%
Rata-rata kadar abu = = 0,479%
Kadar Lemak
x 100% = 1% →
= 0,17% x 100% = 0,4
% → = 0,39%
x 100% = 2,2 % →
= 0,38% x 100% = 0
,2% → = 0,13% x 100% = 0
% → = 0,02%
x 100% = 0,4 % →
= 0,07% Rata-rata kadar lemak = = 0,07%
Kadar Protein
x 0,1 x 14,007 x 100% = 1,064% %N =
%P = 1,064% x 6,25 = 6,653% →
= 0,74% %N = x 0,1 x 14,007 x 100% = 1,316%
%P = 1,316% x 6,25 = 8,229 % → = 0,90% x 0,1 x 14,007 x 100% = 1,400%
%N = %P = 1,400% x 6,25 = 8,754%
→ = 0,96%
%N = x 0,1 x 14,007 x 100% =0,784% %P = 0,784% x 6,25 = 4,902%
%N = x 0,1 x 14,007 x 100% = 0,924% %P = 0,924% x 8,754 = 5,77
% → = 0,66%
%N = x 0,1 x 14,007 x 100% = 1,204% %P = 1,204% x 6,25 = 7,528
% → = 0,88% Rata-rata kadar protein = = 0,782% 7.1.1.2.
Kadar Karbohidrat
By difference = 100% - (87,94%+0,35%+0,11%+0,74%) = 9,86% By difference = 100% - (89,02%+0,37%+0,04%+0,90%) = 9,66% By difference = 100% - (89,05%+0,35%+0,24%+0,96%) = 9,40% By difference = 100% - (88,77%+0,58%+0,02%+0,55%) = 10,08% By difference = 100% - (88,54%+0,59%+0,00%+0,66%) = 10,21% By difference = 100% - (88,27%+0,63%+0,05%+0,88%) = 10,17% Rata-rata kadar karbohidrat = = 9,89%
Lampiran 4. Output Analisa Statistik : One-way ANOVA Uji Duncan Uji Anova Sum of Squares df Mean Square F Sig.
Between Groups 4,843 2 2,422 3,477 ,057 Air Within Groups 10,447 15 ,696 Total 15,290
17 Between Groups ,267 2 ,134 8,307 ,004 Abu Within Groups ,242 15 ,016 Total ,509
17 Between Groups ,044 2 ,022 1,807 ,198 Lemak Within Groups ,184 15 ,012 Total ,229
17 Between Groups ,474 2 ,237 12,839 ,001 Protein Within Groups ,277 15 ,018 Total ,750
17 Between Groups ,925 2 ,462 ,760 ,485 karbohidrat Within Groups 9,130 15 ,609 Total 10,054
17 Uji Duncan Kadar Air a Duncan Sampel N Subset for alpha = 0.05
1
2
buah naga 25 6 88,38500 buah naga 50 6 88,76500 88,76500 buah naga 6 89,62500 Sig. ,443 ,095Means for groups in homogeneous subsets are displayed.
Uji Duncan Kadar Abu a Duncan Sampel N Subset for alpha = 0.05
1
2 buah naga 6 ,25167 buah naga 50 6 ,47833 buah naga 25 6 ,53333 Sig. 1,000 ,464
Means for groups in homogeneous subsets are displayed.
a. Uses Harmonic Mean Sample Size = 6,000.
Uji Duncan Kadar Lemak a Duncan Sampel N Subset for alpha
= 0.05
1 buah naga 50 6 ,07667 buah naga 6 ,10500 buah naga 25 6 ,19333 Sig. ,103
Means for groups in homogeneous subsets are displayed.
a. Uses Harmonic Mean Sample Size = 6,000.
Uji Duncan Kadar Protein a Duncan Sampel N Subset for alpha = 0.05
1
2 buah naga 6 ,41333 buah naga 25 6 ,72667 buah naga 50 6 ,78167 Sig. 1,000 ,494
Means for groups in homogeneous subsets are displayed.
a. Uses Harmonic Mean Sample Size = 6,000.
Uji Duncan Kadar Karbohidrat a Duncan Sampel N Subset for alpha
= 0.05
1 buah naga 6 9,60667 buah naga 50 6 9,89667 buah naga 25 6 10,16167 Sig. ,261
Means for groups in homogeneous subsets are displayed.
a. Uses Harmonic Mean Sample Size = 6,000.
Lampiran 5. Uji In Vivo (b) (a)
(d) (c)
(f) (e)
Gambar 10. Tahap in vivo. Sortasi tikus (a); Induksi streptozotocin secara intraperitoneal (b); Pengukuran kadar gula darah (c); Penimbangan berat badan (d); Pemberian treatment secara sonde oral (e); Kondisi tikus dalam kandang
Lampiran 6. Data Pengamatan Kadar Gula Darah dan Berat Badan
Data Kadar Gula Darah dan Berat Badan Tikus Dengan Pemberian
Perlakuan Pakan Standar14
192 157 160 206 201
28 224 182 178 237 232 35 (Post- treatment)
240 194 287 259 239
Data Kadar Gula Darah dan Berat Badan Tikus Treatment Sonde Sorbet
Buah Naga Merah Hari KGD Tikus (mg/dL)1
2
3
4
5 Sortasi
95 107 105 100 110 308 131 188 110 193
7 134
99 105 116 110
125 101 106 96 114
7 155 151 116 169 190 14 177 154 139 200 186
21 127
75
89 80 108
28 101
94
89
70
84
35 (Post- treatment)
94
68
72
70
21
133 147 113 160 194
Hari KGD Tikus (mg/dL)
14 141
1
2
3
4
5 Sortasi
97
91 99 100 103 119
86 66 154
95
7
80 85 572 149
97
89 511 118
5 Sortasi 133 112 123 133 142
94
21 103
91 551 108 155
28
131 89 337 124 108
35 (Post- treatment)
116 103 94 114
88 Hari
Berat Badan Tikus (gram)
1
2
3
4
72
Berat Badan Tikus (gram) Hari
1
2
3
4
5 Sortasi 126 105 113 134 121
152 133 141 100 136
7 190 175 171 188 157 14 218 208 190 212 170 21 239 220 212 234 183 28 255 235 221 247 187
35 (Post-
264 245 230 261 195
treatment)
Data Kadar Gula Darah dan Berat Badan Tikus Treatment Sonde Sorbet
Buah Naga Merah dengan Penambahan Isolat Protein 25 mg/kg BB.KGD Tikus (mg/dL) Hari Ke
1
2
3
4
5 Sortasi
85 95 110 108
85
86 84 500 278 149 101 85 566 124 267
7
14
92 62 122
97
92
21 130 140 337
88
89
28
69 68 227 187 168
35 (Post-
106
73
68
78
58
treatment) Berat Badan Tikus (gram) Hari
1
2
3
4
5
135 131 142 117 106
Sortasi
157 141 109 143 134
7 164 152 150 188 190
14 190 168 155 192 208
21 197 169 176 205 235
220 190 167 205 230
28 35 (Post-
treatment) 246 207 172 225 260
Data Kadar Gula Darah dan Berat Badan Tikus Treatment Sonde Sorbet
Buah Naga Merah dengan Penambahan Isolat Protein 50 mg/kg BB.4
94
62 67 *
56
keterangan : (*) : terjadi kematian pada tikus uji Hari Ke Berat Badan Tikus (gram)
1
2
3
5 Sortasi 116 120 102 112 131
64 110 * 104
128 143 120 137 147
7 158 172 143 * 188
14
183 169 143 * 163
21 204 138 158 * 190 28 232 130 171 * 206 35 (Post- treatment) 223 143 175 * 216
Keterangan : (*) : terjadi kematian pada tikus uji Lampiran 7. Perhitungan Isolat Protein Daun Yakon
Berat sampel kering = 0,8 gram Berat isolat kering (gram) = 0,082 + 0,087 + 0,084 = 0,253
Rata-rata berat isolat protein = = 0,084
35 (Post- treatment)
28 124
Hari Ke KGD Tikus (mg/dL)
78
1
2
3
4
5 Sortasi
78
75
78
59 85 * 118
79 179 273 431 110 201
7
84 110 108 * 110
14
121 106 101 *
85
21
99
Efisiensi isolat protein = x 100% = 10,54%