b. Pembuatan Vitamin C - Pengaruh Pemberian Vitamin C Dan E Terhadap Gambaran Histologis Ginjal Mencit(Mus musculus L.) Yang Dipajankan Monosodium Glutamat (MSG)
LAMPIRAN A. Prosedur Pembuatan Bahan Uji
a. Pembuatan Monosodium Glutamat (MSG) Serbuk MSG ditimbang dimasukkan ke dalam beaker glass ditambahkan aquadest diaduk menggunakan spatula Larutan MSG b. Pembuatan Vitamin C
Serbuk Vit C ditimbang dimasukkan ke dalam beaker glass ditambahkan aquadest diaduk menggunakan spatula Larutan Vit C c. Pembuatan Vitamin E
Vit E
diambil menggunakan spit
dilarutkan dengan minyak jarak (oleum ricini) Larutan VitLAMPIRAN B. Pembuatan Preparat Histologis Ginjal
Ginjal Dibilas dengan NaCl 0,9% Difiksasi dalam BOUIN selama 1 malam Washing dalam alkohol 70% Dehidrasi dengan alkohol konsentrasi bertingkat mulai 70%, 80%, 96%, dan 100% Clearing dalam xylol Infiltrasi Embedding (penanaman) organ dalam cetakan kemudian dituangkan paraffin murni, dibiarkan hingga didapatkan blok paraffin
Blok Paraffin Cutting (pemotongan) menggunakan mikrotom sehingga didapatkan pita-pita paraffin Pita Paraffin Attaching (penempelan) pita paraffin pada object glass Deparafinasi dengan mencelupkan objek dalam xylol Dealkoholisasi dalam alkohol menurun dari 100%, 96%, 80%, hingga 70%
Pewarnaan dengan mencelupkan dalam Hematoxilin selama 3-7 menit kemudian dibilas dengan air mengalir. Selanjutnya dicelupkan dalam alkohol 70%, lalu dicelupkan dalam Eosin selama 1-3 menit.
Mounting yaitu menutup preparat dengan gelas penutup yang sebelumnya diberi Canada balsam Diberi label dan siap diamati
Hasil
LAMPIRAN C. Alat Dan Bahan Penelitian Tempat pemeliharaan Castrol Oil Vitamin E hewan percobaan Hewan percobaan Dissecting set Bak bedah (Mus musculus L.)
Oven Mikrotom rotary Tempat fiksasi organ ginjal
K+
K+ .300 5 .161 .833 5 .146 P1 .227 5 .200
0.30 0.26 0.28±
0.02 P4
0.29
0.25
0.26
0.26 0.25 0.26±
0.01 Hasil Uji Statistik Berat Ginjal Mencit (Mus musculus L.)
Tests of Normality
Kel_Per lakuan Kolmogorov-Smirnov
a
Shapiro-Wilk Statistic Df Sig. Statistic df Sig. Berat_Ginjal K- .292 5 .188 .877 5 .294
5 .468 P2 .276 5 .200
0.26
5 .203 P3 .265 5 .200
5 .154 P4 .348 5 .047 .779
5 .054
a. Lilliefors Significance Correction *. This is a lower bound of the true significance.
Test of Homogeneity of Variance
Levene Statistic df1 df2 Sig.Berat_Ginjal Based on Mean 2.240
5 24 .083 Based on Median .810
5 24 .554 Based on Median and with adjusted df
.810 5 9.581 .569 Based on trimmed mean 2.157
5 24 .093
0.27
0.31
LAMPIRAN D. Data dan Analisis Statistik Berat Ginjal Mencit (Mus musculus
L.)Rataan Berat Ginjal Mencit (Mus musculus L.) Setelah Pemberian Perlakuan
Vitamin C dan E Setelah Dipajankan Monosodium glutamat (MSG) Perlakuan Ulangan x±SD0.26
1
2
3
4
5 K-
0.23
0.21
0.23
0.27 0.33 0.26±0.05
0.23
0.26
0.03 P3
0.25 0.25 0.25±
0.01 P1
0.25
0.22
0.20
0.23 0.20 0.22±
0.02 P2
0.26
0.21
0.26
0.29 0.29 0.26±
- .910
- .853
- .836
Oneway
ANOVA
Berat_Ginjal Sum of Squares Df Mean Square F Sig. Between Groups .010 5 .002 2.482 .060 Within Groups .019 24 .001 Total .029
29
LAMPIRAN E. Data dan Analisis Statistik Kerusakan Histologis Tubulus
Proksimal Ginjal Mencit (Mus musculus L.)Persentase Jumlah Kerusakan Histologis Tubulus Proksimal Ginjal Mencit (Mus
musculus L.) Setelah Pemberian Vitamin C dan E Setelah Dipajankan
Monosodium Glutamat (MSG)
Perlakuan Ulangan x±SD
1
2
3
4
5 K-
33.3
35.8
35.64
37.22 37.38 35.87±1.64 K+
29.4
32.25
35.35
36.85 40.3 34.83±4.20 P1
71.21
71.65
72.26
59.67 65.49 69.95±5.10 P2
55
66.66
73.65
62.51 56.36 61.70±5.55 P3
50.18
62.44
60.98
61.38 64.97 59.99±5.70 P4
45.1
45.85
52.5
49.7 48.85 48.40±3.00
Hasil Uji Statistik Kerusakan Tubulus Proksimal Ginjal Mencit (Mus musculus
L.)Tests of Normality
Tests of Normality
aKolmogorov-Smirnov Shapiro-Wilk Kel_Perlakuan Statistic df Sig. Statistic df Sig.
- Kerusakan_Histolog K- .245
5 .200 .889 5 .351 is_Ginjal
- K+ .149
5 .200 .991 5 .984 P1 .398 5 .010 .724
5 .017 P2 .322 5 .097 .805
5 .089 P3 .369 5 .025 .798
5 .078
- P4 .202
5 .200 .947 5 .718
a. Lilliefors Significance Correction *. This is a lower bound of the true significance.
Test of Homogeneity of Variance
Levene Statistic df1 df2 Sig.Kerusakan_Histologis Based on Mean 1.499
5 24 .227 _Ginjal
Based on Median .421
5 24 .830 Based on Median and .421 5 15.466 .827 with adjusted df Based on trimmed mean 1.278
5 24 .306
Kruskal-Wallis Test
Ranks
Kel_Perlakuan N Mean Rank Kerusakan_Histologis_Ginjal K-
5
6.20 K+
5
4.80 P1
5
26.70 P2
5
21.90 P3
5
20.20 P4
5
13.20 Total
30
a,b Test Statistics
Kerusakan_Histologis_Ginjal Chi-Square
25.480 df
5 Asymp. Sig. .000
a. Kruskal Wallis Test
b. Grouping Variable: Kel_Perlakuan
Mann-Whitney Test
Ranks
Kel_Perlakuan N Mean Rank Sum of Ranks Kerusakan_Histologis_Ginjal K-
5
6.20
31.00 K+
5
4.80
24.00 Total
10
b Test Statistics
Kerusakan_Histologis_Ginjal Mann-Whitney U
9.000 Wilcoxon W
24.000 Z
- .731 Asymp. Sig. (2-tailed)
.465
a
Exact Sig. [2*(1-tailed Sig.)] .548 a. Not corrected for ties.
b. Grouping Variable: Kel_Perlakuan
Mann-Whitney Test
Ranks
Kel_Perlakuan N Mean Rank Sum of Ranks Kerusakan_Histologis_Ginjal K-
5
3.00
15.00 P1
5
8.00
40.00 Total
10
b Test Statistics
Kerusakan_Histologis_Ginjal Mann-Whitney U
.000 Wilcoxon W
15.000 Z
- 2.611 Asymp. Sig. (2-tailed)
.009
a
Exact Sig. [2*(1-tailed Sig.)] .008 a. Not corrected for ties.
b. Grouping Variable: Kel_Perlakuan
Mann-Whitney Test
Ranks
Kel_Perlakuan N Mean Rank Sum of Ranks Kerusakan_Histologis_Ginjal K-
5
3.00
15.00 P2
5
8.00
40.00 Total
10
b Test Statistics
Kerusakan_Histologis_Ginjal Mann-Whitney U
.000 Wilcoxon W
15.000 Z
- 2.611 Asymp. Sig. (2-tailed)
.009
a
Exact Sig. [2*(1-tailed Sig.)] .008 a. Not corrected for ties.
b. Grouping Variable: Kel_Perlakuan
Mann-Whitney Test
Ranks
Kel_Perlakuan N Mean Rank Sum of Ranks Kerusakan_Histologis_Ginjal K-
5
3.00
15.00 P3
5
8.00
40.00 Total
10
b Test Statistics
Kerusakan_Histologis_Ginjal Mann-Whitney U
.000 Wilcoxon W
15.000 Z
- 2.611 Asymp. Sig. (2-tailed)
.009
a
Exact Sig. [2*(1-tailed Sig.)] .008 a. Not corrected for ties.
b. Grouping Variable: Kel_Perlakuan
Mann-Whitney Test
Ranks
Kel_Perlakuan N Mean Rank Sum of Ranks Kerusakan_Histologis_Ginjal K-
5
3.00
15.00 P4
5
8.00
40.00 Total
10
b Test Statistics
Kerusakan_Histologis_Ginjal Mann-Whitney U
.000 Wilcoxon W
15.000 Z
- 2.611 Asymp. Sig. (2-tailed)
.009
a
Exact Sig. [2*(1-tailed Sig.)] .008 a. Not corrected for ties.
b. Grouping Variable: Kel_Perlakuan
Mann-Whitney Test
Ranks
Kel_Perlakuan N Mean Rank Sum of Ranks Kerusakan_Histologis_Ginjal K+
5
3.00
15.00 P1
5
8.00
40.00 Total
10
b Test Statistics
Kerusakan_Histologis_Ginjal Mann-Whitney U
.000 Wilcoxon W
15.000 Z
- 2.611 Asymp. Sig. (2-tailed)
.009
a
Exact Sig. [2*(1-tailed Sig.)] .008 a. Not corrected for ties.
b. Grouping Variable: Kel_Perlakuan
Mann-Whitney Test
Ranks
Kel_Perlakuan N Mean Rank Sum of Ranks Kerusakan_Histologis_Ginjal K+
5
3.00
15.00 P2
5
8.00
40.00 Total
10
b Test Statistics
Kerusakan_Histologis_Ginjal Mann-Whitney U
.000 Wilcoxon W
15.000 Z
- 2.611 Asymp. Sig. (2-tailed)
.009
a
Exact Sig. [2*(1-tailed Sig.)] .008 a. Not corrected for ties.
b. Grouping Variable: Kel_Perlakuan
Mann-Whitney Test
Ranks
Kel_Perlakuan N Mean Rank Sum of Ranks Kerusakan_Histologis_Ginjal K+
5
3.00
15.00 P3
5
8.00
40.00 Total
10
b Test Statistics
Kerusakan_Histologis_Ginjal Mann-Whitney U
.000 Wilcoxon W
15.000 Z
- 2.611 Asymp. Sig. (2-tailed)
.009
a
Exact Sig. [2*(1-tailed Sig.)] .008 a. Not corrected for ties.
b. Grouping Variable: Kel_Perlakuan
Mann-Whitney Test
Ranks
Kel_Perlakuan N Mean Rank Sum of Ranks Kerusakan_Histologis_Ginjal K+
5
3.00
15.00 P4
5
8.00
40.00 Total
10
b Test Statistics
Kerusakan_Histologis_Ginjal .000
Mann-Whitney Test
Mann-Whitney U Wilcoxon W
15.000 Z
- 2.611 Asymp. Sig. (2-tailed)
.009
a
Exact Sig. [2*(1-tailed Sig.)] .008 a. Not corrected for ties.
b. Grouping Variable: Kel_Perlakuan
Mann-Whitney Test
Ranks
Kel_Perlakuan N Mean Rank Sum of Ranks Kerusakan_Histologis_Ginjal P1
5
7.40
37.00 P2
5
3.60
18.00 Total
10
b Test Statistics
Kerusakan_Histologis_Ginjal Mann-Whitney U
3.000 Wilcoxon W
18.000 Z
- 1.984 Asymp. Sig. (2-tailed)
.047
a
Exact Sig. [2*(1-tailed Sig.)] .056 a. Not corrected for ties.
b. Grouping Variable: Kel_Perlakuan
Mann-Whitney Test
Ranks
Kel_Perlakuan N Mean Rank Sum of Ranks Kerusakan_Histologis_Ginjal P1
5
7.30
36.50 P3
5
3.70
18.50 Total
10
b Test Statistics
Kerusakan_Histologis_Ginjal Mann-Whitney U
3.500 Wilcoxon W
18.500 Z
- 1.886 Asymp. Sig. (2-tailed)
.059
a
Exact Sig. [2*(1-tailed Sig.)] .056 a. Not corrected for ties.
b. Grouping Variable: Kel_Perlakuan
Mann-Whitney Test
Ranks
Kel_Perlakuan N Mean Rank Sum of Ranks Kerusakan_Histologis_Ginjal P1
5
8.00
40.00 P4
5
3.00
15.00 Total
10
b Test Statistics
Kerusakan_Histologis_Ginjal Mann-Whitney U
.000 Wilcoxon W
15.000 Z
- 2.611 Asymp. Sig. (2-tailed)
.009
a
Exact Sig. [2*(1-tailed Sig.)] .008 a. Not corrected for ties.
b. Grouping Variable: Kel_Perlakuan
Mann-Whitney Test
Ranks
Kel_Perlakuan N Mean Rank Sum of Ranks Kerusakan_Histologis_Ginjal P2
5
6.30
31.50 P3
5
4.70
23.50 Total
10
b Test Statistics
Kerusakan_Histologis_Ginjal Mann-Whitney U
8.500 Wilcoxon W
23.500 Z
- .838 Asymp. Sig. (2-tailed)
.402
a
Exact Sig. [2*(1-tailed Sig.)] .421 a. Not corrected for ties.
b. Grouping Variable: Kel_Perlakuan
Mann-Whitney Test
Ranks
Kel_Perlakuan N Mean Rank Sum of Ranks Kerusakan_Histologis_Ginjal P2
5
8.00
40.00 P4
5
3.00
15.00 Total
10
b Test Statistics
Kerusakan_Histologis_Ginjal Mann-Whitney U
.000 Wilcoxon W
15.000 Z
- 2.611 Asymp. Sig. (2-tailed)
.009
a
Exact Sig. [2*(1-tailed Sig.)] .008 a. Not corrected for ties.
b. Grouping Variable: Kel_Perlakuan
Mann-Whitney Test
Ranks
Kel_Perlakuan N Mean Rank Sum of Ranks Kerusakan_Histologis_Ginjal P3
5
7.80
39.00 P4
5
3.20
16.00 Total
10
b Test Statistics
Kerusakan_Histologis_Ginjal Mann-Whitney U
1.000 Wilcoxon W
16.000 Z
- 2.402 Asymp. Sig. (2-tailed)
.016
a
Exact Sig. [2*(1-tailed Sig.)] .016 a. Not corrected for ties.
b. Grouping Variable: Kel_Perlakuan