Efek Imunomodulator Fraksi n-Heksan Daun Mahkota Dewa (Phaleria macrocarpa (Scheff) Boerl) Terhadap Respon Hipersensitivitas Dan Titer Antibodi Sel Imun Mencit Jantan

Lampiran 1. Surat Hasil Identifikasi Tumbuhan

Universitas Sumatera Utara

Lampiran 2. Surat Ethical Clearance

Universitas Sumatera Utara

Lampiran 3. Bagan Alur Penelitian
Uji Hipersensitivitas

25 ekor mencit jantan
Diinduksi dengan SDMS 1% sebanyak 0,1 ml secara i.p

FHDMD :
•
•
•

Dosis 25 mg/kg bb
(5 ekor)

Dosis 50 mg/kg bb
(5 ekor)
Dosis 100 mg/kg bb
(5 ekor)

Kontrol negatif :

Kontrol positif :

CMC Na 0,5 %

Levamisol 25 mg/kg bb

(5 ekor)

(5 ekor)

Diberikan perlakuan selama 7 hari secara per oral
Diberikan tanda batas pada sendi kaki mencit sebelah kanan
Dilakukan pengukuran volume kaki mencit (V0) pada kaki

yang telah diberi tanda batas
Diinjeksikan dengan 0,1 ml SDMS 1% secara intraplantar
pada telapak kaki sebelah kanan
Dilakukan pengukuran volume pembengkakan kaki mencit
pada hari kedelapan (setelah 24 jam) (Vt)

Hasil

Universitas Sumatera Utara

Lampiran 3. Lanjutan
Uji Titer Antibodi

25 ekor mencit jantan
Diinduksi dengan SDMS 1% sebanyak 0,1 ml secara i.p

•
•
•

FHDMD :
Dosis 25 mg/kg bb
(5 ekor)
Dosis 50 mg/kg bb
(5 ekor)
Dosis 100 mg/kg bb
(5 ekor)

Kontrol negatif :
CMC Na 0,5 %
(5 ekor)

Kontrol positif :
Levamisol 25 mg/kg bb
(5 ekor)

Diberikan perlakuan selama 7 hari secara per oral
Diambil darah melalu vena ekor dan masukkan ke
dalammicrotube
Disentrifus dengan kecepatan 1900 rpm selama 10 menit

Diambil serumnya sebanyak 25 µl dan diteteskan ke dalam
lubang microtitration plate 96
Ditambahkan larutan PBS dan SDMS 1% sebanyak 25 µl
Dilakukan pengenceran sebanyak dua kali (1:2; 1:4; 1:8; 1:16;
1:32; 1:64; 1:128; 1:256; 1:512; 1:1024; 1:2048; 1:4096)
Diamati penggumpalan yang terjadi
Ditentukan nilai titer antibodi berdasarkan pengenceran yang
terakhir
Ditransformasikan nilai titer antibodi dengan [2log(titer)+1]

Hasil

Universitas Sumatera Utara

Lampiran 4. Perhitungan Serbuk Levamisol Yang ditimbang
Bobot 20 tablet

= 5,97 g

Kandungan Levamisol / Tablet

= 29,46 mg

Serbuk yang ditimbang setara 25 mg Levamisol :
=

25 mg

20 x 29,46 mg

x 5,97 g = 0,25 gr

Universitas Sumatera Utara

Lampiran 5. Contoh Perhitungan Dosis untuk dosis 100 mg/kg bb
Misalnya BB mencit = 25 gram
100 ��

= 10��/��

-

Konsentrasi EHDMD :

-

Jumlah yang diberikan :

-

Jumlah suspensi yang diberikan :

10 ��

100 ��
1000 �

� 25 � = 2,5 ��
2,5 ��

10 �� /��

= 0,25 ��

Universitas Sumatera Utara

Lampiran 6. Perhitungan federer
(n-1)(t-1) ≥ 15
(n-1)(5-1) ≥ 15
(n-1) 4 ≥ 15
4n-1 ≥ 15
4n ≥ 16
16
n≥
4
n≥4
n=4
artinya dimulai dari 4 ekor.

Universitas Sumatera Utara

Lampiran 7. Gambar alat-alat

(oral sonde & spuit)

(pipet mikro)

(timbangan hewan)

Universitas Sumatera Utara

Lampiran 7. (Lanjutan)

(centrifuge PLC series)

(plethysmometer digital)

Universitas Sumatera Utara

Lampiran 8. Gambar Hewan Percobaan

(mencit jantan)

Universitas Sumatera Utara

Lampiran 9. Pembengkakan Kaki Mencit dan Hemaglutinasi

(sebelum di induksi antigen)

(24 jam setelah di induksi antigen)

Universitas Sumatera Utara

Lampiran 9. (Lanjutan)
Hemaglutinasi CMC Na 0,5 %

Hemaglutinasi (+)

(-)

-

Hemaglutinasi Levamisol 25 mg/kg bb

Hemaglutinasi (+)

Universitas Sumatera Utara

Lampiran 9. (Lanjutan)
Hemaglutinasi FHDMD 25 mg/kg bb

Hemaglutinasi (+)

Hemaglutinasi FHDMD 50 mg/kg bb

Hemaglutinasi (+)

Hemaglutinasi FHDMD 100 mg/kg bb

Hemaglutinasi (+)

Universitas Sumatera Utara

Lampiran 10. Uji Statistik
Tests of Normality
Kolmogorov-Smirnova
Kelompok
VolumeKakiMencit

Statistic

df

Shapiro-Wilk

Sig.

Statistic

df

Sig.

CMC-Na 0,5%

.415

4

.

.716

4

.017

Dosis25mg/kgbb

.245

5

.200

*

.892

5

.368

*

.923

5

.551

*

Dosis50mg/kgbb

.270

5

.200

Dosis100mg/kgbb

.167

5

.200

.967

5

.855

Levamisol25mg/kgbb

.317

5

.111

.833

5

.146

a. Lilliefors Significance Correction
*. This is a lower bound of the true significance.

Test of Homogeneity of Variances
VolumeKakiMencit
Levene Statistic

df1

.459

df2
4

Sig.
19

.765

Descriptives
VolumeKakiMencit
95% Confidence Interval for Mean
N

Mean

Std. Deviation

Std. Error

Lower Bound

Upper Bound

Minimum

Maximum

CMC-Na 0,5%

4

.0900

.09381

.04690

-.0593

.2393

.03

.23

Dosis25mg/kgbb

5

.2560

.08473

.03789

.1508

.3612

.17

.36

Dosis50mg/kgbb

5

.5260

.08173

.03655

.4245

.6275

.44

.64

Dosis100mg/kgbb

5

.7280

.09338

.04176

.6121

.8439

.62

.86

Levamisol25mg/kgbb

5

.5640

.12876

.05758

.4041

.7239

.46

.77

24

.4471

.24403

.04981

.3440

.5501

.03

.86

Total

ANOVA
VolumeKakiMencit
Sum of Squares
Between Groups
Within Groups
Total

df

Mean Square

1.187

4

.297

.183

19

.010

1.370

23

F
30.794

Sig.
.000

Universitas Sumatera Utara

Multiple Comparisons
Dependent Variable:VolumeKakiMencit
95% Confidence Interval

Mean Difference

Tukey HSD

(I) Kelompok

(J) Kelompok

CMC-Na 0,5%

Dosis25mg/kgbb

-.16600

.06584

.127

-.3640

.0320

Dosis50mg/kgbb

-.43600

*

.06584

.000

-.6340

-.2380

Dosis100mg/kgbb

*

-.63800

.06584

.000

-.8360

-.4400

Levamisol25mg/kgbb

-.47400*

.06584

.000

-.6720

-.2760

.16600

.06584

.127

-.0320

.3640

*

.06208

.003

-.4567

-.0833

*

.06208

.000

-.6587

-.2853

*

-.30800

.06208

.001

-.4947

-.1213

.43600*

.06584

.000

.2380

.6340

Dosis25mg/kgbb

*

.27000

.06208

.003

.0833

.4567

Dosis100mg/kgbb

-.20200*

.06208

.030

-.3887

-.0153

Levamisol25mg/kgbb

-.03800

.06208

.971

-.2247

.1487

CMC-Na 0,5%

.63800*

.06584

.000

.4400

.8360

Dosis25mg/kgbb

.47200

*

.06208

.000

.2853

.6587

Dosis50mg/kgbb

*

.20200

.06208

.030

.0153

.3887

Levamisol25mg/kgbb

.16400

.06208

.102

-.0227

.3507

CMC-Na 0,5%

.47400*

.06584

.000

.2760

.6720

Dosis25mg/kgbb

.30800*

.06208

.001

.1213

.4947

Dosis50mg/kgbb

.03800

.06208

.971

-.1487

.2247

Dosis100mg/kgbb

-.16400

.06208

.102

-.3507

.0227

Dosis25mg/kgbb

-.16600*

.06584

.021

-.3038

-.0282

Dosis50mg/kgbb

-.43600

*

.06584

.000

-.5738

-.2982

*

.06584

.000

-.7758

-.5002

*

.06584

.000

-.6118

-.3362

*

.06584

.021

.0282

.3038

Dosis50mg/kgbb

*

-.27000

.06208

.000

-.3999

-.1401

Dosis100mg/kgbb

-.47200*

.06208

.000

-.6019

-.3421

*

.06208

.000

-.4379

-.1781

Dosis25mg/kgbb

CMC-Na 0,5%

Dosis100mg/kgbb

Levamisol25mg/kgbb

LSD

CMC-Na 0,5%

Dosis25mg/kgbb

-.27000

Dosis100mg/kgbb

-.47200

CMC-Na 0,5%

Dosis100mg/kgbb

-.63800

Levamisol25mg/kgbb

-.47400

CMC-Na 0,5%

Levamisol25mg/kgbb
Dosis50mg/kgbb

Std. Error

Dosis50mg/kgbb

Levamisol25mg/kgbb
Dosis50mg/kgbb

(I-J)

.16600

-.30800

Sig.

Lower Bound

Upper Bound

CMC-Na 0,5%

.43600

*

.06584

.000

.2982

.5738

Dosis25mg/kgbb

.27000*

.06208

.000

.1401

.3999

*

.06208

.004

-.3319

-.0721

Dosis100mg/kgbb

-.20200

Universitas Sumatera Utara

Dosis100mg/kgbb

Levamisol25mg/kgbb

-.03800

.06208

.548

-.1679

.0919

CMC-Na 0,5%

.63800

*

.06584

.000

.5002

.7758

*

.06208

.000

.3421

.6019

*

.06208

.004

.0721

.3319

*

.06208

.016

.0341

.2939

CMC-Na 0,5%

*

.47400

.06584

.000

.3362

.6118

Dosis25mg/kgbb

.30800*

.06208

.000

.1781

.4379

Dosis50mg/kgbb

.03800

.06208

.548

-.0919

.1679

*

.06208

.016

-.2939

-.0341

Dosis25mg/kgbb

Levamisol25mg/kgbb

.47200

Dosis50mg/kgbb

.20200

Levamisol25mg/kgbb

.16400

Dosis100mg/kgbb

-.16400

*. The mean difference is significant at the 0.05 level.

Homogeneous Subsets
VolumeKakiMencit
Subset for alpha = 0.05
Kelompok
Tukey HSDa,,b

N

1

2

3

CMC-Na 0,5%

4

.0900

Dosis25mg/kgbb

5

.2560

Dosis50mg/kgbb

5

.5260

Levamisol25mg/kgbb

5

.5640

Dosis100mg/kgbb

5

Sig.

.5640
.7280

.108

.974

.115

Means for groups in homogeneous subsets are displayed.
a. Uses Harmonic Mean Sample Size = 4,762.
b. The group sizes are unequal. The harmonic mean of the group sizes is used. Type I error levels are not
guaranteed.

Universitas Sumatera Utara

Tests of Normality
a

Kolmogorov-Smirnov
Kelompok
TiterAntibodi

Statistic

df

Shapiro-Wilk

Sig.

Statistic

df

Sig.

CMC-Na 0,5%

.367

5

.026

.684

5

.006

Dosis25mg/kgbb

.367

5

.026

.684

5

.006

Dosis50mg/kgbb

.349

5

.046

.771

5

.046

Dosis100mg/kgbb

.367

5

.026

.684

5

.006

Levamisol25mg/kgbb

.367

5

.026

.684

5

.006

a. Lilliefors Significance Correction

Test of Homogeneity of Variances
TiterAntibodi
Levene Statistic

df1

df2

14.595

4

Sig.
20

.000

Descriptives
TiterAntibodi
95% Confidence Interval for Mean

Std.
N

Mean

Deviation

Std. Error

Lower Bound

Upper Bound

Minimum

Maximum

CMC-Na 0,5%

5

2.5700

.32863

.14697

2.1619

2.9781

2.21

2.81

Dosis25mg/kgbb

5

3.7700

.32863

.14697

3.3619

4.1781

3.41

4.01

Dosis50mg/kgbb

5

4.8500

.53666

.24000

4.1837

5.5163

4.01

5.21

Dosis100mg/kgbb

5

5.6900

.98590

.44091

4.4658

6.9142

4.61

6.41

Levamisol25mg/kgbb

5

3.7700

.32863

.14697

3.3619

4.1781

3.41

4.01

25

4.1300

1.20000

.24000

3.6347

4.6253

2.21

6.41

Total

Kruskal-Wallis Test
Ranks
Kelompok
TiterAntibodi

N

Mean Rank

CMC-Na 0,5%

5

3.00

Dosis25mg/kgbb

5

10.80

Dosis50mg/kgbb

5

18.80

Universitas Sumatera Utara

Dosis100mg/kgbb

5

21.60

Levamisol25mg/kgbb

5

10.80

Total

25

Test Statisticsa,b
TiterAntibodi
Chi-Square

20.718

df

4

Asymp. Sig.

.000

a. Kruskal Wallis Test
b. Grouping Variable:
Kelompok

MANN WHITNEY
a. Kelompok CMC-Na dan dosis 25 mg/kgbb
Ranks
Kelompok
TiterAntibodi

N

Mean Rank

Sum of Ranks

CMC-Na 0,5%

5

3.00

15.00

Dosis25mg/kgbb

5

8.00

40.00

Total

10

Test Statisticsb
TiterAntibodi
Mann-Whitney U

.000

Wilcoxon W

15.000

Z

-2.694

Asymp. Sig. (2-tailed)
Exact Sig. [2*(1-tailed Sig.)]

.007
.008a

a. Not corrected for ties.
b. Grouping Variable: Kelompok

b. Kelompok CMC-Na dan dosis 50 mg/kgbb
Ranks
Kelompok
TiterAntibodi

N

Mean Rank

Sum of Ranks

CMC-Na 0,5%

5

3.00

15.00

Dosis50mg/kgbb

5

8.00

40.00

Universitas Sumatera Utara

Ranks
Kelompok
TiterAntibodi

N

Mean Rank

Sum of Ranks

CMC-Na 0,5%

5

3.00

15.00

Dosis50mg/kgbb

5

8.00

40.00

Mean Rank

Sum of Ranks

Total

10

Test Statisticsb
TiterAntibodi
Mann-Whitney U

.000

Wilcoxon W

15.000

Z

-2.685

Asymp. Sig. (2-tailed)

.007
a

Exact Sig. [2*(1-tailed Sig.)]

.008

a. Not corrected for ties.
b. Grouping Variable: Kelompok

c. Kelompok CMC-Na dan dosis 100 mg/kgbb
Ranks
Kelompok
TiterAntibodi

N

CMC-Na 0,5%

5

3.00

15.00

Dosis100mg/kgbb

5

8.00

40.00

Total

10

b

Test Statistics

TiterAntibodi
Mann-Whitney U

.000

Wilcoxon W

15.000

Z

-2.694

Asymp. Sig. (2-tailed)
Exact Sig. [2*(1-tailed Sig.)]

.007
a

.008

a. Not corrected for ties.
b. Grouping Variable: Kelompok

Universitas Sumatera Utara

d. Kelompok CMC-Na dan levamisol 25 mg/kgbb
Ranks
Kelompok
TiterAntibodi

N

Mean Rank

Sum of Ranks

CMC-Na 0,5%

5

3.00

15.00

Levamisol25mg/kgbb

5

8.00

40.00

Total

10

b

Test Statistics

TiterAntibodi
Mann-Whitney U

.000

Wilcoxon W

15.000

Z

-2.694

Asymp. Sig. (2-tailed)

.007
.008a

Exact Sig. [2*(1-tailed Sig.)]
a. Not corrected for ties.
b. Grouping Variable: Kelompok

e. Kelompok dosis 25 mg/kgbb dan dosis 50 mg/kgbb
Ranks
Kelompok
TiterAntibodi

N

Mean Rank

Sum of Ranks

CMC-Na 0,5%

5

3.00

15.00

Levamisol25mg/kgbb

5

8.00

40.00

Total

10

Test Statisticsb
TiterAntibodi
Mann-Whitney U

.000

Wilcoxon W

15.000

Z

-2.694

Asymp. Sig. (2-tailed)
Exact Sig. [2*(1-tailed Sig.)]

.007
a

.008

a. Not corrected for ties.
b. Grouping Variable: Kelompok

Universitas Sumatera Utara

f. Kelompok dosis 25 mg/kgbb dan dosis 100 mg/kgbb
Ranks
Kelompok
TiterAntibodi

N

Mean Rank

Sum of Ranks

Dosis25mg/kgbb

5

3.00

15.00

Dosis100mg/kgbb

5

8.00

40.00

Total

10

b

Test Statistics

TiterAntibodi
Mann-Whitney U

.000

Wilcoxon W

15.000

Z

-2.694

Asymp. Sig. (2-tailed)

.007
.008a

Exact Sig. [2*(1-tailed Sig.)]
a. Not corrected for ties.
b. Grouping Variable: Kelompok

g. Kelompok dosis 25 mg/kgbb dan levamisol 25 mg/kgbb
Ranks
Kelompok
TiterAntibodi

N

Mean Rank

Sum of Ranks

Dosis25mg/kgbb

5

5.50

27.50

Levamisol25mg/kgbb

5

5.50

27.50

Total

10

b

Test Statistics

TiterAntibodi
Mann-Whitney U

12.500

Wilcoxon W

27.500

Z
Asymp. Sig. (2-tailed)
Exact Sig. [2*(1-tailed Sig.)]

.000
1.000
1.000a

a. Not corrected for ties.
b. Grouping Variable: Kelompok

Universitas Sumatera Utara

h. Kelompok dosis 50 mg/kgbb dan dosis 100 mg/kgbb
Ranks
Kelompok
TiterAntibodi

N

Mean Rank

Sum of Ranks

Dosis50mg/kgbb

5

4.40

22.00

Dosis100mg/kgbb

5

6.60

33.00

Total

10

Test Statisticsb
TiterAntibodi
Mann-Whitney U

7.000

Wilcoxon W

22.000

Z

-1.193

Asymp. Sig. (2-tailed)

.233
.310a

Exact Sig. [2*(1-tailed Sig.)]
a. Not corrected for ties.
b. Grouping Variable: Kelompok

i. Kelompok dosis 50 mg/kgbb dan levamisol 25 mg/kgbb
Ranks
Kelompok
TiterAntibodi

N

Mean Rank

Sum of Ranks

Dosis50mg/kgbb

5

7.70

38.50

Levamisol25mg/kgbb

5

3.30

16.50

Total

10

b

Test Statistics

TiterAntibodi
Mann-Whitney U

1.500

Wilcoxon W

16.500

Z

-2.410

Asymp. Sig. (2-tailed)
Exact Sig. [2*(1-tailed Sig.)]

.016
.016a

a. Not corrected for ties.
b. Grouping Variable: Kelompok

Universitas Sumatera Utara