Efek Imunomodulator Ekstrak Etil Asetat Daun Mahkota Dewa (Phaleria macrocarpa (Scheff) Boerl.) Terhadap Respon Hipersensitivitas Dan Titer Antibodi Sel Imun Mencit Jantan

Lampiran 1. Surat Identifikasi Tumbuhan

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Lampiran 2. Surat Ethical Clearence

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Lampiran 3. Bagan Alur Penelitian
Uji Hipersensitivitas

25 ekor mencit jantan
Diinduksi dengan 0,1 ml sel darah merah
sapi (SDMS) 1%

EEADMD
- Dosis 50 mg/kg BB
(5 ekor)
- Dosis 100 mg/kg BB

(5 ekor)

Kontrol pembawa :

Kontrol positif :

CMC Na 0,5 %

Levamisol 25 mg/kg BB

(5 Ekor)

(5 Ekor)

- Dosis 200 mg/kg BB
(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 tanpa 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

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Lampiran 3. Lanjutan
Uji Titer Antibodi

25 ekor mencit jantan
Diinduksi dengan 0,1 ml sel darah
merah sapi (SDMS) 1%

EEADMD
- 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)

- Dosis 200 mg/kg BB
(5 ekor)

Diberikan perlakuan selama 7 hari secara per oral

Diambil darah melalui vena ekor dan masukkan ke dalam
microtube
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
Diamati penggumpalan yang terjadi
Ditentukan nilai titer antibodi berdasarkan pengenceran
yang terakhir
Ditransformasikan nilai titer antibodi dengan [2log(titer)+1]
Hasil

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Lampiran 4. Perhitungan Serbuk Levamisol yang Ditimbang
Bobot 20 tablet

= 5,97 g

Kandungan Levamisol/tablet

= 29,46 mg

Serbuk sampel yang ditimbang setara 25 mg Levamisol :

x 5,97 g

=

= 0,25 g

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Lampiran 5. Contoh Perhitungan Dosis untuk dosis 50 mg/kg BB

Misalnya BB mencit = 27 g


Jumlah yang diberikan :

x 27 g = 1,35 mg



Konsentrasi EEADMD :

= 5 mg/ml



Jumlah suspensi yang diberikan :

= 0,27 ml

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Lampiran 6. Gambar Alat-alat

Pipet mikro

Sentrifuge

Plethismometer Digital

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Lampiran 7. Gambar Hewan Percobaan

Mencit Percobaan

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Lampiran 8. Pembengkakan Kaki Mencit dan Hemaglutinasi
Pembengkakan kaki mencit

Pembengkakan awal (Vo)

Pembengkakan akhir (Vt)

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Lampiran 8. Lanjutan
Hemaglutinasi

CMC

LEVAMISOL

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EEADMD

DOSIS 50 mg

DOSIS 100mg

DOSIS 200mg

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Lampiran 9. Hasil Analisis Statistik

Tests of Normality
Kolmogorov-Smirnova
Kelompok

Statistic

df

Shapiro-Wilk

Sig. Statistic

df

Sig.

Volume CMC-Na 0,5%
kaki
Dosis50mg/kgbb
mencit Dosis100mg/kgbb

.233

5 .200*

.961

5 .813

.234

5 .200*

.928

5 .585

.188

5 .200*

.968

5 .859

Dosis200mg/kgbb

.214

4

.

.963

4 .798

5 .200

*

.862

5 .235

Levamisol25mg/kgbb

.226

a. Lilliefors Significance Correction
*. This is a lower bound of the true significance.
Test of Homogeneity of Variances
Volumekakimencit
Levene
Statistic

df1

df2

6.057

4

Sig.
19

.003

Descriptives
Volumekakimencit

Std.
Deviati
Mean
on

N

95% Confidence
Interval for
Mean
Std.
Error

Lower Upper Mini Maxi
Bound Bound mum mum

CMC-Na 0,5%

5

.1180 .08228 .03680

.0158

.2202

.02

.23

Dosis50mg/kgbb

5

.5100 .02915 .01304

.4738

.5462

.47

.54

Dosis100mg/kgbb

5

.6580 .04207 .01881

.6058

.7102

.61

.72

Dosis200mg/kgbb

4

.7875 .02217 .01109

.7522

.8228

.76

.81

Levamisol25mg/k
gbb

5

.5560 .10574 .04729

.4247

.6873

.42

.65

24

.5150 .23587 .04815

.4154

.6146

.02

.81

Total

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ANOVA
Volumekakimencit
Sum of
Squares
Between Groups
Within Groups
Total

df

Mean Square

1.196

4

.299

.084

19

.004

1.280

23

F

Sig.

67.820

.000

Multiple Comparisons
Dependent Variable:Volumekakimencit
95%
Confidence
Interval

(I) Kelompok
Tukey CMC-Na 0,5
HSD

Mean
Differenc Std.
Lower Upper
e (I-J) Error Sig. Bound Bound

(J) Kelompok

-.39200* .04199 .000 -.5183

-.2657

Dosis100mg/kgbb -.54000* .04199 .000 -.6663

-.4137

Dosis200mg/kgbb -.66950* .04454 .000 -.8034

-.5356

Levamisol25mg/k -.43800* .04199 .000 -.5643
gbb

-.3117

Dosis50mg/kgbb

Dosis50mg/kg CMC-Na 0,5%
.39200* .04199 .000 .2657 .5183
bb
Dosis100mg/kgbb -.14800* .04199 .017 -.2743 -.0217
Dosis200mg/kgbb -.27750* .04454 .000 -.4114
Levamisol25mg/k
gbb

-.1436

-.04600 .04199 .807 -.1723

.0803

.54000* .04199 .000 .4137

.6663

.14800* .04199 .017 .0217

.2743

Dosis200mg/kgbb

-.12950 .04454 .061 -.2634

.0044

Levamisol25mg/k
gbb

.10200 .04199 .150 -.0243

.2283

.66950* .04454 .000 .5356

.8034

.27750* .04454 .000 .1436

.4114

Dosis100mg/k CMC-Na 0,5%
gbb
Dosis50mg/kgbb

Dosis200mg/k CMC-Na 0,5%
gbb
Dosis50mg/kgbb

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Dosis100mg/kgbb
Levamisol25mg/k
gbb

*

.2634

.23150 .04454 .000 .0976

.3654

.43800* .04199 .000 .3117

.5643

.04600 .04199 .807 -.0803

.1723

-.10200 .04199 .150 -.2283

.0243

Dosis200mg/kgbb -.23150* .04454 .000 -.3654

-.0976

-.39200* .04199 .000 -.4799

-.3041

Levamisol25m CMC-Na 0,5%
g/kgbb
Dosis50mg/kgbb
Dosis100mg/kgbb
LSD

.12950 .04454 .061 -.0044

CMC-Na 0,5% Dosis50mg/kgbb

*

-.4521

*

Dosis200mg/kgbb -.66950 .04454 .000 -.7627

-.5763

Levamisol25mg/k -.43800* .04199 .000 -.5259
gbb

-.3501

Dosis100mg/kgbb -.54000 .04199 .000 -.6279

Dosis50mg/kg CMC-Na 0,5%
.39200* .04199 .000 .3041 .4799
bb
Dosis100mg/kgbb -.14800* .04199 .002 -.2359 -.0601
Dosis200mg/kgbb -.27750* .04454 .000 -.3707
Levamisol25mg/k
gbb
Dosis100mg/k CMC-Na 0,5%
gbb
Dosis50mg/kgbb

-.1843

-.04600 .04199 .287 -.1339

.0419

.54000* .04199 .000 .4521

.6279

*

.14800 .04199 .002 .0601

.2359

Dosis200mg/kgbb -.12950* .04454 .009 -.2227

-.0363

Levamisol25mg/k
gbb
Dosis200mg/k CMC-Na 0,5%
gbb
Dosis50mg/kgbb

*

.10200 .04199 .025 .0141

.1899

.66950* .04454 .000 .5763

.7627

.27750* .04454 .000 .1843

.3707

*

Dosis100mg/kgbb

.12950 .04454 .009 .0363

.2227

Levamisol25mg/k
gbb

.23150* .04454 .000 .1383

.3247

.43800* .04199 .000 .3501

.5259

.04600 .04199 .287 -.0419

.1339

Levamisol25m CMC-Na 0,5%
g/kgbb
Dosis50mg/kgbb

*

Dosis100mg/kgbb -.10200 .04199 .025 -.1899

-.0141

Dosis200mg/kgbb -.23150* .04454 .000 -.3247

-.1383

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

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

N

1

2

3

CMC-Na 0,5%

5

Dosis50mg/kgbb

5

.5100

Levamisol25mg/kgbb

5

.5560

Dosis100mg/kgbb

5

Dosis200mg/kgbb

4

Sig.

4

.1180
.5560
.6580
.7875
1.000

.820

.167

1.000

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.
Tests of Normalityb
Kolmogorov-Smirnova
Kelompok

Statistic df

Shapiro-Wilk

Sig.

Statistic

df

Sig.

Titer
Dosis 50mg/kgbb
antibodi Dosis100mg/kgbb

.367

5

.026

.684

5 .006

.367

5

.026

.684

5 .006

Dosis200mg/kgbb

.367

5

.026

.684

5 .006

Levamisol25mg/kgbb

.367

5

.026

.684

5 .006

a. Lilliefors Significance Correction
b. Titerantibodi is constant when Kelompok = CMC-Na0,5%. It has been
omitted.

Test of Homogeneity of Variances
Titerantibodi
Levene
Statistic
19.006

df1

df2
4

Sig.
19

.000

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Descriptives
TiterAntibodi
N

Mean

Std.
Deviation

Std.
Error

Minimu
m
Maximum

CMC-Na 0,5%

4 2.8100

.00000 .00000

2.81

2.81

Dosis50mg/kgbb

5 3.6500

.32863 .14697

3.41

4.01

Dosis100mg/kgbb

5 5.5760

.33411 .14942

5.21

5.82

Dosis200mg/kgbb

5 6.7760

.33411 .14942

6.41

7.02

Levamisol25mg/kgbb

5 3.7700

.32863 .14697

3.41

4.01

24 4.5164

.32548 .59278

2.20

7.02

Total
Kruskal-Wallis Test

Ranks
Kelompok

N

Titerantibodi CMC-Na0,5%

Mean Rank
4

2.50

Dosis 50mg/kgbb

5

9.00

Dosis100mg/kgbb

5

17.00

Dosis200mg/kgbb

5

22.00

Levamisol25mg/kgbb

5

10.00

Total

24

Test Statisticsa,b
Titerantibodi
Chi-Square
df
Asymp. Sig.

21.460
4
.000

a. Kruskal Wallis Test
b. Grouping Variable:
Kelompok

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MANN WHITNEY
a. Kelompok CMC-Na dan Dosis 50 mg/kgbb
Ranks
Kelompok

N

Titerantibodi CMC-Na0,5%

Mean Rank

Sum of Ranks

4

2.50

10.00

Dosis 50mg/kgbb

5

7.00

35.00

Total

9

Test Statisticsb
Titerantibodi
Mann-Whitney U

.000

Wilcoxon W

10.000

Z

-2.619

Asymp. Sig. (2-tailed)

.009

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

.016a

a. Not corrected for ties.
b. Grouping Variable: Kelompok
b. Kelompok CMC-Na dan Dosis 100 mg/kgbb
Ranks
Kelompok

N

Titerantibodi CMC-Na0,5%

Mean Rank Sum of Ranks
4

2.50

10.00

Dosis100mg/kgbb

5

7.00

35.00

Total

9

Test Statisticsb
Titerantibodi
Mann-Whitney U

.000

Wilcoxon W

10.000

Z

-2.619

Asymp. Sig. (2-tailed)

.009

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

.016a

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a. Not corrected for ties.
b. Grouping Variable: Kelompok
c. Kelompok CMC-Na dan Dosis 200 mg/kgbb
Ranks
Kelompok

N

Mean Rank Sum of Ranks

Titerantibodi CMC-Na0,5%

4

2.50

10.00

Dosis200mg/kgbb

5

7.00

35.00

Total

9

Test Statisticsb
Titerantibodi
Mann-Whitney U

.000

Wilcoxon W

10.000

Z

-2.619

Asymp. Sig. (2-tailed)

.009

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

.016a

a. Not corrected for ties.
b. Grouping Variable: Kelompok
d. Kelompok CMC-Na dan Levamisol 25 mg/kgbb
Ranks
Kelompok

N

Titerantibodi CMC-Na0,5%

Mean Rank Sum of Ranks
4

2.50

10.00

Levamisol25mg/kgb
b

5

7.00

35.00

Total

9

Test Statisticsb
Titerantibodi
Mann-Whitney U

.000

Wilcoxon W

10.000

Z

-2.619

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Asymp. Sig. (2-tailed)

.009

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

.016a

a. Not corrected for ties.
b. Grouping Variable: Kelompok
e. Kelompok dosis 50 mg/kg bb dan dosis 100 mg/kg bb
Ranks
Kelompok

N

Mean Rank Sum of Ranks

Titerantibodi Dosis 50mg/kgbb

5

3.00

15.00

Dosis100mg/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)

.007

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

.008a

a. Not corrected for ties.
b. Grouping Variable: Kelompok
f. Kelompok dosis 50 mg/kgbb dan dosis 200 mg/kgbb
Ranks
Kelompok

N

Mean Rank Sum of Ranks

Titerantibodi Dosis 50mg/kgbb

5

3.00

15.00

Dosis200mg/kgbb

5

8.00

40.00

Total

10

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Test Statisticsb
Titerantibodi
Mann-Whitney U

.000

Wilcoxon W

15.000

Z

-2.694

Asymp. Sig. (2-tailed)

.007

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

.008a

a. Not corrected for ties.
b. Grouping Variable: Kelompok
g. Kelompok dosis 50 mg/kg bb dan Levamisol 25 mg/kg bb
Ranks
Kelompok

N

Titerantibodi Dosis 50mg/kgbb
Levamisol25mg/kgbb
Total

Mean Rank Sum of Ranks
5

5.00

25.00

5

6.00

30.00

10

Test Statisticsb
Titerantibodi
Mann-Whitney U

10.000

Wilcoxon W

25.000

Z

-.600

Asymp. Sig. (2-tailed)

.549

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

.690a

a. Not corrected for ties.
b. Grouping Variable: Kelompok
h. Kelompok dosis 100 mg/kg bb dan dosis 200 mg/kg bb
Ranks
Kelompok

N

Mean Rank Sum of Ranks

Titerantibodi Dosis100mg/kgbb

5

3.00

15.00

Dosis200mg/kgbb

5

8.00

40.00

Total

10

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Test Statisticsb
Titerantibodi
Mann-Whitney U

.000

Wilcoxon W

15.000

Z

-2.694

Asymp. Sig. (2-tailed)

.007

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

.008a

a. Not corrected for ties.
b. Grouping Variable: Kelompok
i. Kelompok dosis 100 mg/ kg bb dan Levamisol 25 mg/kg bb
Ranks
Kelompok

N

Titerantibodi Dosis100mg/kgbb
Levamisol25mg/kgbb
Total

Mean Rank Sum of Ranks
5

8.00

40.00

5

3.00

15.00

10

Test Statisticsb
Titerantibodi
Mann-Whitney U

.000

Wilcoxon W

15.000

Z

-2.694

Asymp. Sig. (2-tailed)

.007

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

.008a

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

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j. Kelompok dosis 200 mg/kg bb dan Levamisol 25 mg/kg bb
Ranks
Kelompok

N

Titerantibodi Dosis200mg/kgbb
Levamisol25mg/kgbb
Total

Mean Rank Sum of Ranks
5

8.00

40.00

5

3.00

15.00

10

Test Statisticsb
Titerantibodi
Mann-Whitney U

.000

Wilcoxon W

15.000

Z

-2.694

Asymp. Sig. (2-tailed)

.007

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

.008a

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

Homogeneous Subsets
Subset
Perlakuan

N

1

2

3

CMC Na 0,5%

4

EADMD 50mg/kg BB

5

3.6500

Levamisole 25 mg/kg BB

5

3.7700

EADMD 100 mg/kg BB

5

EADMD 200mg/kg BB

5

4

2.8110

5.5760
6.7760

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