Uji Toksisitas Akut Ekstrak Etanol Buah Adas Manis (Foeniculum vulgare Mill.) Pada Mencit

(1)

(2)

Lampiran 2. Gambar tumbuhan dan buah adas manis (

Foeniculum vulgare)

Tumbuhan adas manis

(3)

(4)

Lampiran 4. Bagan kerja penelitian

Ekstrak kental etanol

buah adas manis

Ekstrak kering buah adas

manis

Skrining fitokimia untuk

pemeriksaan :

a.

Alkaloida

b.

Flavanoida

c.

Glikosida

d.

Antrakinon glikosida

e.

Saponin

f.

Tanin

g.

Steroida / triterpenoida

Uji toksisitas akut

Hasil

Dikeringkan dengan

freeze dryer

pada suhu -40

o

C

500g buah adas manis

Dibersihkan

Dimaserasi dengan pelarut etanol 80%

Maserat

(5)

Lampiran 5. Bagan penelitian uji toksisitas ekstrak etanol buah adas manis

40 ekor mencit jantan

Aklimatisasi (7 hari)

Randomisasi

Kontrol P1

+ CMC

Na 1 %

Gejala klinis, konsumsi makanan dan minuman, berat badan, kematian,

makropatologi organ hewan coba (14 hari)

+ EEBA

M 5 mg/kg

bb

P2 P3

+ EEBAM

50 mg/kg

bb

+ EEBAM

300 mg/kg

bb

+ EEBAM

1000 mg/kg

bb

+ EEBAM

2000 mg/kg

bb

+ EEBAM

4000 mg/kg

bb

+ EEBAM

8000 mg/kg

bb

(6)

Lampiran 6. Contoh perhitungan dosis

Berat badan mencit 25 kg dengan dosis EEBAM 1000 mg/kg bb

Dosis = 1000 mg/kg bb

= (25 : 1000 kg) x 1000 mg/kg

= 25 mg

Konsentrasi ekstrak yang dibuat 8 % = 8 g/ 100 ml

= 8000 mg/100 ml

= 80 mg/ml

Jumlah obat yang disuntik

= 25 mg : 80 mg/ml

= 0,31 ml

(7)

Lampiran 7. Hasil pengukuran berat badan

1. Berat badan pada minggu pertama

Perlakuan

Berat badan hari ke- (g)

0

3

7 kontrol I

_25,78

_28,08

_28,02

kontrol II

_27,09

_29,48

_28,37

kontrol III

_29,98

_29,89

_29,17

kontrol IV

_30,16

_30,79

_29,93

kontrol V

_30,84

_30,81

_30,66

Dosis 5 mg/kg bb

_27,86

_28,05

_28,28

Dosis 5 mg/kg bb

_28,19

_28,16

_29,25

Dosis 5 mg/kg bb

_28,55

_29,48

_30,56

Dosis 5 mg/kg bb

_29,68

_29,99

_31,39

Dosis 5 mg/kg bb

_30,52

_31,57

_31,77

Dosis 50 mg/kg bb

_27,25

_27,55

_28,78

Dosis 50 mg/kg bb

_27,68

_28,87

_29,69

Dosis 50 mg/kg bb

_28,46

_29,61

_29,84

Dosis 50 mg/kg bb

_30,28

_29,67

_30,56

Dosis 50 mg/kg bb

_31,98

_31,55

_31,03

Dosis 300 mg/kg bb

_30,72

_32,14

_33,18

Dosis 300 mg/kg bb

_28,69

_29,09

_29,24

Dosis 300 mg/kg bb

_29,55

_29,73

_29,83

Dosis 300 mg/kg bb

_29,5

_31,01

_31,13

Dosis 300 mg/kg bb

_29,96

_30,17

_30,59

Dosis 1000 mg/kg bb

31,02

31,19

33,11

Dosis 1000 mg/kg bb

28,97

28,63

28,77

Dosis 1000 mg/kg bb

29,28

31,27

28,15

Dosis 1000 mg/kg bb

25,94

27,42

29,55

Dosis 1000 mg/kg bb

29,03

30,43

29,03

Dosis 2000 mg/kg bb

30,97

32,84

31.69 Dosis 2000 mg/kg bb

28,93

29,01

29,29

(8)

Lampiran 7. (lanjutan)

1. Berat badan pada minggu pertama

Perlakuan

Berat badan hari ke- (g)

0

3

7 Dosis 2000 mg/kg bb

29,05

29,65

30,41

Dosis 2000 mg/kg bb

29,09

30,95

30,59

Dosis 4000 mg/kg bb

32,65

31,33

33,98

Dosis 4000 mg/kg bb

28,95

29,45

28,65

Dosis 4000 mg/kg bb

31,09

31,66

31,93

Dosis 4000 mg/kg bb

31,02

30,16

31,13

Dosis 4000 mg/kg bb

30,96

30,86

32,39

Dosis 8000 mg/kg bb

31,26

31,93

33,19

Dosis 8000 mg/kg bb

29,21

30,68

29,26

Dosis 8000 mg/kg bb

30,47

30,54

31,38

Dosis 8000 mg/kg bb

30,81

30,89

31,37

(9)

Lampiran 7. (lanjutan)

2. Berat badan pada minggu kedua

Perlakuan

Berat badan hari ke- (g)

0

10

14 kontrol I

_25,78

_28,89

_29,07

kontrol II

_27,09

_29,02

_29,13

kontrol III

_29,98

_29,16

_29,98

kontrol IV

_30,16

_30,38

_31,13

kontrol V

_30,84

_31,45

_31,24

Dosis 5 mg/kg bb

_27,86

_29,17

_29,97

Dosis 5 mg/kg bb

_28,19

_29,43

_30,66

Dosis 5 mg/kg bb

_28,55

_30,98

_30,74

Dosis 5 mg/kg bb

_29,68

_31,08

_32,64

Dosis 5 mg/kg bb

_30,52

_32,14

_32,69

Dosis 50 mg/kg bb

_27,25

_29,29

_29,86

Dosis 50 mg/kg bb

_27,68

_29,42

_30,19

Dosis 50 mg/kg bb

_28,46

_29,68

_30,99

Dosis 50 mg/kg bb

_30,28

_31,65

_31,18

Dosis 50 mg/kg bb

_31,98

_31,66

_31,68

Dosis 300 mg/kg bb

_30,72

_33,16

_34,16

Dosis 300 mg/kg bb

_28,69

_29,33

_30,93

Dosis 300 mg/kg bb

_29,55

_31,09

_32,21

Dosis 300 mg/kg bb

_29,5

_31,98

_33,14

Dosis 300 mg/kg bb

_29,96

_31,58

_32,64

Dosis 1000 mg/kg bb

31,02

32,24

33,72

Dosis 1000 mg/kg bb

28,97

29,19

30,49

Dosis 1000 mg/kg bb

29,28

30,99

32,37

Dosis 1000 mg/kg bb

25,94

30,91

32,43

Dosis 1000 mg/kg bb

29,03

30,88

32,54

Dosis 2000 mg/kg bb

30,97

33,25

33,48

Dosis 2000 mg/kg bb

28,93

29,86

30,44

(10)

Lampiran 7. (lanjutan)

2. Berat badan pada minggu kedua

Perlakuan

Berat badan hari ke- (g)

0

10

14 Dosis 2000 mg/kg bb

29,05

30,63

31,61

Dosis 2000 mg/kg bb

29,09

31,75

32,11

Dosis 4000 mg/kg bb

32,65

34,12

34,17

Dosis 4000 mg/kg bb

28,95

30,91

31,75

Dosis 4000 mg/kg bb

31,09

32,06

33,15

Dosis 4000 mg/kg bb

31,02

31,82

32,98

Dosis 4000 mg/kg bb

30,96

32,66

34,68

Dosis 8000 mg/kg bb

31,26

33,18

33,68

Dosis 8000 mg/kg bb

29,21

32,19

32,78

Dosis 8000 mg/kg bb

30,47

31,54

31,82

Dosis 8000 mg/kg bb

30,81

32,19

32,61

(11)

Lampiran 8. Perhitungan karakterisasi simplisia buah adas manis

a.

Penetapan kadar air

%

Volume air akhir

Volume air

sebenarnya

1 5,0000

1,4000

1,7500

0,3500

2 5,1550

2,0000

2,4500

0,4500

(12)

Lampiran 8. (Lanjutan)

b.

Penetapan kadar sari larut etanol

%

Berat cawan Berat cawan + sari larut

etanol

Berat sari larut

etanol

1 5,0180

29,6493

29,8053

0,1560

2 5,0270

25,0725

25,2349

0,1624

(13)

Lampiran 8. (Lanjutan)

c.

Penetapan kadar sari larut air

(14)

Lampiran 8. (Lanjutan)

d.

Penetapan kadar abu total

%

100 sampel

berat

abu

berat

abu total

kadar

%





1. Berat sampel

= 2,0700 g

Berat abu

= 0,1570 g

Kadar abu

=

100 %

7 ,

5845

%

0700

,

2 1570

,

0 



2. Berat sampel

= 2,1530 g

Berat abu

= 0,1730 g

Kadar abu

=

100 %

8 ,

0350

%

1530

,

2 1730

,

0 



3. Berat sampel

= 2,0520 g

Berat abu

= 0,1490 g

Kadar abu

=

100 %

7 ,

2612

%

0520

,

2 1490

,

0 



Rata-rata kadar abu total:

7 ,

6269

%

3 2612

,

7 0350

,

8 5845

,

7 



NO.

Berat krus

Berat sampel (g)

Berat krus + abu

Berat abu

1 53,6220

2,0700

53,7790

0,1570

2 50,1090

2,1530

50,2820

0,1730

(15)

Lampiran 8. (Lanjutan)

e.

Penetapan kadar abu tidak larut asam

%

tidak larut asam

Berat abu tidak

larut asam

1 2,0700

53,6220

53,6520

0,0300

2 2,0520

50,1090

50,1300

0,0210

(16)

Lampiran 9. Perhitungan karakterisasi ekstrak etanol buah adas manis

a.

Penetapan kadar air

%

100 sampel

berat

air

volume

air

kadar





No.

Beratsampel (g)

Volume air 1

Volume air 2

Volume air

1 5,0253

2,1

2,5

0,4000

2 5,1725

2,5

2,9

0,4000

3 5,0623

2,9

3,4

0,4000

1. Berat sampel

= 5,0253 g

Volume air

= 0,4000 ml

Kadar air

=

100 %

7 ,

9597

%

0253

,

5 4000

,

0 



2. Berat sampel

= 5,1725 g

Volume air

= 0,4000 ml

Kadar air

=

100 %

7 ,

7332

%

1725

,

5 4000

,

0 



3. Berat sampel

= 5,0623 g

Volume air

= 0,4000 ml

Kadar air

=

100 %

7 ,

9015

%

0623

,

5 4000

,

0 



Rata-rata kadar air:

7 ,

8648

%

3 9015

,

7 7332

,

7 9597

,

7 



(17)

Lampiran 9. (Lanjutan)

b.

Penetapan kadar abu total

1. Berat sampel

= 2,0938 g

Berat abu

= 0,0224 g

Kadar abu

=

100 %

1 ,

0698

%

0938

,

2 0224

,

0 



2. Berat sampel

= 2,0082 g

Berat abu

= 0,0146 g

Kadar abu

=

100 %

0 ,

7270

%

0082

,

2 0146

,

0 



3. Berat sampel

= 2,0179 g

Berat abu

= 0,0177 g

Kadar abu

=

100 %

0 ,

8771

%

0179

,

2 0177

,

0 



Rata-rata kadar abu total:

0 ,

8913

%

3 8771

,

0 7270

,

0 0698

,

1 



No

Berat sampel

(g)

Berat krus

Berat krus + abu Berat abu total

1 2,0938

63,5387

63,5611

0,0224

2 2,0082

58,2095

58,2241

0,0146

3 2,0179

57,2289

57,2466

0,0177

%

100 sampel

berat

abu

berat

abu total

(18)

Lampiran 9. (Lanjutan)

c.

Penetapan kadar abu tidak larut asam

(19)

Lampiran 10. Hasil analisis data

SPSS berat badan mencit

a.

Berat badan mula-mula

Descriptives

Interval for Mean

Min Max

Test of Homogeneity of Variances

Levene Statistic

df1

df2

Sig.

2.098

7

32 .073

Tukey

(I) perlakuan (J) perlakuan

(20)

Mean Difference

(I-J)

Std.

Error Sig.

95% Confidence Interval Lower Bound

Upper Bound dosis 8000 mg/ kg bb -.89600 .92316 .975 -3.8864 2.0944

dosis 1000 mg/ kg bb

kontrol .07800 .92316 1.000 -2.9124 3.0684 dosis 5 mg/ kg bb -.11200 .92316 1.000 -3.1024 2.8784 dosis 50 mg/ kg bb -.28200 .92316 1.000 -3.2724 2.7084 dosis 300 mg/ kg bb -.83600 .92316 .983 -3.8264 2.1544 dosis 2000 mg/ kg bb -.56600 .92316 .998 -3.5564 2.4244 dosis 4000 mg/ kg bb -2.08600 .92316 .346 -5.0764 .9044 dosis 8000 mg/ kg bb -1.73200 .92316 .576 -4.7224 1.2584

kontrol .64400 .92316 .996 -2.3464 3.6344 dosis 5 mg/ kg bb .45400 .92316 1.000 -2.5364 3.4444 dosis 50 mg/ kg bb .28400 .92316 1.000 -2.7064 3.2744 dosis 300 mg/ kg bb -.27000 .92316 1.000 -3.2604 2.7204 dosis 1000 mg/ kg bb .56600 .92316 .998 -2.4244 3.5564 dosis 4000 mg/ kg bb -1.52000 .92316 .720 -4.5104 1.4704 dosis 8000 mg/ kg bb -1.16600 .92316 .906 -4.1564 1.8244

kontrol 2.16400 .92316 .302 -.8264 5.1544 dosis 5 mg/ kg bb 1.97400 .92316 .414 -1.0164 4.9644 dosis 50 mg/ kg BB 1.80400 .92316 .526 -1.1864 4.7944 dosis 300 mg/ kg BB 1.25000 .92316 .871 -1.7404 4.2404 dosis 1000 mg/ kg bb 2.08600 .92316 .346 -.9044 5.0764 dosis 2000 mg/ kg bb 1.52000 .92316 .720 -1.4704 4.5104 dosis 8000 mg/ kg bb .35400 .92316 1.000 -2.6364 3.3444

Kontrol 1.81000 .92316 .522 -1.1804 4.8004 dosis 5 mg/ kg bb 1.62000 .92316 .653 -1.3704 4.6104 dosis 50 mg/ kg bb 1.45000 .92316 .763 -1.5404 4.4404 dosis 300 mg/ kg bb .89600 .92316 .975 -2.0944 3.8864 dosis 1000 mg/ kg bb 1.73200 .92316 .576 -1.2584 4.7224 dosis 2000 mg/ kg bb 1.16600 .92316 .906 -1.8244 4.1564 dosis 4000 mg/ kg bb -.35400 .92316 1.000 -3.3444 2.6364

Lampiran 10. (Lanjutan)

onfide

r

Uppe

29.5200

.48461 28.1745 30.8655

29.8500

.64020 28.0725 31.6275

29.7150

.51149 28.2949 31.1351

30.6110

.60183 28.9401 32.2819

29.7550

.65056 27.9488 31.5612

30.5970

.51271 29.1735 32.0205

31.1540

.61529 29.4457 32.8623

31.2650

.42772 30.0775 32.4525

40 30.3084

.20693 29.8898 30.7269

5

7

32 .979

oups

16.475

7

2.354 oups

50.323

32

1.573

(21)

Lampiran 10. (Lanjutan)

b.

Berat badan minggu I

Descriptives

Interval for Mean

Min Max

Test of Homogeneity of Variances

Levene Statistic

df1

df2

Sig.

.215

7

32 .979

Tukey

(22)

Mean Difference

(I-J)

Std.

Error Sig.

kontrol .23500 .79312 1.000 -2.3341 2.8041 dosis 5 mg/ kg bb -.09500 .79312 1.000 -2.6641 2.4741 dosis 50 mg/ kg bb .04000 .79312 1.000 -2.5291 2.6091 dosis 300 mg/ kg bb -.85600 .79312 .957 -3.4251 1.7131 dosis 2000 mg/ kg bb -.84200 .79312 .960 -3.4111 1.7271 dosis 4000 mg/ kg bb -1.39900 .79312 .647 -3.9681 1.1701 dosis 8000 mg/ kg bb -1.51000 .79312 .558 -4.0791 1.0591

kontrol 1.07700 .79312 .869 -1.4921 3.6461 dosis 5 mg/ kg bb .74700 .79312 .979 -1.8221 3.3161 dosis 50 mg/ kg bb .88200 .79312 .949 -1.6871 3.4511 dosis 300 mg/ kg bb -.01400 .79312 1.000 -2.5831 2.5551 dosis 1000 mg/ kg bb .84200 .79312 .960 -1.7271 3.4111 dosis 4000 mg/ kg bb -.55700 .79312 .996 -3.1261 2.0121 dosis 8000 mg/ kg bb -.66800 .79312 .989 -3.2371 1.9011

kontrol 1.63400 .79312 .460 -.9351 4.2031 dosis 5 mg/ kg bb 1.30400 .79312 .721 -1.2651 3.8731 dosis 50 mg/ kg BB 1.43900 .79312 .615 -1.1301 4.0081 dosis 300 mg/ kg BB .54300 .79312 .997 -2.0261 3.1121 dosis 1000 mg/ kg bb 1.39900 .79312 .647 -1.1701 3.9681 dosis 2000 mg/ kg bb .55700 .79312 .996 -2.0121 3.1261 dosis 8000 mg/ kg bb -.11100 .79312 1.000 -2.6801 2.4581

Kontrol 1.74500 .79312 .378 -.8241 4.3141 dosis 5 mg/ kg bb 1.41500 .79312 .635 -1.1541 3.9841 dosis 50 mg/ kg bb 1.55000 .79312 .526 -1.0191 4.1191 dosis 300 mg/ kg bb .65400 .79312 .990 -1.9151 3.2231 dosis 1000 mg/ kg bb 1.51000 .79312 .558 -1.0591 4.0791 dosis 2000 mg/ kg bb .66800 .79312 .989 -1.9011 3.2371 dosis 4000 mg/ kg bb .11100 .79312 1.000 -2.4581 2.6801

Lampiran 10. (Lanjutan)

onfide

r

Uppe

29.5200

461 28.1745 30.8655

29.8500

.64020 28.0725 31.6275

29.7150

.51149 28.2949 31.1351

30.6110

.60183 28.9401 32.2819

50 .65056 27.9488 31.5612

30.5970

.51271 29.1735 32.0205

31.1540

.61529 29.4457 32.8623

31.2650

.42772 30.0775 32.4525

40 30.3084

.20693 29.8898 30.7269

5

7

32 .979

oups

16.475

7

2.354

50.323

32

573

(23)

Lampiran 10. (Lanjutan)

c.

Berat badan minggu 2

Descriptives

Interval for Mean

Min Max (I) perlakuan (J) perlakuan

Mean

Test of Homogeneity of Variances

Levene

Statistic

df1

df2

Sig.

(24)

Mean Difference

(I-J)

Std.

Error Sig.

kontrol 1.63100 .68533 .285 -.5890 3.8510 dosis 5 mg/ kg bb .62600 .68533 .983 -1.5940 2.8460 dosis 50 mg/ kg bb 1.01600 .68533 .811 -1.2040 3.2360 dosis 300 mg/ kg bb -.44600 .68533 .998 -2.6660 1.7740 dosis 2000 mg/ kg bb .01500 .68533 1.000 -2.2050 2.2350 dosis 4000 mg/ kg bb -1.25400 .68533 .606 -3.4740 .9660 dosis 8000 mg/ kg bb -.91400 .68533 .879 -3.1340 1.3060

kontrol 1.61600 .68533 .295 -.6040 3.8360 dosis 5 mg/ kg bb .61100 .68533 .985 -1.6090 2.8310 dosis 50 mg/ kg bb 1.00100 .68533 .822 -1.2190 3.2210 dosis 300 mg/ kg bb -.46100 .68533 .997 -2.6810 1.7590 dosis 1000 mg/ kg bb -.01500 .68533 1.000 -2.2350 2.2050 dosis 4000 mg/ kg bb -1.26900 .68533 .592 -3.4890 .9510 dosis 8000 mg/ kg bb -.92900 .68533 .870 -3.1490 1.2910

kontrol -1.88500 .68533 .360 .6650 5.1050 dosis 5 mg/ kg bb 1.88000 .68533 .146 -.3400 4.1000 dosis 50 mg/ kg BB -1.27000 .68533 .420 .0500 4.4900 dosis 300 mg/ kg BB .80800 .68533 .932 -1.4120 3.0280 dosis 1000 mg/ kg bb 1.25400 .68533 .606 -.9660 3.4740 dosis 2000 mg/ kg bb 1.26900 .68533 .592 -.9510 3.4890 dosis 8000 mg/ kg bb .34000 .68533 1.000 -1.8800 2.5600

Kontrol -1.54500 .68533 .320 .3250 4.7650 dosis 5 mg/ kg bb 1.54000 .68533 .352 -.6800 3.7600 dosis 50 mg/ kg bb 1.93000 .68533 .126 -.2900 4.1500 dosis 300 mg/ kg bb .46800 .68533 .997 -1.7520 2.6880 dosis 1000 mg/ kg bb .91400 .68533 .879 -1.3060 3.1340 dosis 2000 mg/ kg bb .92900 .68533 .870 -1.2910 3.1490 dosis 4000 mg/ kg bb -.34000 .68533 1.000 -2.5600 1.8800

Lampiran 10. (Lanjutan)

onfide

r

Uppe

29.5200

.48461 28.1745 30.8655

29.8500

020 28.0725 31.6275

29.7150

.51149 28.2949 31.1351

30.6110

.60183 28.9401 32.2819

29.7550

.65056 27.9488 31.5612

.5970

.51271 29.1735 32.0205

31.1540

.61529 29.4457 32.8623

31.2650

.42772 30.0775 32.4525

40 30.3084

.20693 29.8898 30.7269

5

7

32 .979

oups

16.475

7

2.354 oups

50.323

32

1.573

(25)

Lampiran 11. Hasil analisis data

SPSS konsumsi makanan mencit

a.

Berat Makanan minggu I

Descriptives

Interval for Mean

(26)

(27)

Lampiran 11. (Lanjutan)

b.

Berat makanan minggu II

Descriptives

Interval for Mean

(28)

(29)

Lampiran 12. Hasil analisis data

SPSS konsumsi minuman mencit

a.

Volume minuman minggu I

Descriptives

Interval for Mean

(30)

(31)

Lampiran 12. (Lanjutan)

b.

Volume minuman minggu II

Descriptives

Interval for Mean

(32)

(33)

Lampiran 13. Gambar makropatologi organ hati dan ginjal mencit

1. Organ hati

CMC Na 1%

EEBAM 5 mg/kg bb

EEBAM 50 mg/kg bb

EEBAM 300 mg/kg bb

EEBAM 1000 mg/kg bb

EEBAM 2000 mg/kg bb

(34)

Lampiran 13. (Lanjutan)

2. Organ ginjal

CMC Na 1%

EEBAM 5 mg/kg bb

EEBAM 50 mg/kg bb

EEBAM 300 mg/kg bb

EEBAM 1000 mg/kg bb

EEBAM 2000 mg/kg bb