63
64
65
Lampiran 3. Karakteristik Tumbuhan Pecut Kuda
66
Ekstrak etanol pecut kuda
67
Lampiran 4. Hasil Pemeriksaan Mikroskopik
Gambar: Serbuk simplisia pecut kuda
Keterangan: 1. Rambut
2. Rambut kelenjar 3. Jaringan palisade
4. Pembuluh kayu dengan penebalan jala 5. Fragmen mesofil
6. Epidermis
6 4 3
5
68
Lampiran 5. Perhitungan karakterisasi simplisia dan ekstrak pecut kuda Perhitungan penetapan kadar air dari serbuk simplisia pecut kuda
No Berat Sampel (g) Volume Awal (ml) Volume Akhir (ml)
69
% Kadar sari larut dalam air rata-rata =
3
70
% Kadar sari larut dalam etanol rata-rata =
3
71
Perhitungan Penetapan Kadar Abu tidak Larut Asam dari Serbuk Simplisia pecut kuda (Lanjutan)
No Berat Sampel (g) Berat Abu (g) Perhitungan penetapan kadar air ekstrak pecut kuda (Lanjutan)
72 Perhitungan penetapan kadar abu total ekstrak pecut kuda (Lanjutan)
73
Perhitungan penetapan kadar abu tidak larut asam ekstrak pecut kuda (Lanjutan) No Berat Sampel (g) Berat Abu (g)
% Kadar abu yang tidak larut dalam asam rata-rata
74
Lampiran 6. Alat yang digunakan
Modifikasi alat penampung urin
Alat AAS
75
Lampiran 7. Data volume urin (ml), pH urin, kadar natrium, dan kadar kalium (meq/l) setelah pemberian ekstrak dan furosemid (pembanding) No Parameter Kontrol
CMC Na 0,5%
Furosemid 10 mg/kg
bb
76
Lampiran 8. Tabel konversi dosis hewan dengan manusia Konversi dosis antara jenis hewan dengan manusia (Laurence and Bacharach, 1964).
Mencit 20 g
Tikus 200 g
Marmut 400 g
Kelinci 1,2 kg
Kera 4 kg
Anjing 12 kg
Manusia 70 kg Mencit
20g
1,0 7,0 12,25 27,8 64,1 124,2 387,9
Tikus 200g
0,14 1,0 1,74 3,9 9,2 17,8 56,0
Marmut 400 g
0,08 0,57 1,0 2,25 5,2 10,2 31,5
Kelinci 1,2 kg
0,04 0,25 0,44 1,0 2,4 4,5 14,2
Kera 4 kg
0,016 0,11 0,19 0,42 1,0 1,9 6,1
Anjing 12 kg
0,008 0,06 0,10 0,22 0,52 1,0 3,1
Manusia 70 kg
77
Lampiran 9. Contoh perhitungan dosis kontrol CMC Na 0,5% - Pembuatan CMC 0,5%
= 500 mg/ 100 ml = 5 mg/ml
- Perhitungan CMC Na 0,5% pada tikus 200 g = 1% × BB Tikus
=
ml 100
g 1
× 200 g
= 2 ml
78
Lampiran 10. Contoh Perhitungan Dosis Furosemid Furosemid yang digunakan = 40 mg
Dosis Furosemid 10 mg/kg bb untuk tikus 200 g Furosemid yang diberikan =
g 1000
bb mg/kg 10
× 200 g
= 2 mg Volume yang diberikan =
mg 40
mg 2
× 10 ml
= 0,5 ml
79
Lampiran 11. Contoh perhitungan dosis dari ekstrak etanol pecut kuda 1%
1. Untuk dosis 100 mg/kg bb pada tikus 180,2 g =
g 1000
g 180,2
x 100 mg/kg bb = 18,02 mg
Volume yang diberikan =
mg/ml 10
mg 18,02
= 1,802 ml
2. Untuk dosis 150 mg/kg bb pada tikus 179,5 g =
g 1000
g 179,5
x 150 mg/kg bb = 26,92 mg
Volume yang diberikan =
mg/ml 10
mg 26,92
= 2,692 ml
3. Untuk dosis 200 mg/kg bb pada tikus 188,9 g
=
g 1000
g 188,9
x 200 mg/kg bb = 37,78 mg
Volume yang diberikan =
mg/ml 10
mg 37,78
80
Lampiran 12. Hasil pengujian AAS dari volume urin untuk kadar Na+ dan K+
No Kelompok pengujian Tikus Konsentrasi
Na+ K+
1. Kontrol CMC-Na 0,5%
1 1,5384 3,5221
2 1,8014 3,7006
3 1,4531 4,0022
4 1,8664 3,7044
5 1,9220 4,5113
2. Furosemid dosis 10 mg/kg bb
1 3,7151 6,4331
2 4,2884 8,3544
3 4,1803 7,9225
4 4,5468 7,5239
5 4,8321 8,5321
3. EEPK dosis 100 mg/kg bb
1 2,7504 4,4252
2 3,0080 4,6997
3 3,2051 5,1220
4 2,7671 4,8150
5 3,6228 5,9921
4. EEPK dosis 150 mg/kg bb
1 2,9204 4,7308
2 3,1005 5,5321
3 2,9884 5,6682
4 3,4260 5,1595
5 3,8846 6,8311
5. EEPK dosis 200 mg/kg bb
1 3,4821 5,0330
2 4,0111 6,8766
3 3,3550 5,8850
4 4,1250 6,9236
81
Lampiran 13. Contoh perhitungan kadar elektrolit (Lanjutan)
Kadar Na =
x 500
Pembacaan pada AAS = 1,5384 mcg/ml
Kadar Na =
ml 1
mcg/ml 1,5384
x 500 = 769,2 mcg/ml
= 0,7692 mg/ml
meq/l =
x 1 meq
=
23 mg/ml 0,7692
x 1 meq = 0,03344 meq/ml
82
Lampiran 14. Data kalibrasi natrium dengan AAS, perhitungan persamaan garis regresi dan koefisien korelasi
No. Konsentrasi (µg/ml) (X)
15,0000 2,1147 7,7676 55,0000 1,09708249 = 2,5 = 0,35245 Dengan nilai r sebagai berikut :
83
Lampiran 15. Data kalibrasi kalium dengan AAS, perhitungan persamaan garis regresi dan koefisien korelasi
No. Konsentrasi (µg/ml) (X) 30,0000 1,2709 9,366 220,0000 0,39881489
= 5 = 0,211817 Dengan nilai r sebagai berikut :
84
Lampiran 16. Skema pembuatan ekstrak etanol pecut kuda dan uji aktivitas diuretik
Maserasi dengan etanol 96% selama 5 hari
Didiamkan selama 2 hari
Diuapkan dengan Rotary Evaporator
Dikeringkan dengan Freeze Dryer
Uji aktivitas diuretik
Dikondisikan selama 2 minggu
Dipuasakan selama ± 12 jam
Diberi ekstrak etanol Pecut kuda
(EEPK) dengan dosis berbeda
Diukur Diukur dengan pH meter dengan AAS
Nilai pH
Serbuk simplisia herba pecut kuda
Ampas Maserat
Ekstrak kental Etanol
Ekstrak etanol pecut kuda (EEPK)
Tikus
Urin
Volume urin
Maserat
Elektrolit
85
Lampiran 17. Hasil data statistik ANAVA dengan Uji Tukey Jam ke 1- 5 ANOVA
Volume_Urin_Jam_1
Sum of Squares df Mean Square F Sig. Between Groups 5.286 4 1.321 12.779 .000 Within Groups 2.068 20 .103
Total 7.354 24
Post Hoc Tests
Multiple Comparisons
Volume_Urin_Jam_1 Tukey HSD
(I)
Kelompok_Peng ujian
(J)
Kelompok_Pengujian
Mean Differenc
e (I-J) Std. Error Sig.
95% Confidence Interval Lower
Bound
Upper Bound Kontrol CMC Na
o.5%
Furosemid 10 mg/kg bb -1.3800* .2034 .000 -1.989 -.771
EEPK 100 mg/kg bb -.9800* .2034 .001 -1.589 -.371 EEPK 150 mg/kg bb -.5800 .2034 .006 -1.189 .029 EEPK 200 mg/kg bb -.8800* .2034 .003 -1.489 -.271 Furosemid 10
mg/kg bb
Kontrol CMC Na o.5% 1.3800* .2034 .000 .771 1.989 EEPK 100 mg/kg bb .4000 .2034 .017 -.209 1.009 EEPK 150 mg/kg bb .8000* .2034 .007 .191 1.409 EEPK 200 mg/kg bb .5000 .2034 .141 -.109 1.109 EEPK 100
mg/kg bb
Kontrol CMC Na o.5% .9800* .2034 .001 .371 1.589
Furosemid 10 mg/kg bb -.4000 .2034 .017 -1.009 .209 EEPK 150 mg/kg bb .4000 .2034 .017 -.209 1.009 EEPK 200 mg/kg bb .1000 .2034 .987 -.509 .709 EEPK 150
mg/kg bb
Kontrol CMC Na o.5% .5800 .2034 .006 -.029 1.189
86
mg/kg bb Furosemid 10 mg/kg bb -.5000 .2034 .141 -1.109 .109 EEPK 100 mg/kg bb -.1000 .2034 .987 -.709 .509 EEPK 150 mg/kg bb .3000 .2034 .589 -.309 .909 *. The mean difference is significant at the 0.05 level.
ANOVA
Volume_Urin_Jam_Ke_2
Sum of Squares df Mean Square F Sig. Between Groups 11.254 4 2.813 13.086 .000
Within Groups 4.300 20 .215
Total 15.554 24
Post Hoc Tests
Multiple Comparisons
Volume_Urin_Jam_Ke_2 Tukey HSD
(I)
Kelompok_Pen
gujian (J) Kelompok_Pengujian
Mean Difference
(I-J)
Std. Error Sig.
95% Confidence Interval Lower
Bound
Upper Bound Kontrol CMC
na 0.5%
Furosemid 10 mg/kg bb -2.1000* .2933 .000 -2.978 -1.222
EEPK 100 mg/kg bb -1.1800* .2933 .005 -2.058 -.302 EEPK 150 mg/kg bb -.9400* .2933 .032 -1.818 -.062 EEPK 200 mg/kg bb -1.2000* .2933 .005 -2.078 -.322 Furosemid 10
mg/kg bb
Kontrol CMC na 0.5% 2.1000* .2933 .000 1.222 2.978
EEPK 100 mg/kg bb .9200* .2933 .037 .042 1.798 EEPK 150 mg/kg bb 1.1600* .2933 .006 .282 2.038 EEPK 200 mg/kg bb .9000* .2933 .053 .022 1.778 EEPK 100
mg/kg bb
Kontrol CMC na 0.5% 1.1800* .2933 .005 .302 2.058
87
mg/kg bb Furosemid 10 mg/kg bb -1.1600* .2933 .006 -2.038 -.282 EEPK 100 mg/kg bb -.2400 .2933 .922 -1.118 .638 EEPK 200 mg/kg bb -.2600 .2933 .899 -1.138 .618 EEPK 200
mg/kg bb
Kontrol CMC na 0.5% 1.2000* .2933 .005 .322 2.078
Furosemid 10 mg/kg bb -.9000* .2933 .053 -1.778 -.022 EEPK 100 mg/kg bb .0200 .2933 1.000 -.858 .898 EEPK 150 mg/kg bb .2600 .2933 .899 -.618 1.138 *. The mean difference is significant at the 0.05 level.
ANOVA
Volume_Urin_Jam_Ke_3
Sum of Squares df Mean Square F Sig. Between Groups 19.728 4 4.932 22.459 .000 Within Groups 4.392 20 .220
Total 24.120 24
Post Hoc Tests
Multiple Comparisons
Volume_Urin_Jam_Ke_3 Tukey HSD
(I)
Kelompok_Peng
ujian (J) Kelompok_Pengujian
Mean Differenc
e (I-J)
Std.
Error Sig.
95% Confidence Interval Lower
Bound
Upper Bound Kontrol CMC na
0.5%
Furosemid 10 mg/kg bb -2.6400* .2964 .000 -3.527 -1.753
EEPK 100 mg/kg bb -1.3200* .2964 .002 -2.207 -.433 EEPK 150 mg/kg bb -1.2000* .2964 .005 -2.087 -.313 EEPK 200 mg/kg bb -2.0400* .2964 .000 -2.927 -1.153 Furosemid 10
mg/kg bb
Kontrol CMC na 0.5% 2.6400* .2964 .000 1.753 3.527
88
mg/kg bb Furosemid 10 mg/kg bb -1.3200* .2964 .002 -2.207 -.433 EEPK 150 mg/kg bb .1200 .2964 .994 -.767 1.007 EEPK 200 mg/kg bb -.7200 .2964 .148 -1.607 .167 EEPK 150
mg/kg bb
Kontrol CMC na 0.5% 1.2000* .2964 .005 .313 2.087
Furosemid 10 mg/kg bb -1.4400* .2964 .001 -2.327 -.553 EEPK 100 mg/kg bb -.1200 .2964 .994 -1.007 .767 EEPK 200 mg/kg bb -.8400 .2964 .069 -1.727 .047 EEPK 200
mg/kg bb
Kontrol CMC na 0.5% 2.0400* .2964 .000 1.153 2.927 Furosemid 10 mg/kg bb -.6000 .2964 .291 -1.487 .287 EEPK 100 mg/kg bb .7200 .2964 .148 -.167 1.607 EEPK 150 mg/kg bb .8400 .2964 .069 -.047 1.727 *. The mean difference is significant at the 0.05 level.
ANOVA
Volume_Urin_Jam_Ke_4
Sum of Squares df Mean Square F Sig. Between Groups 21.956 4 5.489 27.146 .000 Within Groups 4.044 20 .202
Total 26.000 24
Post Hoc Tests
Multiple Comparisons
Volume_Urin_Jam_Ke_4 Tukey HSD
(I)
Kelompok_Penguj ian
(J)
Kelompok_Pengujian
Mean Differenc
e (I-J)
Std.
Error Sig.
95% Confidence Interval Lower
Bound
Upper Bound Kontrol CMC na
0.5%
Furosemid 10 mg/kg bb -2.7600* .2844 .000 -3.611 -1.909
EEPK 100 mg/kg bb -1.3000* .2844 .002 -2.151 -.449 EEPK 150 mg/kg bb -1.3400* .2844 .001 -2.191 -.489 EEPK 200 mg/kg bb -2.2000* .2844 .000 -3.051 -1.349 Furosemid 10
mg/kg bb
89
EEPK 150 mg/kg bb 1.4200* .2844 .001 .569 2.271 EEPK 200 mg/kg bb .5600 .2844 .316 -.291 1.411 EEPK 100 mg/kg
bb
Kontrol CMC na 0.5% 1.3000* .2844 .002 .449 2.151
Furosemid 10 mg/kg bb -1.4600* .2844 .000 -2.311 -.609 EEPK 150 mg/kg bb -.0400 .2844 1.000 -.891 .811 EEPK 200 mg/kg bb -.9000* .2844 .035 -1.751 -.049 EEPK 150 mg/kg
bb
Kontrol CMC na 0.5% 1.3400* .2844 .001 .489 2.191
Furosemid 10 mg/kg bb -1.4200* .2844 .001 -2.271 -.569 EEPK 100 mg/kg bb .0400 .2844 1.000 -.811 .891 EEPK 200 mg/kg bb -.8600* .2844 .047 -1.711 -.009 EEPK 200 mg/kg
bb
Kontrol CMC na 0.5% 2.2000* .2844 .000 1.349 3.051 Furosemid 10 mg/kg bb -.5600 .2844 .316 -1.411 .291 EEPK 100 mg/kg bb .9000* .2844 .035 .049 1.751 EEPK 150 mg/kg bb .8600* .2844 .047 .009 1.711 *. The mean difference is significant at the 0.05 level.
ANOVA
Volume_Urin_Jam_Ke_5
Sum of Squares df Mean Square F Sig. Between Groups 26.528 4 6.632 32.257 .000 Within Groups 4.112 20 .206
Total 30.640 24
Post Hoc Tests
Multiple Comparisons
Volume_Urin_Jam_Ke_5 Tukey HSD
(I)
Kelompok_Peng
ujian (J) Kelompok_Pengujian
Mean Differenc
e (I-J)
Std.
Error Sig.
95% Confidence Interval Lower
Bound
Upper Bound Kontrol CMC na
0.5%
Furosemid 10 mg/kg bb -2.8200* .2868 .000 -3.678 -1.962
90
EEPK 150 mg/kg bb -1.5200* .2868 .000 -2.378 -.662 EEPK 200 mg/kg bb -2.6800* .2868 .000 -3.538 -1.822 Furosemid 10
mg/kg bb
Kontrol CMC na 0.5% 2.8200* .2868 .000 1.962 3.678 EEPK 100 mg/kg bb 1.5400* .2868 .000 .682 2.398 EEPK 150 mg/kg bb 1.3000* .2868 .002 .442 2.158 EEPK 200 mg/kg bb .1400 .2868 .988 -.718 .998 EEPK 100
mg/kg bb
Kontrol CMC na 0.5% 1.2800* .2868 .002 .422 2.138
Furosemid 10 mg/kg bb -1.5400* .2868 .000 -2.398 -.682 EEPK 150 mg/kg bb -.2400 .2868 .916 -1.098 .618 EEPK 200 mg/kg bb -1.4000* .2868 .001 -2.258 -.542 EEPK 150
mg/kg bb
Kontrol CMC na 0.5% 1.5200* .2868 .000 .662 2.378
Furosemid 10 mg/kg bb -1.3000* .2868 .002 -2.158 -.442 EEPK 100 mg/kg bb .2400 .2868 .916 -.618 1.098 EEPK 200 mg/kg bb -1.1600* .2868 .005 -2.018 -.302 EEPK 200
mg/kg bb
91
Lampiran 18. Hasil data statistik ANAVA dengan Uji Tukey Hasil perhitungan nilai pH urin secara ANAVA
ANOVA
Nilai_pH
Sum of Squares df Mean Square F Sig. Between Groups .018 4 .004 .289 .881 Within Groups .304 20 .015
Total .322 24
Post Hoc Tests
Multiple Comparisons
Nilai_pH Tukey HSD
(I)
Kelompok_Peng ujian
(J)
Kelompok_Pengujian
Mean Difference
(I-J)
Std.
Error Sig.
95% Confidence Interval Lower
Bound
Upper Bound CMC-Na 0,5% Furosemid 10 mg/kg bb -.0200 .0780 .999 -.253 .213
EEPK 100 mg/kg bb .0200 .0780 .999 -.213 .253 EEPK 150 mg/kg bb -.0200 .0780 .999 -.253 .213 EEPK 200 mg/kg bb -.0600 .0780 .936 -.293 .173 Furosemid 10
mg/kg bb
CMC-Na 0,5% .0200 .0780 .999 -.213 .253
EEPK 100 mg/kg bb .0400 .0780 .985 -.193 .273 EEPK 150 mg/kg bb .0000 .0780 1.000 -.233 .233 EEPK 200 mg/kg bb -.0400 .0780 .985 -.273 .193 EEPK 100
mg/kg bb
CMC-Na 0,5% -.0200 .0780 .999 -.253 .213
Furosemid 10 mg/kg bb -.0400 .0780 .985 -.273 .193 EEPK 150 mg/kg bb -.0400 .0780 .985 -.273 .193 EEPK 200 mg/kg bb -.0800 .0780 .840 -.313 .153 EEPK 150
mg/kg bb
CMC-Na 0,5% .0200 .0780 .999 -.213 .253
92
EEPK 200 mg/kg bb -.0400 .0780 .985 -.273 .193 EEPK 200
mg/kg bb
CMC-Na 0,5% .0600 .0780 .936 -.173 .293
Furosemid 10 mg/kg bb .0400 .0780 .985 -.193 .273 EEPK 100 mg/kg bb .0800 .0780 .840 -.153 .313 EEPK 150 mg/kg bb .0400 .0780 .985 -.193 .273
Hasil perhitungan kadar natrium secara ANAVA ANOVA
Kadar_Natrium
Sum of Squares df Mean Square F Sig. Between Groups 9578.435 4 2394.609 32.305 .000 Within Groups 1482.481 20 74.124
Total 11060.916 24
Post Hoc Tests
Multiple Comparisons
Kadar_Natrium Tukey HSD
(I)
Kelompok_Peng ujian
(J)
Kelompok_Pengujian
Mean Difference
(I-J)
Std.
Error Sig.
95% Confidence Interval Lower
Bound
Upper Bound CMC-Na 0,5% Furosemid 10 mg/kg bb -56.61400* 5.44515 .000 -72.9079 -40.3201
EEPK 100mg/kg bb -29.61800* 5.44515 .000 -45.9119 -13.3241 EEPK 150 mg/kg bb -33.19000* 5.44515 .000 -49.4839 -16.8961 EEPK 200 mg/kg bb -49.12800* 5.44515 .000 -65.4219 -32.8341 Furosemid 10
mg/kg bb
CMC-Na 0,5% 56.61400* 5.44515 .000 40.3201 72.9079
EEPK 100mg/kg bb 26.99600* 5.44515 .001 10.7021 43.2899 EEPK 150 mg/kg bb 23.42400* 5.44515 .003 7.1301 39.7179 EEPK 200 mg/kg bb 7.48600 5.44515 .650 -8.8079 23.7799 EEPK 100mg/kg
bb
CMC-Na 0,5% 29.61800* 5.44515 .000 13.3241 45.9119
93
EEPK 150 mg/kg bb -3.57200 5.44515 .963 -19.8659 12.7219 EEPK 200 mg/kg bb -19.51000* 5.44515 .014 -35.8039 -3.2161 EEPK 150
mg/kg bb
CMC-Na 0,5% 33.19000* 5.44515 .000 16.8961 49.4839
Furosemid 10 mg/kg bb -23.42400* 5.44515 .003 -39.7179 -7.1301 EEPK 100mg/kg bb 3.57200 5.44515 .963 -12.7219 19.8659 EEPK 200 mg/kg bb -15.93800 5.44515 .057 -32.2319 .3559 EEPK 200
mg/kg bb
CMC-Na 0,5% 49.12800* 5.44515 .000 32.8341 65.4219
Furosemid 10 mg/kg bb -7.48600 5.44515 .650 -23.7799 8.8079 EEPK 100mg/kg bb 19.51000* 5.44515 .014 3.2161 35.8039 EEPK 150 mg/kg bb 15.93800 5.44515 .057 -.3559 32.2319 *. The mean difference is significant at the 0.05 level.
Hasil perhitungan kadar kalium secara ANAVA ANOVA
Kadar_Kalium
Sum of Squares df Mean Square F Sig. Between Groups 1634.840 4 408.710 16.945 .000 Within Groups 482.386 20 24.119
Total 2117.225 24
Post Hoc Tests
Multiple Comparisons
Kadar_Kalium Tukey HSD
(I)
Kelompok_Peng ujian
(J)
Kelompok_Pengujian
Mean Difference
(I-J)
Std.
Error Sig.
95% Confidence Interval Lower
Bound
Upper Bound CMC-Na 0,5% Furosemid 10mg/kg bb -24.14400* 3.10608 .000 -33.4385 -14.8495
94
10mg/kg bb EEPK 100 mg/kg bb 16.93400* 3.10608 .000 7.6395 26.2285 EEPK 150 mg/kg bb 13.29200* 3.10608 .003 3.9975 22.5865 EEPK 200 mg/kg bb 8.64000 3.10608 .076 -.6545 17.9345 EEPK 100
mg/kg bb
CMC-Na 0,5% 7.21000 3.10608 .009 -2.0845 16.5045 Furosemid 10mg/kg bb -16.93400* 3.10608 .000 -26.2285 -7.6395 EEPK 150 mg/kg bb -3.64200 3.10608 .766 -12.9365 5.6525 EEPK 200 mg/kg bb -8.29400 3.10608 .095 -17.5885 1.0005 EEPK 150
mg/kg bb
CMC-Na 0,5% 10.85200* 3.10608 .017 1.5575 20.1465
Furosemid 10mg/kg bb -13.29200* 3.10608 .003 -22.5865 -3.9975 EEPK 100 mg/kg bb 3.64200 3.10608 .766 -5.6525 12.9365 EEPK 200 mg/kg bb -4.65200 3.10608 .576 -13.9465 4.6425 EEPK 200
mg/kg bb
CMC-Na 0,5% 15.50400* 3.10608 .001 6.2095 24.7985
