Toxicity Tests

Sukamto S M

Types of Toxicity Test

LD50

Miller & Tainter Method (1944)

Reed & Muench Method (1938)
Karber Method (1931)
Irritation

Test

Corrosive

Test

Miller & Tainter Method

Miller & Tainter Method

Results of the lethal doses of thymoquinone for the
determination of LD50 after intraperitoneal injection in
rats (n=10)
Group

Dose
(mg/kg
)

Log
dose

Dead

% Dead

*Correct
ed %

Probi
t

25

1.4

2.5

3.04

50

1.7

40

40

4.75

75

1.88

70

70

5.52

100

90

90

6.28

150

2.18

10

100

97.5

6.96

Rumus koreksi :
Untuk mati 0 % = 100 (0,25/n)
Untuk mati 100 % = 100 [(n-0.25)/n]
Di mana n = jumlah hewan dalam satu
kelompok
*

Tabel Harga Probit Sesuai Dengan Persentasenya

Probit

Persentase

2,67

2,95

3,12

3,25

3,36

3,45

3,52

3,59

3,66

10

3,72

3,77

3,82

3,87

3,92

3,96

4,01

4,05

4,08

4,12

20

4,16

4,19

4,23

4,26

4,29

4,33

4,36

4,39

4,42

4,45

30

4,48

4,50

4,53

4,56

4,59

4,61

4,64

4,67

4,69

4,72

40

4,75

4,77

4,80

4,82

4,85

4,87

4,90

4,92

4,95

4,97

50

5,00

5,03

5,05

5,08

5,10

5,13

5,15

5,18

5,20

5,23

60

5,25

5,28

5,31

5,33

5,36

5,39

5,41

5,44

5,47

5,50

70

5,52

5,55

5,58

5,61

5,64

5,67

5,71

5,74

5,77

5,81

80

5,84

5,88

5,92

5,95

5,99

6,04

6,08

6,13

6,18

6,23

90

6,28

6,34

6,41

6,48

6,55

6,64

6,75

6,88

7,05

7,33

0,0

0,1

0,2

0,3

0,4

0,5

0,6

0,7

0,8

0,9

7,33

7,37

7,41

7,46

7,51

7,58

7,65

7,75

7,88

8,09

99

Miller & Tainter Method

8
7
6

f(x) = 5.07x - 3.97

5
Probits 4
3
2
1
0
1.3 1.4 1.5 1.6 1.7 1.8 1.9 2 2.1 2.2 2.3
Log dose

Miller & Tainter Method

y = 5.0658x 3.9705
Probit 50% = 5
5 = 5.0658x 3.9705
x = (5 + 3.9705)/5.0658
x = 1.77 (nilai dalam Log-dose)
x = 10^1.77 = 58.88 mg/kg nilai LD50

Standard Error (SE) of LD50

SE of LD50 = (LD84 - LD16) /
2n
y = 5.0658x 3.9705

Probit 84 = 5.99 log LD84 (x) = (5.99 + 3.9705) / 5.0658

= 1.97
Antilog 1.97 = 93.32
Probit 16 = 4.01 log LD16 (x) = (4.01 + 3.9705) / 5.0658
= 1.58
Antilog 1.58 = 38.02
SE of LD50 = (93.32 38.02) / 20

So.......

LD50 = 58.88 12.37

LD50 of thymoquinone when given
intraperitoneally is 58.8812.37, with 95%
confidence
interval of 46.51 71.25

Miller & Tainter Method

Dosis

Kons.

(mg/kg)

(mg/ml)

1000

40

10

100

97.5

6.97

700

28

60

60

5.25

500

20

40

40

4.75

350

14

10

2.5

3.04

Klp

*5

Mati Hidup

mati

terkorek Probit

250koreksi10
0
10
Rumus
:
Untuk mati 0 % = 100 (0,25/n)
Untuk mati 100 % = 100 [(n0.25)/n]
Di mana n = jumlah hewan dalam satu
kelompok

si

Karbers Method

Karbers Method

LD50 = LD100 - (a x b) / n
a = the difference between two successive doses of
administered extract
b = the average number of dead animals in two
successive doses.
n = total number of animal in a group.

LD50 = LD100 - (a x b) / n
a = the difference between two successive doses of
administered extract
b = the average number of dead animals in two successive
doses.
nIf=ntotal
number
of animal
in a group.
= 8 rats
per group,
determine
LD50 by using Karbers
method!
Dead

Dose
differen
ce (a)

Mean
(b)

330

300

30

7.5

225

280

20

120

250

30

120

220

30

60

120

100

0.5

50

Group

Dose
(mg/kg)

Product
(a x b)

Total

LD50 = 330 (575/8) = 330 71.88 =

575

Potency of toxic chemicals

(Gosselin, Smith, Hodge)

Toxicity rating
Dosage
Practically non toxic >15 g/kg
Slightly toxic
5-15 g/kg
Moderately toxic
0.5-5 g/kg
Very toxic
50-500 mg/kg
Extremely toxic
5-50 mg/kg
Super toxic
< 5mg/kg

Thank you