CALCULATION OF LD50 or LC50 USING PROBIT ANALYSIS

Dr.Alpha Raj.M, MVSc, PhD Assistant Professor, Veterinary Pharamcology & Toxicology, SVVU, India
Instructions
This spread sheet caters upto 20 groups and can handle zero values or gaps in between cells
Dose/ Conc. 1 is invariably Zero / Control; If control was not used, leave total and no. dead blank and start from dose
Delete all values of example and enter Dose/Conc. units, Dose/ Conc., Total subjects and No. dead in each group and
Read the respective LD/LC values and 95% Fiducial confidence limits.
For steps involved in calucation refer to Sheet 2. In case of errors or corrections please email at alpharajm@gmail.com

Dose Calculating LD50/LC50 using Pr

Units= ppm

Dose/Conc. Total Dead 5.00f(x) = 2.3967732989x + 13.6710923485 5.00

Mortality(%) in Probits
R² = 0.6108229874
Control 0.00 10 0
2 0.0002 10 5
3 0.0004 10 5
4 0.0006 10 9
5 0.0008 10 10
6
7
-3.80 -3.70 -3.60 -3.50 -3.40
8 Log10 Dose
9
10 Calculating LD50/LC50 using Probit Analysis
11 Group Log10 Dose
12 2 -3.70
13 3 -3.40
14 4 -3.22
15
16
17
18
19
20
(Original values used for curve fitting)
OBIT ANALYSIS
ology & Toxicology, SVVU, India. alpharajm@gmail.com

between cells
and no. dead blank and start from dose/conc. 2
bjects and No. dead in each group and hit enter

s please email at alpharajm@gmail.com LD/LC LD/LC

(%) ppm
Calculating LD50/LC50 using Probit Analysis
6.28 7.00 LD47 0.000
6.00
LD48 0.000
.00f(x) = 2.3967732989x + 13.6710923485 5.00 LD49 0.000
R² = 0.6108229874 5.00
LD50 0.000
4.00 LD51 0.000
3.00 LD52 0.000
LD53 0.000
2.00
LD54 0.000
1.00 LD55 0.000
0.00 LD56 0.000
3.70 -3.60 -3.50 -3.40 -3.30 -3.20 -3.10
Log10 Dose LD57 0.000
LD58 0.000
LC50 using Probit Analysis CURVE_FITTING (Scroll down for LC/LC values)
Empirical Probits Slope 2.397
5.00 Intercept 13.671
5.00 SD (σ) 0.417
6.28 SE 0.093
R^2 0.611
Chi-test (χ2) Sig 0.946
df 1
Chi-Test NON-SIG
Fitting GOOD FIT

WORK._PROBIT_CURVE
Slope 3.098
Intercept 16.141
R^2 0.757
d for curve fitting)
 95%Fiducial CI
Lower Upper

0.000 0.000
0.000 0.000
0.000 0.000
0.000 0.000
0.000 0.000
0.000 0.000
0.000 0.000
0.000 0.000
0.000 0.000
0.000 0.000
0.000 0.000
0.000 0.000
n for LC/LC values)
CALCULATION OF LD50 or LC50 USING PROBIT ANALYSIS
Dr.Alpha Raj.M, MVSc, PhD Assistant Professor, Veterinary Pharamcology & Toxicology, SVVU, India
Instructions
This spread sheet caters upto 20 groups and can handle zero values or gaps in between cells
Dose/ Conc. 1 is invariably Zero / Control; If control was not used, leave total and no. dead blank and start from dose
Delete all values of example and enter Dose/Conc. units, Dose/ Conc., Total subjects and No. dead in each group and
Read the respective LD/LC values and 95% Fiducial confidence limits.
For steps involved in calucation refer to Sheet 2. In case of errors or corrections please email at alpharajm@gmail.com

Dose Calculating LD50/LC50 using Pr

Units= ppm

Dose/Conc. Total Dead 5.00f(x) = 2.3967732989x + 13.6710923485 5.00

Mortality(%) in Probits
R² = 0.6108229874
Control 0.0000 10 1
2 0.0002 10 2
3 0.0004 10 3
4 0.0006 10 4
5 0.0008 10 5
6
7
-3.80 -3.70 -3.60 -3.50 -3.40
8 Log10 Dose
9
10 Calculating LD50/LC50 using Probit Analysis
11 Group Log10 Dose
12 2 -3.70
13 3 -3.40
14 4 -3.22
15 5 -3.10
16
17
18
19
20
(Original values used for curve fitting)
OBIT ANALYSIS
ology & Toxicology, SVVU, India. alpharajm@gmail.com

between cells
and no. dead blank and start from dose/conc. 2
bjects and No. dead in each group and hit enter

s please email at alpharajm@gmail.com LD/LC LD/LC

(%) ppm
Calculating LD50/LC50 using Probit Analysis
6.28 7.00 LD50 0.001
6.00
LD51 0.001
.00f(x) = 2.3967732989x + 13.6710923485 5.00 LD52 0.001
R² = 0.6108229874 5.00
LD53 0.001
4.00 LD54 0.001
3.00 LD55 0.001
LD56 0.001
2.00
LD57 0.001
1.00 LD58 0.001
0.00 LD59 0.001
3.70 -3.60 -3.50 -3.40 -3.30 -3.20 -3.10
Log10 Dose LD60 0.001
LD61 0.001
LC50 using Probit Analysis CURVE_FITTING (Scroll down for LC/LC values)
Empirical Probits Slope 1.389
4.12 Intercept 9.223
4.45 SD (σ) 0.720
4.72 SE 0.150
4.97 R^2 0.980
Chi-test (χ2) Sig 0.034
df 2
Chi-Test NON-SIG
Fitting GOOD FIT

WORK._PROBIT_CURVE
Slope 1.388
Intercept 9.220
R^2 0.980
d for curve fitting)
 95%Fiducial CI
Lower Upper

0.000 0.002
0.000 0.002
0.001 0.002
0.001 0.002
0.001 0.002
0.001 0.002
0.001 0.002
0.001 0.002
0.001 0.002
0.001 0.003
0.001 0.003
0.001 0.003
n for LC/LC values)
STEPS FOLLOWED IN THE CALCUATION OF LD50 or LC50
The following steps are used in the calculation of LD or LC in this spread sheet
1. Converting doses to log (10) doses (x)
2. Converting mortality to proportions
3. The proportions are corrected for control mortality if its is more than 10% using Schneider-Orelli’s (1947) formu

% Responded – % Responded in Control

Corrected mortality (p) = ________________________________ x 100
100 - Responded % in Control

4. Converting corrected proportions (p) to empirical probits (y).

5. A dose response curve is drawn using the log(10) doses (x) and empirical probits (y) and the regression equatio
Empirical probits less than 1 and more than 7 are ignored as they have little and no significance in the estimation
y=5+(x-µ)/s

6. From the equation of the curve and log10 doses, the expected probits (Yi) are derived
7. From the expected probits (Yi), expected mortality proportion followed by expected no.of animals are derived
8. The original mortality (Observed) and derived mortality (Expected) are used to calculate the Chi-Square test wi
If the Chi-square test is non-significant, it indicates good curve fitting.
9. Z value is derived using the formula

Z=1/(v2p)e(-1/2(Yi-5)^2)
where Yi = Expected probits

9. The weighting coefficents (W) are derived using the formula

W=Z^(2 )/PQ,
where P = Expected proportion
Q=(1-P)

10. The weighted coefficients were used to calculate the standard error

SE= s/v?nW
where s = Standard deviation (1/slope)
n= number of animals in each group
W= Weighting coefficient

11. Working probits (Yw) are derived from the regression equation as follows

Yw = Yi-(P/Z)-p/Z
Y = Expectedl probits
P = Expected Proportion
p = Observed proportion
11. The LD or LC values are derived from the curve drawn using working probits and log doses. Antilog of the dose

12. 95% Fiducial confidence limits are calculated using the formula

Fiducial Limits =Antilog (Log10 Dose ± 1.96 (SE)

(Your comments and corrections are welcome to make this spread sheet better: alpharajm@gmail.com)

References
Busvine J.r. 1971. A critical review of the techniques for testing insecticides. Commonwealth Agricultural
Bureaux, London, ISBN 0851980309
Finney, D. J., Ed. (1952). Probit Analysis. Cambridge, England, Cambridge University Press.
Finney, D. J. and W. L. Stevens (1948). "A table for the calculation of working probits and weights in probit analysi
Greenberg, B. G. (1980). "Chester I. Bliss, 1899-1979." International Statistical Review / Revue Internationale de S
Hayes, W. J. and C. L.Kruger(eds).2014 Handbook of Haye's principles and methods of toxicology, 6th Edition, CRC
g Schneider-Orelli’s (1947) formula:

its (y) and the regression equation is derived.

no significance in the estimation of LD or LC (Hayes, 2014).

ected no.of animals are derived

calculate the Chi-Square test with (No. of log doses used -2) degrees of freedom.
 nd log doses. Antilog of the dose corresponding to respective probit value.

alpharajm@gmail.com)

monwealth Agricultural

bits and weights in probit analysis." Biometrika 35(1-2): 191-201.

view / Revue Internationale de Statistique 8(1): 135-136.
ds of toxicology, 6th Edition, CRC Press, Boca Raton, NY