CALCULATION OF LD50 or LC50 USING PROBIT ANALYSIS

Dr.Alpha Raj.M, MVSc, PhD Assistant Professor, Veterinary Pharamcology & Toxicology, S
INSTRUCTIONS
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Dose Calculating LD50/LC

Units= ppm

Don’t change Dose/Conc. Total Dead f(x) = 1.0403624971x + 5.42

Mortality(%) in Probits
R² = 0.9817355421
4.48
Control Control 0.00 10 0
3.72
Conc. 2 0.03 10 1
3 0.10 10 3
4 0.30 10 5
5 1.00 10 7
6 3.00 10 8
7 10.00 10 10
-2.00 -1.50 -1.00
8 Lo
9
10 Calculating LD50/LC50 using Probit Analy
11 Group Log10 Dose
12 2 -1.52
13 3 -1.00
14 4 -0.52
15 6 0.48
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Values used for curve fitting

OBIT ANALYSIS
ology & Toxicology, SVVU, India. alpharajm@gmail.com
How to cite:
rsion for calculations. D.J.Finney (1952) Probit Analysis (2nd Ed),
Journal of the Institute of Actuaries, 78 (3): 388-390.

LD/LC LD/LC
(%) ppm
Calculating LD50/LC50 using Probit Analysis
7.00 LD46 0.298
5.84 LD47 0.315
6.00
5.00
f(x) = 1.0403624971x + 5.4269954909 LD48 0.332
R² = 0.9817355421
4.48 5.00
LD49 0.351
3.72 4.00 LD50 0.371
3.00 LD51 0.392
LD52 0.414
2.00
LD53 0.437
1.00 LD54 0.462
0.00 LD55 0.488
-1.50 -1.00 -0.50 0.00 0.50 1.00
Log10 Dose LD56 0.516
LD57 0.545
LC50 using Probit Analysis CURVE_FITTING (Scroll down for LC/LC values)
Empirical Probits Slope 1.060
3.72 Intercept 5.457
4.48 SD (σ) 0.943
5.00 SE 0.176
5.84 R^2 0.982
Chi-test (χ2) Sig 0.994
df 3
Chi-Test NON-SIG
Fitting GOOD FIT

WORK._PROBIT_CURVE
Slope 1.053
Intercept 5.454
R^2 0.973
nalysis (2nd Ed),
Actuaries, 78 (3): 388-390.

95%Fiducial CI
Lower Upper

0.135 0.658
0.142 0.695
0.150 0.735
0.159 0.776
0.168 0.820
0.177 0.866
0.187 0.915
0.198 0.966
0.209 1.021
0.221 1.079
0.233 1.140
0.247 1.205
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) fo

% 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 equ
Empirical probits less than 1 and more than 7 are ignored as they have little and no significance in the estimati
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 deriv
8. The original mortality (Observed) and derived mortality (Expected) are used to calculate the Chi-Square tes
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
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 ana
Greenberg, B. G. (1980). "Chester I. Bliss, 1899-1979." International Statistical Review / Revue Internationale d
Hayes, W. J. and C. L.Kruger(eds).2014 Handbook of Haye's principles and methods of toxicology, 6th Edition,
 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

o calculate the Chi-Square test with (No. of log doses used -2) degrees of freedom.

and 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