Professional Documents
Culture Documents
Pharmaceutical Industry
Kate Arnot
Overview
The drug development process
General recommendations
Examples
Assay
Dissolution test for an immediate release tablet
Identity
Identification test for a drug substance
Impurities
Quantitative test residual solvents in a
drug substance
Limit test residual solvents in a tablet
10
11 12 13
Start
14
15
16
Approval
Clinical Trials
Initial
research
Drug
discovery
Phase 1
Phase 2
Phase 3
50-100
people
100-200
people
500-5000
people
Phase 4
Full Scale
Manufacture
Clinical
Development
Cost
2M
10,000 compounds
150M
300M
1 new medicine
3
General recommendations
Design your validation studies to reflect
the way that the method will be applied
in routine use as far as possible
Ensure that you have considered the
specification you want to achieve in
designing your studies
Know your method before you start to
validate!
5
Dissolution apparatus
100
90
80
70
60
% dissolved
D
i
s
s
o
l
u
t
i
o
n
50
40
30
20
10
0
0
15
30
45
60
75
time (mins)
Time (mins)
Specificity
Linearity
Accuracy and range
Precision
Repeatability
Intermediate
precision
Robustness
10
Specificity
Demonstrate absence of interference from
excipients and reagents at the monitoring
wavelength
For example calculate the absorbance of a
placebo solution as a percentage of the
absorbance of a standard solution
Typically results of less that 2% would be
considered acceptable
Specificity may also be inferred from linearity
and accuracy data
11
Linearity
Range studied 0 to 150% of nominal (7 solutions,
including a blank solution)
Linearity experiments conducted in the presence and
absence of excipients
Determine equation of the line, correlation coefficient,
95% confidence interval of the slope and the
intercept. Verify that the 95% confidence interval of
the intercept includes zero, or that the result is
insignificant in the context of the experiment
Check that the response for both experiments is the
same calculate the slopes as a % of each other
Typically results of 98 to 102% would be considered
acceptable
12
y = 42.983x + 0.004
r = 0.99989
95% CI of slope 0.0730
95% CI of intercept 0.016
1.5
A
b
s
o
r
b
a
n
c
e
1.4
1.3
1.2
1.1
Ab
0.9
sor
ban
0.8
ce
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
0
0.002
0.004
0.006
0.008
0.01
0.012
0.014
0.016
0.018
0.02
0.022
0.024
0.026
0.028
0.03
0.032
0.034
0.036
0.038
concentration (g/100ml)
Concentration (g/100ml)
13
y = 43.033x + 0.006
r = 0.99990
95% CI of slope 0.695
95% CI of intercept 0.009
1.4
1.3
1.2
1.1
1
0.9
Absorbance
A
b
s
o
r
b
a
n
c
e
0.8
0.7
0.6
Comparison of slopes:
(42.983/43.033) x 100
= 99.9%
0.5
0.4
0.3
0.2
0.1
0
0
0.002
0.004
0.006
0.008
0.01
0.012
0.014
0.016
0.018
0.02
0.022
0.024
0.026
0.028
0.03
0.032
0.034
0.036
0.038
concentration (g/100ml)
Concentration (g/100ml)
14
Experiment 2
50% 99.3
85% 99.4
100%99.6
115%98.7
125%98.9
Mean99.2
sd
0.5
16
Experiment 2
Samples stirred continuously
Time
1
Recovery
98.8%
(hours) 4
99.1%
21
99.2%
27
99.3%
17
Accuracy/Linearity how do
you prepare your samples?
Prepare samples containing drug substance
plus an appropriate quantity of ground
placebo tablet
Prepare samples containing drug substance
plus appropriate quantities of an excipient mix
Prepare samples by using different weights
from a ground active tablet preparation,
making a correction for nominal concentration
Make sure that you are confident in your
sample extraction technique
18
Precision - Repeatability
One analyst conducting multiple replicates of
the same sample, on the same day using the
same equipment, and applying the method as
written.
For dissolution experiments correcting for
tablet weight may be appropriate
Determine the mean, the range, the standard
deviation and the relative standard deviation.
19
Robustness
A number of attributes were evaluated prior to
validation:
Standard and sample solution stability
Effect of pH and the requirement to buffer
and/or degass the dissolution medium
Reagents from different suppliers were
evaluated
Sampling tubes, filters and sampling and
filtration speeds were evaluated
Single point vs profile testing
21
Specificity
Demonstrate specificity between drug substance and
other potentially interfering compounds
eg. starting materials, isolated intermediates and
other materials manufactured or formulated in the
same plant
Consideration should also be given to
enantiomers, counter ions/salts, solvates and
polymorphs if necessary.
24
1.0
1.3
1.2
1.1
82968I01
Drug substance
0.9
0.8
1.0
0.7
0.9
0.6
Kubelka-Munk
0.8
Kubelka-Munk
Development
compound
0.7
0.6
0.5
0.4
0.5
0.3
0.4
0.3
0.2
0.2
0.1
0.1
0.0
4000
3500
3000
2500
2000
1500
1000
3500
500
3000
0.85
0.70
0.65
0.60
0.55
Isolated
intermediate
0.55
0.50
0.45
0.40
0.50
1500
1000
500
Compound formulated
in same plant
0.35
0.45
Kubelka-Munk
Kubelka-Munk
2000
0.60
0.80
0.75
2500
W avenumbers (cm-1)
Wavenumbers (cm-1)
0.40
0.35
0.30
0.30
0.25
0.20
0.25
0.15
0.20
0.10
0.15
0.10
0.05
0.05
-0.00
0.00
-0.05
4000
-0.05
3500
3000
2500
2000
Wavenumbers (cm-1)
1500
1000
500
4000
3500
3000
2500
2000
1500
1000
Wavenumbers (cm-1)
25
500
Specificity
Linearity
Accuracy
Precision
Reproducibility
Intermediate precision
Range
Limits of detection and
quantitation
Robustness
27
Specificity
Demonstrate method is specific for all
the solvents used in the registered
synthetic process
Can use linearity data for solvents in the
presence and absence of the drug
substance to show that the drug
substance does not interfere
28
22000
0.0
1.0
28000
2.0
Time (min)
D im e th y ls u lp h o x id e
A c e to n itrile
Tolu e n e
uV
26000
T rie th y la m in e
E th y l A c e ta te
P ro p a n -2 -o l
24000
Specificity chromatogram
3.0
29
Linearity
Range studied 0 to 150% of nominal (9 solutions,
including a blank solution and a solution at the LOQ)
Drug substance required without solvent, or low
levels of solvent and then make a correction
Determine equation of the line, correlation coefficient,
95% confidence interval of the slope and the
intercept. Verify that the 95% confidence interval of
the intercept includes zero, or that the result is
insignificant in the context of the experiment
30
Accuracy
Calculate recoveries from the linearity
experiments by analysing the solutions
against an external standard as per the
method.
Typically results in the range 80 to 120%
would be considered acceptable
The overall standard deviation and relative
standard deviation may be used as an
estimate of precision
31
Table 1
Concentration ethyl
acetate % nominal
(with respect to
standard
concentration)
Ethyl acetate
added
theoretical
(l/100 ml)
Ethyl acetate
theoretical
(mg/100 ml)a
Ethyl acetate
determined
(mg/100 ml)
Recovery
(% w/w)
0b
NA
10
1.0
0.902
0.844
93.5
15
1.5
1.353
1.289
95.3
25
2.5
2.255
2.017
89.4
50
5.0
4.510
4.495
99.7
75
7.5
6.765
6.874
101.6
100
10.0
9.020
8.991
99.7
125
12.5
11.275
11.435
101.4
150
15.0
13.530
13.616
100.6
Mean
97.7
Standard deviation
4.4
% RSD
4.5
This figure is corrected for the density of ethyl acetate (0.902 g/ml)
The 0 level solution represents a test solution of drug substance to which no ethyl acetate was
added. A correction for the level of ethyl acetate found in this solution was made for all other test
solutions prepared.
NA Not applicable
b
32
1
1
1
1
2
1
2
2
3
1
2
1
4
1
1
2
5
2
1
1
6
2
2
2
7
2
2
1
8
2
1
2
Range
Determined from the linearity, accuracy and
precision data
34
35
18000
0.0
2.762 DMSO
0.809 EtOAc
20000
22000
1.086 ACN
uV
3.061 NMP
24000
LOQ chromatogram
1.0
2.0
Time (min)
3.0
36
Robustness
A number of attributes were evaluated prior to
validation using an experimental design
approach:
Injector temperature
Detector temperature
Initial temperature
Ramp rate
Hold time
Split flow
Column head pressure
37
Limit of quantitation
Linearity
39
References
ICH www.ICH.org
International Pharmaceutical Product Registration
Aspects of Quality, Safety and Efficacy, eds
Cartwright and Matthews, Ellis Horwood, 1994,
Chapter 8, Analytical validation
The validation of analytical methods for drug
substances and drug products in UK pharmaceutical
laboratories, G S Clarke, J. Pharm. Biomed. Anal.,
12. 643 652 (1994)
FDA www.FDA.gov
Technical review guide: Validation of chromatographic
methods
40