25/6/2014

Tuan Amran Tuan Abdullah, PhD

Institute of Hydrogen Economy
Universiti Teknologi Malaysia

SCREENING- FULL & FRACTIONAL

DESIGN
1

Screening
Aim identify significant factors
(variables)
A factor is significant if its influence
is greater than the noise level
(experimental error)
Usually carry out screening using
reduced designs such as factorial or
Plackett-Burman designs
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The trials in a factorial design can be represented

as points on an n-dimensional cube (n=3 in this case)

1,-1,1

1,1,1

11,-1

1,-1,-1

-1,-1,1
-1,1,1

-1,-1,-1

-1,1,-1

Slide 3

Case Study HPLC method

Aim: to optimise the separation of
peaks in a HPLC analysis

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Define the Response

The CRF (chromatographic response function) is used to quantify

separation of peaks. This function thus gives a single number to the
quality of a chromatogram. The aim is thus to maximise the CRF

Slide 5

Define the Factors

The factors studied in this study
were levels in the eluent of: Acetic Acid
Methanol
Citric Acid

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Experimental Domain
Low

High

Acetic Acid
(mol/L)

0.004

0.01

% Methanol

70

80

Citric Acid (g/L)

Slide 7

Factorial design (Coded form)

This design gives all combinations of the factors at 2 levels
+, high -, low

Run
Number

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Acetic
Acid

Methanol

Citric Acid CRF

Slide 8

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Factorial design (Uncoded)

This table shows the actual levels of the variables used in the experiments. Normally
the order of experiments is randomised but we will keep it in this structured forms so
you can see the patterns

Run
Number

1
2
3
4
5
6
7
8

Acetic
Acid

0.004
0.01
0.004
0.01
0.004
0.01
0.004
0.01

Methanol

70
70
80
80
70
70
80
Slide80
9

Citric Acid CRF

2
2
2
2
4
4
4
4

Results are inserted

here when the
experiments
are performed

Factorial design (Uncoded)

The CRF values are now inserted after the experiments
(chromatographic runs) are carried out

Run
Number

1
2
3
4
5
6
7
8

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Acetic
Acid

0.004
0.01
0.004
0.01
0.004
0.01
0.004
0.01

Methanol

70
70
80
80
70
70
80
Slide 80
10

Citric Acid CRF

2
2
2
2
4
4
4
4

10
9.5
11
10.7
9.3
8.8
11.9
11.7

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Analysis of the results - Excel

Calculate Main Effects this
calculates the effect on the
response solely due to one factor
Main effects are the difference
between average response at high
level of the factor average
response at low level

Slide 11

Calculation of Main Effects

Acetic
Acid
-1

Average of the high

values of CRF for each
variable e.g for AA =
(9.5+10.7+8.8+11.7)/4

Average of low
values of CRF for each
variable e.g for AA =
(10+11+9.3+11.9)/4

Methanol

Citric
Acid

CRF

-1

-1

10

+1

-1

-1

9.5

-1

+1

-1

11

+1

+1

-1

10.7

-1

-1

+1

9.3

+1

-1

+1

8.8

-1

+1

+1

11.9

+1

+1

+1

11.7

10.18

11.33

10.43

10.55

9.40

10.30

-0.37

1.93

0.13

The main effects

can also be
calculated by
multiplying the
variable column by
the CRF column
pairwise , adding up
the column and
then dividing by 4

The Main Effect is the difference between the high

and low average e.g for AA = (10.19-10.55)

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Calculation of Interactions
Interactions coefficients found by multiplying the appropriate variable columns
Acetic
Acid

Methanol

Citric
Acid

AA*M

AA*CA

M*CA

CRF

AA*M*CRF

-1

-1

-1

+1

+1

+1

10

+10

+1

-1

-1

-1

-1

+1

9.5

-9.5

-1

+1

-1

-1

+1

-1

11

-11

+1

+1

-1

+1

-1

-1

10.7

+10.7

-1

-1

+1

+1

-1

-1

9.3

+9.3

+1

-1

+1

-1

+1

-1

8.8

-8.8

-1

+1

+1

-1

-1

+1

11.9

-11.9

+1

+1

+1

+1

+1

+1

11.7

+11.7
Sum = 0.5

0.125

0.025

0.825

0.5/4 =
0.125

Interactions calculated by multiplying the CRF column and the

appropriate variable interactions column. To get the interaction effect
add up the column and divide by 4
Slide 13

Factorial Calculations using

Minitab

The program Minitab can be used to carry out

calculations as follows:-

To set up the design:

Stat > DOE > Factorial > Create Factorial Design
Type of Design: 2 level factorial design 9default
generators)
Number of factors: 3
Designs: Full Factorial
Factors see screen dump on next slide
Options: do not randomize (normally should randomize but for
the tutorial not randomizing makes it easier to see patterns in the layout)
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Slide 15

The generated FFD design as it

should appear in Minitab
Type the CRF responses here
after performing the experiments

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Slide 17

Factorial Calculations using

Minitab
To analyse the design:

Stat > DOE > Factorial > Analyse factorial Design

Click on C8 CRF as the response . Accept the default

values

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Factorial Calculations - Minitab

31/07/2007 11:26:30

Welcome to Minitab, press F1 for help.

Results for: Worksheet 2

Factors: 3 Base Design:

3, 8
Runs: 8 Replicates:
1
Blocks: 1 Center pts (total): 0

All terms are free from aliasing.

Design Table

Run A B C
1 - - 2 + - 3 - + 4 + + 5 - - +
6 + - +
7 - + +
8 + + +

Full Factorial Design

Minitab Output

Slide 19

Factorial Fit: CRF versus Acetic Acid, Methanol, Citric Acid

Estimated Effects and Coefficients for CRF (coded units)
Term
Effect Coef
Constant
10.3625
Acetic Acid
-0.3750 -0.1875
Methanol
1.9250 0.9625
Citric Acid
0.1250 0.0625
Acetic Acid*Methanol
0.1250 0.0625
Acetic Acid*Citric Acid
0.0250 0.0125
Methanol*Citric Acid
0.8250 0.4125
Acetic Acid*Methanol*Citric Acid 0.0250 0.0125

Note , however,
coefficients are simply
twice the effects so
no new information

Main Effects

Interactions
Coefficients from fitting to a second order equation
Y = bo+b1*x1+b2*x2+b3*x3+b12*x1*x2+b13*x1*x3+b23*x2*x3+b123*x1*x2*x3
Where x1 is acetic acid , x2 is methanol and x3 is citric acid
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What do the results tell us?

The main effects tell us which variable has the
strongest effect on the response (CRF) in this
case methanol has the strongest effect on CRF

A negative effect means the response is reduced

as the variable increases. The negative effect for
acetic acid means that as we increase the
concentration of acetic acid, the CRF gets
smaller (and hence our separation is worse)

Slide 21

What about interactions?

An interaction effect is where the
effect on the response of one
variable depends on the level of
another variable.
In this study methanol and citric
acid seem to have the largest
interaction.
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Main Effects and Interactions

Plots
Main effects plots help to visually display the
variable effect. They graph the average response
at the high and low levels. The steeper the
graph, the stronger the effect
The plots can be drawn in Miniab as follows: Stat > DOE >Factorial > factorial Plots
Tick Main Effects Plots
Setup > Select CRF as the response and choose
all 3 variables (>>)
The Interactions plots can be produced similarly
(just select Interactions instead of Main Effects
Plots)
Slide 23

CRF average at high methanol

Main Effects Plot (data means) for CRF

Acetic Acid

11.5

Methanol

11.0
10.5

Mean of CRF

10.0
9.5
0.004

0.010
Citric Acid

11.5

70

80

CRF average at low methanol

11.0
10.5

Note that Methanol has the steepest

slope, indicating the strongest effect

10.0
9.5
2

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Interaction Plot (data means) for CRF

70

80

11
A cetic A cid

Acetic
Acid
0.004
0.010

10
9

11
Methanol

Methanol
70
80

10

The plots show there is

an interaction effect with
methanol and citric acid
at high methanol CA has a
Positive effect but at low
methanol it has a negative
effect on CRS

C itr ic A cid

Slide 25

Conclusions
Methanol has the largest effect on
CRF
The Methanol effect strongly
depends on the Citric Acid level.
Citric acid has a positive effect at
high Methanol but a negative effect
at low Methanol
All 3 variables do seem to affect the
result. Citric acid has the smallest
main effect but large interaction
effect
Hence probably cant screen out
any of these variables from further
study
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Significance Normal Probability

Plots
Normal Probability Plots are used to test
whether data is normally distributed.
In our case, we can use such a plot to test for
significance of the effects/coefficients
If the effects are not significant we expect
variations just to be due to random error and
this can be tested with the plots. It is only a
guide, however, as we have no real estimate of
the experimental error
In Minitab the plot can be generated:
Stat > DOE >factorial > Analyse Factorial Design
> Graphs and Select Effects Plots (Normal)
27

Normal Probability Plot of the Effects

(response is CRF, Alpha = .05)

99

Effect Type
Not Significant
Significant

95
B

90

Percent

80

BC

70
60
50
40
30

F actor
A
B
C

N ame
A cetic A cid
M ethanol
C itric A cid

Effects due to random errors should be

on a straight line. This plot indicates
Methanol and the Methanol/Citric Acid
interaction are significant effects

20
10
5
1

-0.5

0.0

0.5

1.0

1.5

2.0

Effect
Lenth's PSE = 0.1875

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Problems
We have not replicated any experiments
so no determination of error. We cannot
tell if the coefficients (effects) overall are
significant (although normal probability
plots help). We can only compare them
to see which is the most significant
We also cannot test for curvature i.e
are the effects of the variables linear. A
non-linear effect can be when the
response at the high and low levels is
similar but at intermediate values is much
higher or lower. pH effects are often nonlinear
Slide 29

Solution?
Add centre points!!
Centre points are
experiments with all
variables set at 0
(coded) i.e. mid values
Replication of the
centre point allows
determination of error

Coded

Uncoded

Acetic
Acid

0.007

Methanol

75

Citric Acid

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Acetic Acid

Methanol

Citric Acid

-1
1
-1
1
-1
1
-1
1
0
0

-1
-1
1
1
-1
-1
1
1
0
0

-1
-1
-1
-1
1
1
1
1
0
0

CRF
10.0
9.5
11
10.7
9.3
8.8
11.9
11.7
10.2 Results added
for the centre points
10.4

Slide 31

Estimated Effects and Coefficients for CRF (coded units)

Term
Effect Coef
SE Coef
T
P
Constant
10.362 0.05000 207.25 0.003
Acetic Acid
-0.3750 -0.1875 0.05000 -3.75 0.166
Methanol
1.9250 0.9625 0.05000 19.25 0.033
Citric Acid
0.1250 0.0625 0.05000 1.25
0.430
Acetic Acid*Methanol
0.1250 0.0625 0.05000 1.25
0.430
Acetic Acid*Citric Acid
0.0250 0.0125 0.05000 0.25
0.844
Methanol*Citric Acid
0.8250 0.4125 0.05000 8.25
0.077
P is the probability a coefficient is not significantly
different from zero i.e no effect on CRF. A low probability
(< 0.05 at the 5% level) indicates high significance.
The methanol effect is the only significant one at the 5% level
although the methanol-citric acid effect is just above the 5% level

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Main Effects Plot (data means) for CRF

Acetic Acid

11.5

Methanol

Point Type
Corner
Center

11.0
10.5

Mean of CRF

10.0
9.5
0.004

0.007

0.010

70

75

80

Citric Acid

11.5
11.0

The centre point responses are all

on the linear response line. Thus
no curvature is indicated.

10.5
10.0
9.5
2

Slide 33

Normal Probability Plot of the Standardized Effects

(response is CRF, Alpha = .05)

99

Effect Type
Not Significant
Significant

95
B

90

Percent

80
70

F actor
A
B
C

N ame
A cetic A cid
M ethanol
C itric A cid

The Normal Plot shows also that

Methanol is the only significant
effect but Methanol/CA interaction is
probably also significant

60
50
40
30
20
10
5

-5

5
10
Standardized Effect

15

20

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Fractional Factorial Design in

Minitab
The method for creating fractional
factorial very similar to the method
for creating full factorial design

35

Example Fractional design

StdOrder RunOrder CenterPt
1
1
1
2
2
1
3
3
1
4
4
1
5
5
1
6
6
1
7
7
1
8
8
1
9
9
1
10
10
1
11
11
1
12
12
1
13
13
1
14
14
1
15
15
1
16
16
1

Blocks
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1

A
-1
1
-1
1
-1
1
-1
1
-1
1
-1
1
-1
1
-1
1

B
-1
-1
1
1
-1
-1
1
1
-1
-1
1
1
-1
-1
1
1

C
-1
-1
-1
-1
1
1
1
1
-1
-1
-1
-1
1
1
1
1

D
-1
-1
-1
-1
-1
-1
-1
-1
1
1
1
1
1
1
1
1

Rate
45
71
48
65
68
60
80
65
43
100
45
104
75
86
70
96
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Normal Plot of the Standardized Effects

(response is Rate, Alpha = 0.05)

99

95

90

F actor
A
B
C
D

AD

80

Percent

Effect Type
Not Significant
Significant

70

60
50
40
30

N ame
A
B
C
D

20
10

AC

-5

0
Standardized Effect

10

41

Half Normal Plot of the Standardized Effects

(response is Rate, Alpha = 0.05)

Effect Type
Not Significant
Significant

98

F actor
A
B
C
D

95
A

Percent

90
85

AC

80
AD

70

60

50
40
30
20
10
0

N ame
A
B
C
D

2
3
4
5
6
7
Absolute Standardized Effect

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Pareto Chart
Pareto Chart of the Standardized Effects
(response is Rate, Alpha = 0.05)

2.571

The
significant
rank order
A>D>C>B

F actor
A
B
C
D

A
AC
AD

N ame
A
B
C
D

Term

D
C
B
BC
CD
BD
AB
0

4
5
6
Standardized Effect

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