1 Experimental Designs I Stat 162 1

Preface
This lecture syllabus was a compilation of lectures notes written over decades for the
course Experimental Design I (currently with course no. STAT 162) at the Institute of
Statistics (InStat), University of the Philippines Los Bafios. The materials used were
collected from the writer's file amassed from designing and analyzing experimental
data while he was working at the International Rice Research Institute (IRRI),
Philippine Sugar Commision (tben Philippine Sugar Institute and now the Sugar
Regulatory
Administration) and also at the Statistical Consulting Group of INSTAT.
Parts of the lecture notes were first distributed as handouts. Others were written in
longhand on transparencies and shown on overhead projectors. These notes were refined
as lecture papers used in the numerous computer-based training workshops on statistics
conducted by the INSTAT.
Computations illustrating the statistical analyses were done on SAS (SAS Institute, Inc.)
and SPSS (SPSS Inc.) as well as using the calculator. This lecture syllabus is being
supplemented with exercises during the laboratory session of the course. These
sessions are computer-based using statistical softwares to carry out the computations.
This lecture syllabus has not yet undergone any editing by a competent editor. Many
typographical, grammatical, syntax and other editorial errors may be found. These errors
are also concerns later but the immediate concern now is to reproduce this syllabus for a
handy reference on the course.
Although this syllabus is, intended for STAT 162, it may also come in handy for
researchers and students who are conducting experiments for their researches.
Other INSTAT faculty who have contributed to this syllabus are: Dr. Arturo Y.
Pacificador, Jr, Prof. Lina A. Catahan, Dr. Concorcia E. Reafio and Prof. Mae
Solivas
Cababasay. The writer also thank Dr. Santiago M. Alviar from whom he first learned of
design and analysis of experiment.
Gratitude are given to Ms. Lita O. Averion and Mr. Sonny E. Nerpio who took care of all
the works to the University Publishing Center of UP Los Bafios.

Table of Contents
Cha ter To ics Pa e
Introductory Concepts 1-1
The role ofStatistics in research 1-1
Basic concepts on statistically designing an experiment 1-2
Steps in conducting an experiment 1-2
Definition of common terms used in experimental design 1-3
Princi les of ex erimental desi 1-4
2 One-way Classification Design: CRD 2-1
Randomization and layout 2-1
Data presentation and linear model Anal 2-1
sis of data 2-2
3 Assumptions Underlying the Analysis of Variance 3-1
Homogeneity of error variances 3-1
Normality of experimental errors 3-2
Independence of experimental errors 3-3
Additivity of effects 3-3
Some remedial measures 3-5
4 Multiple Comparisons Among Means 4-1
Pairwise mean comparisons 4-1
Group comparisons Trend 4-8
com arisons 4-12
5 CRD With Subsampling 5-1
Data presentation and linear models 5-1
Model of Effects 5-2
Anal sis of data 5-2
6 Two-way Classification Designs 6-1
The Randomized Complete Block Design 6-1
Data presentation and linear model 6-2
Analysis of data 6-3
The Latin Square Design 6-6
7 Two-Factor Factorial Experiments 7-1
Kind offactorial treatment effects 7-1
Two-Factor Factorial Experiment in RCBD 7-4
Three-Factor Factorial Experiment in RCBD 7-11
Table of Contents
Cha ter To ics Page
8 Split-plot Designs 8-1
Split-plot in RCBD 8-1
Strip-plot design 8-6
Randomization and layout
Data presentation 8-7
Strip-split-plot design 8-10
Data presentation 8-1 1
Analvsis of data 8-1 1
9 Regression and Correlation Analysis 9-1
Simple linear regression 9-2
Pearson correlation coefficient 9-6
10 Analysis of Covariance 10-1
ANACOVA in RCBD 10-2
The ANACOVA linear model
References
A endix Tables
Experiment"/ Devizns I Page I-I
CHAPTER 1. INTRODUCTORY CONCEPTS
THE ROLE OF STATISTICS IN RESEARCH

Research is a studbus, cntjca! investjgation into the nature of, reasons for, and
the consequences of a set of circumstances.
Objective. discover/ of new facts and at revising accepted ±.eories taws in the
light of newly discovered facts.
E/emen&. observations, analysis and reasoning.
In scientific research, statistics deals with:

Designing experiments and surveys
Collecting and summarizing data
Describing the data and the variables Estimating population parameters
Testing hypotheses about the populations
Studying relationships among variables
Data
Collection
Data
Data
Examination
Data
Analysis
Fig. 1; Statistical methods in research (Rag 1989).

Statistics, when proper/y used, makes efficient research. However, it is not a
remedy for poor/y conceived and badly executed research. Thus, it is essential
that d user ofstatistics dear/y understands the techniques he employs.
Terry S. Solivas, T-CAS, UP Los

Page 1-2
Chapter l. Inntroductory Concepts
BASIc CONCEPTS ON STATISTICALLY DESIGNING AN

EXPERIMENT
A researcher conducts an experiment to obtain data. Based on results of data analysis,

conclusions are stated and inferences are made regarding the population.
There is element of random error in the result of the experiment; hence, perfect
generalization of the result can not be made.
However, if the experiment is performed using appropriate principles, design and
techniques of experimentation, then the degree of uncertainb/ can be measured in
terms of probability, and so probabilistic statements and conclusions can be made.
• Experimental design - the set of rules, plans and course of action taken in the
conduct of an experiment.
It is needed to :
• Ensure cost effective collection of appropriate date
• Provide an appropriate and valid analysis of data

• Provide reliable conclusions leading to reliable inferences.
Ronald A. Fisher "To consult a statistician after an experiment is finished is often
merely to ask him to conduct a post mortem
examination. He can perhaps say what the experiment died
of. "@
Steps in Conducting an Experiment
1. State the problem of study.

•population and variables of the study
• factors influencing the variables,
• objectives of the problem.
2. Formulate the hypotheses.
3. Define the data to collect, the statistical analyses of data.
4. Devise the experimental technique and design.
Terry S. Solivas, INSTA T-CAS, UP Los Banos
Page 1-3
• define the treatments, experimentai units, size of the experiment.

• draw the experimental layout.
5. Conduct the experiment.
6. Analyze the data obtained from the experiment.
7. Summarize the results, make conclusions and inferences.
8. Evaluate the whole study in relation to similar investigations.
E.xperimental Designs i
Definition ofCommon Terms Used in Experimental
Treafrnents are procedures or conditions whose effects are to be measUred

and compared.
Examples: Kinds of fertilizer, rates of irrigation, age of harvest, different locations,

food formulations, marketing strategies, teaching methods.
Experimental unit is a group of experimental materials or individuals to which a

sing:e treatment is applied once.
Examples: A plot of land, a plant, a portion of a leaf, a dish of culture medium, a

production run, a class of students, a cage of birds, a marketing day.
Response variable is a characteristic used to measure the treatment effects.

Examples: Yield of the crop, height of the piant, degree of infestation, biomass
production, gain in weight, volume of sales, GPA.
Sampling unitis a portion of the experimental unit on which the

response variabie is observed and measured.
Examples: A tiller in a hill, a plant in a pot, a seedling in a seedbed, a tree_in a

group of trees, a branch of a tree, a farmer in a barangay, a sample in a
batch.
Note: Sampling unit may be the whole experimental unit.
Experimental error is the measure of variations among experimental units

treated alike.
Some sources of experimenta/ errors.
Terry S. Solivas, INSTA T-CAS, UP Los

Page 1-4
Inherent variability of the experimentai materials, e Errors in

experimentation
lack of uniformity in the conduct of the experiment
failure to standardize the experimental technique
Errors in observations and measurements
Combined effects of all extraneous factors
0
Sampling en•oris the measure of variations among sampiing units within
experimental unit.
Buyios
Chapter I. Inntroductory Concepts
Precision denotes repeatability of measurements. It is measured by the

variance.
Some ways ofincreasing precision are:
Increase the number of samples or replications,
Skillful grouping of experimental materials,
Proper selection of treatments.
+ Accuracy denotes unbiasedness or the closeness of the average values of the

measurements to the true value.
Some ways ofihcreasing accuracy are.
Refinement of experimental technique and

Proper selection of treatments.
Layout refers to the final arrangement of treatments over the whole

experimental area.
Principles ofExperimental Design
A valid experimental design must have these three principles: replication,

randomization and /oca/ control.
Replication is the repetition of the application of treatments on a number of

experimental units.

Page 1-5
Functions ofreplication
To provide an estimate of thé experimental error.
to enable test of significance and interval estimation.

To increase the precision of estimates. e To increase the scope of the
experiment. Factor affecting the number ofreplications e Degree of
precision required.
more replications for higher precision desired

Uniformity of experimental units.
more replications for less uniform experimental units

Page
Experimental Designs I-S
Number of treatments.
For a required precision, smaller number of treatments is compensated by more

replications.
Experimental design.
Ttme allotment for the experiment.
Cost and availability of resources.
Randomization is the allocation of treatments to the experimental units by

means of a chance device sush that every treatment has equal chance of
being assigned to any experimental unit.
Functions of randomization:
Provide a random sample of observations.

To satisfy the assumption of independence of observations.
To eliminate systematic bias in assigning the treatments.
Local control orerror control is any process or technique used to minimize

the experimental error.
Common Techniques for Local Centro!
Proper choice of experimental design,
Proper choice of size and shape of experimental units,

Use of concomittant variable,
Refinement of experimental technique o Uniform application of treatments and

management o More control over external influences o Devising unbiased
measurement technique o Preventing gross errors.
Bafios

Experimental Designs I Page 2-1
CHAPTER 2. ONE-WAY CLASSIFICATION DESIGN: CRD
CRD is a design wherein the allocation of treatments is done by randomizing the

treatments completely over the entire experimental units (eu's) without any
restriction imposed on the units and there is only one criterion for data
classification.
CRD is commonly used when:
1. The eu's are sufficiently homogenous (like dishes of culture medium) and
2. Effective local control is assured (as those in laboratories, greenhouse).
> Randomization and layout
Suppose there are t = 3 treatments, T1, T2 and T3, which are replicated rl = 2, r2 = 3
and = 4 times, respectively; hence, the nos. of eu's required is n = rt + r2 + = 9. The
randomization (using random no. generator key on calculator) may be as follows:
I. Label the eu's consecutively from 1 to n = 9.
2. Obtain a sequence of n = 9 random numbers. Rank the numbers in increasing
order. Using the sequence of ranks as a randomization of the eu's, assign the first rl
= 2 eu's to Tl, the next r2 = 3 eu's to T2 and the last = 4 eu's to T3.
Random no. . .739 .218 .781 .396 .527 .324 .463 .064 .942 rank(eu
no.): 7 2 8 4 6 3 5 1
9
The corresponding /ayoutfor this randomization randomization procedure is:
1 2 3
4 5 6
7 8 9
> Data Presentation: (One-way classification)

Observations Reps Total Mean
Treatment Yi.
Irl
22 2r2 2.
t.
t2 trt

Page 2-2
Chapter 2. One-Way Classification Design
Linear Model:
Let Yij = observation on the response variable from the jth experimental unit with
the ith treatment.
Yij + tij;
where: = genera! mean of ai! possible observations,

= the effect of the ith treatment,
= random error associated with the jth eu with the ith
treatment, = + ti, the true mean of the ith treatment.
> Analysis ofData:

Illustration 1:
Three methods of soil analysis, Ml, M2, M3, were tried by a research institute. Twelve
uniform soil samples were taken from a certain farm and Ml was randomly assigned to
5 of the soil samples, M2 to 3 sampies and M3 to 4 samples. The researcher was
iinterested in the time to complete the soil analysis. The data on the times
(in hours) of the soil analyses were summarized as follows:
Method Time to com lete anal sis hrs Re s Total Mean

2.4 3.8 2.9 4.6 3.1 5 16.8 3.36
M 4.8 1.6 0.2 3 6.6 2.20
7.2 5.3 2.9 3.5 4 18.9 4.72
12 42.3 3.35
Terry S. Solivas, UP Los

At the level of significance test the hypothesis that there is no difference in the
u = 5%,
3. State the decision rule

Reject Ho and accept Ha if FC Ftab; else, accept Ho.
Alternatively, reject Ho and accept Ha if Prob(Fc Ftab) < cc, else, accept Ho.
4. Construct the ANOVA tab/e outlined as fo//ows:
sv df MS
Due to treatments t-1 TrSS MSTr MSTr/MSE
Ex erimentai error n-t ESS MSE
TOTAL n-l TSS
Computations:
Let Yo = jth observation on the ith treatment,
= total of observations on the ith treatment, Y..
= grand total of all observations.
then compute
a. the sums ofsquares
CF = (42.3)2 = 149.11 n 12
2
+ + 3.52] - 149.11 = 36.50
2
18.92

+ 3.8
Page 2-4
6.6 - 140.11 = 11.16
3
4
E
SS = TSS - TrSS = 36.50 - 11.16 = 25.34
b. the mean squares:

MSTr = TrSS/(t-1) = 11.16/2 = 5 58
MSE = ESS/(n-t) = 25.34/9 = 2.82
c. the test statistics:
• FC = MStr/MSE = 5.58/2.82 = 1.98 and the
critical value: hab = = 4.26

Page 2-5
2. One-Way Clmssification Design
Chapter
d. Set up the ANOVA table:

sv df MS
Treatment 2 11.16 5.58 1.98ns 4.26
Error 9 25.34 2.82
TOTAL 11 36.50
ns — not significant at 5% level.
5. State the decision and make the conclusion.

It is concluded that there is no significant differences in the mean time to
complete the soil analysis for the three methods .
Note: The observed differences among the means in the experiment are not
large enough to warrant that in the population (or in genera!) there are
at least two means which are different.
Compute other summary statistics

Experimental Designs I Page
1. Coeficient of variation, CV(%) is the experimental error expressed as percentage
of the mean. It measures the degree of precision of the experiment or as an index
of the reliability of the experiment. The higher the CV, the lower is the reliability
of the experimental results.
MSE 2.82
x 100 = x 100 = 47.60%
3.52
2. Standard error ofa treatment mean, s.e.(ül ) is the measure of the average error in
estimating the true treatment mean. It measures the degree of precision of Y, as
the estimate of the true treatment mean.
MSE
s.e.(Y, ) =
3. Standard error of the difference between two treatment means, s.e.(f', -t. ) is a
measure of the average error in estimating the difference between two treatment
means. It measures the degree of precision of (t-t. ) as the estimate of the difference
between the true means of treatment i and treatment i'.
MSE —+—
s.e.(V, = 1
2-5
Estimates of treatment means and effec&:
1. Estimate of true mean of treatmentl, g,

For treatment I: = Yl- = 3.36
For treatment 2: = — 2.20
For treatment 3: = 4 72
For general mean: p = Y- = 3.35
b. Estimate of the effect of treatment i; = -Y
For treatment = 1: - Y = 3.36 - 3.35

= 0.01 For treatment 2: = =
2.20 - 3.35 = -1.15 For treatment
3: = 4.72 - 3.35 = 1.37
Summarize the treatment means and effects:

Method Means Effects
3.36 0.01
2.20 -1.15
3 4.72 1.37
Mean 3.35 0.00
Note: Since the ANOVA test showed no significant effects, the observed effects
can not be generalized to be significant.
3-1
CHAPTER 3. ASSUMPTIONS UNDERLYING THE ANALYSIS OF
VARIANCE
Inferences based on the ANOVA are meaningful only when the data set satisifed certain
characteristics, called assumptions underlying the ANOVA. These are:
1. Homogeneity of variance of the experimental errors,

2. Norma/ distribution of the experimental errors,
3. Independence of the experimental errors, and
4. Additivityof the treatment effects and environmental effects.
@ In practice, many experimental data violates some of these assumptions. Such
violations affect the level of significance and the sensitivity of the F-test. Analyzing
such data with thp usual statistical techniques can produce results that lead to dubious
or meaningless interpretations.
Homogeneity ofEnor Variances

@ The most serious consequences on the resulte happen when the experimental error
variance is not constant across groups or population (also called heteroscedasticity).
This situation occurs when some treatments are erratic in their effects; there are
outliers due to the nature of the experimental materials. Some kind of data (e.g.
binomial, Poisson and other count and percentage data) likewise gives heterogeneous
variances.
Using the Bartlett's Test
a. Ho:the variances are homogenious

Ha:
the variances are heterogenious
b. The Bartlett's test statistic is

U = 2.30269 h where: q = (n-
t)log10Sp —E (ri —1)
log10Si ;
Si2 = sample variance of the ith treatment (group) n-t
30-1)
c. Decision rule: Rejected Ho (and accept Ha) if U > or Prob(U ) where a is the
prescribed level of significance.

Page 3-4
Chapter 3. Assumptions Underlying the ANOVA
Independence ofExperimental Errors

Responses on neighboring plots tend to be correlated, adjacent plant have some form
of symbiotic relationship or competition for light, water and other environmental
factors. Neighboring households are closely related.
@ Independence of errors is also violated in experiments where the experimental units or

replications are arranged systematica'ly.
& The remedy is proper randomization of treatments to the experimental units to give
independent errors.
& Plot of the Residua/
A residual plot.may indicate the presence of correlation of the experimental errors.
Independent errors Correlated errors
Other procedures.
Durbin-Watson test for seria/ correlation

Runs test for randomness.
Additivity of Effects
€9 A common departure from this assumption in research is one where the treatment
and the environmental effects are multiplicative. Multiplicative effect is encountered
in disease studies because the effects of disease organisms are usually in multiples of
the numbers present.
Table 1. Data set A. Data with additive effect.

Treatment Block 1 Block2 Block
effect
100 150 50
250 300 50
Treatment effect 150 150
Terry S. Solivas, T-CAS, UP Los Banos

The effects of treatments across blocks are uniform (150) and the
effects of blocks across treatments are also uniform (50).
Table 2. Data set B. Data with multiplicative effect .
Treatment Biock 1 Block2 Block

effect
100 150 50
250 375 125
Treatment 150 60% 250 60%
effect
The effects of treatments across blocks are not uniform and
the the effects of blocks across treatments are also not
uniform but the effects are uniform if expressed in
percentage.
Table 3. Log?rithmic transformation of data set B.
Treatment Block 1 Block2 Block

effect
2.0 2.2 0.2
2.4 2.6 0.2
Treatment 0.4 0.4
effect
-s When a set of data has multiplicative effects, a logarithmic
transformation will convert the data to have additive effects.
The Tukey's test ofaddivity of effects
Some Remedial Measures

The usual remedy for some violations of the assumptions of ANOVA is to find an
appropriate data transformation. The simple transformations that have been
found useful are the logarithmic, square root and arcsine transformation. The
general one is by the Box-Cox transformation.
Terry S. Solivas, UP Los Banos

Page 3-6
1. Logarithmic transformation
• This is appropriate for data where its standard

deviation is proportional to its mean as in data with
greatly skewed distribution and in data with
multiplicative effects. Count data usually do not
satisfy the additivity assumption.
• The transformation is y; = log(YiJ +1)

The 1 is added when the data contain o's and very small values.
INSTAT-CAS,
Chapter 3. Assumptions Underlying the ANOVA
2. Square root transformation
This is appropriate for data where its variance is proportional to its mean as in
data with Of small whole numbers like counts of rare events (e.g. Poisson data) and
frequency count data.
The transformation is
Y,} = Y!+O.05
The 0.5 is added when the data contain small values (say less than 10) or when o's
are present.
3. Arcsine transformation
This is appropriate for proportion or percentage data (e.g. binary data).
The transformation is
After transformation the values are checked for possible improvement. The ANOVA
and subsequent analyses wilt be done on the transformed values.


CHAPTER 4. MULTIPLE COMPARISONS AMONG

MEANS
When the F-test in the ANOVA is significant at the prescribed level of significance,
it reveals that at least two treatment means are significantly different. The test,
however, does not locate the specific means which are significantly different.
Thus, the next question is "which means are different and by how much are the
differences?" This can be answered by testing multiple comparisons among the
means to locate which among the means are significantly different from each
other.
The three kinds of multiple comparisons among treatment means are:

painvise means comparisons, group comparisons and trend comparisons.
I. Pairwise Mean Comparisons

The pairwise mean comparisons compares pairs of treatment means and tests if they are
significantly different at a prescribed level of significance.
The statistical hypotheses are:
Ho: - Eli, = 0 The treatment means, and are not different Ha:
0 The treatment means, and are different
The commonly used tests for pairwise mean comparisons are:

1. Least significant difference test (LSD) test
2. Duncan's multiple range test (DMR7)
3. Student-Newman-Keul (SNR) test
4. Honest significant difference (HSD) test,
5. Scheffe's (S) test.
Other tests are obtained from statistical softwares.
& For illustration, consider the results from an experiment conducted to measure the
effects of six organic fertilizer treatments and a control treatment on the yield of
soybean.
ANOVA
sv df MS
Treatment 6 5 587 174 931 2.57
196
Error 21 1 990 238 94
773
TOTAL 7 577 412
= 15 1 se(Yi - ) = 153.9 se(d) = 217.7
Page 4-2
Chapter 4. Multiple Comparisions Among Means
Treatment means:
INSTA
test
• The /eastsignificant difference (LSD)
Some features and when to use:

1. It is used only when the F-test in the ANOVA is significant.
2. It may not be used when there are more than four tretments to test. But
may be is used for preplanned comparisons regardless of the number of
treatments.
3. It is used when the experiment is with equal or unequai replications.
4. It is the least conservative among painuise comparisons test..
5. It uses the Student t table.
Test procedure:
1. Test statistic:
LSD = ta x se(d) - = = 453
(error df)
2
2 Decision rule:
Reject Ho: gl - = 0 (and accept Ha: - P, O) if ( t - t ) 2 LSD
3. Perform the pairwise comparisons ofthe means
a. Sort the means by arranging them from largest to smallest.
b. Compare the largest mean to each of the smaller means starting from
the next smaller mean. Connect nonsignificant comparisons by a sideline.
c. Compare the next largest means as in (b).
d. Repeat (b) and (c) until all the means have been compared.
Sorted Treatment Means Significance
org C 2,678
org D 2,552

org E 2,128 Org
A 2,127 org B 1,796
Org F 1,681
Control 1,316
e. Label the line groups alphabetically
4. Summarize the result ofcomparisons:
Treatment Means*
Or A 2 127 b
Or B 1 796 c
Or C 2 678 a
Or D 2 552 ab
Or E 2 128 bc
1 681 cd
Control 1 316 d
* Any two means having a common letter(s) are not
significantiy different; otherwise, they are significantly
different (at 5% level using LSD test).
The Duncan's Multi /e Ran e Test DMRT

1. The DMRT is more sensitive than the F-test in the ANOVA. Thus it may still be
used even if the F-test is not significant.
2. It is used to compare differences between all possible pairs of means.
3. It is a sequentiai test; that is, it requires a series of values of the test statistic,
each corresponding to a specific set of pair comparisons.
4. it is used when the experiment is with equal replications.
5. It uses the new studentized range table.
Ø Test procedure:
1. Test statistic
Let p = range of the number of means in a particular comparison where
counting starts from the higher mean to the iower mean; p = 2, 3

Page 4-4
Correponding to each p values, at the a level of significance and error df,
obtain the rp values from the new studentized range table.
For each rp, compute Rp as the values of the test statistic by
Rp = rp x se(Yi )
In the illustrative example: Using cc = 5%

p 2 3 4 5 6 7
2.95 3.10 3.18 3.25 3.30 3.34
454 477 489 500 508 514
2. Decision rule:
Reject Ho: - y, = 0 (and accept Ha: - p, * 0) if (t 2 Rp
3. Pefform the pain-vise comparisons ofthe means
org C 2,678 org

D 2,552
org E 2,128
Org A 2,127
Org B 1,796
Org F 1,681
Control 1,316
d
e. Label the line groups alphabetically
4. Summarize the result of comparisons:
Treatment Means*
Or A 2 127 b
Or B 1 796 c
Or C 2 678 a
Or D 2 552 ab

Or E 2 128 bc
1 681 cd
Control 1 316 d
* Any MO means having a common letter(s) are not
significantly different; otherwise, they are
significantly different (at 5% level using DMRT).

Page 4-6
• The Student-Newmann-Keu/:s ($NK) Test
1. It is used only when the ANOVA F-test is significant.

2. It requires equal replications.
3. It accounts for the number of treatments in the experiment.
4. It is also a sequential test.
5. It makes use of the studentized ranged table.
rest procedure:
1. Test statistic:
• This test proceeds as in the DMRT. Corresponding to each p values, at

level of significance and error df, obtain the qp values from the studentized
range table.
For each qp, compute Wp as the values of the test statistic by
Wp = qp X se(Yi )
• In the illustrative example: Using cc = 5%
p 2 3 4 5 6 7
2.95 3.58 3.96 4.23 4.45 4.62
w 454 551 609 651 685 711
3. Decision rue:
Reject Ho: - Eli, = 0 (and accept Ha: - O) if (Yi -t ) Wp

3. Perform the painuise comparisons of the means
(Assignment)
(Assignment)

The Tuke 's Honest Si niFcant Difference HSD Tes
Some features and when to Use:
1. It is used on!y when the F-test in the ANOVA is significant.
2. It is an exact test for any number of means to compare.
3. There is only one critical value of the test statistic to use.

4. It is more conservative than LSD and DMRT.
5. It is used when the experiment is with equai number of replications.
Test procedure:
1. Test statistic:
• Let qp be the tabular value of the studentized range table at (1 level of
significance, error df and p = t,
Compute the value of the test statistic HSD as:
HSD = qp x sect)
• In the illustrative example: At the = 5%

Page 4-8
HSD = = 711.0
2. Decision ru/e:
Reject Ho: - = 0 (and accept Ha: - 0) if HSD

3. Perform the painv;se comparisons of the means
org C 2,678 org D

2,552 org E 2,128
Org A 2,127 org B
1,796 org F 1,681
Control 1,316
c
Label the line groups alphabetically
Treatment Means*
Or A 2 127 ab
Or B 1 796 bc
Or C 2 678 a
Or D 2 552 ab
Or E 2 128 ab
Or F 1 681 bc
Control 1 316 c
* Any Ovo means having a common letter(s) are not
significantly different; otherwise, they are significantly
different (at 5% level using HSD).
The Scheffe's S Test
1. It is based on an F-tabular values.

2. It can be used to test any mean comparisons.
3. It can be used for treatment with equal and unequal reps
4. It is the most conservative of all the pairwise tests.

1. Test statistic:
• Let Ft be the tabular value of the F table at level of significance with the df's
(trt df, error df). Compute the value of the test statistic S as:
(t-1)Ft x se(d)
• In the illustrative example: At the a = 5%
x (217.7) = 841
2. Decision ru/e:
Reject Ho: - = 0 (and accept Ha: - * 0) if (Y. - t.) 2 S

Perform the painvise comparisons ofthe means. (As/gament)
4. Summarize the resu/t ofcomparisons. (Assignment)
Il. Group Comparison

When the F-test in the ANOVA is significant; the next step in the analysis may be to
compare specific groups of treatments.
• Linearcomparison. Let P2, ... , be t population means. The linear function expressed as
= Cl!ll +C2P2 + ... + Ctyt
is a linear comparison (or linear contrast) among the t population means if
where the coefficients q,'s are fixed constants.
The sample contrast analogous to is the linear function L of treatment means

as
The meaning of the contrast is defined by specifying the qts.

Page 4-10
• O"hogonalcontrasts. Two contrasts, and

= Cllktl +0-1412 + ... + = +C22ß2 + +
are said to be orthogona/if
= CllC21 + C12C22 + + CltC2t = O
• Set of Orthogonal contrasts. The set of t-1 contrasts, 11, 12, ...t is a set of orthogona/
contrasts if they are all pairwise orthogonal.
Ilfustration:
Consider the following results from an experiment conducted to determine the
yield (tons/ha) response of a certain variety of rice to different kinds of fertilizer.
Trt Fertilizer and a lication Reps Total Means
T Control no fertilizer a !ication 4 15.32 3.83
Complete fertilizer (14-1+14) 4 31.20 7.80
Or anic fertiiizer A, single application 4 26.84 6.71
Or anic fe@lizer A, split application 4 29.20 7.30
O anic fertilizer b single application 4 16.12 4.03
Or anic fertilizer B s lita iication 4 19.68 4.92
ANOVA
sv df MS
Treatments 5 59.45 11.89 49.54* *
Error 18 4.30 0.24
TOTAL 23 63.95
Since the ANOVA shows that the treatment means are significantly different, we may
compare groups of these means.
A set of meaningful comparisons (contrasts) are:
1. Comparing the mean of the control with the means of afi treated. This comparison is
defined by specifying the q's as

and the sample contrast Ll is
5Y1 - + % + Y4 + Ys + % ) contro/vsfreated
2. Comparing the mean of complete fertilizer with the means of organic ferti!izers
and the sample contrast is
-(+++) --5 complete vs organic
3. Comparing the means of organic fertilizer A with the means of organic fertilizer B.
and the sample contrast L3 is

Chapter 4. Multiple Comparisions Among Means Page 4-12
4. Comparing organic fertilizers A single with A spiit.
and the sample contrast 1.4 is
u = - 94 -z A single vs A split
5. Comparing organic fettilizers B single with B spiit.
and the sample contrast 1-5 is
6. B sing/e vs B sp/it
Other meaningful contrasts are:
1. Comparing single against split applications of organic fertilizers.
2. Comparing A single against B single applications of organic fertilizers.
3, Comparing A split against B sp!it applications of organic fertilizers.
• Testing the significance oflinear contrast

Given the sample linear contrast for the contrast to
We want to test the hypotheses:
Ho: o vs Ha: x
s/ The test statistic is given by
SSE
MSE ' where: SSL = for equal replications
Terry
S. Solivas, INSTA T-CAS, UP Los Banos

4- I I
Example:
To test if I-I contrast is significant, the data give:

5(3.83) - (7.80 + 6.71 + 7.30 + 4.03 + 4.92) = -11.61
SSLI = = 17.97
17.97
MSE - 0.24 = 74.88 vs F = 4.41, F = 8.28
Thus the decision is to reject Ho: = 0.
Conclusion: There is significant difference between the controi mean and the treated
means.
The tests for the other contrasts in the orthogonal set are summarized as follows:
ANOVA with Group Comparisons

sv df MS
Treatments 5 59.45 11.89
Control vs treated 1 17.97 17.97
Complete vs 13.5 13.58
inorganic 1 8 26.50
Organic A vs organic 1 26.50 0.70 56.58**
B 1 0.70 1.58
A single vs A split 1.58
B sin le vs Bs lit
1 6.58*
Error 18 4.30 0.24
TOTAL 23 63.75
* ** - significant at 5% and 1%, respectively; ns — not significant
Assignment:
1. Complete the ANOVA for another set of orthogonal contrasts for the example.
sv df MS
Treatments 1 59.45 11.89 74.88**
Control vs treated 17.97 17.97
Complete vs
1 13.58 13.58
inorganic 1
Single vs split 1
A single vs B single
As lit vs Bs lit 1
Error 18 4.30 0.24
TOTAL 23 63.75
2. Discuss the results.
Page
Ill. Trend Comparisons

When the treatments are in quantitative levels, we may want to determine the trend Of the
response instead of comparing the treatment means. For this we find ? functional
relationship between the treatments end the response variable by trend comparisons
which fit the orthogonal polynomial trends such as linear, quadratic, cubic, quartic, and
other higher degree of polynomial trends cf the response.
We consider only the case where the quantitative level of the treatments are equally
spaced.
Illustration:
Consider the results from an experiment conducted in CRD to determine the yield response
of corn to a newly formulated organic fertilizer:
Fertilizer Observations Total Mean

(kg/ha)
4.8 4.79 4.65 4.47 18.80
9 4.70
50 5.08 5.19 4.89 4.92 20.08 5.02
100 5.25 5.18 5.26 5.23 20.92 5.23
150 5.38 5.37 5.46 5.31 21.52 5.38
200 5.55 5.34 5.30 5.25 21.44 5.36
250 5.29 5.22 5.40 5.21 21.12 5.28
123.88 5.16
ANOVA
sv df MS Prob
Treatments 5 1.356 0.271 0.0000
Error 18 0.249 0.014
TOTAL 23 1.605
Since the ANOVA shows that the treatment effects are significantly, we determine the
trend of response by trend comparisons.

The orthogona/ polynomial contrast:
The responses to fit in the trend comparisons are called the orthogonal polynomials of
different degrees: Linear, quadratic, cubic, quartic, quintic and other higher degree
polynomials.
The polynomial responses are defined in terms of contrast where the coefficients of the
contrast are given in the following table:

Designs I
Ewerimental Page 4-13
Table 4. Coefficients for sets of orthogonol polynomial comparisons for equally

spaced treatments.
• Testing the significance oflinear contrast

Given a sample polynomial contrast
for the contrast X, we want to test
Ho: Ha:
The test statistic is given by

SSL where: SSL = for equal replications
MSE'
SSL = for unequal replications
Decision rule: Reject Ho (and accept Ha) if FC
In the illustrative example, the seguent/@/ tests for the different degree polynomial
(linear, quadratic, cubic, etc) comparisons are follows:
Page 4-14
For the linear trend, I-I •

Experimenlal Designs I
L - 4.07
SSL=
SSResidual = Trtss - SSLI = 0.409

ANOVA with trend comparisons
sv df MS Prob
Treatments 5 1.356 0.271 0.0000
Linear 0.947 0.947 0.0000
Residual 1 0.409 0.102 0.0011
4
Error 18 0.249 0.014
TOTAL 23 1.605
Note: If the residua/ is signifiqnt, we test the next degree po/vnomia/.
trend, 1-2
= -2.92
SSResidual = TrtSS - SSLI - SSI-2 = 0.003

ANOVA with trend comparisons
sv df MS prob
Treatments 5 1.356 0.271 0.0000
Linear 0.947 0.947 0.0000
Quadratic
1 0.406 0.406 0.0000
Residual 1 0.003 0.001 29.00** 0.9807
3 0.07ns
Error 18 0.249 0.014
TOTAL 23 1.605
Since the residual is no longer significant then the remaining polynomials are no
longer significant.
Conc/us/on: The yield response to this fertilizer formulation is quadratic.
The next step of the analysis is to fit a quadratic regression equation of the fertilizer
levels on the treatment means.
Page 5-1

CHAPTER 5. CRD WITH SUBSAMPLING
CRD with egval €uksmpliag: We have an experiment laid out in CRD with t
treatments and r replicates per treatment (i.e. equal replications). If we make more
than one observations, say s observations on each experimentai unit, then we have
CRD with equal subsampling,
Example. An experiment is conducted to determine the sugar content of three

varieties of sugarcane. At maturity, four canes of each variety are available for
refractometer brix test. From each cane three determinations are made: at the
base, middle and top portions. The refractometer brix readings are recorded for
analysis.
Data presentation:
Let Yijk = kth observation on the response variable from the jth experimental unit
with the ith treatment.
The data may be presented as follows:

Treatment Sampling Ex erimental unit Trt Trt
unit Total mean
121 131 141
111
122 132 142
Y112
14
11. 12. Y13. 14.
Total
211 221 241
212 222 242
232
Total 21. 22. Y2..

321 331 341
311
322
Y312
Total Y31. 32.
Linear Model:
Yijk = + Eij + öijk 1 2 .., t; j =
where: = general mean ot all possible observations,
= the effect of the ith treatment,
Eij = random error associated with the jth eu with the ith
treatment, öijk — - random error associated with the kth su in the
jth eu with ith trt, = + (i, the true mean of the ith treatment.
Assumptions: 1. tij and öijk NID(O, ,02s )
2. The €ij's are independent of the öijk's

Chapter CRD With Subsamplirg
Page S-2
f.
—+ Estimation ofparameters
The estimates of the parameters:
= Y, ; = Y ; and =t-Y
Model of Effects
Experiments may be classified according to the kind of their effects:
1, MQCe/-U-cÉxeU-e@ct$-EQde0 is assumed when the interested is on the effects of

particular treatments in the experiment.
Example: An experiment is conducted to compare three varieties of corn: var 1, var
2 and var 3. There are other varieties but the researcher is not interested on them.
2. Mode/ 11 or random effects mode/ is assumed when the interest is on a popuiaticn

of treatments from which a random sample of t treatments is drawn for the
experiment. The population of treatments will then be described in terms of the t
treatments.
Example: A researcher wants to characterize the height variability of the upland rice
varieties. Say, there are two thousand upland rice varieties in a germplasm bank.
The researcher randomly selected on!y ten varieties to use in the experiment.
Analysis of Data
1. Perform diagnostic checking of the satisfaction of the ANOVA assumptions.
2. Testing of hypothesis
a. Construct the ANOVA table as outline below:
ANOVA
sv df MS Ex ected Mean uares
Model I Model 11
Treatments t-1 TrSS MSTr
+ 52 +
t(r-l)
Expt'i error ESS MSE 52
Sampling error tr(s-l) DSS MSD 2
TOTAL trs - 1 TSS

where: - variance componentof the populations
5-3
b. First test the hypothesis on the variability of the eu's.

Hypotheses:
Ho: =0 There is no variation among the eu's.

—Y Same precision is obtained if no subsamples are taken.
Ha: There is variation among the eu's.

Greater precision is obtained when subsamples are taken.
v/ Test statistic:
MSE
MSD VS Ft F tr(s-l))
Reject Ho (and accept Ha) if F 2 Ft ; else, fail to reject Ho or

equivalently, reject Ho (accept Ha) if Prob(Fc) s a.
c. Next test the hypothesis on the treatment means.

Hypotheses:
Ho: = = ... = no differences among the treatment means,
Ha: at least two treatment means are different.
v/ Test statistic:
• Case 1: When Ho: = 0 is rejected.

MSTr
VS Ft = Fac(t-I), t(r-l)) MSE
• Case 2: When Ho: = O is accepted.

Experimental Designs Page
MSTr
VS Ft = t(r-l)]
MSE'
ESS + DSS
where: MSE' =
t(rs-l )
Reject Ho (and accept Ha) if F 2 Ft ; else, fail to reject Ho

or equivalently, reject Ho (accept Ha) if Prob(Fc) a.
S.
Measures ofprecision:
1. The standard error of a treatment mean
MSE
s.e.(t ) = rs
2. The standard error of the difference between two treatment means
2MS
s.e.(ü ) = E
rs
3. The CV of the experiment.
Note: The MSE' may be used as the case mav be.
Estimates of the Variance Componen%

In the Model Il experiment, the interest of the researcher is to estimate the variance
components as measures of variability.
= MSD,

MSE-
MSD
s
MSTr - MSE
rs
Note: The MSE' mav be used as the case may be.

I 5-5
Illustrative example:
An experiment was conducted to determine the levels of phosphorus (ppm) of 3
types of soil: A = Maahas clay loam, B = Luisiana clay loam and C = Lipa clay
loam. For each soil type, 4 locations were selected at random and within each
location, two samples were taken. The phosphorus concentration was determined
for each sample. The following data were collected:
Soil Soil Locations Soil type Soil type
sam ies 2 3 4 total mean
1
Maahas 1 9.1 7.3 7.3 10.7
2 7.3 9.0 8.9 12.7
Sub-total 16.4 16.3 16.2 23.4 72.3 9.04
Luisiana 1 12.6 9.1 10.9 8.0
2 14.5 10.8 12.8 9.8
Sub-total 27.1 19.9 23.7 17.8 88.6 11.06
Lipa 1 7.3 6.6 5.2 5.3
9.0 8.4 6.8 6.8
2
Sub-total 16.3 15.0 12.1 12.1 55.5 6.94
216.3 9.01
Analysis:
Identi6,' the treatments, experimenta/ units, sampling units and response variable.
0
Treatments - soil types: A, B, C
0
Expttl units - locations within soil types r = 4 0 Sampl'g
units - samples within locations s = 2. 0 Response var. -
phosphorus concentration (ppm).
> Test ofsignificance

Note: The tests ofsignificance are done in sequence. First, test for the variations
or differences among the experimenta/ units. Then, based on the result of
this first test, test the differences among the treatments.
1. Construct the ANOVA table.
Computations:
(216.3)2 = 1949.40
CF =
trs 3-4-2
S

TSS = -CF = [9.12 + 7.32 + ... + 6.82 ] -CF

= 2089.21 - 1949.40 = 137.812
INSTAT-CAS,
Page
S.
! [72.37 + 88.52 + 55.32] -CF

= 2017.49 1949.40 68.07
Location within soil type SS =
1
[16.402 + 16.33+ ... + 12.12] - 2017.49 = 51.08
2068.56 = 18.65
Samples within location ss
- — E E Y} = 2087.21
ANOVA of the phophorus concentrations (ppm)
df MS Fc.os
sv
Amon soil e 2 68.09 34.04 5.99 4.26
Locations within soil 9 51.08 5.68 3.66 2.86
Sam les within location 12 18.65 1.55
TOTAL 23 137.81
= 26.45%; ) = 0.84; se(d) i 19
2. Test ifphosporus concentration vanes among different /ocations within so,7
types.
Statistical hypotheses
Terry Solivas, T-CAS, UP Los

I 5-7
3. Next test if treatment means are different.
Hypotheses:
Ho: = = no differences among the soil type means,
Ha: at least soil type means are different.
Test statistic:
• Since Ho: = O is rejected, the teststatistic is
MSTr 34.04
VS Ft = F =
4.26 MSE 5.68
Decision: Reject Ho and accept Ha.
Conclusion: At least two soil types have significantly different mean phosporus
concentration.

> Estimates ofthe soi/ type means and effects.
Estimate of treatment
Treatments mans( ) Effect( )
Soil 9.04 0.03
Soil 11.06 2.05
Soil 6.94 -2.07
Perform the pairwise mean comparisons using LSD at a
(Assignment)
INSTA T-CAS,
Terry Solivas, T-CAS, UP Los

CHAPTER 6. TWO-WAY CLASSIFICATION DESIGNS
The use Of one-way classifcation or CRD requires that the eu•s are
homogeneous. sometimes this requirement is not satisfied since the eu's are
markedly heterogeneous with respect to some criteria of classification. For
instance, plots differ in fertility, trees differ in age or height, analysts differ in
efficiency, etc.
The differences among the eu's is a major source of experimental error. When the
eu's are heterogeneous, the appropriate design must account' for these
heterogeneity. These design are the randomized complete block destgn'(RCBD)
and the /atin square desgn,
THE RANDOMIZED COMPLETE BLOCK DESIGN (RA

As a local control, one way of increasing the precision of an experiment is by
proper grouping or blocking of the experimental units.
The eu's are grouped into r blocks in such a way that the differences between the
units among different blocks are greater than the differences between ,the units
within each block.
Likewise, the blocking should be done in such a way that the blocks cut across or
are perpendicular to the direction of the eu's gradient.
This way, if there are differences among the blocks, the variability is removed from the
experimental error thereby improving the precision of the experiment.
Randomization and layout

Suppose there are t = 4 treatments (T1, T2, T2 and T4) to be replicated r = 3
times; thus the total number of eu's needed is tr = 12. The randomization
procedure (say by using draw lots) may be as follows:
1. Group the eu's into r 3 blocks.
2. Divide each block into t = 4 eu's.
3. For each block, allocate the treatments into the eu's at random and independently of
other blocks as follows:
a. Label the eu's consecutively from 1 tot = 4.

b. Obtain a sequence oft = 4 numbers using draw lots.
Page 6-2
c. i
Using the sequence of draws as the treatment numbers and the drawn numbers
as the eu numbers assign the treatments to the respective eus.
Chapter 6. Two-Way Classification Designs
1 2 3 4 = treatment no.
Ex: For block 1, draw sequence :
drawn numbers: 2 4 1 3 = eu number
= treatment no.
For block 2, draw sequence : 1 2 3 4
= treatment no.
For block 3, draw sequence : 1 2 3 4
The corresponding layoutis:
1 1 1
2 2
3 3 3
4 4 4
Block 1 Block 2
> Data Presentatiuon:

Let Yij = observation on the response variable from the jth experimental unit
with the ith treatment.
Block Total Mean
Treatmen
t 1 2
1 11 12 1.
ir
Y21 22
Total
> Linear Model:
Yij = + pj +

where: = general mean of all possibie obserations,
> Analysis ofResul&
To illustrate the analysis of results, we consider the following data obtained from
an experiment conducted to study the effects of four varieties of mongo. The
experiment was laid out in RCBD with five farms as the blocks. In each farm, four
uniform plots were chosen on which the four varieties were randomly assigned.
Varietie Farms Variety
s 2 3 4 5 Totals
32.3 34.0 34.3 35.0 36.5 172.1
33.3 33.0 36.3 36.8 34.5 173.9
30.8 34.3 35.3 32.3 35.8 168.5
c
D 27.0 26.0 28.9 28.0 28,8 138.7
Totals 123.4 127.3 134.8 132.1 135.6 653.2
At = 5%, test if the different varieties have different yields.
1. Testing significance of treatments via ANOVA F-test
Statisticai hypotheses
Ho: All the variety means are not different.
Ha: At least two variety means are different.
•z Test statistic and critical value at = 5%,
MSTr
Test statistic: FC = ; critical value: Ft = = 3.49 MSE
Decision rule: Reject Ho (and accept Ha) if FC Ft; else, accept Ho.
Construct the ANOVA table outlined as follows:
ANOVA
sv df MS

Page 6-4
Block r-l RSS MSR MS MSE
Treatment t-1 TrSS MSTr MSTr/MSE
Error r-l t- ESS MSE
1
tr-l
TOTAL TSS
Computation:
Let Yij = observation on the jth block with the ith

treatment = total of observations with the ith treatment
Y j = total of observations on the jth

block = grand total of all observations
INSTAT-CAS,

Page 6-4
Chapter 6. Two-Way Classification Designs

1 Experimental Designs I Stat 162 1

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Preface

the works to the University Publishing Center of UP Los Bafios.

CHAPTER 1. INTRODUCTORY CONCEPTS

THE ROLE OF STATISTICS IN RESEARCH

E/emen&. observations, analysis and reasoning.

In scientific research, statistics deals with:

Fig. 1; Statistical methods in research (Rag 1989).

Terry S. Solivas, T-CAS, UP Los

Chapter l. Inntroductory Concepts

BASIc CONCEPTS ON STATISTICALLY DESIGNING AN

A researcher conducts an experiment to obtain data. Based on results of data analysis,

• Provide an appropriate and valid analysis of data

Steps in Conducting an Experiment

1. State the problem of study.

• define the treatments, experimentai units, size of the experiment.

7. Summarize the results, make conclusions and inferences.

8. Evaluate the whole study in relation to similar investigations.

Definition ofCommon Terms Used in Experimental

Treafrnents are procedures or conditions whose effects are to be measUred

Examples: Kinds of fertilizer, rates of irrigation, age of harvest, different locations,

Experimental unit is a group of experimental materials or individuals to which a

Examples: A plot of land, a plant, a portion of a leaf, a dish of culture medium, a

Response variable is a characteristic used to measure the treatment effects.

Sampling unitis a portion of the experimental unit on which the

Examples: A tiller in a hill, a plant in a pot, a seedling in a seedbed, a tree_in a

Note: Sampling unit may be the whole experimental unit.

Experimental error is the measure of variations among experimental units

Some sources of experimenta/ errors.

Terry S. Solivas, INSTA T-CAS, UP Los

Inherent variability of the experimentai materials, e Errors in

Precision denotes repeatability of measurements. It is measured by the

+ Accuracy denotes unbiasedness or the closeness of the average values of the

Some ways ofihcreasing accuracy are.

Refinement of experimental technique and

Layout refers to the final arrangement of treatments over the whole

Principles ofExperimental Design

A valid experimental design must have these three principles: replication,

Replication is the repetition of the application of treatments on a number of

Terry S. Solivas, INSTA T-CAS, UP Los Banos

To provide an estimate of thé experimental error.

to enable test of significance and interval estimation.

experiment. Factor affecting the number ofreplications e Degree of

more replications for higher precision desired

Terry S. Solivas, INSTA T-CAS, UP Los

For a required precision, smaller number of treatments is compensated by more

Cost and availability of resources.

Randomization is the allocation of treatments to the experimental units by

Provide a random sample of observations.

Local control orerror control is any process or technique used to minimize

Common Techniques for Local Centro!

Proper choice of experimental design,

Proper choice of size and shape of experimental units,

Refinement of experimental technique o Uniform application of treatments and

Terry S. Solivas, INSTA T-CAS, UP Los

CRD is a design wherein the allocation of treatments is done by randomizing the

> Randomization and layout

The corresponding /ayoutfor this randomization randomization procedure is:

> Data Presentation: (One-way classification)

Terry S. Solivas, INSTA T-CAS, UP Los Banos

Chapter 2. One-Way Classification Design

where: = genera! mean of ai! possible observations,

> Analysis ofData:

Method Time to com lete anal sis hrs Re s Total Mean