SAMPLE SIZE

CALCULATION

Melchor V.G. Frias, IV

Clinical Epidemiology Unit
Angelo King Medical Research Center
De La Salle Health Sciences Institute
 Learning Objectives:
At the end of this session, learners should be able to:
1. Explain the concept/importance of sample size,
2. Explain and apply the concept of hypothesis
testing,
3. Apply sample size formulas for descriptive and
analytic studies,
4. Identify the requirements for sample size
calculation ,
5. Apply OPEN EPI/EPIINFO for sample size
calculation for cross-sectional, cohort, case-
control and experimental studies.
 How many subjects are to be
included in the sample?
 SAMPLE SIZE CALCULATION
 Why calculate?
 for planning purposes
 for “power” of the study (low power – it will
have little chance of giving a statistically
significant difference).
 meaningful results (small sample - the study
will have failed to establish that the
intervention has no appreciable effect).
How do we calculate sample size?

♦ Using formulas
♦ Using tables of sample sizes
♦ Using statistical calculators
(StatCalc of EpiInfo, Open EPI)
 Sample size calculation
 Things to know:
 type of the study: descriptive or analytic?
 proportions or means? usual values?
 amount of deviation from the true value?
Clinically important difference?
 confidence level? power?
 one-tailed or two-tailed hypotheses
 Hypotheses testing

The first thing to do when given a claim is to

write the claim mathematically (if possible),
and decide whether the given claim is the null
or alternative hypothesis.
 Hypotheses testing

If the given claim contains equality, or a

statement of no change from the given or
accepted condition, then it is the null
hypothesis, otherwise, if it represents change, it
is the alternative hypothesis.
 Hypotheses testing
 hypothesis -- a statement about the
population
 null hypothesis (Ho) -- equality
 alternative hypothesis (Ha) --
 two-tailed -- not equal
 one-tailed -- one is greater than the other
Hypotheses testing
"He's dead, Jim," said Dr. McCoy to
Captain Kirk.
 Hypotheses testing

Mr. Spock, as the science officer, is put in charge

of statistically determining the correctness of
Bones‘ statement and deciding the fate of the crew
member (to vaporize or try to revive)
 Hypotheses testing
• His first step is to arrive at the hypothesis to be
tested.
• Does the statement represent a change in
previous condition?

 Yes, there is change, thus it is the alternative

hypothesis, H1
 No, there is no change, therefore it is the null
hypothesis, H0
 Hypotheses testing

 The correct answer is that there is change.

Dead represents a change from the accepted
state* of alive. The null hypothesis always
represents no change. Therefore, the
hypotheses are:

 H0 : Patient is alive.
 H1 : Patient is not alive (dead).
 Hypotheses testing

 Possible states of nature (Based on H0)

 Patient is alive (H0 true - H1 false )

 Patient is dead (H0 false - H1 true)
 Hypotheses testing

Decisions are something that you have

control over. You may make a correct
decision or an incorrect decision. It depends
on the state of nature as to whether your
decision is correct or in error.
 Hypotheses testing

 Possible decisions (Based on H0 ) /

conclusions (Based on claim )

 Reject H0 / "Sufficient evidence to say

patient is dead"
 Fail to Reject H / "Insufficient evidence
0
to say patient is dead"
 Hypotheses testing

 There are four possibilities that can occur

based on the two possible states of nature
and the two decisions which we can make.
 Hypotheses testing

 Statisticians will never accept the null

hypothesis, we will fail to reject. In other
words, we'll say that it isn't, or that we don't
have enough evidence to say that it isn't, but
we'll never say that it is, because someone
else might come along with another sample
which shows that it isn't and we don't want
to be wrong.
 Hypotheses testing -
Statistically speaking:
State of Nature

Decision H0 True H0 False

Reject H0 Patient is alive, Patient is dead,

Sufficient Sufficient evidence of
evidence of death death

Fail to reject H0 Patient is alive, Patient is dead,

Insufficient Insufficient evidence of
evidence of death death
 Hypotheses testing – In
English (or Clingon?)
State of Nature

Decision H0 True H0 False

Reject H0 Vaporize a live Vaporize a dead person

person

Fail to reject H0 Try to revive a Try to revive a dead

live person person
 Hypotheses testing – Were
you right?
State of Nature

Decision H0 True H0 False

Reject H0 Type I Error Correct Assessment

alpha

Fail to reject H0 Correct Type II Error

Assessment beta
 Hypotheses testing
Which of the two errors is more serious?
Type I or Type II ?

State of Nature

Decision H0 True H0 False

Reject H0 Type I Error Correct Assessment

alpha

Fail to reject H0 Correct Assessment Type II Error

beta
 Hypotheses testing
State of Nature

Decision H0 True H0 False

Reject H0 Patient is alive, Sufficient Correct Assessment

evidence of death:
vaporize a live person
Fail to reject Correct Assessment
H0
Patient is dead,
Insufficient evidence of
death: revive a dead person

Which of the two errors is more serious?

Type I or Type II ?
 Hypotheses testing

Disease actually present

Diagnosis No Yes

Disease present Mis-diagnosis Correct diagnosis

Disease absent Correct diagnosis Missed diagnosis

 Hypotheses testing

Assumption of innocence

Judgment True False

Pronounced guilty Serious error in Correct judgment

judgment

Pronounced not Correct judgment Error in judgment

guilty
 Hypotheses testing
 Since Type I is the more serious error (usually),
that is the one we concentrate on.

 We usually pick alpha to be very small

(0.05, 0.01). Note: alpha is not a Type I error.

 Alpha is the probability of committing a Type

I error. Likewise beta is the probability of
committing a Type II error.
 Hypotheses testing
Conclusions
 Conclusions are sentence answers which
include whether there is enough evidence or
not (based on the decision), the level of
significance, and whether the original claim
is supported or rejected.
 Hypotheses testing
Conclusions
 Conclusions are based on the original claim,
which may be the null or alternative
hypotheses. The decisions are always based
on the null hypothesis
 Hypotheses testing -
Conclusions
Original Claim
H0 H1
Decision "REJECT" "SUPPORT"
Reject H0 There is sufficient There is sufficient evidence at
"SUFFICIENT" evidence at the alpha the alpha level of significance
level of significance to to support the claim that (insert
reject the claim that original claim here)
(insert original claim
here)
Fail to reject H0 There is insufficient There is insufficient evidence
"INSUFFICIENT" evidence at the alpha at the alpha level of
level of significance to significance to support the
reject the claim that claim that (insert original claim
(insert original claim here)
here)
 Definitions
 Null Hypothesis ( H0 )
 Statement of zero or no change.
 If the original claim includes equality (<=, =, or
>=), it is the null hypothesis.
 If the original claim does not include equality (<,
not equal, >) then the null hypothesis is the
complement of the original claim.
 The null hypothesis always includes the equal
sign. The decision is based on the null
hypothesis.
 Definitions
 Alternative Hypothesis ( H1 or Ha )
 Statement which is true if the null hypothesis is
false.
 The type of test (left, right, or two-tail) is based
on the alternative hypothesis.
 Definitions
 One-Tailed (Sided) Test
 Definitions
 Two-Tailed (Sided) Test
 Definitions
 Type I error
 Rejecting the null hypothesis when it is true
(saying false when true). Usually the more
serious error.
 Type II error
 Failing to reject the null hypothesis when it is
false (saying true when false).
 Definitions
 alpha ( - probability of committing Type I
error
1-  - the confidence level

 beta - probability of committing Type II

error
1-  - power of the study; ability to detect a
true difference
 Definitions
 Significance level ( alpha )
 The probability of rejecting the null hypothesis
when it is true. alpha = 0.05 and alpha = 0.01 are
common.
 If no level of significance is given, use alpha =
0.05.
 The level of significance is the complement of the
level of confidence in estimation.
 Confidence level, Power
 Usual Values:

 = 0.05,
 1-  (confidence level) = .95

 = 0.20,

 1-  (power) = 0.80
 Confidence level, Power
 The easiest ways to increase power are to:

 increase sample size

 increase desired difference (or effect size)

 decrease significance level desired e.g. 10%

 Definitions
 Decision
 A statement based upon the null hypothesis.
 It is either "reject the null hypothesis" or "fail to
reject the null hypothesis".
 We will never accept the null hypothesis.
 Definitions
 Conclusion
 A statement which indicates the level of evidence
(sufficient or insufficient),
 at what level of significance,
 and whether the original claim is rejected (null)
or supported (alternative).
 How do we calculate sample size?
A.J. Dobson’s formula (SIMPLE RANDOM SAMPLE)
 descriptive studies
 population proportion
 population mean
 analytic studies
 comparing two proportions
 comparing two means
Sample size for descriptive studies
1. Estimation of a population proportion

p(100  p )
n  f (1   )
2

where
n = computed sample size
p = estimate of the proportion
 = the desired width of the confidence interval
1-  = confidence level
Sample size for descriptive studies
1. Estimation of a population proportion
Table 1 Values for f(1-) for various
confidence levels 100 (1-) %
(1-) 0.8 0.9 0.95 0.99
f(1-)* 1.642 2.706 3.842 6.635

* f(1-) is the square of the upper 1/2  point of the std.

Normal Distribution
 Sample size for descriptive studies
1. Estimation of a population proportion
A researcher wants to estimate the smoking prevalence
in high school students . What is the sample size if it is
expected that the smoking prevalence is 15%, and a 95%
confidence interval will be used for an interval of 4%
(11-19%)?

p(100  p )
n  f (1   )
2
Sample size for descriptive studies
1. Estimation of a population proportion
Table 1 Values for f(1-) for various
confidence levels 100 (1-) %
(1-) 0.8 0.9 0.95 0.99
f(1-)* 1.642 2.706 3.842 6.635

* f(1-) is the square of the upper 1/2  point of the std.

Normal Distribution
 Sample size for descriptive studies
1. Estimation of a population proportion
A researcher wants to estimate the smoking prevalence
in high school students . What is the sample size if it is
expected that the smoking prevalence is 15%, and a 95%
confidence interval will be used for an interval of 4%
(11-19%)?

p(100  p )
n  f (1   )
2

15(100  15)
n 2
 3.842
4
n  306
Sample size for descriptive studies
2. Estimation of a population mean

2
s
n  2  f (1   )

where
n = computed sample size
s = estimate of the standard deviation of the
observations
 = the desired width of the confidence interval
1-  = confidence level
Sample size for descriptive studies
2. Estimation of a population mean
Table 1 Values for f(1-) for various
confidence levels 100 (1-) %
(1-) 0.8 0.9 0.95 0.99
f(1-)* 1.642 2.706 3.842 6.635

* f(1-) is the square of the upper 1/2  point of the std.

Normal Distribution
 Sample size for descriptive studies
2. Estimation of a population mean
A researcher wants to estimate the mean serum
cholesterol level (mg/100ml) in a group of men. How
many men should be included if he wants to be 90%
confident that the estimate of the mean will fall within
10mg/100ml of the true value and standard deviation is
estimated to be 40mg/100ml?

2
s
n  2  f (1   )

Sample size for descriptive studies
2. Estimation of a population mean
Table 1 Values for f(1-) for various
confidence levels 100 (1-) %
(1-) 0.8 0.9 0.95 0.99
f(1-)* 1.642 2.706 3.842 6.635

* f(1-) is the square of the upper 1/2  point of the std.

Normal Distribution
 Sample size for descriptive studies
2. Estimation of a population mean
A researcher wants to estimate the mean serum
cholesterol level (mg/100ml) in a group of men. How
many men should be included if he wants to be 90%
confident that the estimate of the mean will fall within
10mg/100ml of the true value and standard deviation is
estimated to be 40mg/100ml?
2
s
n  2  f (1   )

2
40
n  2  2.706  43
10
 Sample size for analytic studies
1. Hypothesis testing between two proportions

p1(100  p1)  p 2(100  p 2)

n  f ( ,  )
( p1  p 2) 2

where
n = computed sample size
p1, p2 = estimate of the sample proportion for each group
1-  = confidence level
1-  = power of the test
Sample size for analytic studies
1. Hypothesis testing between two proportions
Table 2 Values for f(,)*
Significance level, 
Power, one-tailed two-tailed
1-
0.05 0.01 0.05 0.01
0.5 2.71 5.41 3.84 6.63
0.8 6.18 10.04 7.85 11.68
0.9 8.56 13.02 10.51 14.88
* f(,) is the square of the sum of the upper tail  and
the upper tail  point (for one tailed test) or 1/2  point
(for two-tailed test) of the std. Normal distribution
 Sample size for analytic studies
1. Hypothesis testing between two proportions
A new antibiotic is to be compared to a standard drug
with respect to cure rate of urinary tract infection. The
new drug will be considered better than the standard
drug if it shows a 5% difference from the cure rate of
80%. How many patients are needed if the investigator
wants 90% power and 95% confidence?

p1(100  p1)  p 2(100  p 2)

n  f ( ,  )
( p1  p 2) 2
Sample size for analytic studies
1. Hypothesis testing between two proportions
Table 2 Values for f(,)*

Significance level, 
Power, one-tailed two-tailed
1-
0.05 0.01 0.05 0.01
0.5 2.71 5.41 3.84 6.63
0.8 6.18 10.04 7.85 11.68
0.9 8.56 13.02 10.51 14.88
* f(,) is the square of the sum of the upper tail  and
the upper tail  point (for one tailed test) or 1/2  point
(for two-tailed test) of the std. Normal distribution
 Sample size for analytic studies
1. Hypothesis testing between two proportions
A new antibiotic is to be compared to a standard drug
with respect to cure rate of urinary tract infection. The
new drug will be considered better than the standard
drug if it shows a 5% difference from the cure rate of
80%. How many patients are needed if the investigator
wants 90% power and 95% confidence?

p1(100  p1)  p 2(100  p 2)

n  f ( ,  )
( p1  p 2) 2

80(100  80)  85(100  85)

n  8.56  984
(80  85) 2
Sample size for analytic studies
2. Hypothesis testing between two means
2
2s
n  f ( ,  )
 2

where
n = computed sample size
s = estimate of the standard deviation of the
observations, assuming it is the same for
each group
 = the true difference between the means
1-  = confidence level
1-  = power
Sample size for analytic studies
1. Hypothesis testing between two means
Table 2 Values for f(,)*

Significance level, 
Power, one-tailed two-tailed
1-
0.05 0.01 0.05 0.01
0.5 2.71 5.41 3.84 6.63
0.8 6.18 10.04 7.85 11.68
0.9 8.56 13.02 10.51 14.88
* f(,) is the square of the sum of the upper tail  and
the upper tail  point (for one tailed test) or 1/2  point
(for two-tailed test) of the std. Normal distribution
 Sample size for analytic studies
2. Hypothesis testing between two means

To determine whether an antihypertension therapy can

reduce the average blood pressure of some group by 5
mmHg when the standard deviation is 10 mmHg, how
many patients are needed for a two-tailed test at the 5%
significance level, and power of 90%?

2
2s
n  2  f ( ,  )

Sample size for analytic studies
2. Hypothesis testing between two means
Table 2 Values for f(,)*
Significance level, 
Power, one-tailed two-tailed
1-
0.05 0.01 0.05 0.01
0.5 2.71 5.41 3.84 6.63
0.8 6.18 10.04 7.85 11.68
0.9 8.56 13.02 10.51 14.88
* f(,) is the square of the sum of the upper tail  and
the upper tail  point (for one tailed test) or 1/2  point
(for two-tailed test) of the std. normal distribution
 Sample size for analytic studies
2. Hypothesis testing between two means

To determine whether an antihypertension therapy can

reduce the average blood pressure of some group by 5
mmHg when the standard deviation is 10 mmHg, how
many patients are needed for a two-tailed test at the 5%
significance level, and power of 90%?

2
2s
n   f ( ,  )
 2

2
2(10)
n 2
 10.51  84
5
Sample size calculation using EPI-Info6
http://www.cdc.gov/epiinfo/Epi6/ei6.htm
 STATCALC program
 Sample size for analytic studies
2. Hypothesis testing between two means

To compare two antianemia treatment groups

in terms of outcome of hemoglobin level. What
is the sample size needed if expected mean hgb
level after treatment for group A is 132.86 with
standard deviation of 15.34 and the mean
hemoglobin level for group B is 127.44 with sd
of 18.23?
http://www.openepi.com/Menu/OpenEpiMenu.htm
 Sample size for analytic studies

Case Control Study

Research question: Is there an association between
receiving HRT and development of breast CA among
women in Dasmarinas, Cavite?

Odds of exposure among diseased = 175/75 = 2.3

Odds of exposure among non-diseased = 25/225 = 0.11

Odds Ratio = 21
You need to have an estimate of the percentage of exposure among the controls
and either the odds ratio or the percentage of exposure among cases
 Sample size for analytic studies

Cohort Study
Research question: Is Hib vaccine associated with the
development of leukemia among children in
Dasmarinas, Cavite ?

Incidence of disease among exposed = 150/500 = 0.3

Incidence of disease among unexposed = 400/500 = 0.8

Relative Risk = 0.375

You need to know the percentage of outcome among the unexposed, and either
an OR, RR or the percentage of the outcome among the exposed.
 Calculate sample size: RCT
Example: Efficacy of flubendazole compared to mebendazole in
the treatment of trichiuriasis among pediatric patients.

Objective: To compare resolution of trichiuriasis for pediatric

patients given flubendazole and those given mebendazole.

+ resolution
Flubendazole
group
(-) resolution
(Exposed)

+ resolution
Mebendazole
group (-) resolution
(Unexposed)
 Calculate sample size: RCT
Example: Efficacy of flubendazole compared to mebendazole in
the treatment of trichiuriasis among pediatric patients.

Objective: To compare resolution of trichiuriasis for pediatric

patients given flubendazole and those given mebendazole.

+ resolution 75%
Flubendazole
group
(-) resolution
(Exposed)

+ resolution 50%
Mebendazole
group (-) resolution
(Unexposed)
50% with resolution in 75% with resolution
Mebendazole group in flubendazole group
 General comments on estimation of sample size
 Compute the sample size as early as possible
during the design phase, (to estimate the
resources required and the feasibility of the
study.
 The rarer the condition being investigated, the
larger the sample size, all other things being
equal.
 Complex data analysis generally requires larger
samples than simple analysis.
 In general, longitudinal studies require a larger
sample size than case-control and cross
sectional studies.
 General comments on estimation of sample size

 The higher the level of accuracy and precision

desired for the resulting estimates, the larger
the sample size necessary.
 When more than 1 item or outcome are to be
studied, sample sizes are estimated separately
for each item. The final sample size will be a
compromise between the largest n and the
resources to conduct the study.
 Summary
 Explained the concept/importance of sample size,
 Explained and applied the concept of hypothesis
testing,
 Applied sample size formulas for descriptive and
analytic studies,
 Identified the requirements for sample size
calculation ,
 Introduced OPEN EPI/EPIINFO for application in
sample size calculation for cross-sectional, cohort,
case-control and experimental studies.
 Summary
 Statistical inference allows us to generalize
sample results to the target population
 sample size is based on the
 research objectives/design
 sample estimates, variability from previous
studies
 power, level of confidence
 operational constraints (time, resources)