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Repeated measurements

Descriptive statistics

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 Objective of this lesson

• At the end of this lesson, the student must be able to:

– Calculate the mean, range, relative standard deviation
– Calculate the standard deviation, the confidence limits for
both large data sets and small data sets
– Be able to distinguish between confidence interval and
confidence limits
– Be able to know the properties of the normal distribution
curve of repeated measurements
– Apply confidence limits to assess the presence of systematic
errors

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 Statistics
• Are a tool to answer questions:
– How accurate is the analyses?
– How many analyses do I have to make to
overcome problems of inhomogeneity or
imprecision?
– Does the product meet the specification?
– With what confidence is the limit exceeded?

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 Repeated measurements

• Why do we make repeated measurements of a

quantity?
– Estimate true value of that quantity
– Establish the reliability of this estimate
– Have confidence on the reported results
– Be able to estimate random errors and assess their
significance
– Detect presence systematic errors if any
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 Analytical Results Vary
• Because of unavoidable deviation during
measurement

• Because of inhomogeneity between the subsamples

• It is very important to differentiate between these

two cases

• Because the measures to be taken can be different

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 Variations
322 322,3 322,29
322 322,5 322,49
322 322,3 322,28
322 322,2 322,17
• Are always present 322
322
321,7
321,8
321,67
321,76
322 321,7 321,75
322 322,2 322,17
322 321,6 321,58
• If there seems to be 322
322
322,4
322,4
322,36
322,40

none, the resolution

322 321,5 321,53
322 322,4 322,42
322 322,1 322,05
is not high enough 322
322
322,2
322,5
322,16
322,57
322 321,7 321,73
322 321,8 321,76
322 321,9 321,85
322 322,4 322,41

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 Some definitions

• Sample – A finite set of measurements which represent

the population, n.
• Population – A collection of an infinite number of
measurements, N, i.e., as n à infinity
• True value (true mean)- (µ), mean value for the
population.
• Observed Mean – ( x ), mean value of the sample set

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 Distribution of repeated
measurements

• In a any given population,

many of the values will cluster
about the most common
value
• This is defined as measures of
central tendency
• Populations tend to have a
Gaussian distribution

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 Properties of the normal distribution
curve
• The Normal or Gaussian distribution
n General properties:
n Symmetrical about the
given by: exp{- ( x - µ ) 2 / 2s 2 }
y= mean, µ
s 2p
n The greater the value of σ,

the greater the spread

n 68% of population values

lies with ± 1σ
n 95% of population values

lies with ± 2σ
n 99.7% of population values

lies with ± 3σ
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Figure 2.4 Properties of the normal distribution: (i) approximately 68% of values lie
within ±1s of the mean; (ii) approximately 95% of values lie within ±2s of the mean; (iii)
approximately 99.7% of values lie within ± 3s of the mean
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 Arithmetic mean and range

– Defined as the sum of all measurements divided by

the number of measurements
– The equation:
x=
å xi
n
– Range is the difference between the highest and the
lowest values
– Range is used to measure the spread of the
measurements and hence precision

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 Standard Deviation

– Standard deviation also measures how spread the

values are from their mean
– It measures variability
– Standard deviation is likened to the range but in this
case all values are utilised in the calculation
– The smaller the standard deviation, the more closely
the data are clustered about the mean
– Obtained as the sum of squared deviation of the
values from the mean divided by the number of
observations
– Standard deviation nkoanebm@ub.ac.bw
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 Standard Deviation - equation

• For an infinite number of measurements (σ)

N
1
s=
N
å
i -1
( x i - x ) 2

• For a finite number of measurements (s)

1 N
• Variance
s= å i
N - 1 i -1
( x - x ) 2

– The square of the standard deviation, s2

– Necessary when looking at propagation of errors

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 Relative Standard Deviation

• Measure of spread also known as coefficient of

variance (CV)
• Standard deviation expressed as a percentage,
therefore has units of per cent (%)
• It is an example of relative error
• RSD, is often reported as %RSD, is used in
comparison of precision of results with different
units or magnitudes
• The equation: RSD = 100s
x
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 Confidence Intervals

• From a data set, it is very difficult to find the true

population mean, µ or the true standard deviation, σ.
What we can determine are and s
x
• The confidence interval is an expression stating that
the true mean is likely to lie within a certain distance
from the measured mean
• Extreme values are known as confidence limits
• The size will depend on how certain we want to be that
the interval includes the true value

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 Confidence Intervals

• If distribution is normal, then

95% of the sample means lie
within the range given by the
confidence interval of the
mean:

x - 1.96(s / n ) < µ < x + 1.96(s /( n )

• The 95% confidence limits are: Figure 2.6: The sampling distribution of the
mean, showing the range within which 95% of
x ± 1.96(s /( n ) sample means lie

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 Confidence Limits – large numbers

• In practice we are unlikely to know the exact σ. However,

when dealing with large number of replicates, the standard
deviation can be used in place of σ:
• Other confidence
x ± 1.96(limits
s / n ) may be used especially at 99% or
99.7%. For large samples, the confidence limits are given
by:
– The z values depends on degree of confidence. For:
x ±limits,
95% confidence zs / z=1.96
n
99% confidence limits, z=2.58
99.7% confidence limits, z= 2.97

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 Confidence Limits – small numbers
• As the sample gets smaller, s becomes less reliable as an
estimate of σ
• Therefore, for small data sets, the confidence limits of
the mean is given by:
• t-students t is a statisticalx tool
± t n -1 stonexpress confidence
levels and to compare results form different
experiments
• Value of t depends on degrees of freedom (n-1) and
degrees of confidence
• Tabulated t- values
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 Student t-values

Degrees of freedom 95 % confidence level 99% confidence level

1 12.71 63.66
2 4.30 9.92
3 3.18 5.84
4 2.78 4.60
5 2.57 4.03
10 2.23 3.17
20 2.09 2.85
30 2.04 2.75
50 2.01 2.68
100 1.98 2.63
∞ 1.96 2.58
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 Other uses of confidence limits

• To test for presence of systematic errors

• Example:
– The absorbance scale of a spectrometer is tested at a particular
wavelength with a standard solution which has an absorbance given as
0.470.
– Ten measurements of the absorbance with the spectrometer give an
average 0.461 and s= 0.003
– Find the 95% confidence interval for the mean absorbance as measured
by the spectrometer and decide whether a systematic error is present
– Confidence limits (tn-1=9=2.26):
x ± t n -1 s n = 0.461 ± 2.26 ´ 0.003 / 10 = 0.461 ± 0.002
– The confidence limits do not include 0.470 and therefore it is likely that
systematic error has occurred
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 Remember: Reporting data
• When reporting data, quote the mean as the estimate
of the quantity measured and the standard deviation as
the estimate of precision
• Always state the value of n (the number of
measurements taken)
• Quote as significant figures all the digits that are certain
(depends on the precision of the experiment) and the
first uncertain one i.e., x ± s
• Example for 10.09, 10.11, 10.09, 10.10 and 10.12
x ± s = 10.10 ± 0.01(n = 5)
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 Home exercise

• The carbohydrate content of a glycoprotein is

determined to be 12.6, 11.9, 13.0, 12.7, and
12.5g carbohydrate per 100g glycoprotein in
replicate analyses.
• Find the 90 % and the 99 % confidence
intervals for the carbohydrate content

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