Comparison of Results

The comparison of values obtained from a set of results with either (a) the true value or (b) other
set of data makes it possible to determine whether the analytical procedure has been accurate and
/ or precise, or if it is superior to another method.
There are two common methods for comparing results (a) Student’s t-test (b) the variance ratio
test (F-test).
These methods of test require a knowledge of what is known as the number of degree of
freedom.
In statistical terms, this is the number of independent values necessary to determine the statistical
quantity.
Student’s t-test
This is a test used for small samples; its purpose is to compare the mean from a sample with
some standard value and to express some level of confidence in the significance of the
comparison. It is also used to test the difference between the means of two set of data and .
The value of t is obtained from the equation;
t= where µ is the true value.
It is then related to a set of t-tables in which the probability (P) of the t-value falling within
certain limits is expressed, either as a percentage or as a function of unity, relative to the number
of degree of freedom.
Example. T-test when the true mean is known.
If the mean (of 12 determinations = 8.37 and the true mean value (µ) = 7.91, say whether or not
this result is significant if the standard deviation is 0.17
Solution
t= t= = 9.4
From t-tables for eleven degrees of freedom (one less than those used in the calculation)
For P = 0.10(10 percent) 0.05(5 percent) 0.01(1 percent)
t= 1.80 2.20 3.11
And as the calculated value for t is 9.4 the result is highly significant. The t-table tells us that the
probability of obtaining the difference of 0.46 between the experimental and true result is less
than 1 in 100. This implies that some particular bias exists in the laboratory procedure. If the
calculated t is less than the t table value at the level of probability given, then, there would not be
a significant difference in the results.
F-test
This is used to compare the precisions of two sets of data for example, the results of two
different analytical methods or the results from two different laboratories. It is calculated from
the equation:
F = S2A / S2B
The larger value of s is always used as the numerator so that the value of F is always greater than
unity. The value obtained for F is then checked for its significance against values in the F- table
calculated from an F- distribution corresponding to the umbers of degree of freedom for the two
sets of data.
Example: F-test comparison of precisions
The standard deviation from one set of 11 determinations was sA = 0.210 and the standard
deviation from another 13 determinations was sB = 0.641. Is there any significant difference
between the precision of these two sets of results?
Solution
F = S2A / S2B
F = (0.641)2 / (0.210)2 = 0.411 / 0.044 = 9.4
F table at different level of Probability
P = 0.10 0.05 0.01
F = 2.28 2.91 4.71
The first value (2.28) corresponds to 10 % probability, the second value (2.91) to 5% probability,
and the third value (4.71) to 1% probability.
Under these conditions there is less than one chance in 100 that these precisions are similar. To
put it another way, the difference between the two sets of data is highly significant. Had the
value of F turned out to be less than 2.28 then it would have been possible to say that there was
no significant difference between the precisions at the 10% level.
Comparison of the Means of two Samples
When a new analytical method is being developed it is usual practice to compare the values of
the mean and precision of the new (test) method with those of an established (reference)
procedure. The value of t when comparing two sample means and is given by the expression;
t= where sp the pooled standard deviation, is calculated from the two sample standard
deviation s1 and s2, as follows; S22
sp =
It should be stressed that there must not be a significant difference between the precisions of the
methods. Hence the F-test is applied prior to using the t-test in equation.
Example: Comparison of two sets of data
The following results were obtained in a comparison between a new method and standard
method for the determination of the percentage nickel in a special steel.
New method Standard method
Mean =7.85% = 8.03%
SD s1 = ±0.130% s2 = ±0.095%
Number of samples n1 = 5 n2 = 6
Test at the 5 per cent probability value if the new method means is significant difference between
the precisions of the two methods.
F = S2A / S2B = = 1.87
The F-value (P = 5 percent) the table for four and five degree of freedom respectively for sA and
sB = 5.19
Thus, the calculated value of F (1.87) is less than the tabulated value; therefore the methods
have comparable precisions (standard deviation) and so the t-test can be used with confidence.
sp = = ±0.112
and from equation
t = = t = = = 2.66

At the percent level, the tabulated value of t for (n1 +n2-2), i.e, nine degrees of freedom, is 2.26.
Since t calculated 2.66> t tabulated 2.26, there is a significant difference, at the specified probability,
between the mean results of the two methods.
Paired t-test
Another method of validating a new procedure is to compare the results using samples of varying
composition with the values obtained by an accepted method.
Example. The t-test using samples of differing composition (the paired-test). Two different
methods, A and B, were used for the analysis of five different iron compounds.
Sample 1 2 3 4 5
Method A 17.6 6.8 14.2 20.5 9.7%Fe
Method B 17.9 7.1 13.8 20.3 10.2%Fe

Solution
It should be apparent that in this example it would not be correct to attempt the calculation by the
method described previously.
In this case the differences (d) between each pair of results are calculated and, the mean of the
difference, is obtained. The standard deviation sd of the differences is then evaluated. The results
are tabulated as follows.
Method A Method B d d- (d - 2
17.6 17.9 +0.3 0.2 0.04
6.8 7.1 +0.3 0.2 0.04
14.2 13.8 -0.4 0.5 0.25
20.5 20.3 -0.2 -0.3 0.09
9.7 10.2 +0.5 0.4 0.16
= 0.5 =0.58
= 0.1

Recall; t =
In this example, we are determining whether there is significant difference in the two methods.
t= = = 0.589
The tabulated value of t is 2.78 (P = 0.05) and since the calculated value is less than this, there
is no significant difference between the methods.

Correlation
When using instrumental methods, it is often necessary to carry out a calibration procedure by
using a series of Samples (standards) each having a known concentration of the analyte to be
determined. A calibration curve is constructed by measuring the instrumental signal for each
standard and plotting this response against concentration. Provided the same experimental
conditions are used for the measurement of the standards and for the test (unknown) sample, the
concentration of the latter may be determined from the calibration curve by graphical
interpolation.
There are two statistical tests that should be applied to a calibration curve
(a) To ascertain if the graph is linear, or in the form of a curve
(b) To evaluate the best straight line (or curve) throughout the data points.
Correlation coefficient. In order to establish whether there is a linear relationship between
two variables x1 and y1 the Pearson’s correlation coefficient r is used.
r= where n is the number of data points.

The value of r must lie between +1 and -1; the nearer it is to +1, or in the case of negative
correlation to -1, then the greater the probability that a definite linear relationship exists
 between the variables x and y. values of r that tend towards zero indicate that x and y are
not linearly related (they may be related in a non-linear fashion).
Example
Quinine may be determined by measuring the fluorescence intensity in 1M H2SO4
solution. Standard solutions of quinine gave the following fluorescence values. Calculate
the correlation coefficient r.
Concentration of quinine (x1) 0.00 0.10 0.20 0.20 0.40 µgmL-1
Fluorescence intensity (y1) 0.00 5.20 9.90 15.30 19.10 arbitrary unit

Solution

x1 y1 x12 y12 x1y1

0.00 0.00 0.00 0.00 0.00
0.10 5.20 0.01 27.04 0.52
0.20 9.90 0.04 98.01 1.98
0.30 15.30 0.09 234.09 4.59
0.40 19.10 0.16 364.81 7.64
= 1.00 =49.5 = 0.30 = 723.95 = 14.73

Therefore,
= 1.00; = 2450.25; n= 5

Substituting the above values in

r= ,r=

r= = 0.9987

Hence, there is a very strong indication that a linear relation exits between fluorescence intensity
and concentration (over a given range of concentration).