Week 2

CHAPTER 1

Sample preparation-continued
14 SEPT 2015

Week 2

Preparing solutions
How to prepare a solution?
How to prepare a solution with a
certain concentration?
Expression of concentration
Concentration is the quantity of solute in a known amount of
volume or mass of solution or solvent
Concentration = Amount of solute
Amount of solution

Unit for concentrations

Molarity (M)
Formality (F)
Normality (N) (not common)
Molality (m)
Part per thousand (ppt)
Parts per million (ppm)
Parts per billion (ppb)
Percent concentration (%w/w, %w/v, %v/v)
Week 2

 Molarity (M) = number of moles of solute

liters of solution
concentration of a particular chemical species

Molality (m) = number of moles of solute

kilograms of solution
Formality (F) = number of moles of solute
liters of solution
substance's total concentration without regard to its specific chemical form

Week 2

For substances that DO NOT ionize in solution, such as glucose, molarity and
formality are the SAME
For substances that ionize in solution, such as NaCl, molarity and formality are
DIFFERENT.
NaCl (aq)
initial 0.1 mol final
0.0

Na+

Cl

0.1 mol

0.1 mol

The molarity of NaCl zero since there is no undissociated NaCl in solution. The
solution, instead, is 0.1 M in Na+ and 0.1 M in Cl. When we state that a solution is
0.1 M NaCl we understand it to consist of Na+ and Cl ions.
The formality of NaCl 0.1 F because it represents the total amount of NaCl in
solution.
The unit of formality is used only when it provides a clearer description
of solution chemistry.
4

Week 2

 Normality concentration in terms of an equivalent, which is the amount of

one chemical species reacting stoichiometrically with another chemical
species
Normality (N) = number of equivalents of solute
liters of solution

Week 2

Concentration in terms of
percent composition
The concentration of substance in commercial aqueous reagents, organic
solvents and commercial household products are usually expressed in
percent composition

Weight percent = Weight of solute

(%w/w)
Weight of solution

x 100%

Volume percent = Volume of solute

x 100%
(%v/v)
Volume of solution

Weight-volume =
(%w/v)

Weight of solute
x 100%
Volume of solution

Example: 37% is labeled on a HCl reagent bottle. This means that it contains 37 g
HCl per 100 g solution
Week 2

The w/w % unit may also be expressed as a fraction

e.g. 37% (w/w) can be expressed as 37 parts per hundred
How about smaller fractions??

Parts per thousand

Or

Parts per million (ppm)

Or

=
=
=
=

Both of these
units are for
SOLID SAMPLE

g of solute
103 g solution
mg/g
g solute
106 g solution
g/g

When the sample is in liquid form, we can use w/w % in

volume unit as well. How?
As the density of aqueous solution often very close to 1.00 g/L, we usually
correlates 1 g water with 1 mL water (approximation)

Therefore, 1 ppm = 1 g/g can also be converted to 1 g/mL = 1

mg/L
Week 2

For trace analysis in liquid sample

~Analysis of very very very little amount of analytes

Parts per billion (ppb)

g solute
109 g solution

Therefore,1 ppb = 1 ng/g = 1 ng/mL = 1 g/L

Parts per trillion (ppt)

g solute
1012 g solution

Therefore,1 ppt = 1 pg/g = 1 pg/mL = 1 ng/L

Week 2

Converting percent composition to molar concentration

-converting commercial term to easier laboratory term-

Example:
A commercial aqueous ammonia (NH3 ) bottle is labeled as 28.0% w/w,
and has a density of 0.899 g/mL and MW of 17.03 g/mol. Let say you want
to convert the concentration into molarity for easier calculation in your lab
book, how can you do that?
=

28 g/100 g

0.2517 g/mL (part per thousand)

0.2517 g/mL

0.01478 mol/mL

14.8 mol/L

0.899 g/mL

17.031 g/mol
x

density

MW

1000

or 14.8 M
9

Week 2

Preparation of solution
Example: How to prepare 250 mL of 0.100 M NaOH from NaOH
solid? [MW: NaOH = 40]
1. Calculate the weight (g) of NaOH pellet using MW to produce 0.1 M
solution
Calculations ???
2. Weigh ??? g of solid (generally 0.1 mg, ie up to 4 decimal places in
grams)
3. Dissolve in water, transfer (quantitatively with rinsing) to a 250 mL
volumetric flask, and dilute to the mark

Calculate

Weigh

Week 2

Dissolve and
transfer

10

Dilution of solution
The moles solute in concentrated (1) solution equals the
moles in dilute (2) solution

M1 V1 = M2 V2
M1 : Initial concentration of solution
V1 : Volume of concentrated solution transferred
M2 : Concentration of final solution (diluted)
V2 : Volume of concentrated solution transferred

Example: Describe the preparation of 50 mL of 0.100 M NaOH

solution from a 0.5 M solution
Calculations
How to ???
Glassware requirement: ?? mL pipet and ?? mL volumetric flask

Week 2

11

Week 2

12

Week 2+3
CHAPTER 2
Data analysis and statistics

Week 2

13

DATA ANALYSIS
Normal phrases in describing results of an analysis
pretty sure
Replaced by using statistical tests important to
very sure
understand the significance of data and therefore to
most likely
set limitations on each step of analysis.
probably

Errors in chemical analysis

Is there such a thing as ERROR FREE ANALYSIS?
Impossible to eliminate errors.
What is the maximum error that we
can tolerate?
Can only be approximated to an
acceptable precision.

Week 2

How reliable
is our data??

14

ACCURACY
degree of agreement between measured value
and the true value (which may not be known!)
Therefore, it is the degree of agreement
between measured value and the accepted true
value

Accuracy is how close you

get to the bullseye. Precision
is how close the repetitive
shots are one to another. It
is nearly impossible to have
accuracy without good
precision

PRECISION
Degree of agreement between replicate
measurements of the same quantity;
repeatability of a result.
Expressed by standard deviation, the coefficient
of variation, the range of the data or as
confidence interval (e.g. 95%) about the mean
value
How similar are values obtained in exactly the
same way?
Useful for measuring deviation from the mean.

di x i x
Week 2

15

High Precision
High accuracy

High precision
Low accuracy

Low precision
Low accuracy

Low precision
High accuracy

x
Week 2

16

Types of error in chemical analysis

Accuracy is expressed in terms of error
A. Systematic (determinate) Error
Determinable and that presumably can be either avoided or corrected.
Can be constant (e.g uncalibrated weight used in all weighing) or variable (e.g.buret whose
volume readings are in error by different amount at different volumes)
Readings all too high or too low that can affect accuracy.
How to detect?analysis of reference sample or determine recovery after adding known
standard
B. Random/accidental (indeterminate) Error
Represents experimental uncertainty occurred in any measurement
Random and cannot be avoided. Affects precision, can only be
controlled
Revealed by small differences in successive measurements made by
the same analyst under identical conditions cant be
predicted/estimated but can be dealt with statistics
Data scattered approximately symmetrically about a mean value.
C. Gross Errors
Serious but very seldom occur in analysis.
Usually obvious - give outlier readings.
Detectable by carrying out sufficient replicate measurements.
Experiments must be repeated.
e.g. Instrument faulty, contaminated reagent
Week 2

17

SOURCES OF SYSTEMATIC ERROR

Instrumental Error
1. All measuring apparatus contribute to systematic error; therefore need frequent calibration
pipettes, volumetric flasks, burettes
2. Also for electronic devices such as spectrometers, pH meter
3. Examples:
a) Temperature changes
Method Error
1. Due to inadequacies in physical or chemical behaviour of reagents or reactions (e.g. slow or
incomplete reactions, instability of reacting species, occurrence of side
reaction/interference).
2. Difficult to detect and the most serious systematic error (can be changed if the conditions of
determination are altered)
3. Correction by running
a) reagent blank- analysis on the added reagent only
b) 2 independent methods or SRM
Personal Error
1. Sources: Physical handicap, prejudice, not competent
2. Examples:
a) Insensitivity to colour changes
b) Transfer of solutions
c) Incomplete drying of samples
d) Tendency to estimate scale readings to improve precision
e) Math error in calculations
Week 2
18

Significant figures (SF)

The SF of a number are those digits that carry meaning contributing to its precision.
Zero is significant only when:
A. It occurs in the middle of a number. Examples:
a) 401
- 3 SF
b) 6.0015
- 5 SF
B. It is the last number to the right of the decimal point. Examples:
a) 3.00
- 3 SF
b) 6.00 102
- 3 SF
c) 0.0500
- 3 SF
Rounding off in the step of addition & subtraction
Use the same number of decimal places (dp) as the number with the fewest
decimal places. Example:
12.2 + 0.365 + 1.04 = 13.605 = 13.6 (1 dp)
(1 dp) (3 dp)
(2 dp)
SF in multiplication &Division
Use the same number of digits as
the number with the fewest number
of digits.
Week 2

40.1 0.1633

3.21 10

204 . 228
19

(3 sf )

Application of statistics in data analysis

Defining the confidence interval (RANGE) of values around a set mean (x)
within which the population mean () can be expected with a given
probability
Determining the number of replicates required to assure (at a desired
probability) that an experimental mean (x) falls within a predicted interval
of values around the population mean ().
Estimating the probability that the experimental mean (x) and true value
() are different or two experimental mean are different (t test).
Estimating the probability that data from two experiments are different
in precision (F-test).
Deciding when to accept/reject outliers among replicates (Q-test/
Dixons).
Treating calibration data.

Week 2

20

Statistical treatment of random error

Sample Mean/Average
N

xi

x i 1
N

If for large data set,

it is called population
mean ()

Xi = individual values of x
N = number of replicate measurements
Range
The difference between the highest
and lowest result.

Week 2

Median
Data in the middle if the number is odd,
arranged in ascending order.
For even numbers, the mean of the median
pair is used.
used when a set of data contains an outlier
Errors
Absolute error = measured value true value
E.g. if a 2.72 g sample is analysed to be 2.62g,
the absolute error is -0.10g
Relative error is the absolute error expressed
as % of the true value
E.g. (-0.10g/2.72 g) x 100% = -3.7%

21

Standard Deviation (SD)

a very important precision indicator
Each set of analytical results should be accompanied by an indication of the precision of the
analysis standard deviation which is measuring of the precision of a population of data.
Can be calculated using Excel spreadsheet
Small sample size (N=20)

xi x
N1

Population (N= >20)

xi

Xi = individual values of x
Xi = individual values of x
= mean
x = mean
N = number of replicate measurements N = number of replicate measurements

Smaller SD means more precise the analysis is

Week 2

22

Varian, V

Relative Standard Deviation (RSD)

also a very important precision
indicator

The square of standard deviation.

For sample, V = s2

s
RSD 100%
x

s 2 (V )

2
(
x

x
)
i
i 1

Acceptable RSD % depends on the

type of analysis. I.e.
Trace analysis, RSD % should be <5%
Environmental analysis <15%
Pharmaceutical analysis <10 %

N 1

Week 2

23

Week 2

24

Examples:
HPLC chromatogram of toxins
1

Can you guess which result (peak)

with higher precision?

2
Replicate 1
1

example sd rsd.xlsx

2
Replicate 2

Replicate 3
Week 2

25

Confidence limits (CL) and confidence

interval (CI) how sure are you?
Calculation of SD for a set of data provides indication of the precision inherent in
particular procedure.
If for large data set, it doesn't by itself give any info about how close the
experimentally determined mean ( x ) to the true mean value ().
Statistic allows to estimate range within which true value might fall, within a given
probability, defined by the experimental mean and sd.
This range confidence interval and the limits of this range confidence limit

The likelihood that the true value falls within the range is called
probability or confidence level, usually expressed as %

Week 2

26

Confidence interval for small data set (N = 20)

ts
CI ( ) x
N
N=Number of
measurements/replicates

Values of t depend on degree of freedom, v (N - 1) and

confidence level (from Table).
t also known as students t and will be used in
hypothesis test.

t=Students t values (constant and

given)
s= SD

Week 2

27

Values of t at various confidence level

Degree
of
Freedom,
v

Confidence Level

11

1.80

2.20

3.11

12

1.78

2.18

3.06

63.66

13

1.77

2.16

3.01

4.30

9.92

14

1.76

2.14

2.98

2.35

3.18

5.84

15

1.75

2.13

2.95

2.13

2.78

4.60

16

1.75

2.12

2.92

2.02

2.57

4.03

17

1.74

2.11

2.90

1.94

2.45

3.71

18

1.73

2.10

2.88

1.90

2.26

3.50

19

1.73

2.09

2.86

1.86

2.31

3.36

20

1.72

2.09

2.85

1.83

2.26

3.25

infinity

1.64

1.96

2.58

10

1.81

2.23

3.17

90 %

95 %

99 %

6.31

12.70

2.92

Week 2

28

Example:
Data for the analysis of calcium in rock are given by 14.35%, 14.41%, 14.40%, 14.32% and
14.37%. Within what range are you 95% confident that the true value lies??

ts
CI ( ) x
N

Solution:
Mean, x = 14.37
SD, s = 0.037
From the table, at 95 % confidence level, N - 1 = 4, t = 2.78.
Therefore, CI is :

2.78 0.037
x
5
14.37% 0.05%

So you are 95% confidence that (in the absence of determinate error),
the true value falls within 14.32% to 14.42%

Week 2

29

At different confidence level,

Confidence Level

Confidence Interval

90%

= 14.37% 0.04

95%

= 14.37% 0.05

99%

= 14.37% 0.08

Summary:
If the confidence level increased, the confidence interval (CI)
also increased. The probability of the true mean value ()
appeared in the interval will increase

Week 2

30

Confidence interval for large data set (N >20)

since the exact value of population mean () cannot be determined, one must
use statistical theory to set limits around the measured mean, that probably
contain .
CI only having meaning with the measured standard deviation, s, is a good
approximation of the population standard deviation, , and there is no bias in
the measurement.
CI when is known (population),

z
x
N

For the standard normal

distribution, P(-1.96 < Z < 1.96) = 0.95,
i.e., there is a 95% probability that a
standard normal variable, Z, will fall
between -1.96 and 1.96

N = Number of measurements/replicates
z= the value from the standard normal
distribution for the selected confidence
level
Week 2

31

Table 1 Values for z at various confidence levels

Confidence Level, %

50

0.67

68

1.00

80

1.29

90

1.64

95

1.96

96

2.00

99

2.58

99.7

3.00

99.9

3.29

Week 2

32

Examples:
At 90% confidence level, z 1.64,
x 1.64

At 95% confidence level, z 1.96,

x 1.96
N
At 99% confidence level, z 2.58,

x 2.58
or,
x
N

Week 2

2.58
x 2.58
N
N
33

Other usage of confidence interval (CI)

To determine number of replicates needed for the mean to be within the
confidence interval.
To determine systematic error.
A. To determine number of replicates
If is known (s ),

z
x
N

z
N
x

If is unknown

ts
N

Week 2

ts
N

x
34

B. To determine systematic error

1) Calculation method 1
Example
A standard solution gave an absorption reading of 0.470 at a particular wavelength.
Ten measurements were done on a sample and the mean gave a value of 0.461,
with standard deviation (s) of 0.003. Show whether systematic error exists in the
measurements at 95% confidence level.
Solution
At 95% confidence level, N = 10, t = 2.26,
Does the true mean 0.470
belong to the interval?

ts
CI ( ) x
N
0.461 2.26

0.003

Does systematic error present?

10

0.461 0.002
The calculation gives confidence limit of,
0.459 < < 0.463

Week 2

35

Values of t at various confidence level

Degree
of
Freedom,
v

Confidence Level

11

1.80

2.20

3.11

12

1.78

2.18

3.06

63.66

13

1.77

2.16

3.01

4.30

9.92

14

1.76

2.14

2.98

2.35

3.18

5.84

15

1.75

2.13

2.95

2.13

2.78

4.60

16

1.75

2.12

2.92

2.02

2.57

4.03

17

1.74

2.11

2.90

1.94

2.45

3.71

18

1.73

2.10

2.88

1.90

2.26

3.50

19

1.73

2.09

2.86

1.86

2.31

3.36

20

1.72

2.09

2.85

1.83

2.26

3.25

infinity

1.64

1.96

2.58

10

1.81

2.23

3.17

90 %

95 %

99 %

6.31

12.70

2.92

Week 2

36

B. To determine systematic error

2) Calculation method 2
Example
A standard solution gave an absorption reading of 0.470 at a particular wavelength.
Ten measurements were done on a sample and the mean gave a value of 0.461,
with standard deviation (s) of 0.003. Show whether systematic error exists in the
measurements at 95% confidence level.
Solution
At 95% confidence level, N = 10, t = 2.26,

t x

N
s

0.461 0.470

The tcalc >ttable

10
0.003

Does systematic error

present?

t 9.49

Week 2

37

Testing a hypothesis
Observations

Hypothesis Model
NO
Valid?

Reject

YES
Basis for further experiments

Week 2

38

Significant tests
Approach tests whether the difference between the two results is significant (due to
systematic error) or not significant (merely due to random error).

Hypothesis Data Theory

Normally, the measured data is not always the same and seldom agree.
We use statistics to test the disagreement.

Week 2

39

Null hypothesis, ho
The values of two measured quantities do not differ (significantly) UNLESS we can
prove it that the two values are significantly different.
Innocent until proven guilty
The calculated value of a parameter from the equation is compared to the
parameter value from the table.
If the calculated value is smaller than the table value, the hypothesis is
accepted and vice-versa.
Can be used to compare:

Week 2

40

Values of t at various confidence level

Degree
of
Freedom,
v

Confidence Level

11

1.80

2.20

3.11

12

1.78

2.18

3.06

63.66

13

1.77

2.16

3.01

4.30

9.92

14

1.76

2.14

2.98

2.35

3.18

5.84

15

1.75

2.13

2.95

2.13

2.78

4.60

16

1.75

2.12

2.92

2.02

2.57

4.03

17

1.74

2.11

2.90

1.94

2.45

3.71

18

1.73

2.10

2.88

1.90

2.26

3.50

19

1.73

2.09

2.86

1.86

2.31

3.36

20

1.72

2.09

2.85

1.83

2.26

3.25

infinity

1.64

1.96

2.58

10

1.81

2.23

3.17

90 %

95 %

99 %

6.31

12.70

2.92

Week 2

41

t-test
Comparison between experimental mean and true mean
( x and )
To check the presence of systematic error
Can be used to compare replicate measurement
Can be used to compare individual difference

Week 2

42

Steps in t-test
1) Comparing two mean values x and
i) If is not known,

ii) Calculate t or z (tcalc) from the data.

ts
x
N

iii) Compare tcalc and ttable

iv) If tcalc > ttable

t x -

N
s

Reject Null Hypothesis (Ho) i.e.

The difference is due to systematic error.

If is known,

N
z x

Week 2

v) If tcalc < ttable

Accept Null Hypothesis (Ho) i.e.

( x)

The different is due to random error.

43

Example:
The sulphur content in a sample of kerosene was found to be 0.123%. A new method
was used on the same sample and the following data is obtained:
%Sulphur : 0.112; 0.118; 0.113; 0.119
Show whether systematic error is present in the new method.
Null Hypothesis, Ho : =

x = 0.116%, = 0.123%, s = 0.0032

t x

N
s

0.116 0.123
4.38 (t calc )
Week 2

4
0.0032

t table = 3.18 (95 %, N-1 = 3)

Since tcalc> ttable, Ho is rejected and the
two means are significantly different and
thus systematic error is present.

44

Other Solution:

x 0.116 - 0.123
0.007 (experimental data)
x

ts
N
3.18 0.0032
4

0 .0051 ( Table value )

Since,

Ho is rejected, the difference ( x ) is

significant and there is systematic error in
the measurement.

x calculated x - table

ts
N

Week 2

(i.e. 0.007 > 0.0051)

45

2.Comparing two mean values

x 1 and x 2

Normally used to determine whether the two samples are identical or not.
The difference in the mean of two sets of the same analysis will provide information on
the similarity of the sample or the existence of random error.
Data:
Ho :

x1

x2

and s1, s2

x1= x 2

We want to test whether

x 1- x 2

=0

x1
1

x2
2

Week 2

ts

N
ts

46

 1 2 and 1 2

Assume,
Calculate the value of t;

t calc

x1 - x 2

sp

N1N2
N1 N2

Compare tcalc with ttable; if tcalc< ttable, Ho is accepted.

The pooled standard deviation, sp is calculated using:

(N1 - 1 )s12+(N 2 - 1 )s22

s p=
N1 + N 2 2
where,
N1, N2 are numbers of data in sets 1 and 2

Week 2

47

Example;

Since tcalc < ttable, Ho is accepted

and the source of the urine
statistically is the same.

Week 2

48

Values of t at various confidence level

Degree
of
Freedom,
v

Confidence Level

11

1.80

2.20

3.11

12

1.78

2.18

3.06

63.66

13

1.77

2.16

3.01

4.30

9.92

14

1.76

2.14

2.98

2.35

3.18

5.84

15

1.75

2.13

2.95

2.13

2.78

4.60

16

1.75

2.12

2.92

2.02

2.57

4.03

17

1.74

2.11

2.90

1.94

2.45

3.71

18

1.73

2.10

2.88

1.90

2.26

3.50

19

1.73

2.09

2.86

1.86

2.31

3.36

20

1.72

2.09

2.85

1.83

2.26

3.25

infinity

1.64

1.96

2.58

10

1.81

2.23

3.17

90 %

95 %

99 %

6.31

12.70

2.92

Week 2

49

F-test
Comparing the precision of two measurements
Is Method A more precise than Method B?
Is there any significant difference between both methods?

Do you remember?
V= s2

v1
F

v2

2
s1
2
s2

nominator
denominator

With the degree of freedom = N 1,

Ho : the precision is identical; s1 = s2
Then, if Fcalc < Ftable , Ho is accepted.
Since the values of F (from table) are always greater than 1, the smaller
variance (the more precise) always become the denominator.
V1 > V2, so F 1
Week 2

50

F values

s 21

v1
F
2
v2 s 2

Week 2

51

Example:
The determination of CO in a mixture of gases using the standard procedure
gave an s value of 0.21 ppm. The method was modified twice giving s1 of
0.15 (10 degrees of freedom (N-1)) and s2 of 0.12 (10 degrees of freedom
(N-1))
Are the two modified methods more precise than the standard?
Solution:

Ho : s1 = sstd and Ho : s2 = sstd

s 2std 0.212
F1 2
1.96
2
s1
0.15

F2

s 2std
s 22

0.212

3.06
2
0.12

In the standard method, s and the degrees of freedom becomes infinity.

Refer the F table:
Numerator = , and denominator = 10; giving the critical: Ftable= 2.54
Since F1 < Ftable, Ho is accepted, while F2>Ftable, Ho is rejected.

Week 2

52

Conclusions

Ho : s1 = sstd.
and
Ho : s2 = sstd.
Week 2

53

The Dixon test (Q test)

A way of detecting outlier a data that does not belong to the set.

Example:
Data: 10.05, 10.10, 10.15, 10.05, 10.45, 10.10
By inspection,10.45 seem to be out of the data normal range.
It is easier to see it when the numbers are arranged in a decreasing or
increasing order.
10.05, 10.05, 10.10, 10.10, 10.15, 10.45
At 95% confidence limit, should this data be eliminated??
If this data (10.45) is eliminated, the mean will
change from the original value!
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Q exp t

x q - xn
w

where,
xq = the questionable data
xn = its nearest neighbour
w = the difference between the highest and the lowest value (range).

The Qexpt or Qcalc will be compared with the Qcritical or Qtable,

and the Ho is checked.

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Values of Q

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Solution:
Q exp t
Q exp t

xq - xn
w

10.45 - 10.15

10.45 - 10.05

= 0.75
Qcritical (95%, n = 6) = 0.625

Qexpt > Qcritical

Data (10.45) can be
rejected.
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Example:
An analysis on calcite gave the following percentage of CaO:
55.45, 56.04, 56.23, 56.00, 55.08
Q: Is there any that data should be
rejected at 95% confidence level?
Solution:
Arrange data
55.08, 55.45, 56.00, 56.04, 56.08, 56.23
Suspected data: 55.45 OR 56.23
Qtable from 5 determinations, 95% = 0.710

Q calc

56.23 - 56.08
56.23 - 55.45

Qcalc<Qtable. Data cannot be rejected.

0.19

Q calc

55.45 - 56.00

56.23 - 55.45
0.71

Week 2

Qcalc = Qtable. Data cannot be rejected.

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