Introduction

1.1 The Scope of Analytical Chemistry:

Analytical chemistry has bounds which are amongst the widest of any technological discipline.
An analyst must be able to design, carry out, and interpret his measurements within the context of the
fundamental technological problem with which he is presented. The selection and utilization of
suitable chemical procedures requires a wide knowledge of chemistry, whilst familiarity with and the
ability to operate a varied range of instruments is essential. Finally, an analyst must have a sound
knowledge of the statistical treatment of experimental data to enable him to gauge the meaning and
reliability of the results that he obtains.
When an examination is restricted to the identification of one or more constituents of sample, it is
known as qualitative analysis, while an examination to determine how much of a particular species
is present constitutes a quantitative analysis. Sometimes information concerning the spatial
arrangement of atoms in a molecule or crystalline compounds is required or confirmation of the
presence or position of certain organic functional groups is sought. Such examinations are described
as structural analysis and they may be considered as more detailed forms of analysis. Any species
that are the subjects of either qualitative or quantitative analysis are known as anlayte.

1.1.1 The Function of Analytical Chemistry:

Some of the major areas of application are listed below.
(a) Fundamental research:
The first steps in unraveling the details of an unknown system frequently involve the
identification of its constituents by qualitative chemical analysis. Follow up investigations usually
require structural information and quantitative measurements. This pattern appears in such diverse
areas as the formulation of new drugs, the examination of meteorites, and studies on the results of
heavy ion bombardment by nuclear physicists.

(b) Product development:

The design and development of a new product will often depend upon establishing a link
between its chemical composition and its physical properties or performance. Typical examples are
the development of alloys and of polymer composites.

(c) Product quality control:

Most manufacturing industries require a uniform product quality. To ensure that this requirement
is met, both raw materials and finished products are subjected to extensive chemical analysis. On the
other hand, the necessary constituents must be kept at the optimum levels, while on the other
impurities such as poisons in foodstuffs must be kept below the maximum allowed by law.
(d) Monitoring and control of pollutants:
Residual heavy metals and organo-chlorine pesticides represent two well known pollution
problems. Sensitive and accurate analysis is required to enable the distribution and level of a
pollutant in the environment to be assessed and routine chemical analysis is important in the control
of industrial effluents.
(e) Assay:
In commercial dealings with raw materials such as ores, the value of the ore is set by its metal
content. Large amounts of material are often involved, so that taken overall small differences in
concentration can be of considerable commercial significance. Accurate and reliable chemical
analysis is thus essential.
(f) Medical and Clinical Studies:
The level of various elements and compounds in body fluids are important indicators of
physiological disorders. A high sugar content in urine indicating a diabetic condition and lead in
blood are probably the most well-known examples.

Table (1.1): A general classification of important analytical methods

Group

Property measured

gravimetric

weight of pure analyte or of a stoichiometric compound containing it

volumetric

volume of standard reagent solution reacting with the analyte

spectrometric

intensity of electromagnetic radiation emitted or absorbed by the

analyte

electrochemical

electrical properties of analyte solutions

chromatographic

physico-chemical properties of individual analytes after separation

1.1.2 Glossary of Terms

Accuracy:
The agreement between a measured value and the accepted true value.
Precision:
The degree of agreement between replicate measurements of the same quantity. That is, it is the
repeatability of result. Good precision does not assure good accuracy.
Analyte:
Constituent of the sample which is to be studied by quantitative measurements or identified
qualitatively.
Blank:
A measurement or observation in which the sample is replaced by simulated matrix, the conditions
otherwise being identical to those under which a sample would be analyzed. Thus, the blank can be
used to correct for background effects and to take account of analyte other than that present in the
sample which may be introduced during the analysis, e.g. from reagents.

Primary standard:
A substance whose purity and stability are particularly well-established and with which other
standards may be compared.

Requirements of titration:
1-

The reaction must be stoichiometric. That is, there must be well-defined and known reaction

between the analyte and titrant. In the titration of acetic acid in vinegar with sodium hydroxide, for
example, a well defined reaction takes place:
CH3COOH + NaOH CH3COONa + H2O
2- The reaction should be rapid.
3-

There should be no side reactions, and the reaction should be specific.

4-

There should be a marked change in some property of the solution when the reaction is complete.

This may be change in the color of the solution or in some electrical or other physical property of the
solution. In the titration of acetic acid with sodium hydroxide, there is a marked increase in the pH of
the solution when the reaction is complete. A color change is usually brought about by addition of an
indicator whose color is dependent on the properties of the solution, for example, the pH.
5- The point at which an equivalent or stoichiometric amount of titrant is added is called the
equivalence point. The point at which the reaction is observed to be complete is called the end point,
that is, when a change in some property of the solution is detected. The end point should coincide
with the equivalence point.
6- The reaction should be quantitative. That is, the equilibrium of the reaction should be far to the right
so that a sufficiently sharp change will occur at the end point to obtain the desired accuracy.
Requirements of primary standard:
1- It should be 100% pure.
2- It should be stable to drying temperatures.
3- It should be readily available.
4- It should have a high formula weight (to minimize weighing error).

5- It should posses the properties required for a titration (soluble and react rapidly ....).

1.3 Classification of volumetric methods:

There are four general classes of volumetric methods.
1- Acid-Base:
Many compounds, both inorganic and organic, are either acids or bases and can be titrated with a
standard solution of a strong base or a strong acid. The reactions involve the combination of
hydrogen and hydroxide ions to form water. The end points of these titrations are easy to detect,
either by means of indicator or by following the change in pH with a pH meter. The acidity and
basicity of many organic acids and bases can be enhanced by titrating in nonaqueous solvent, so the
weaker acids and bases can be titrated.
2- Precipitation:
In the case of precipitation, the titrant forms an insoluble product with the analyte. An example is
the titration of chloride ion with silver nitrate solution.
3- Complexometric:
In complexometric titrations, the titrant is a complexing agent and forms a water-soluble complex
with the analyte, a metal ion. The titrant is often a chelating agent.
4- Reduction-oxidation:
The "redox" titrations involve the titration of an oxidizing agent with a reducing agent, or vice
versa. An oxidizing agent gains electrons and a reducing loses electrons in a reaction between them.

1.3.1 Expressions of concentration:

1- Concentration of solutions:
Standard solutions are expressed in terms of molar concentrations or molarity (M).
Molar solution is defined as one that contains one mole of substance in each liter of a solution.
Molarity of a solution is expressed as moles per liter or as millimoles per milliliters.
moles = (moles/liter) x liters = molarity x liters
millimoles = molarity x milliliters

mmole = M x ml
Molarity (M) = (moles of solute) / (volume of solution in liters)

2- The equivalent weight:

The equivalent weight is that weight of a substance in grams that will furnish one mole of the
reacting unit. Thus, for HCl, the equivalent weight is equal to the formula weight:
eq . wt . HCl

Formula wt. HCl (g / mol)

1 (eq / mol)

The milliequivalent weight is one thousandth of the eq.wt.

A normal solution contains one gram eq.wt of solute in one liter of solution:
N

No. of gram - equivalents

No. of liters

No. milligram - equivalents

No. of milliliters

By rearrangement of these equations we obtain the expression for calculating other quantities:
No of gram eq. = N x No. of liters
No. of milligram eq. = N x ml = N
3- Formality (F):
Is the number of gram formula weight of solute dissolved in liter of solution?
F =

number FWs solute

liters solution

4- Normality:
It is often convenient to calculate the titer of the titrant.
The titer is the weight of analyte that is chemically equivalent to 1 ml of titrant, usually expressed in
milligrams. For example, if 1 ml of a hydrochloric acid solution will exactly neutralize 4 mg of
sodium hydroxide, the titer is 4 mg/ml.
Titer (T) can be easily converted to normality as seen from the following equations:

mg
ml

N=

mg
ml x Eq. wt

Thus T= N x Eq.wt
In the example, if the titer of hydrochloric acid solution is 4 mg/ml of sodium hydroxide, the normality
is obtained upon dividing by 40 mg/meq (the equivalent weight of sodium hydroxide) giving a
normality of 0.1 meq/ml.
5- Weight, Volume, and Weight-to-Volume Ratios
Weight percent (% w/w), volume percent (% v/v) and weight-to-volume percent (% w/v)
express concentration as units of solute per 100 units of sample. A solution in which a solute has a
concentration of 23% w/v contains 23 g of solute per 100 mL of solution.
Parts per million (ppm) and parts per billion (ppb) are mass ratios of grams of solute to one million or
one billion grams of sample, respectively. For example, a steel
that is 450 ppm in Mn contains 450 g of Mn for every gram of steel. If we approxi- mate the density
of an aqueous solution as 1.00 g/mL, then solution concentrations can be expressed in parts per
million or parts per billion using the following relationships.
PPm = mg = g
Liter

mL

PPb = g

= ng

Liter

mL

For gases a part per million usually is a volume ratio. Thus, a helium concentration of 6.3 ppm
means that one liter of air contains 6.3 L of He.

1.3.2 The pH scale:

Sorensen suggested a scale based on the exponent of the hydrogen ion concentration but with sign
reversed. The concentration of H+ or OH- in aqueous solutions can vary over wide range, from 1 M
Or greater to 10-14 M or less.
The pH of a solution is defined as:
pH = - log [H+]
A similar definition is made for the hydroxyl ion concentration:
pOH = - log [OH-]

The following summary indicates the relationship: Table 2

H

10

10

0
n

pH

pO

1
H

10

10

10

10

10

10

10-

10

10-

10-

1
4

10

11

12

13

14

13

12

11

10

4
Acidic

N.

pH scale of water
Kw = [H+] [OH-]
- log Kw = - log [H+] [OH-] = - log [H+] - log [OH-]
pKw = pH + pOH
A t 25C

10-

10-

14 = pH + pOH

The reaction of any aqueous solution at room temperature is defined:

pH < 7 < pOH acidic reaction
pH = 7 = pOH

neutral reaction

pH > 7 > pOH

basic reaction

Example:
i-Find the pH of a solution of which [H+] = 4.0x 10-5.
pH = - log [H+] - log (4x10-5) = 5 - log 4 = 5 - 0.602 = 4.398
ii- Calculate the pH of a 2.0 x 10-3 M solution of HCl ?
HCl is completely ionized, so
[H+] = 2.0 x 10-3 M
pH = - log (2.0 x 10-3) = 3 - log 2.0 = 3 - 0.30 = 2.70

Alkaline

Calculation of pH:
1- pH of strong acid and bases:
Ionization of strong acids and strong bases is complete. The concentration of H+ or OH- is
determined readily from the concentration of the acid or base.
pH = - log [H+]
pOH = - log [OH-]
2- pH of weak acids:
HA

+ A

weak acid

[H ] [A - ]
Ka = acidity constant =
........................................................ (5)
[HA]
.. [H+] = [A-]
Eq (1) becomes:

[ H ]2
Ka =
................................................................................................ (6)
[ HA ]
If the molecular concentration of acid is Ca:
undissociated part = [Ca - [H+]]
Eq. (2) becomes:
Ka =

[ H ]2
.......................................................................................... (7)
C a - [H + ]

[H+]2 = Ca . Ka
[H+] Ca . Ka
pH = pCa + pKa
3- pH of weak bases:
+

BOH

weak base

+ OH

[ B ] [OH - ]
Kb =
......................................................................................... (8)
[ BOH]
.. [B+] = [OH-]
Eq (1) becomes:

[OH ]2
Kb =
.......................................................................................... (9)
[BOH]
If the molecular concentration of base is Cb and undissociated part is very small,
Eq. (9) becomes:
Kb =

[OH ]2
Cb

[OH-]2 = Ca . Kb
[OH-] Cb . Kb
pOH = pCb + pKb
But since pOH = pKw - pH
pKw - pH = pCb + pKb
and pH = pKw - PCb - pKb
4- pH of salts solution:
When an acid in solution is exactly neutralized with base, the resulting solution corresponds
to a solution of the salt of the acid-base pair. This is a situation which frequently arises in analytical
procedures and the calculation of the exact pH of such a solution may be of considerable importance.
The neutralization point or end point in an acid-base titration is a particular example.
Salts may in all cases be regarded as strong electrolytes so that a salt AB derived from acid AH
and base BOH will dissociate completely in solution. If the acid and base are strong, no further
reaction is likely to occur and the solution pH remains unaffected by the salt. However, if either or
both acid and base are weak a more complex situation will develop. It is convenient to consider three
separate cases: (a) weak acid - strong base, (b) strong acid - weak base and (c) weak acid - weak
base.

10

(a) Weak acid - strong base solution:

The conjugate base A- will react with water and undergo hydrolysis,
-

A + H2O

AH + OH ............................................... (10)

Prodrug undissociated acid and hydroxyl ions with an accompanying rise in pH. The equilibrium
constant for this reaction is known as the hydrolysis constant Kh.

[AH ] [OH - ]
Kh
............................................................................... (11)
[A ] [H 2 O]
(The solvent term [H2O] is by convention omitted. Kb is simply related to Kw and Ka by equation
(12) and as such is a redundant constant whose use should be discouraged).

[H + ] [OH ] [AH]

K h ............................ (12)
[H + ]
[A ]

K w [AH ] [OH - ]

Ka
[A ]

Equation (10) shows that the amounts of AH and OH- generated in the hydrolysis are equal.
Furthermore, if it is assumed that only a small amount of the salt is hydrolyzed, the concentration C Aof the salt dissolved is approximately the same as the concentration of A-. Then from (12):
[ OH ] 2 K w

Kb
CA
Ka

[OH-] =

Kw
.C A C A K w / K a
Ka

1/ 2

.................................................. (13)

pOH = pKw - pKa + pCA- = pKw - pH

pH = pKw + pKa - PCs
The pH of the solution will be dependent upon both pKa for the acid HA and on the
concentration of the salt dissolved in the solution. For example, the pH of solutions of sodium
cyanide may be calculated as follows:

11

pKa for HCN = 9.22

pH = 7.0 + 4.6 + log CA= 11.6 + log CAThus when:
C= 1 mole dm-3

pH = 11.6

C= 0.1 mol dm-3

pH = 11.1

C= 2 mol dm-3

pH = 11.8

(b) Strong acid - weak base solution:

Similar reasoning shows hydrolysis leading to the production of hydrogen ions and drop in pH,
B- + H2O

BOH + H+

and enables an analogous expression for pH to be derived, i.e.

pH = pKw - pKb + pC ....

1.3.3 Buffers:
A buffer is defined as a solution that resists change in pH when a small amount of an acid or
base is added or when the solution is diluted. A buffer solution consists of a mixture of a weak acid
and its conjugate base or a weak base and its conjugate acid at predetermined concentrations and
ratios. Typical mixtures are acetic acid and sodium acetate, ammonia and ammonium chloride, boric
acid and borate ...etc. The pH values of such mixtures will lie at point on rather flat regions of the
titration graphs, pH vs. milliters, for the various weak acids or bases.
Consider an acetic acid - acetate buffer. The acid equilibrium that governs this system is:
HOAc

OAc ......................................................... (21)

Since we have added a supply of acetate ions to the system (from sodium acetate for example), the
hydrogen ion concentration is no longer equal to the acetate ion concentration. The hydrogen ion
concentration is:

12

[H ] K a

[ HOAc ]
............................................................................... (22)
[ OAc ]

Taking the negative logarithm of each side of this equation we have

log[ H ] log K a log

[ HOAc ]
................................................... (23)
[ OAc ]
pH pK a log

[ HOAc ]
[ OAc ]

upon inverting the last log term, it becomes positive:

[OAc ]
pH pK a log
[ HOAc]
This terms of the ionization constant equation is called the Henderson-Hasselbach equation. It is
useful for calculating the pH of a weak acid solution containing its salt. A general form can be
written for a weak acid HA that ionizes to its salt, A-, and H+:
HA

+ A ..................................................................... (24)

[A ]
pH pK a log
...................................................................... (25)
[ HA]
pH pK a log

[ conjugate base ]
[ acid ]

The mixture of a weak acid and its salt may also be obtained by mixing an excess of weak acid
and some strong base to produce the salt by neutralization or by mixing an excess of salt with strong
acid to produce the weak acid component of the buffer.
Mechanism of buffer for a mixture of a weak acid and its salt can be explained as follows. The
pH is governed by the logarithm of the ratio of the salt and acid.

pH cons tan t + log

13

[A - ]
[HA]

If the solution is diluted, the ratio remains constant, and so the pH of the solution does not
change. If a small amount of a strong acid is added, it will combine with an equal amount of the A
to convert it to HA. That is, in the equilibrium HA H+ + A, Le Chtelier's principle indicates that
add H+ will combine with A to form HA, with equilibrium lying far to the left if there is an excess
of A. The change in the ratio [A]/[HA] is small and hence the change in pH is small. If a small
amount of a strong base is added, it combine with part of the HA to form an equivalent amount of
A. Again, the change in the ratio is small.
The amount of acid or base that can be added without causing a large change in pH is governed
by the buffering capacity of the solution. The buffering capacity increases with concentrations of the
buffering species. In addition to concentration, the buffering capacity is governed by the ratio of HA
to A. It is maximum when the ratio is unity, that is, when the pH = pKa.
pH pK a log

[1]
pK a ...................................................................... (26)
[1]

In general the buffering capacity is satisfactory over a pH range of pKa 1.

Examples:
Calculate the pH of the solution produced by adding 10.0 ml of N hydrochloric acid to 1 liter
of a solution which is 0.1 N in acetic acid and 0.1 N in sodium acetate (Ka= 1.82 x 10-5).
pH pK a log

[Salt ]
[ Acid ]

Neglecting the volume change from 1000 to 1010 ml, the hydrochloric acid reacts with acetate ion
forming practically undissociated acetic acid.
+

CH3COO

CH3COOH

[CH3COO] = 0.1 - 0.01 = 0.09

[CH3COOH] = 0.1 - 0.01 = 0.11
pH = 4.74 + log

0.09
= 4.71 - 0.09 = 4.65
0.11

14

Hence on adding the strong acid, the pH changes only by 4.74-4.65 = 0.09 pH unit, whereas, if
10 ml of N-hydrochloric acid were added to 1 liter of pure water (pH= 7), the pH would have
changed

from

to

- log (0.01) = 2, i.e., by 5 pH units. This illustrates the action of the acetic acid sodium acetate buffer.
Similar calculation applies for mixtures of weak bases and its salt. We can consider the equilibrium
between the base B and its conjugate acid BH+ and write a pKa for the conjugate (Brnsted) acid:
BH+ = B + H+ ....................................................................................... (27)

Ka

[ B][ H ] K w

.................................................................................... (28)
Kb
[ BH ]

The logarithmic Henderson-Hasselbach form exactly as above:

[H ] Ka .

[ BH ] K w [ BH ]

.
[ B]
K b [ B]

Kw
[ BH ]
[ BH ]
log[ H ] log K a log
log
log
[ B]
Kb
[ B]

pH pK a log

pH pK a log

[ B]
[ B]
( pK w pK b ) log

[ HB ]
[ BH ]

[Pr oton accep tor ]

[Pr oton accep tor ]
( pK w pK b ) log
[Pr oton donor ]
[Pr oton donor ]

Since pOH = pKw - pH, we can also write:

pOH pK b log

[ BH ]
[Pr oton donor ]
pK b + log
[ B]
[Pr oton acceptor ]

The alkaline buffering capacity is maximum at pH = pKa = 14 - pKb or pOH = pKb with a usual
range of pKa 1.
We must select a buffer with pKa value near the desired pH i.e. a weak acid and its salt give the
best buffering in acid solution and a weak base and its salt give the best buffering in alkaline
solution. Table 3 shows some typical buffer solutions.

15

Table 1.3.4.1: Some typical buffer solutions

Solutions

pH range

Phthalic acid and potassium hydrogen phthalate

2.2-4.2

Citric acid and sodium citrate

2.5-7.0

Acetic acid and sodium acetate

3.8-5.8

Sodium dihydrogen phosphate and disodium hydrogen phosphate

6.2-8.2

Ammonia and ammonium chloride

8.2-10.2

Borax and sodium hydroxide

9.2-11.2

1.3.4 Acid-Base Indicators:

Acid-base indicators are substances whose presence during a titration renders the end-point
visible. Thus, at a certain pH very near, or at the equivalence point of the titration the indicator
produces in the system a change which is easily perceptible to the eye, and may consist of:
a-

Sharp transformation from one color to another or to colorless.

b- Formation of a turbidity in a clear solution, or clearing up of a turbidity.

c- Development or disappearance of a fluorescence.
Most of the color acid base indicators of practical value are organic in nature. As the color
changes of these indicators depend on the change of the pH, they must themselves be acids or bases.
Table 1.3.5.1 : A range of visual indicators of acid-base titrations

Indicator

Low pH

High pH

color

color

pKIn

Experimental
color change
range/pH

cresol red

ca. 1

red

yellow

0.2-1.8

thymol blue

1.7

red

yellow

1.2-2.8

bromo-phenol blue

4.0

yellow

blue

2.8-4.6

methyl orange

3.7

red

yellow

3.1-4.4

16

methyl red

5.1

red

yellow

4.2-6.3

bromo-thymol blue

7.0

yellow

blue

6.0-7.6

phenol red

7.9

yellow

red

6.8-8.4

phenolphthalein

9.6

colorless

red

8.3-10.0

The well known indicator phenolphthalein (below) is a diprotic acid and its colorless. It
dissociates first to a colorless form and then, on losing the second hydrogen to an ion with a
conjugated system; a red color results.
HO

HO

OH

OH

+ H 2O
+ H 3O

OH
O
Phenolphthalein
colorless, H2In

COO
COO

Phenolphthalein
colorless, HIn-

Phenolphthalein
Red, In2-

Methyl orange, another widely used indicator, is a base and is yellow in the molecular form.
Addition of a hydrogen ion gives a cation which is pink in color.

+-

Na O3S

N(CH3)2 + H3O+

N N
In, Yellow

H
+-

N N

Na O3S

N(CH3)2 + H2O

In , Pink

Increasing the concentration of indicators has a serious effect on one-color indicators such as
phenolphthalein.

1.3.5Back titration:

17

In back-titration, a known number of millimoles of reactant are taken in excess of the analyte.
The unreacted portion is titrated for example, in the titration of antacid tablets with a strong acid such
as HCl.

1.4 Gravimetric Methods of Analysis:

Gravimetry encompasses all techniques in which we measure mass or a change in mass.
When you step on a scale after exercising you are making, in a sense, a gravimetric determination of
your mass. Measuring mass is the most fundamental of all analytical measurements, and gravimetry
is unquestionably the oldest analytical technique.

1.4.1 Types of Gravimetric Methods:

1- Precipitation Gravimetry
Precipitation gravimetry is based on the formation of an insoluble compound fol- lowing
the addition of a precipitating reagent, or precipitant, to a solution of the analyte. In most methods
the precipitate is the product of a simple metathesis reac- tion between the analyte and precipitant;
however, any reaction generating a pre- cipitate can potentially serve as a gravimetric method. Most
precipitation gravimet- ric methods were developed in the nineteenth century as a means for
analyzing ores. Many of these methods continue to serve as standard methods of analysis.The
indirect determination of PO3-2 by precipitating HgCl2 is representative example, as is the direct
determination of Cl by precipitating AgCl
2-electrogravimetry:
In electrogravimetry the analyte is deposited as a solid film on one electrode in an
electrochemical cell. The oxidation of Pb+2 and its deposition as pbo2 on a Pt anode is one example
of electrogravimetry. Reduction also may be used in electro- gravimetry. The electrodeposition of Cu
on a Pt cathode, for example, provides a direct analysis for Cu+2
3- volatilization Gravimetry :

18

When thermal or chemical energy is used to remove a volatile species, we call the method
volatilization gravimetry. In determining the moisture content of food, thermal energy vaporizes
the H2 O. The amount of carbon in an organic com- pound may be determined by using the chemical
energy of combustion to convert C to CO2
4- particulate gravimetry:
In particulate gravimetry the analyte is determined following its re- moval from the
sample matrix by filtration or extraction. The determination of sus- pended solids is one example of
particulate gravimetry.
1.4.2 Important OF Gravimetry:
Gravimetry is one of only a small number of techniques whose measurements require only base SI units, such as mass and moles, and defined constants, such as Avogadros
number and the mass of 12C.* The result of an analysis must ultimately be traceable to methods,
such as gravimetry, that can be related to fundamental physical properties. Most analysts never
use gravimetry to validate their methods. Verifying a method by analyzing a standard reference
material, however, is com- mon. Estimating the composition of these materials often involves
a gravimetric analysis.

1.5 Volumetric Precipitation Titrations (Precipitimetry)

Precipitation titrations are volumetric methods based on the formation of a slightly soluble
precipitate. They are in many ways simpler than gravimetric methods. The precipitate needs not be
separated, and needs not be pure, as long as the impurity does not consume titrant. The substance is
determined simply by converting it into an insoluble form of known composition by adding a
standard solution of the titrant. The equivalence point is reached when an equivalent amount of the
titrant has been added. From the volume of the latter, the amount of the substance is calculated. The

19

precipitate must be sufficiently insoluble to ensure completion of the reaction and to ensure a marked
change in the concentration of the ions of precipitate at the equivalence point of the titration
.
Table:1.5.1 Substances determined by precipitation titrations with Ag+.
AsO43-, Br-, CNO-, CO32-, CrO42-, CN-, Cl-, C2O42-, I-, PO43-, SCN-, S2-, fatty
acids
Table 1.5.2: Miscellaneous precipitation titrations
Analyte

Reagent

Precipitate

Cl-, Br-

Hg2(NO3)2

Hg2Cl2, Hg2Br2

SO42-, MoO42-

Pb(NO3)2

PbSO4, PbMoO4

Zn2+

K4Fe(CN)6

K2Zn3[Fe(CN)6]2

PO34-, C2O42-

Pb(OAc)2

Pb3(PO4)2, PbC2O4

1.5.2 Limitations of volumetric precipitation titrations:

Volumetric precipitation reactions have several limitations combining those of the
titrimetric methods in general, and some of those of the gravimetric methods. In particular, few
points need here be stressed in comparing volumetric precipitation reactions with the other
volumetric reactions, where no precipitates form.

1- The rate of reaction:

Particularly in dilute solutions, is often slow so that a long wait is necessary for each addition
of titrant to react completely. As the equivalence point is approached, a high degree of
supersaturation will not exist, and the rates of precipitation and attainment of solubility equilibrium
become too slow for convenience of titration. The most suitable alternative, and often the only one, is
to take advantage of a more rapid precipitation in the reverse direction, i.e. to add a measured excess
of titrant and back-titrate.

20

2- The lack of suitable indicators for many precipitation titrations imposes another limitations in
such titrations.

1.5.3 Limitations of argentometric titrations:

1-

Reducing agents, such as, sulphur dioxide interfere by reducing the silver ions, and must be

removed by previous oxidation.

2-

Coloured compounds of any sort obscure the end point, which is taken as the faintest ting of

colour detectable on the precipitated silver halide, or in solution, as the case may be.
3-

Silver halides are sensitive to photodecomposition, and the titration should be carried out in

diffused daylight, or artifical light.

4-

Most cations except the alkalies and alkaline earths interfere in several ways. (a) Some, such as

Fe3+ form insoluble coloured hydroxide in neutral or slightly acid medium; (b) Some, such as Al 3+,
hydrolyse to insoluble basic salts in neutral or slightly acid solution, showing a tendency to
coprecipitate chloride; (c) Hg2+ form soluble complexes with halides of the type [HgI4]2-.

1.5.4 Solubility product:

Ionic reactions are actually complete when any change occurs that lowers the concentrations of
ions to very small values. The factor governing the completeness of a precipitation reaction is the
solubility of the precipitate formed. The more insoluble the precipitate, the more complete is the
reaction at the equivalence point of the titration, and the larger is the change in concentration of the
reacting ions. The equilibrium constant expressing the solubility of a precipitate is the familiar
solubility product constant.
Let us consider what happens when the sparingly soluble salt AB comes in contact with water.
Some of the salt dissolves in water and, assuming this compound to be an ionic solid, dissociates into
its ions A+ and B-. This reaction can be represented in its simplest form (i.e. without considering
hydration of the ions) by the equation:
AB A+ + B- ................................................................................... (1)

21

However, as the salt AB dissolves, more and more A+ and B- are in solution, with the net result
that the chance of their recombining to form AB increases; that is, the equilibrium represented
simply by equation (2) for the saturated solution is established.
AB (solid)

A+ + B- (solution)

(2)

The equilibrium constant for this reaction is:

[A ][ B ]
[AB]

Since the concentration of AB is constant as long as the temperature remains constant and there
is some solid AB in contact with the solution, this equilibrium expression becomes.
K X const. = [A+] [B-] = SAB = Solubility product constant.
Let us consider in the same way the saturated solution of the sparingly soluble salt X m Yn
which dissociates into m cation, Xn+ and n anions, Ym-. The equilibrium for this saturated solution
can be represented by the equation:

[Xn+]m [Ym-]n = SXm Yn

1.6 spectrometric Methods of Analysis:

Electromagnetic radiation, or light, is a form of energy whose behavior is described by the
properties of both waves and particles. The optical properties of electromag- netic radiation, such as
diffraction, are explained best by describing light as a wave. Many of the interactions between
electromagnetic radiation and matter, such as ab- sorption and emission, however, are better
described by treating light as a particle, or photon.

22

1.6.1 Wave Properties of Electromagnetic Radiation :

Wave Properties of Electromagnetic Radiation Electromagnetic radiation consists of
oscillating electric and magnetic fields that propagate through space along a lin- ear path and with a
constant velocity (Figure). In a vacuum, electromagnetic.
In a vacuum, electromagnetic radiation travel at the speed of lightc,which 2.99*108 m/s. .
Electromagneticradiation moves through a medium other than a vacuum with a velocity, v, less than
that of the speed of light in a vacuum.
The difference between v and c is small enough (0.1%) that the speed of light to three
significant figures ,3.00 108 m/s, is sufficiently accurate for most purposes.
Oscillations in the electric and magnetic fields are perpendicular to each other,and to the
direction of the waves propagation. Figure 10.1 shows an example of plane-polarized
electromagnetic radiation consisting of an oscillating electric field and an oscillating magnetic
field, each of which is constrained to a single plane. Normally, electromagnetic radiation is
unpolarized, with oscillating electric and magnetic fields in all possible planes oriented
perpendicular to the direction of propagation.
The interaction of electromagnetic radiation with matter can be explained using either
the electric field or the magnetic field. For this reason, only the electric field component is shown in
Figure 10.2. The oscillating electric field is described by a sine wave of the form

E =Ae sin(2t + )
where E is the magnitude of the electric field at time t, Ae is the electric fields maxi-mum
amplitude, is the frequency, or the number of oscillations in the electric field per unit time,
and is a phase angle accounting for the fact that the electric fields magnitude need not be zero at t
= 0. An identical equation can be written for the magnetic field, M

M= Am sin(2t + )

23

where Am is the magnetic fields maximum amplitude.An electromagnetic wave, therefore,

is characterized by several fundamental properties, including its velocity, amplitude, frequency,
phase angle, polarization, and direction of propagation. Other properties, which are based on
these funda-mental properties, also are useful for characterizing the wave behavior of electromagnetic radiation. The wavelength of an electromagnetic wave, , is defined as the distance
between successive maxima, or successive minima (see Figure.1.6.1).

1.6.3 Particle Properties of Electromagnetic Radiation

When a sample absorbs electro- magnetic radiation it undergoes a change in energy. The
interaction between the sample and the electromagnetic radiation is easiest to understand if we
assume that electromagnetic radiation consists of a beam of energetic particles called photons. When
a photon is absorbed by a sample, it is destroyed, and its energy acquired by the sample. The
energy of a photon, in joules, is related to its frequency, wave- length, or wavenumber by the
following equations

24

where h is Plancks constant, which has a value of 6.62606896(33)1034 Js

Wavelength
(m)

10

Frequency (s1)

Type of transition
Spectral region

1012

1010

108

106

104

102

100

102

1
4

1022

1020

1018

Nuclear
spin

Molecular
rotations;
electron
spin
Radio wave
Microwave

1016

1014

Valence
Molecular electrons
vibrations
UV

1012

1010

Corelevel
electron
s
X-

IR

Visible

Wavelength (nm)

480

580

680

780

8
0

Violet

Blue Green Yellow Orange

Figure 1.6.2

The electromagnetic spectrum

showing the colors of the
visible spectrum.

25

Red

108

Nuclear

-ray

26

27

2.1 Chromatography
The term "chromatography" is derived from the original use of this method for
separating yellow and green plant pigments. Chromatography has since evolved into
a very general separation method for many types of mixtures. Examples of the
application of chromatographic methods are (i) the purification of reaction mixtures
in chemical synthesis, (ii) the purification of bio-molecules such as proteins for
pharmaceutical research, (iii) the analysis of complex sample mixtures such as
those

obtained

in forensics (body fluids, paints etc) and (iv) the analysis of

environmental samples.

Key terms:
Adsorbtion:

Interaction of solute molecules (or atoms or ions) with the

surface of the stationary phase (note that it is different from
absorption where the molecules fill the pores of a solid).

Eluent:

The mobile phase (usually for solvents)

Elution:

Motion of the mobile phase through the stationary phase

Elution time:

The time taken for a solute to pass through the system. A

solute with a short elution time travels through the stationary
phase rapidly, i.e. it elutes fast.

Mobile phase:

The part of the chromatography system that is mobile.

Commonly a solvent mixture (as in column chromatography or thin
layer chromatography or a gas (as in gas chromatography).

Normal phase:

Unmodified stationary phase where polar solutes interact

28

strongly and run slowly

Reverse phase:

Modified stationary phase where polar solutes run fast i.e.

reverse order

Degree of separation of different solutes. In principle, resolution

Resolution:

can be improved by using a longer stationary phase, finer stationary

phase (e.g. column packing or TLC plate coating) or slower elution.
Stationary phase:

The part of the chromatography system that is fixed in place. Most

commonly a solid e.g. the packing in column chromatography or gas
chromatography or the coating on a chromatographic plate.

Rf value

distance travelled by solute

distance travelled by solvent

Rf = retardation factor. The Rf value has to be between 0 and 1 (by definition),

and it is typically reported to two decimal places.

2.1.1 Description
Chromatography is based on selective adsorption of compounds on a solid (or
liquid) with high surface area (the stationary phase). As the solute mixture passes
over the solid, the components are adsorbed and then released from the surface at
differing rates.

This means that the solutes are continuously partitioned between the

adsorbent and the mobile phase, either a gas or a solvent mixture.

The stronger the interaction of the solute with stationary phase, the slower the
solute will progress. The motion of the solute and solvent through the stationary
phase in called elution.

The process is analogous to fractional distillation or

29

extraction, in which different compounds are partitioned between liquid and vapour,
or between two immiscible liquids, respectively.

Chromatography can be carried out in many ways. In column chromatography

and in thin layer chromatography (TLC) a solution of the mixture flows over a solid
adsorbent.

Separation occurs as molecules are adsorbed and desorbed while

passing over the surface.

In paper chromatography applications, the mixture is

partitioned between water molecules adsorbed on the paper and a solvent that moves
over the paper.

In gas chromatography (GC, also called vapour phase

chromatography), a mixture of volatile compounds is separated by passing the vapour

over an adsorbent packing in a long, heated tube.
.

Chromatographic methods have high "resolving power", i.e. they are capable of
sharp separations of closely related compounds, particularly when very small samples
are used. Both GC and column chromatography can be carried out with instruments
that detect extremely small amounts of compounds in the gas or liquid stream, as it
leaves the chromatographic column. In GC, the detector responds to the thermal
conductivity of the gas stream or the ionisation of the gas as it passes through a
flame.
This gas stream can be introduced straight into a mass spectrometer (MS) to
give the very important technique of GCMS which facilitates the separation and
identification of samples within a mixture.

In liquid (column) chromatography

instruments, the detector senses changes in the refractive index or uv-visible

absorption of the solution.
Signals from the detector corresponding to each component in the mixture

30

and proportional to the amount of the compound are recorded automatically on a

chart. These instruments thus provide powerful methods for quantitative analysis.

There are four main types of chromatography

Gas Chromatography

Liquid Chromatography

Thin - layer Chromatography

Paper Chromatography

2.2 Analysis of a Mixture by Gas Chromatography:

GC is carried out using an instrument containing a long but very thin metal
tube filled with an inert support (usually silica) as the stationary phase and a stream
of helium gas as the mobile phase. The coiled tube, the "column" is heated in a
thermostatically controlled oven. A dilute solution of the liquid
sample is typically prepared in a volatile solvent such as diethyl ether. A small
amount of this sample solution (maybe 1 L) is then injected onto the column and is
carried forward by the helium carrier gas. The oven temperature is typically
gradually increased up to about 270oC over a 10 to 20 minute period. In general, the
lower the boiling point of the liquid, the quicker it will be carried through the column

31

(so it has a short retention time). When the sample exits the column, the liquid is
detected by the detector and the amount of liquid is measured, this information is
usually plotted as a "trace" with x-axis as time and the y-axis the response of the
detector as the abundance (i.e. the amount detected).

Therefore the area of the peak is directly proportional to the amount of material
it represents. A typical GC trace is shown below. This sample contained three
compounds. Based on the peak areas we see a 49 : 48 : 3 ratio. The minor impurity
(retention time 11.48 minutes) is a high boiling contaminant.

Figur 2.1

32

2.2.1Use of Gas Chromatography :

Is used in airports to detect bombs and is used is forensics in many different
ways. It is used to analyze fibers on a persons body and also analyze blood found at a
crime scene. In gas chromatography helium is used to move a gaseous mixture through
a column of absorbent material.

2.3 Liquid Chromatography

2.3.1 Brife History and Deffintion:

Liquid chromatography was defined in the early 1900s by the work of the
Russian botanist, Mikhail S. Tswett. His pioneering studies focused on separating
compounds (leaf pigments), extracted from plants using a solvent, and in a column
packed with particles
Tswett filled an open glass column with particles. Two specific materials that he
found useful were powdered chalk (calcium carbonate) and alumina. He poured his
sample (solvent extract of homogenized plant leaves) into the column and allowed it to
pass into the particle bed.This was followed by pure solvent.
As the sample passed down through the column by gravity, different colored
bands could be seen separating because some components were moving faster than
others.He related these separated, different-colored bands to the different compounds
that were originally contained in the sample.

33

He had created an analytical separation of these compounds based on the

differing strength of each compounds chemical attraction to the particles. The
compounds that were more strongly attracted to the particles slowed down, while other
compounds more strongly attracted to the solvent moved faster. This process can be
described as follows:
The compounds contained in the sample distribute, or partition differently
between the moving solvent, called the mobile phase, and the particles, called the
stationary phase. This causes each compound to move at a different speed, thus
creating a separation of the compounds.
Tswett coined the name chromatography (from the Greek words chroma,
meaning color, and graph, meaning writing literally, color writing) to describe his
colorful experiment. (Curiously, the Russian name Tswett means color).

Today, liquid chromatography, in its various forms, has become one of the most
powerful tools in analytical chemistry
.

Figure (2.2) Tswetts Experiment

2.4 Liquid Chromatography (LC) Techniques

34

Liquid chromatography can be performed using planar (Techniques 1 and 2) or

column techniques (Technique 3). Column liquid chromatography is the most powerful
and has the highest capacity for sample. In all cases, the sample first must be dissolved
in a liquid that is then transported either onto, or into, the chromatographic device

2.5Technique (1): Thin - layer Chromatography

TLC is carried out on glass plates or strips of plastic or metal coated on one
side with a thin layer of adsorbent. The adsorbent contains a small amount of gypsum
(CaSO4) which acts as a binder to give an adherent coating.

For routine work, small TLC plates can be prepared by dipping microscope
slides in a slurry of the adsorbent in chloroform..
More uniform plates are obtained by mixing the adsorbent with an equal weight
of water, spreading the mixture on a glass plate and allowing it to set dry.Precoated
TLC plates are commercially available with various adsorbents in very uniform
layers.A series of photographs are provided at the end of this document to show the
steps in running a TLC plate.

To "load" the plate, very small samples of the sample mixture in some volatile
solvent (e.g. diethyl ether or chloroform) are applied as spots near one end and the
volatile application solvent is allowed to evaporate. The plate is then placed, sample
end down, in a closed vessel containing a shallow pool of the "developing solvent".

The solvent rises on the plate by capillary action, passing over the sample and
causing the compounds to move at varying rates depending on their relative affinities
for the adsorbent and the solvent. When the solvent front has risen to just below the
top of the plate, the plate is removed and the solvent is allowed to evaporate.

The

zones or spots containing the various components of the mixture are then detected at

35

various points along the plate. If the compounds are colourless, they are made visible
by treating the plate with a reagent, such as iodine vapour, that causes colour to
develop.

2.5.1 Identification by TLC:

The main uses of TLC are for quick observation of the number of compounds
present in a sample, and for qualitative detection of a given compound in a mixture.
For the latter purpose, a sample of the compound in question is placed in one position
at the bottom of the plate and a sample of the mixture is placed in an adjacent position.
With a plate 3 to 4 cm wide, it is possible to place samples in as many as five or six
"lanes".
If identical compounds are present in two or more lanes, they should appear at
the same height after the plate has been developed with solvent. The position of the
spot relative to the solvent front, called the Rf value (distance travelled by spot /
distance travelled by solvent), depends on the thickness of the coating, the amount of
sample and the temperature, and may vary from one plate to the next. Comparison of
two samples on the same plate is therefore essential.

In addition to the position of the known sample and that of a spot in the mixture,
the appearance and / or colour of the compound on the developed plate
May greatly strengthen the identification if it is distinctive after visualisation. If
there is a reagent specific for the compound of interest, this is sprayed in a fine mist
over the surface.
A general reagent such as iodine will cause brown or black spots with
practically all compounds; these may be slightly different shades, but the colour is
usually not very characteristic. Another method for visualising spots is illumination of
the plate with an ultraviolet lamp. Many substances, particularly aromatic compounds,

36

will show a bright fluorescence, which may have a characteristic colour. A further
possibility is use of an adsorbent layer that contains a trace of fluorescent dye.
Compounds that are fluorescent still show up as bright spots on a light background;
any others appear as a dark spot since they quench the fluorescence of the background
dye.
2.5.2Preparing to run TLC
Capillaries or TLC applicators for applying the samples to the chromatographic
plate can be prepared by heating and drawing out a soft glass pipette or melting point
or capillary tubes. Soften a 1 cm section in the centre of the glass tube by heating in a
low Bunsen burner flame, then remove the tube from the flame and draw it out to a
thread-like thickness. Allow the glass to cool then carefully break the glass in the thin
portion gives you the capillaries.

The developing jar can be prepared from a beaker or jar with watch glass
as a lid. The developing jar is usuallylined with a piece of filter paper (cut to
shape and size) which helps create a solvent saturated environment.The solvent
system is added to the developing jar which is then carefully turned at an angle

37

to saturate the filter paper The solvent depth should be less than 5mm (typically
2-5 mm)

origin

TLC plates or sheets should be handled by the edges, and you should avoid touching
the coated surface otherwise fingerprints will ruin the experiment. Samples of the test
solutions should be spotted on the coated side about 1 cm from one end of the sheet,
and about 0.5 cm apart, with the outer two spots about 0.5 cm from the edge of the
sheet. It is common practice to draw a light pencil line about 1cm up from the bottom
edge of the plate (this line defines the origin) and then dividethis line at about 5mm
intervals to create the lanes.
Samples of unknowns are typically spotted in the middle lanes of the plate. It is
important to avoid applying too large an amount of sample.

The spot after

application should be about 1 to 2 mm in diameter. It is a good idea first to practice

applying spots to a small scrap of the sheet. You can check to see how well you are
doing by examining the spots under the UV lamp. If the spots are too pale at this
stage, then you will need more applications to the plate, or you should concentrate
your samples.

To apply the samples, touch the end of a capillary tube to the solution and then
touch this gently to the plate at the proper place. When the samples have been applied,
place the sheet, spotted end (origin) down, in a developing jar containing a pool of

38

solvent no more than 5 mm deep. On no account must spots dip below the surface of
the solvent pool when the sheet is placed in the developing jar.
The solvent system used depends on the nature of the samples. Many
experimental procedures will define the solvent system used, but at times a chemist
will need to develop their own system. It is important to handle the plates without
touching the front surface of the silica covered plate or fingerprints will result and the
plate may be spoiled. Use clean forceps for transferring the plates in and out of the
developing and visualising jars. Once the plate is in place, cap the jar securely and
develop the chromatogram until the solvent has risen to within 1 or 2 cm of the top of
the sheet.
When the solvent front is close to the top of the sheet remove the sheet from the
jar, and mark the solvent front with a small scratch (using a spatula or forceps) or a
pencil mark. Recap the developing jar and out the TLC sheet to one side and it allow
it to dry horizontally. In some cases the spots on the plate will be visible to the naked
eye. In other cases the plate will need to be visualised using other methods such as a
UV lamp or with a stain such as iodine.

It is a good idea to draw circles around the spots with a pencil as a permanent
record of thev isualisation. The Rf value (retardation factor) for a spot can now be
calculated and recorded along with the colour (if any) of the spot.
solvent front

y = distance travelled by solvent

x = distance travelled by sample
(measured to the point of
maximum colour intensity)

origin

Rf = x/y

39

When looking at a developed TLC plate it is good idea to review the quality of the
analysis or note if there are any problems with
the plate that means it might need
to be rerun. A poor quality plate would mean that it is difficult to obtain the
desired information from the plate. Some common problems are described below:

Problem

Possible causes

Samples does not show up

Not enough sample, different

visualisation method required

Streaking

This could be due to the nature of the

sample, due to sample overload or a poor
choice of solvent mixture.

Diffuse / blurred spots

The origin line was below the solvent

level or the plate was developed for too long so
the solvent front reached the top of the plate.

Sample lanes are not straight

/ parallel

A problem with the way the solvent ran due

to the plate being crooked in the solvent,
touching the filter paper in the developing jar or
the plate was chipped.

40

Preparing a sample to use on TLC by dissolving

a small amount of the sample in a small

Adding the volatile solvent to dissolve the

sample

Spotting the sample onto the TLC plate with a

microcapillary

41

Handling the TLC plate with forceps in order to place

it in the developing tank.

With the lid on, the TLC plate is developed by allowing

the solvent to run up the plate. Note that the solvent
level is below the pencil line on the plate that marks
the origin (i.e. the line where the samples were
applied).

After development, the TLC plate is removed and

visualised. Here we see it under a UV lamp.

Figur(2.3)

42

Used of Thin-layer Chromatography

2.5.3:
uses an absorbent material on flat glass or plastic plates. This is a simple and rapid
method to check the purity of an organic compound. It is used to detect pesticide or
insecticide residuesin food. Thin-layer chromatography is also used in forensics to
.analyze

the dye composition of fibers

2.6 Techniqe : Paper Chromatography

In Figure(2.4), samples are spotted onto paper (stationary phase). Solvent
(mobilephase) is then added to the center of the spot to create an outward radial flow.
This is a form of paper . (Classic paper chromatography is performed in a manner
similar to that of TLC with linear flow.) In the upper image, the same black FD&C dye
sample is applied to the paper

Figure(2.4): Paper Chromatography

The difference in separation power for this particular paper when compared to
the TLC plate. The green ring indicates that the paper cannot separate the yellow and

43

blue dyes from each other, but it could separate those dyes from the red dyes. In the
bottom image, a green sample, made up of the same yellow and blue dyes, is applied
to the paper. As you would predict, the paper cannot separate the two dyes. In the
middle, apurple sample, made up of red and blue dyes, was applied to the paper. They
are well separated.

Paper Chromatography is one of the most common types of chromatography. It

uses a strip of paper as the stationary phase. Capillary action is used to pull the
solvents up through the paper and separate the solutes.
2.7 Techniqe : Solid-Phase Extraction (SPE)
In this, the most powerful approach, the sample passes through a column or a
cartridge device containing appropriate particles (stationary phase). These particles are
called the chromatographic packing material.
Solvent (mobile phase) flows through the device. In solid-phase extraction (SPE), the
sample is loaded on to the cartridge and the solvent stream carries the sample through
the device.
As in Tswetts experiment, the compounds in the sample are then separated by
traveling at different individual speeds through the device.
Here the black sample is loaded onto a cartridge. Different solvents are used in
each step to create the separation

44

Figure (2.5): Column Chromatography Solid-Phase Extraction (SPE)

When the cartridge format is utilized, there are several ways to achieve flow.
Gravity or vacuum can be used for columns that are not designed to withstand
pressure. Typically, the particles in this case are larger in diameter (> 50 microns) so
that there is less resistance to flow.
Open glass columns (Tswetts experiment) are an example of this. In addition,
small plastic columns, typically in the shape of syringe barrels, can be filled with
packing- material particles and used to perform sample preparation. This is called
solid-phase extraction (SPE). Here, the chromatographic device, called a cartridge, is
used, usuallywith vacuum-assisted flow, to clean up a very complex sample before it
is analyzed further.
Smaller particle sizes (<10 microns) are required to improve separation power.
However, smaller particles have greater resistance to flow, so higher pressures
are needed to create the desired solvent flow rate. Pumps and columns designed

45

to withstand high pressure are necessary. When moderate to high pressure is used to
flow the solvent through the chromatographic column, the technique is called (HPLC).

2.8 High Preformance Liquid Chromatography (HPLC)

The acronym HPLC, coined by the late (Prof. Csaba Horvth for his 1970 Pittcon
paper), originally indicated the fact that high pressure was used to generate the flow
required for liquid chromatography in packed columns. In the beginning, pumps only
had a pressure capability of (500 psi) (35 bars). This was called high pressure liquid
chromatography (HPLC).The early 1970s saw a tremendous leap in technology.
These new HPLC instruments could develop up to 6,000 psi (400 bars) of pressure,
and incorporated improved injectors, detectors, and columns. HPLC really began to
take hold in the mid-to late-1970s. With continued advances in performance during this
time (smaller particles, even higher pressure), the acronym remained the same, but the
name was changed to high-performance liquid chromatography (HPLC)

High-performance liquid chromatography (HPLC) is now one of the most

powerful tools in analytical chemistr

It has the ability to separate, identify, and

quantities the compounds that are present in any sample that can be dissolved in a
liquid. Today, compounds in trace concentrations as low as parts per trillion (ppt) may
easily be identified
HPLC can be, and has been, applied to just about any sample, such as
pharmaceuticals, food, nutraceuticals, cosmetics, environmental matrices, forensic
samples, and industrial chemicals.

Figure (2.9) HPLC Column

2.9 Ultra- High Preformance Liquid Chromatography

46

In 2004, further advances in instrumentation and column technology were made to

achieve very significant increases in resolution, speed, and sensitivity in liquid
chromatography.
Columns with smaller particles (1.7 micron) and instrumentation with
specialized capabilities designed to deliver mobile phase at( 15,000 psi) (1,000 bar)
were needed to achieve a new level of performance. A new system had to be holistically
created to perform ultra-performance liquid chromatography (UPLC technology).

Basic research is being conducted today by scientists working with columns containing
even smaller (1-micron-diameter) particles and instrumentation capable of performing
at (100,000 psi) (6,800 bars). This provides a glimpse of what we may expect in the
future
2.9.1 High-Performance Liquid Chromatograph Work:
.1: The Components Of HPLC System:
The components of a basic high-performance liquid chromatography
(HPLC)

Figure (2.10): High-Performance Liquid Chromatography

HPLC System
2.2: A Resrvoir Holds The Solvent:
Called the mobile phase, because it moves.

47

2.3: A High-PressurePump:
Solvent delivery system or solvent manager) is used to generate and meter a

specified flow rate of mobile phase, typically milliliters per minute.

2.4: An Injector:
Sample manager auto sampler is able to introduce (inject) the sample into the

continuously flowing mobile phase stream that carries the sample into the
HPLC column.

2.5: The Column:

Contains the chromatographic packing material needed to effect the separation.

This packing material is called the stationary phase because it is held in place by the
column hardware. Is needed to see the separated compound bands as they elute from
the HPLC column (most compounds have no color, so we cannot see them with our
eyes).
The mobile phase exits the detector and can be sent to waste, or collected, as
desired.
When the mobile phase contains a separated compound band, HPLC provides
the ability to collect this fraction of the eluate containing that purified compound for
further study. This is called preparative chromatography (discussed in the section on
HPLC Scale).
Notice that high- pressure tubing and fittings are used to interconnect the
pump, injector, column, and detector components to form the conduit for the mobile
phase, sample, and separated compound bands.
2.9.2 Detectors And Computer Date Station:

48

The detector is wired to the computer data station, the HPLC system component
that records the electrical signal needed to generate the chromatogram on its display and
to identify and quantities the concentration of the sample constituents (see Figure 2.11).

Figure (2.11): A Typical HPLC System

2.7: Detectors Types:

Since sample compound characteristics can be very different, several types
of detectors have been developed.
For example: if a compound can absorb ultraviolet light, a UV-absorbance
detector is used. If the compound fluoresces, a fluorescence detector is used. If the
compound does not have either of these characteristics, a more universal type of
detector is used, such as an (evaporative-light-scattering detector) (ELSD). The most
powerful approach is the use multiple detectors in series.
For example: A (UV and or ELSD detector) may be used in combination with a
mass spectrometer (MS) to analyze the results of the chromatographic separation.
This provides, from a single injection, more comprehensive information
about an analyte. The practice of coupling a mass spectrometer to an HPLC system is
called (LC/MS).

49

HPLC Operation

2.9.3 chromatographic Column Works Bands:

A simple way to understand how we achieve the separation of the compounds
contained in a sample is to view the diagram in (Figure 8).
Mobile phase enters the column from the left, passes through the particle bed, and exits
at the right. Flow direction is represented by green arrows.

Figure (2.12): Understanding how a chromatographic column works Bands

First, consider the top image; it represents the column at time zero (the moment
of injection), when the sample enters the column and begins to form a band.
The sample shown here, a mixture of yellow, red, and blue dyes, appears at the
inlet of the column as a single black band. (In reality, this sample could be anything that
can be dissolved in a solvent; typically the compounds would be colorless and the
column wall opaque, so we would need a detector to see the separated compounds as
they elute).
After a few minutes (lower image), during which mobile phase flows
continuously and steadily past the packing material particles, we can see that the
individual dyes have moved in separate bands at different speeds.

50

This is because there is a competition between the mobile phase and the
stationary phase for attracting each of the dyes or analytes.

Notice: The yellow dye band moves the fastest and is about to exit the column. The
yellow dye likes [is attracted to] the mobile phase more than the other dyes. Therefore,
it moves at a faster speed, closer to that of the mobile phase. The blue dye band likes
the packing material more than the mobile phase. Its stronger attraction to the particles

causes it to move significantly slower. In other words, it is the most retained compound
in this sample mixture. The red dye band has an intermediate attraction for the mobile
phase and therefore moves at an intermediate speed through the column. Since each dye
band moves at different speed, we are able to separate it chromatographically.

Peaks AreCreated:
As the separated dye bands leave the column, they pass immediately into the
detector. The detector contains a flow cell that sees (detects) each separated compound
band against a background of mobile phase (see Figure 9).

Figure (2.13): How peaks are created

51

In reality, solutions of many compounds at typical HPLC analytical

concentrations are colorless.

An appropriate detector has the ability to sense the

presence of a compound and send its corresponding electrical signal to a computer data
station.

A choice is made among many different types of detectors, depending upon the
characteristics and concentrations of the compounds that need to be separated and
analyzed, as discussed earlier.

2.10 A chromatogram:
A chromatogram is a representation of the separation that has chemically
(chromatographically) occurred in the HPLC system. A series of peaks rising from a
baseline is drawn on a time axis. Each peak represents the detector response for a
different compound. The chromatogram is plotted by the computer data station (see
Figure H). In Figure (H), the yellow band has completely passed through the detector
flow cell; the electrical signal generated has been sent to the computer data
station. The resulting chromatogram has begun to appear on screen.
Notice: The chromatogram begins when the sample was first injected and starts as a
straight line set near the bottom of the screen. This is called the baseline; it represents
pure mobile phase passing through the flow cell over time. As the yellow analyte band
passes through the flow cell, a stronger signal is sent to the computer.
The line curves, first upward, and then downward, in proportion to the
concentration of the yellow dye in the sample band. This creates a peak in the
chromatogram. After the yellow band passes completely out of the detector cell, the
signal level returns to the baseline; the flow cell now has, once again, only pure m it is
the first peak drawn. A little while later, the red band reaches the flow cell.
The signal rises up from the baseline as the red band first enters the cell, and the
peak representing the red band begins to be drawn. In this diagram, the red band has not
fully passed through the flow cell. The diagram shows what the red band and red peak

52

would look like if we stopped the process at this moment.

Since most of the red band has passed through the cell, most of the peak has
been drawn, as shown by the solid line. If we could restart, the red band would
completely pass through the flow cell and the red peak would be completed (dotted
line). The blue band, the most strongly retained, travels at the slowest rate and elutes
after the red band.
The dotted line shows you how the completed chromatogram would appear if we had let
the run continue to its conclusion. It is interesting to note that the width of the blue
peak will be the broadest because the width of the blue analyte band, while
narrowest on the column, becomes the widest as it elutes from the column.
This is because it moves more slowly through the chromatographic packing
material bed and requires more time (and mobile phase volume) to be eluted
completely.
Since mobile phase is continuously flowing at a fixed rate, this means that the
blue band widens and is more dilute. Since the detector responds in proportion to the
concentration of the band, the blue peak is lower in height, but larger in width.

2.11 Identifying And Quanttating Compounds

2.11.1 Identification:
In Figure (H), three dye compounds are represented by three peaks
separated in time in the chromatogram. Each elutes at a specific location, measured by
the elapsed time between the moment of injection (time zero) and the time when the
peak maximum elutes.

By comparing each peaks retention time (tR) with that of injected reference
standards in the same chromatographic system (same mobile and stationary phase), a
chromatographer may be able to identify each compound.

53

Figure (2.14) Identification

In the chromatogram shown in (Figure 2.14), the chromatographer know that,

under these LC system conditions, the analyte, acrylamide, would be separated and
elute from the column at (2.85) minutes (retention time).

2.11.2Identifying And Quanttating :

Whenever a new sample, which happened to contain acrylamide (name of
a compound under test), was injected into the LC system under the same conditions, a
peak would be presen t at (2.85) minutes (see sample B in Figure I-2). For a better
understanding of why some compounds move more slowly (are better retained) than
others, once identity is established, the next piece of important information is how much
of each compound was present in the sample.

The chromatogram and the related data from the detector help us calculate the
concentration of each compound. The detector basically responds to the concentration
of the compound band as it passes through the flow cell. The more concentrated it is,
the stronger the signal; this is seen as a greater peak height above the baseline.

In (Figure I- 2), chromatograms for Samples A and B, on the same time scale, are
stacked one above the other. The same volume of sample was injected in both runs.

54

Both chromatograms display a peak at a retention time (tR) of (2.85 minutes),

indicating that each sample contains acrylamide. However, Sample A displays a much
bigger peak for acrylamide

Figure (2.15): Identification and Quantitation

The area under a peak (peak area count) is a measure of the concentration of
the compound it represents. This area value is integrated and calculated automatically
by the computer data station.
In this example, the peak for acrylamide in Sample A has 10 times the area of
that for Sample B. Using reference standards, it can be determined that Sample A
contains ( 10 picograms) of acrylamide, which is ten times the amount in Sample B (1
picogram).

Notice: There is another peak (not identified) that elutes at (1.8 minutes)
in both samples. Since the area counts for this peak in both samples are about the same,

55

this unknown compound may have the same concentration in both samples.

2.11.3 Factor s That Determine The OverallL Separtion Power OR Resolution

OF HPLC Column:
The degree to which two compounds are separated is called chromatographic
resolution (RS). Two principal factors that determine the overall separation power or
resolution that can be achieved by an HPLC column are (mechanical separation power),
(created by the column length, particle size, and packed-bed uniformity), (chemical
separation power) and (created by the physicochemical competition for compounds
between the packing material and the mobile phase).
Efficiency

is a measure of mechanical separation power, while selectivity is a

measure of chemical separation power.

2.11.4: Mechanical Separation Power (Efificency ):
If a column bed is stable and uniformly packed, its mechanical separation power
is determined by the column length and the particle size.
Mechanical separation power, also called efficiency, is often measured and
compared by a plate number (symbol = N). Smaller-particle chromatographic beds have
higher efficiency and higher backpressure
For a given particle size, more mechanical separation power is gained by
increasing column length.
However, the trade-offs are longer chromatographic run times, greater solvent
consumption, and higher back pressure. Shorter column lengths minimize all these
variables also reduce mechanical separation power, as shown in

56

(Figure 2.16 ) .

Figure (2.16): Column Length and Mechanical Separating Power (Same Particle Size)

Figure (2.17): Particle Size and Mechanical Separating Power (Same Column Length)

For example: For a given particle chemistry, mobile phase, and flow rate, as shown in
(Figure 13) , a column of the same length and (i.d.) , but with a smaller particle size, will
deliver more mechanical separation power in the same time. However, its back pressure
will be much higher.
2.11.5: Chemical Separation Power (Seectivity ):
The choice of a combination of particle chemistry (stationary phase) and (mobilephase composition), the separation system will determine the degree of chemical
separation power (how we change the speed of each analyte).

57

Optimizing selectivity is the most powerful means of creating a separation; this

may obviate the need for the brute force of the highest possible mechanical efficiency.
To create a separation of any two specified compounds, a scientist may choose among a
multiplicity of phase combinations (stationary phase and mobile phase) and retention
mechanisms (modes of chromatography).

2.12 HPLC Sepration Modes

In general, three primary characteristics of chemical compounds can be used to
create HPLC separations. They are:
Polarity
Electrical Charge
Molecular Size

First, lets consider polarity and the two primary separation modes that exploit this
characteristic: normal phase and reversed-phase chromatography.

2.12.1 Seprations Based On Polarity:

A molecules structure, activity, and physicochemical characteristics are determined

by the arrangement of its constituent atoms and the bonds between them. Within a
molecule, a specific arrangement of certain atoms that is responsible for special
properties and predictable chemical reactions is called a functional group.
This structure often determines whether the molecule is polar or non-polar.
Organic molecules are sorted into classes according to the principal functional group(s)

58

each contains.
Using a separation mode based on polarity, the relative chromatographic
retention of different kinds of molecules is largely determined by the nature and location
of these functional groups. As shown in (Figure 13), classes of molecules can be ordered
by their relative retention into a range or spectrum of chromatographic polarity from
highly polar to highly non-polar.

Figure (13): Chromatographic Polarity Spectrum by Analyte Functional

Group

Water (a small molecule with a high dipole moment) is a polar compound.

Benzene (an aromatic hydrocarbon) is a

(non-polar compound). Molecules with

similar chromatographic polarity tend to be attracted to each other; those with dissimilar
polarity exhibit much weaker attraction, if any, and may even repel one another. This
becomes the basis for chromatographic separation modes based on polarity.
Another way to think of this is by the familiar analogy (oil) (non-polar) and water
(polar) dont mix. Unlike in magnetism where opposite poles attract each other,
chromatographic separations based on polarity depend upon the stronger attraction
between likes and the weaker attraction between opposites. Remember, Like attracts
like in polarity-based chromatography.

Figure (2.18): Proper Combination of Mobile and Stationary Phases

59

2.12.2: Affects An Affect On Separtion Based On Polarity:

To design a chromatographic separation system (see Figure 14), we create
competition for the various compounds contained in the sample by choosing a mobile
phase and a stationary phase with different polarities. Then, compounds in the sample
that are similar in polarity to the stationary phase (column packing material)

will be

delayed because they are more strongly attracted to the particles. Compounds whose
polarity is similar to that of the mobile phase will be preferentially attracted to it and
move faster.
In this way, based upon differences in the relative attraction of each compound
for each phase, a separation is created by changing the speeds of the analytes. In the
figures (R-1), (R-2), and(R-3) display typical chromatographic polarity ranges for
mobile phases, stationary phases, and sample analytes, respectively.
Lets consider each in turn to see how a chromatographer chooses the appropriate
phases to develop the attraction competition needed to achieve a polarity - based HPLC
separation.

Figure (2.19) Mobile Phase Chromatographic Polarity Spectrum

2.12.3MOBILE PHASE CHROMATOGRAPHIC POLARITY SPECTRUM:

A scale, such as that shown in Figure (15), upon which some common solvents
are placed in order of relative chromatographic polarity is called an eluotropic series.
Mobile phase molecules that compete effectively with analyte molecules for the
attractive stationary phase sites displace these analytes, causing them to move faster
through the column (weakly retained).
Water is at the polar end of mobile - phase- solvent scale, while hexane, an

60

aliphatic hydrocarbon, is at the non-polar end. In between, single solvents, as well as

miscible- solvent mixtures (b) ended in proportions appropriate to meet specific
separation requirements), can be placed in order of elution strength.
2.12.4: STATIONARY PHASE PARTICLE CHROMATOGRAPHIC POLARITY
SPECTRUM:
Which end of the scale represents the strongest mobile phase depends upon the
nature of the stationary phase surface where the competition for the analyte molecules
occurs.

Figure (2.20) Stationary Phase Particle Chromatographic Polarity Spectrum

Silica has an active, hydrophilic (water-loving) surface containing acidic

silanol (silicon-containing analog of alcohol) functional groups.
Consequently, it falls at the polar end of the stationary-phase scale shown in Figure

The activity or polarity of the silica surface may be modified selectively by

chemically bonding to it less polar functional groups (bonded phase).
Examples,

shown here include, in order of decreasing polarity, cyanopropylsilyl-

(CN), n-octylsilyl- (C8), and n-octadecylsilyl- (C18, ODS) moieties on silica. The latter
is a hydrophobic (water-hating), very non-polar packing.

61

modes, like attracts like.

Table (2.21) Phase Characteristics for Separations Based on Polarity

2.13Chromatography Used:
Chromatography was developed in Russia in 1906 by an Italian-born botanist named
Mikhail Tswett (sometimes spelled Tsvet; 18721919), who used it for studying plant
pigments such as chlorophyll.
During the 20th century, chemists found chromatography was a superb technique
for studying and separating all kinds of complex mixtures. It's now widely used in
forensic science (for identifying samples taken from crime scenes), in pollution
monitoring (for identifying small concentrations of unknown pollutants in air and water
samples), and for studying complex mixtures in such things as food, perfume,
petrochemical, and pharmaceutical production.
One of chromatography's big advantages is that it works with tiny samples and low
concentrations (particularly helpful when it comes to such things as forensic science
and drug or pollution testing).

62

63

3 Oil Analysis
3.1 Define :
Oil analysis involves sampling and analyzing oil for various
properties and materials to monitor wear and contamination in an
engine, transmission or hydraulic system. Sampling and analyzing on
a regular basis establishes a baseline of normal wear and can help
indicate when abnormal wear or contamination is occurring.
Oil analysis works like this. Oil that has been inside any
moving mechanical apparatus for a period of time reflects the exact
condition of that assembly. Oil is in contact with engine or mechanical
components as wear metallic trace particles enter the oil. These
particles are so small they remain in suspension. Many products of the
combustion process also will become trapped in the circulating oil.
The oil becomes a working history of the machine.
Particles caused by normal wear and operation will mix
with the oil. Any externally caused contamination also enters the
oil. By identifying and measuring these impurities, you get an
indication of the rate of wear and of any excessive
contamination. An oil analysis also will suggest methods to
reduce accelerated wear and contamination.

64

The typical oil analysis tests for the presence of a number

of different materials to determine sources of wear, find dirt and
other contamination, and even check for the use of appropriate
lubricants.

3.2 Oil analysis can detect:

Fuel dilution of lubrication oil

Dirt contamination in the oil

Antifreeze in the oil

Excessive bearing wear

Misapplication of lubricants

Some wear is normal, but abnormal levels of a particular

material can give an early warning of impending problems
and possibly prevent a major breakdown.

33Early detection can:

Reduce repair bills

Reduce catastrophic failures

Increase machinery life

Reduce non-scheduled downtime

Early detection with oil analysis can allow for corrective action
such as repairing an air intake leak before major damage occurs.
Probably one of the major advantages of an oil analysis program

65

is being able to anticipate problems and schedule repair work to

avoid downtime during a critical time of use.

3.4 Oil Analysis

History

The first use of used oil analysis dates back to the early 1940s by
the railway companies in the Western United States. Prompted by the
purchase of a fleet of new locomotives, technicians used simple
spectrographic equipment and physical tests to monitor locomotive
engines.
As steam locomotives gave yield to diesel locomotives, oil
analysis practices by railways caught on. By the 1980s oil analysis
formed the basis of Condition Based Maintenance in most railways in
North America.
Owing to the success of oil analysis in the railways, the American
Navy used spectrometric techniques to monitor jet engines on their
aircraft in the mid 1950s. Around this time Rolls-Royce was also
experimenting with oil analysis for their jet turbines.
Oil analysis began to spread and programs were developed by the
American Army and Air Force throughout the 1950s and early 1960s.
Then commercial oil analysis laboratories first as appeared on the
scene in the early 1960s.

66

A detailed analysis of a sample of engine, transmission and

hydraulic oils is a valuable preventive maintenance tool . In many as it
enables identification of potential problems before a major repair is
necessary, has the potential to reduce the frequencies of oil changes,
and increases the resale value of used equipment.

3.5 Important of oil analysis :

One purpose of oil analysis is to provide a means of predicting
possible impending failure without dismantling the equipment.
A person can "look inside" an engine, transmission or hydraulic
systems without taking it apart.
.3.6 Evaluating Used Equipment:
A complete record of oil analysis may be a great tool for
selling a used piece of equipment. It shows the potential buyers how
the equipment has been maintained and how adjustments were made
during its life. The history is a good indicator of potential future
repairs and overhaul requirements.
One potential use of oil analysis is in the evaluation of used
equipment being considered for purchase. Without knowing the
amount of operation of the oil being analyzed, this test should be
considered conclusive only if it indicates a problem. A good report

67

could result from either no problems or a short length of service of the

oil.

3.7 Physical Tests

Some of the physical properties tested for and usually included in
analysis of an oil sample are:

Antifreeze forms a gummy substance that may reduce oil

flow. It leads to high oxidation, oil thickening, high acidity, and
engine failure if not corrected.

Fuel dilution thins oil, lowers lubricating ability, and might

drop oil pressure. This usually causes higher wear.

Oxidation measures gums, varnishes and oxidation products.

High oxidation from oil used too hot or too long can leave
sludge and varnish deposits and thicken the oil.

Total base number generally indicates the acid-neutralizing

capacity still in the lubricant.

Total solids include ash, carbon, lead salts from gasoline

engines, and oil oxidation.

Viscosity is a measure of an oil's resistance to flow. Oil may

thin due to shear in multi-viscosity oils or by dilution with fuel.
Oil may thicken from oxidation when run too long or too hot.
Oil also may thicken from contamination by antifreeze, sugar
and other materials

68

Karl Fischer measures all forms of water at low levels and is

recommended for industrial equipment. (The Qualitative Crackle
test is used for engine oils.)

Acid Number measures acid buildup, which denotes

oxidation.

FTIR measures chemistry changes in a lubricant, which are

good indicators of oxidation and nitration.

Base Number is for engine oils. It measures the depletion of the

detergent, which neutralizes acids.

Flash Point is a measure of light components, which lower the

viscosity of lubricants.

Particle Counts, by size and amount, are determined with the

use of a laser counter.

Voltammetry measures depletion of antioxidants in lubricants

Tests for equipment condition through the measurement of wear

debris include:

Atomic Emission Spectroscopy measures metals in parts per

million (limited to particles under 10 microns in size).

Ferrous Density, both direct-read ferrogram and particle

quantifier, measures ferrous particles without the size limitation
of emission spectroscopy.

Analytical Ferrography looks at size, shape and color (the

three most important physical characteristics of a particle)to
determine the wear mechanism and severity in machinery. This

69

is the only common oil-analysis test that can justify equipment

shutdown.
.You can determine the value of the Free Fatty Acids (FFA)
in oils and fats with different systems of FoodLab Line:

FoodLab Touch for fats and oils: designed to measure the free
fatty acids (FFA)/Acidity, peroxide value (PV), soaps, Anisidine
value (AnV) in oils and fats

OxiTester: to test the Ferr Fatty Acids (FFA), Peroxide Value (PV)
and Polyphenols/Oil Stability Index (OSI) in vegetable oil like olive
oil, avocado oil, seeds oil etc.

FoodLab Touch: designed for analysis on food like milk, eggs,

tomatoes, vegetable puree, cheese, dairy products and fats.

PalmOilTester: designed to measure acidity (FFA), Iodine Value

(IV), DOBI & carotene content, Peroxide Value (PV), Anisidine
Value (AnV) in palm oil.

WineLab Touch: designed for analysis of Wine and Must,

configurable to determine FFA, Peroxide Value, poliphenols in
olive oil.

MiniFood: to test Free Fatty Acids and Peroxide Value in olive oil.

3.8 Metal Tests

Some of the metals tested for and usually included in analysis of
an oil sample and their potential sources are:

70

Aluminum (Al): Thrust washers, bearings and pistons are

made of this metal. High readings can be from piston skirt
scuffing, excessive ring groove wear, broken thrust washers, etc.

Boron, Magnesium, Calcium, Barium, Phosphorous, and Zinc:

These metals are normally from the lubricating oil additive
package. They involve detergents, dispersants, extreme-pressure
additives, etc.

Chromium (CR): Normally associated with piston rings. High

levels can be caused by dirt coming through the air intake or
broken rings.

Copper (CU), Tin: These metals are normally from bearings

or bushings and valve guides. Oil coolers also can contribute to
copper readings along with some oil additives. In a new engine
these results will normally be high during break-in, but will
decline in a few hundred hours.

Iron (Fe): This can come from many places in the engine such
as liners, camshafts, crankshaft, valve train, timing gears, etc.

Lead (Pb): Use of regular gasoline will cause very high test
results. Also associated with bearing wear, but fuel source
(leaded gasoline) and sampling contamination (use of galvanized
containers for sampling) are critical in interpreting this metal.

Silicon (Si): High readings generally indicate dirt or fine sand

contamination from a leaking air intake system. This would act

71

as an abrasive, causing excessive wear. Silicon is also used as a

anti-foam agent in some oils. more on silicon

Sodium (Na): High readings of this metal normally are

associated with a coolant leak, but can be from an oil additive
package.

3.9 Chemical Test :

Acid Number (ASTM D974) is a good indicator of oil
degradation and whether it is likely to attack other materials.
Flash Point (ASTM D92) is a good indication of volatility and GCMS Analysis can be used to give a breakdown of the organic
components in oil. Small particles in oil can be filtered out and
identified using EDS Analysis. Chlorine content by XRF is an
indicator of the presence of undesirable chlorides which can attack
the interior of hydraulic systems. Water in oil can also corrode
unprotected steel components so moisture content (ASTM E1064)
measurements are also important

3.10 Taking an Oil Sample

It is important to get an oil sample that is representative of all
of the oil in the machine. Remember, your analysis will be based only
on the sample that you send in for analysis. Always have the oil hot and

72

thoroughly mixed before sampling. Handle hot drained oil with care it
could cause serious burns.
The easiest way to obtain a sample may be when the oil is being
drained for an oil change. Sampling at this time usually involves letting
some of the oil drain and then catching a sample in an appropriate
container.
Samples also can be obtained without draining oil by suctioning
out through plastic tubing routed down into the oil reservoir.
In any case, it is important to have an appropriate container and
follow sampling directions thoroughly. Remember, many of the tests are
for measuring materials on a parts per million basis, so safe, effective
sampling is needed.

3.11Cost and Convenience

Cost of oil analysis will vary according to the laboratory and
extent of the analysis. Typical charges are $10 to $30 per analysis. The
expense easily can be justified if it alerts the owner of a major problem
that can be corrected and will help prevent downtime when the machine
is needed.
Several companies have developed oil analysis kits that make oil
analysis convenient. These kits include the sample bottles, suction pump

73

and tubing, and possibly a pre-addressed, postage-paid mailing

container.
The reasonable cost and convenience of oil analysis for use
makes it another management tool that should be considered by anyone
wanting to do preventive maintenance. .

74

Conclusion
This research include three chapters dealing with the oil
analysis.
In chapter one there in an introduction study about analytical
chemistry . which include history of analytical chemistry and
classification was discussed .
In chapter two the study dealt with the Chromatography and
type of Chromatography such as :
Gas Chromatography, liquid Chromatography , Thin
layerChromatography , peper Chromatography and HPLC
.contents and how work it
In chapter three the study dealt oil analysis which include
sample of oil ,physical test , chemical test .In addtion to
important of oil analysis

75

Reference
i.

Modern Analytical Chemistry David Harvey

ii.

www.university .arabsbook,com PDF

iii.

http://www.chem.ucalgary.ca/courses/351/labo
ratory/chromatography.pdf

iv.

http://www.apmtesting.com/testingservices/materials-tested/oil-testing.php

v.

https://en.wikipedia.org/wiki/Oil_analysis

vi.

http://www.explainthatstuff.com/chromatograp
hy.html

vii.

http://www.bobistheoilguy.com/whatisoilanaly
sis.htm

viii.

http://www.maintenancetechnology.com/2011/
01/introduction-to-common-oil-analysis-testsand-how-to-take-a-successful-

76

77