Wet chemistry is a term used to refer to chemistry generally done in the liquid phase.

It
is also known as bench chemistry because many of the tests performed are done at a lab
bench

Materials
Traditionally, it involves the use of laboratory glassware, such as beakers and flasks, and
excludes quantitative chemical analysis using instrumentation. Many high school and
college laboratories teach students basic wet chemistry methods.

History
Before the age of theoretical and computational chemistry it was the predominant form of
scientific discovery in the chemical field. This is why it is sometimes referred to as
classic chemistry or classical chemistry. Because of the high volume of wet chemistry
that must be done in today's society and quality control requirements, many wet
chemistry methods have been automated and computerized for streamlined analysis.

Uses
Wet chemistry techniques can be used for qualitative chemical measurements, such as
changes in color (colorimetry), but often involves more quantitative chemical
measurements, using methods such as gravimetry and titrimetry. Some uses for wet
chemistry include tests for:

• pH (acidity, alkalinity)
• concentration
• conductivity (Specific Conductance)
• cloud point (nonionic surfactants)
• hardness
• solids or dissolved solids
• salinity
• specific gravity
• density
• turbidity
• viscosity
• moisture (Karl Fischer titration)

Wet chemistry is also used in environmental chemistry settings and is used for to test:

• Biochemical Oxygen Demand (BOD)

• Chemical Oxygen Demand
• eutrophication
• coating identification
It can also involve the elemental analysis of samples, e.g., water sources, for items like:

• Ammonia Nitrogen
• Chloride
• Chromium
• Cyanide
• dissolved Oxygen
• Fluoride
• Nitrogen
• Nitrate
• Phenols
• Phosphate
• Phosphorus
• Silica
• Sulfate, Sulfide

Karl Fischer titration

is a classic titration method in analytical chemistry that uses coulometric or volumetric

titration to determine trace amounts of water in a sample. It was invented in 1935 by the
German chemist Karl Fischer.

Coulometric titration
The main compartment of the titration cell contains the anode solution plus the analyte.
The anode solution consists of an alcohol (ROH), a base (B), SO2 and I2. A typical
alcohol that may be used is methanol or diethylene glycol monomethyl ether, and a
common base is imidazole.

The titration cell also consists of a smaller compartment with an anode immersed in the
anode solution of the main compartment. The two compartments are separated by an ion-
permeable membrane.

The Pt anode generates I2 when current is provided through the electric circuit. The net
reaction as shown below is oxidation of SO2 by I2. One mole of I2 is consumed for each
mole of H2O. In other words, 2 moles of electrons are consumed per mole of water.

B·I2 + B·SO2 + B + H2O → 2BH+I− + BSO3

BSO3 + ROH → BH+ROSO3−

The end point is detected most commonly by a bipotentiometric method. A second pair of
Pt electrodes are immersed in the anode solution. The detector circuit maintains a
constant current between the two detector electrodes during titration. Prior to the
equivalence point, the solution contains I- but little I2. At the equivalence point, excess I2
appears and an abrupt voltage drop marks the end point. The amount of current needed to
generate I2 in order to reach the end point can then be used to calculate the amount of
water in the original sample.

Volumetric titration
The volumetric titration is based on the same principles as the coulometric titration
except that the anode solution above now is used as the titrant solution. The titrant
consists of an alcohol (ROH), base (B), SO2 and a known concentration of I2.

One mole of I2 is consumed for each mole of H2O. The titration reaction proceeds as
above, and the end point may be detected by a bipotentiometric method as described
above.

Advantage of analysis
The popularity of the Karl Fischer titration is due in large part to several practical
advantages that it holds over other methods of moisture determination, including:

• High accuracy and precision

• Selectivity for water
• Small sample quantities required
• Easy sample preparation
• Short analysis duration
• Nearly unlimited measuring range (1ppm to 100%)
• Suitability for analyzing:
o Solids
o Liquids
o Gases
• Independence of presence of other volatiles
• Suitability for automation

In contrast, loss on drying will detect the loss of any volatile substance.

Conductivity (electrolytic)
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The conductivity (or specific conductance) of an electrolyte solution is a measure of its

ability to conduct electricity. The SI unit of conductivity is siemens per meter (S/m).

Conductivity measurements are used routinely in many industrial and environmental

applications as a fast, inexpensive and reliable way of measuring the ionic content in a
solution.[1] For example, the measurement of product conductivity is a typical way to
monitor and continuously trend the performance of the water purification systems.

In many cases, conductivity is linked directly to the total dissolved solids (T.D.S.). High
quality deionized water has a conductivity of about 5.5 μS/m, typical drinking water in
the range of 5-50 mS/m, while sea water about 5 S/m[2] (i.e., sea water's conductivity is
one million times higher than deionized water).

Conductivity is traditionally determined by measuring the AC resistance of the solution

between two electrodes.

Please note that multimeters originally designed for electronic use measure
resistance, a related quantity, with DC. (A few measure conductance, which is
more-closely related, also with DC.) However, applying DC to conductance-
measuring electrodes creates undesirable and cumulative electrochemical changes
(electrolysis, primarily) that can seriously invalidate the readings. Measurement
with AC means that these electrochemical effects are reversed every time the
polarity of the AC reverses, effectively nullifying the effects.

Dilute solutions follow Kohlrausch's Laws of concentration dependence and additivity of

ionic contributions. Onsager gave a theoretical explanation of Kohlrausch's law by
extending Debye–Hückel theory.

Units
The SI unit of conductivity is S/m and, unless otherwise qualified, it refers to 25 °C
(standard temperature). Often encountered in industry is the traditional unit of μS/cm.
The numbers in μS/cm are lower than those in μS/m by a factor of 100 (i.e., a solution
with a conductivity of 100 μS/m has a conductivity of 1 μS/cm). Occasionally a unit of
"EC" (electrical conductivity) is found on scales of instruments: 1 EC = 1 μS/cm.
Sometimes encountered is so-called mho (reciprocal of ohm): 1 mho/m = 1 S/m.
Historically, mhos antedate Siemens by many decades; good vacuum-tube testers, for
instance, gave transconductance readings in micromhos.

The commonly used standard cell has a width of 1 cm, and thus for very pure water in
equilibrium with air would have a resistance of about 106 ohm, known as a megohm,
occasionally misspelled as "megaohm". Ultra-pure water could achieve 18 megohms or
more. Thus in the past megohm-cm was used, sometimes abbreviated to "megohm".[3]
Sometimes, a conductivity is given just in "microSiemens" (omitting the distance term in
the unit). While this is an error, it can often be assumed to be equal to the traditional
μS/cm. The typical conversion of conductivity to the total dissolved solids is done
assuming that the solid is sodium chloride: 1 μS/cm is then an equivalent of about 0.6 mg
of NaCl per kg of water.
Molar conductivity has the SI unit S m2 mol−1. Older publications use the unit Ω−1 cm2
mol−1.

[edit] Measurement
Main article: Electrical conductivity meter

A conductivity meter and probe

The electrical conductivity of a solution of an electrolyte is measured by determining the

resistance of the solution between two flat or cylindrical electrodes separated by a fixed
distance.[4] An alternating voltage is used in order to avoid electrolysis. Typical
frequencies used are in the range 1-3 kHz. The dependence on the frequency is usually
small.[5] The resistance is measured by a conductivity meter.

A wide variety of instrumentation is commercially available.[6] There are two types of

cell, the classical type with flat or cylindrical electrodes and a second type based on
induction.[7] Many commercial systems offer automatic temperature correction.

[edit] Definitions
Resistance, R, is proportional to the distance, l, between the electrodes and is inversely
proportional to the cross-sectional area of the sample, A. Writing ρ (rho) for the specific
resistance (or resistivity),

In practice the conductivity cell is calibrated by using solutions of known specific

resistance, ρ*, so the quantities l and A need not be known precisely.[8] If the resistance of
the calibration solution is R*, a cell-constant, C, is derived.

The specific conductivity, κ (kappa) is the reciprocal of the specific resistance.

Conductivity is also temperature-dependant.

Theory
The conductivity of a solution containing one electrolyte depends on the concentration of
the electrolyte. Therefore it is convenient to divide the specific conductivity by
concentration. This quotient is termed molar conductivity, is denoted by Λm

Strong electrolytes

Strong electrolytes are believed to dissociate completely in solution. The conductivity of

a solution of a strong electrolyte at low concentration follows Kohlrausch's Law

where is known as the limiting molar conductivity, K is an empirical constant and c

is the electrolyte concentration. (Limiting here means "at the limit of the infinite
dilution".)

Moreover, Kohlrausch also found that the limiting conductivity of anions and cations are
additive: the conductivity of a solution of a salt is equal to the sum of conductivity
contributions from the cation and anion.

where:

• ν + and ν − are the number of moles of cations and anions, respectively, which are
created from the dissociation of 1 mole of the dissolved electrolyte;
• and are the limiting molar conductivities of the individual ions.

The following table gives values for limiting molar conductivities for selected ions.

Limiting ion conductivity in water at 298 K

Cations λ+0 /mS m2mol-1 anions λ-0 /mS m2mol-1
H+ 34.96 OH- 19.91
Li+ 3.869 Cl- 7.634
Na+ 5.011 Br- 7.84
K+ 7.350 I- 7.68
Mg2+ 10.612 SO42- 15.96
Ca2+ 11.900 NO3- 7.14
Ba2+ 12.728 CH3CO2- 4.09

A theoretical interpretation of these results was provided by the Debye-Hückel-Onsager

equation.[9]

where A and B are constants that depend only on known quantities such as temperature,
the charges on the ions and the dielectric constant and viscosity of the solvent. As the
name suggests, this is an extension of the Debye–Hückel theory, due to Onsager. It is
very successful for solutions at low concentration.

Weak electrolytes

A weak electrolyte is one that is not fully dissociated . Typical weak electrolytes are
weak acids and weak bases. The concentration of ions in a solution of a weak electrolyte
is less than the concentration of the electrolyte itself. For acids and bases the
concentrations can be calculated when the value(s) of the acid dissociation constant(s)
is(are) known.

For a monoprotic acid, HA, with a dissociation constant Ka, an explicit expression for the
conductivity as a function of concentration, c, known as Ostwald's dilution law, can be
obtained.

Higher concentrations

Both Kolrausch's law and the Debye-Hückel-Onsager equation break down as the
concentration of the electrolyte increases above a certain value. The reason for this is that
as concentration increases the average distance between cation and anion decreases, so
that there is more inter-ionic interaction. Whether this constitutes ion-association is a
moot point. However, It has often been assumed that cation and anion interact to form an
ion-pair. Thus the electrolyte is treated as if it were like a weak acid and a constant, K,
can be derived for the equilibrium

A + + B- A+B-; K=[A+][B-]/[A+B-]

Davies describes the results of such calculations in great detail, but states that K should
not necessarily be thought of as a true equilibrium constant, rather, the inclusion of an
"ion-association" term is useful in extending the range of good agreement between theory
and experimental conductivity data.[10] Various attempts have been made to extend
Onsager's treament to more concentrated solutions.[11]

The existence of a so-called conductance minimum has proved to be a controversial

subject as regards interpretation. Fuoss and Kraus suggested that it is caused by the
formation of ion-triplets,[12] and this suggestion has received some support recently.[13][14].

Applications
Notwithstanding the difficulty of theoretical interpretation, conductivity measurements
are used extensively in many industries.[15] For example, conductivity measurements are
used to monitor quality in public water supplies, in hospitals, in boiler water and
industries which depend on water quality such as brewing. This type of measurement is
not ion-specific; it can sometimes be used to determine the amount of total dissolved
solids (T.D.S.) if the composition of the solution and its conductivity behavior are
known.[1]

Sometimes, conductivity measurements are linked with other methods to increase the
sensitivity of detection of specific types of ions. For example, in the boiler water
technology, the boiler blowdown is continuously monitored for "cation conductivity",
which is the conductivity of the water after it has been passed through a cation exchange
resin. This is a sensitive method of monitoring anion impurities in the boiler water in the
presence of excess cations (those of the alkalizing agent usually used for water
treatment). The sensitivity of this method relies on the high mobility of H+ in comparison
with the mobility of other cations or anions.

Conductivity detectors are commonly used with ion chromatography.[16]

Titration
From Wikipedia, the free encyclopedia
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Jump to: navigation, search
Not to be confused with the mathematical notion of tetration.
This article is about volumetric titration. For other uses, see Titration (disambiguation).
Titration setup: the titrant drops from the burette into the analyte solution in the flask. An
indicator present then changes color permanently at the endpoint.

Titration is a common laboratory method of quantitative chemical analysis that is used

to determine the unknown concentration of a known reactant. Because volume
measurements play a key role in titration, it is also known as volumetric analysis. A
reagent, called the titrant or titrator,[1] of a known concentration (a standard solution) and
volume is used to react with a solution of the analyte or titrand,[2] whose concentration is
not known. Using a calibrated burette or chemistry pipetting syringe to add the titrant, it
is possible to determine the exact amount that has been consumed when the endpoint is
reached. The endpoint is the point at which the titration is complete, as determined by an
indicator (see below). This is ideally the same volume as the equivalence point—the
volume of added titrant at which the number of moles of titrant is equal to the number of
moles of analyte, or some multiple thereof (as in polyprotic acids). In the classic strong
acid-strong base titration, the endpoint of a titration is the point at which the pH of the
reactant is just about equal to 7, and often when the solution takes on a persisting solid
color as in the pink of phenolphthalein indicator. There are however many different types
of titrations (see below).

Many methods can be used to indicate the endpoint of a reaction; titrations often use
visual indicators (the reactant mixture changes color). In simple acid-base titrations a pH
indicator may be used, such as phenolphthalein, which becomes pink when a certain pH
(about 8.2) is reached or exceeded. Another example is methyl orange, which is red in
acids and yellow in alkali solutions.

Not every titration requires an indicator. In some cases, either the reactants or the
products are strongly colored and can serve as the "indicator". For example, a redox
titration using potassium permanganate (pink/purple) as the titrant does not require an
indicator. When the titrant is reduced, it turns colorless. After the equivalence point, there
is excess titrant present. The equivalence point is identified from the first faint persisting
pink color (due to an excess of permanganate) in the solution being titrated.

Due to the logarithmic nature of the pH curve, the transitions are, in general, extremely
sharp; and, thus, a single drop of titrant just before the endpoint can change the pH
significantly—leading to an immediate colour change in the indicator. There is a slight
difference between the change in indicator color and the actual equivalence point of the
titration. This error is referred to as an indicator error, and it is indeterminate.

Contents
History and etymology
The word "titration" comes from the Latin word titulus, meaning inscription or title. The
French word titre, also from this origin, means rank. Titration, by definition, is the
determination of rank or concentration of a solution with respect to water with a pH of 7
(which is the pH of pure H2O under standard conditions).

The origins of volumetric analysis are in late-18th-century French chemistry. Francois

Antoine Henri Descroizilles developed the first burette (which looked more like a
graduated cylinder) in 1791. Joseph Louis Gay-Lussac developed an improved version of
the burette that included a side arm, and coined the terms "pipette" and "burette" in an
1824 paper on the standardization of indigo solutions. A major breakthrough in the
methodology and popularization of volumetric analysis was due to Karl Friedrich Mohr,
who redesigned the burette by placing a clamp and a tip at the bottom, and wrote the first
textbook on the topic, Lehrbuch der chemisch-analytischen Titrirmethode (Textbook of
analytical-chemical titration methods), published in 1855.[3]

Preparing a sample for titration

In a titration, both titrant and analyte are required to be in a liquid (solution) form. If the
sample is not a liquid or solution, the samples must be dissolved. If the analyte is very
concentrated in the sample, it might be useful to dilute the sample.

Although the vast majority of titrations are carried out in aqueous solution, other solvents
such as glacial acetic acid or ethanol (in petrochemistry) are used for special purposes.

A measured amount of the sample can be given in the flask and then be dissolved or
diluted. The mathematical result of the titration can be calculated directly with the
measured amount. Sometimes the sample is dissolved or diluted beforehand, and a
measured amount of the solution is used for titration. In this case the dissolving or
diluting must be done accurately with a known coefficient because the mathematical
result of the titration must be multiplied with this factor.
Many titrations require buffering to maintain a certain pH for the reaction. Therefore,
buffer solutions are added to the reactant solution in the flask to maintain the pH of the
solution.

Some titrations require "masking" of a certain ion. This can be necessary when two
reactants in the sample would react with the titrant and only one of them must be
analysed, or when the reaction would be disturbed or inhibited by this ion. In this case
another solution is added to the sample, which "masks" the unwanted ion (for instance by
a weak binding with it or even forming a solid insoluble substance with it).

Some redox reactions may require heating the solution with the sample and titration
while the solution is still hot, in order to increase the reaction rate. For instance, the
oxidation of certain oxalate solutions requires heating the solution to approximately 60
degrees in order to maintain a reasonable rate of reaction.

Procedure
A typical titration begins with a beaker or Erlenmeyer flask containing a precise volume
of the reactant and a small amount of indicator, placed underneath a burette or buretting
syringe containing the reagent. By controlling the amount of reagent added to the
reactant, it is possible to detect the point at which the indicator changes color. As long as
the indicator has been chosen correctly, this should also be the point where the reactant
and reagent neutralize each other, and, by reading the scale on the burette, the volume of
reagent can be measured.

As the concentration of the reagent is known, the number of moles of reagent can be
calculated (since Molarity = numberofmoles / volume(L)). Then, from the chemical
equation involving the two substances, the number of moles present in the reactant can be
found. Finally, by dividing the number of moles of reactant by its volume, the
concentration is calculated.

Titration curves
Main article: Titration curve

A typical titration curve of a diprotic acid, oxalic acid, titrated with a strong base, sodium
hydroxide. Each of the two equivalence points are visible
A titration curve is a curve in the plane whose x-coordinate is the volume of titrant added
since the beginning of the titration, and whose y-coordinate is the concentration of the
analyte at the corresponding stage of the titration (in an acid-base titration, the y-
coordinate is usually the pH of the solution at the corresponding stage). Often it is the
case that the titration curve of a titration reflects the nature of the titration quite well; for
instance, it reflects the nature of all solutions involved in the titration.

In the case of acid-base titrations, titration curves reflect the strength of the corresponding
acid and base. For instance, in a strong acid and strong base titration, the titration curve
will be relatively smooth, although very steep for points near the equivalence point of the
titration. Since in this case, small changes in the volume of the titrant result in large
changes of the pH near the equivalence point, an extensive range of indicators would be
appropriate (for instance litmus, phenolphthalein or bromothymol blue).

On the other hand, if one of the constituents of an acid-base titration is either a weak acid
or a weak base, and the other is either a strong acid or a strong base, the titration curve is
fairly irregular near the equivalence point (and the pH does not change as much due to
the addition of small volumes of titrant). For instance, the titration curve for the titration
between oxalic acid (a weak acid) and sodium hydroxide (a strong base) is depicted in the
image above. Here, the equivalence point occurs at a pH of about 8-10, and thus the
analyte is basic at the equivalence point (more precisely, the sodium salt produced by the
reaction hydrolyses in water to produce hydroxide ions). An indicator such as
phenolphthalein would be appropriate for this particular titration. The titration curve
corresponding to a weak base and strong acid titration is similarly behaved. In this case,
indicators such as methyl orange or bromothymol blue are regularly used.

On the other hand, titration curves corresponding to acid-base titrations in which the
constituents are a weak acid and weak base, are quite irregular in nature. Due to the
nature of such titrations, no definite indicator may be appropriate, and thus pH meters are
often used.

Types of titrations
There are various sorts of titrations whose goals are different to the others. The most
common types of titrations in qualitative work are acid-base titrations and redox
titrations.

[edit] Acid-base titration

Main article: Acid-base titration
Color on Acidic Range of Color Color on Basic
Indicator
Side Change Side
Methyl Violet Yellow 0.0 - 1.6 Violet
Bromophenol Blue Yellow 3.0 - 4.6 Blue
 Methyl Orange Red 3.1 - 4.4 Yellow
Methyl Red Red 4.4 - 6.2 Yellow
Litmus Red 5.0 - 8.0 Blue
Bromothymol Blue Yellow 6.0 - 7.6 Blue
Phenolphthalein Colorless 8.3 - 10.0 Pink
Alizarin Yellow Yellow 10.1 - 12.0 Red

These titrations are based on the neutralization reaction that occurs between an acid and a
base, when mixed in solution. The acid (resp. base) is added to a burette which was
rinsed with the same acid prior to this addition to prevent contamination or diluting of the
acid being measured. The base (resp. acid) is added to a volumetric flask which had been
rinsed with distilled water prior to the addition to prevent contamination or dilution of the
base/alkali being measured. The solution in the volumetric flask is often a standard
solution; one whose concentration is exactly known. The solution in the burette, however,
is the solution whose concentration is to be determined by titration. The indicator used
for such an acid-base titration often depends on the nature of the constituents as described
in the above section. Common indicators, their colours, and the pH range in which they
change colour, are given in the table above. When more precise results are required, or
when the titration constituents are a weak acid and a weak base, a pH meter or a
conductance meter are used.

Redox titration
Main article: Redox titration

These titrations are based on a redox reaction between an oxidizing agent and a reducing
agent. The oxidizing agent (resp. reducing agent) is added to the burette which was rinsed
with the same oxidizing agent. The reducing agent (resp. oxidizing agent) is added to the
conical flask, which had been rinsed with distilled water. Like in an acid-base titration,
the standard solution is often the one in the conical flask, and the solution whose
concentration is to be determined is the one in the burette. The procedure for carrying out
redox titrations is similar to that required for carrying out acid-base titrations.

Most commonly, a potentiometer or a redox indicator are used to determine the end point
of the titration. For example, when one of constituents of the titration is the oxidizing
agent potassium dichromate, the colour change of the solution from orange to green is not
definite and thus an indicator such as sodium diphenylamine is used. The analysis of
wines for their sulfur dioxide content requires the use of iodine as an oxidizing agent. In
this case, starch is used as an indicator; a blue starch-iodine complex is formed once an
excess of iodine is present, thus signalling the endpoint of the titration.

On the other hand, some redox titrations do not require an indicator, due to the intense
colour of some of the constituents. For instance, in a titration where the oxidizing agent
potassium permanganate (permanganometry) is present, a slight faint persisting pink
colour signals the endpoint of the titration, and no particular indicator is therefore
required.

[Complexometric titration
Main article: Complexometric titration

These titrations are based on the formation of a complex between the analyte and the
titrant. The chelating agent EDTA is very commonly used to titrate metal ions in solution.
These titrations generally require specialized indicators that form weaker complexes with
the analyte. A common example is Eriochrome Black T for the titration of calcium and
magnesium ions.

Zeta potential titration

Main article: Zeta potential titration

These titrations characterize heterogeneous systems, such as colloids. Zeta potential plays
role of indicator. One of the purposes is determination of iso-electric point when surface
charge becomes 0. This can be achieved by changing pH or adding surfactant. Another
purpose is determination of the optimum dose of the chemical for flocculation or
stabilization.

Miscellaneous

A form of titration can also be used to determine the concentration of a virus or

bacterium. The original sample is diluted (in some fixed ratio, such as 1:1, 1:2, 1:4, 1:8,
etc.) until the last dilution does not give a positive test for the presence of the virus. This
value, the titre, may be based on TCID50, EID50, ELD50, LD50 or pfu. This procedure is
more commonly known as an assay.

Measuring the endpoint of a titration

Main article: Endpoint (chemistry)

Different methods to determine the endpoint include:

• pH indicator: This is a substance that changes colour in response to a chemical

change. An acid-base indicator (e.g., phenolphthalein) changes colour depending
on the pH. Redox indicators are also frequently used. A drop of indicator solution
is added to the titration at the start; when the colour changes the endpoint has
been reached.
• A potentiometer can also be used. This is an instrument that measures the
electrode potential of the solution. These are used for titrations based on a redox
reaction; the potential of the working electrode will suddenly change as the
endpoint is reached.
 • pH meter: This is a potentiometer that uses an electrode whose potential depends
on the amount of H+ ion present in the solution. (This is an example of an ion-
selective electrode.) This allows the pH of the solution to be measured throughout
the titration. At the endpoint, there will be a sudden change in the measured pH. It
can be more accurate than the indicator method, and is very easily automated.
• Conductance: The conductivity of a solution depends on the ions that are present
in it. During many titrations, the conductivity changes significantly. (For instance,
during an acid-base titration, the H+ and OH- ions react to form neutral H2O. This
changes the conductivity of the solution.) The total conductance of the solution
depends also on the other ions present in the solution (such as counter ions). Not
all ions contribute equally to the conductivity; this also depends on the mobility of
each ion and on the total concentration of ions (ionic strength). Thus, predicting
the change in conductivity is harder than measuring it.
• Colour change: In some reactions, the solution changes colour without any added
indicator. This is often seen in redox titrations, for instance, when the different
oxidation states of the product and reactant produce different colours.
• Precipitation: If the reaction forms a solid, then a precipitate will form during the
titration. A classic example is the reaction between Ag+ and Cl- to form the very
insoluble salt AgCl. This usually makes it difficult to determine the endpoint
precisely. As a result, precipitation titrations often have to be done as "back"
titrations (see below).
• An isothermal titration calorimeter uses the heat produced or consumed by the
reaction to determine the endpoint. This is important in biochemical titrations,
such as the determination of how substrates bind to enzymes.
• Thermometric titrimetry is an extraordinarily versatile technique. This is
differentiated from calorimetric titrimetry by the fact that the heat of the reaction
(as indicated by temperature rise or fall) is not used to determine the amount of
analyte in the sample solution. Instead, the endpoint is determined by the rate of
temperature change.
• Spectroscopy can be used to measure the absorption of light by the solution
during the titration, if the spectrum of the reactant, titrant or product is known.
The relative amounts of the product and reactant can be used to determine the
endpoint.
• Amperometry can be used as a detection technique (amperometric titration). The
current due to the oxidation or reduction of either the reactants or products at a
working electrode will depend on the concentration of that species in solution.
The endpoint can then be detected as a change in the current. This method is most
useful when the excess titrant can be reduced, as in the titration of halides with
Ag+. (This is handy also in that it ignores precipitates.)

Back Titration

The term back titration is used when a titration is done "backwards"; instead of titrating
the original analyte, one adds a known excess of a standard reagent to the solution, then
titrates the excess. A back titration is useful if the endpoint of the reverse titration is
easier to identify than the endpoint of the normal titration. They are also useful if the
reaction between the analyte and the titrant is very slow.

Particular uses
• As applied to biodiesel, titration is the act of determining the acidity of a sample
of WVO by the dropwise addition of a known base to the sample while testing
with pH paper for the desired pH=8.5 reading. By knowing how much base
neutralizes an amount of WVO, we discern how much base to add to the entire
batch.
• Titrations are a very common procedure held in secondary education, to assess a
chemistry student's practical aptitude[citation needed].
• Titrations in the petrochemical or food industry to define oils, fats or biodiesel
and similar substances. An example procedure for all three can be found here: [1].
o Acid number: an acid-base titration with colour indicator is used to
determine the free fatty acid content. See also: pH of fatty acids.
o Iodine number: a redox titration with colour indication, which indicates
the amount of unsaturated fatty acids.
o Saponification value: an acid-base back titration with colour indicator or
potentiometric to get a hint about the average chain length of fatty acids in
a fat.
o Karl Fischer titration: a method to analyse trace amounts of water in a
substance in a lab.

Thin layer chromatography

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Thin layer chromatography

Separation of black ink on a TLC plate

Acronym TLC
Classification Chromatography
Other techniques
 Agarose gel electrophoresis
Related
SDS-PAGE

Thin layer chromatography (TLC) is a chromatography technique used to separate

mixtures.[1] Thin layer chromatography is performed on a sheet of glass, plastic, or
aluminum foil, which is coated with a thin layer of adsorbent material, usually silica gel,
aluminium oxide, or cellulose (blotter paper). This layer of adsorbent is known as the
stationary phase.

After the sample has been applied on the plate, a solvent or solvent mixture (known as
the mobile phase) is drawn up the plate via capillary action. Because different analytes
ascend the TLC plate at different rates, separation is achieved.[2].

Thin layer chromatography can be used to:

• Monitor the progress of a reaction

• Identify compounds present in a given substance
• Determine the purity of a substance

Specific examples of these applications include:

• determination of the components a plant contains

• analyzing ceramides and fatty acids
• detection of pesticides or insecticides in food and water
• analyzing the dye composition of fibers in forensics, or
• assaying the radiochemical purity of radiopharmaceuticals

A number of enhancements can be made to the original method to automate the different
steps, to increase the resolution achieved with TLC and to allow more accurate
quantitation. This method is referred to as HPTLC, or "high performance TLC".

Contents
[hide]

• 1 Plate preparation
• 2 Technique
• 3 Preparative TLC
• 4 Analysis
• 5 Applications

• 6 References

[edit] Plate preparation

TLC plates are usually commercially available, with standard particle size ranges to
improve reproducibility. They are prepared by mixing the adsorbent, such as silica gel,
with a small amount of inert binder like calcium sulfate (gypsum) and water. This
mixture is spread as a thick slurry on an unreactive carrier sheet, usually glass, thick
aluminum foil, or plastic. The resultant plate is dried and activated by heating in an oven
for thirty minutes at 110 °C. The thickness of the adsorbent layer is typically around 0.1 –
0.25 mm for analytical purposes and around 0.5 – 2.0 mm for preparative TLC.[3]

[edit] Technique

Development of a TLC plate, a purple spot separates into a red and blue spot.

Chromatogram of 10 essential oils coloured with vanillin reagent.

The process is similar to paper chromatography with the advantage of faster runs, better
separations, and the choice between different stationary phases. Because of its simplicity
and speed TLC is often used for monitoring chemical reactions and for the qualitative
analysis of reaction products.

To run a TLC, the following procedure is carried out: [4]

• A small spot of solution containing the sample is applied to a plate, about 1.5
centimeters from the bottom edge. The solvent is allowed to completely evaporate
off, otherwise a very poor or no separation will be achieved. If a non-volatile
solvent was used to apply the sample, the plate needs to be dried in a vacuum
chamber.
• A small amount of an appropriate solvent (elutant) is poured in to a glass beaker
or any other suitable transparent container (separation chamber) to a depth of less
than 1 centimeter. A strip of filter paper is put into the chamber, so that its bottom
touches the solvent, and the paper lies on the chamber wall and reaches almost to
the top of the container. The container is closed with a cover glass or any other lid
and is left for a few minutes to let the solvent vapors ascend the filter paper and
 saturate the air in the chamber. (Failure to saturate the chamber will result in poor
separation and non-reproducible results).
• The TLC plate is then placed in the chamber so that the spot(s) of the sample DO
NOT TOUCH the surface of the elutant in the chamber, and the lid is closed. The
solvent moves up the plate by capillary action, meets the sample mixture and
carries it up the plate (elutes the sample). When the solvent front reaches no
higher than the top of the filter paper in the chamber, the plate should be removed
(continuation of the elution will give a misleading result) and dried.

Different compounds in the sample mixture travel at different rates due to the differences
in their attraction to the stationary phase, and because of differences in solubility in the
solvent. By changing the solvent, or perhaps using a mixture, the separation of
components (measured by the Rf value) can be adjusted. Also, the separation achieved
with a TLC plate can be used to estimate the separation of a flash chromatography
column.[5]

Separation of compounds is based on the competition of the solute and the mobile phase
for binding places on the stationary phase. For instance, if normal phase silica gel is used
as the stationary phase it can be considered polar. Given two compounds which differ in
polarity, the more polar compound has a stronger interaction with the silica and is
therefore more capable to dispel the mobile phase from the binding places. Consequently,
the less polar compound moves higher up the plate (resulting in a higher Rf value). If the
mobile phase is changed to a more polar solvent or mixture of solvents, it is more capable
of dispelling solutes from the silica binding places and all compounds on the TLC plate
will move higher up the plate. It is commonly said that "strong" solvents (elutants) push
the analyzed compounds up the plate, while "weak" elutants barely move them. The order
of strength/weakness depends on the coating (stationary phase) of the TLC plate. For
silica gel coated TLC plates, the elutant strength increases in the following order:
Perfluoroalkane (weakest), Hexane, Pentane, Carbon tetrachloride, Benzene/Toluene,
Dichloromethane, Diethyl ether, Ethylacetate, Acetonitrile, Acetone, 2-Propanol/n-
Butanol, Water, Methanol, Triethylamine, Acetic acid, Formic acid (strongest). For C18
coated plates the order is reverse. Practically this means that if you use a mixture of ethyl
acetate and heptane as the mobile phase, adding more ethyl acetate results in higher Rf
values for all compounds on the TLC plate. Changing the polarity of the mobile phase
will normally not result in reversed order of running of the compounds on the TLC plate.
An eluotropic series can be used as a guide in selecting a mobile phase. If a reversed
order of running of the compounds is desired, an apolar stationary phase should be used,
such as C18-functionalized silica.

[edit] Preparative TLC

TLC can also be used on a small semi-preparative scale to separate mixtures of up to a
few hundred milligrams. The mixture is not "spotted" on the TLC plate as dots, but rather
is applied to the plate as a thin even layer horizontally to and just above the solvent level.
When developed with solvent the compounds separate in horizontal bands rather than
horizontally separated spots. Each band (or a desired band) is scraped off the backing
material. The backing material is then extracted with a suitable solvent (e.g. DCM) and
filtered to give the isolated material upon removal of the solvent. For small-scale
reactions with easily separated products, preparative TLC can be a far more efficient in
terms of time and cost than doing column chromatography. Obviously, the whole plate
can not be chemically developed or the product will be chemically destroyed. Thus this
technique is best used with compounds that are coloured, or visible under UV light.
Alternatively, a small section of the plate can be chemically developed e.g. cutting a
section out and chemically developing it, or masking most of the plate and exposing a
small section to a chemical developer like iodine.

[edit] Analysis
As the chemicals being separated may be colorless, several methods exist to visualize the
spots:

• Often a small amount of a fluorescent compound, usually manganese-activated

zinc silicate, is added to the adsorbent that allows the visualization of spots under
a blacklight (UV254). The adsorbent layer will thus fluoresce light green by itself,
but spots of analyte quench this fluorescence.
• Iodine vapors are a general unspecific color reagent
• Specific color reagents exist into which the TLC plate is dipped or which are
sprayed onto the plate[6]
o Potassium permanganate - oxidation
o Iodine
• In the case of lipids, the chromatogram may be transferred to a PVDF membrane
and then subjected to further analysis, for example mass spectrometry, a
technique known as Far-Eastern blotting.

Once visible, the Rf value , or retention factor, of each spot can be determined by dividing
the distance traveled by the product by the total distance traveled by the solvent (the
solvent front). These values depend on the solvent used, and the type of TLC plate, and
are not physical constants. Eluent on the thin layer is put on top of the plate

[edit] Applications
In organic chemistry, reactions are qualitatively monitored with TLC. Spots sampled with
a capillary tube are placed on the plate: a spot of starting material, a spot from the
reaction mixture, and a "co-spot" with both. A small (3 by 7 cm) TLC plate takes a
couple of minutes to run. The analysis is qualitative, and it will show if the starting
material has disappeared, i.e. the reaction is complete, if any product has appeared, and
how many products are generated (although this might be under-estimated due to co-
elution). Unfortunately, TLCs from low-temperature reactions may give misleading
results, because the sample is warmed to room temperature in the capillary, which can
alter the reaction—the warmed sample analyzed by TLC is not the same as what is in the
low-temperature flask. One such reaction is the DIBALH reduction of ester to aldehyde.
As an example the chromatography of an extract of green leaves (for example spinach) in
7 stages of development. Carotene elutes quickly and is only visible until step 2.
Chlorophyll A and B are halfway in the final step and lutein the first compound staining
yellow.

Step 1 Step 2 Step 3 Step 4

Step 5 Step 6

pH meter

A pH meter

A pH meter is an electronic instrument used to measure the pH (acidity or alkalinity) of

a liquid (though special probes are sometimes used to measure the pH of semi-solid
substances). A typical pH meter consists of a special measuring probe (a glass electrode)
connected to an electronic meter that measures and displays the pH reading.
Contents
The probe
The pH probe measures pH as the activity of hydrogen ions surrounding a thin-walled
glass bulb at its tip. The probe produces a small voltage (about 0.06 volt per pH unit) that
is measured and displayed as pH units by the meter. For more information about pH
probes, see glass electrode.

The meter
The meter circuit is no more than a voltmeter that displays measurements in pH units
instead of volts. The input impedance of the meter must be very high because of the high
resistance — approximately 20 to 1000 MΩ — of the glass electrode probes typically
used with pH meters. The circuit of a simple pH meter usually consists of operational
amplifiers in an inverting configuration, with a total voltage gain of about -17. The
inverting amplifier converts the small voltage produced by the probe (+0.059 volt/pH)
into pH units, which are then offset by seven volts to give a reading on the pH scale. For
example:

• At neutral pH (pH 7) the voltage at the probe's output is 0 volts. 0 * 17 + 7 = 7.

• At basic pH, the voltage at the probe's output ranges from +0 to +0.41 volts (7 *
0.059 = 0.41). So for a sample of pH 10 (3 pH units above neutral), 3 * 0.059 =
0.18 volts), the output of the meter's amplifier is 0.18 * 17 + 7 = 10.
• At acid pH, the voltage at the probe's output ranges from -0.41 volts to -0. So for
a sample of pH 4 (3 pH units below neutral), -3 * 0.059 = -0.18 volts, the output
of the meter's amplifier is -0.18 * 17 + 7 = 4.

The two basic adjustments performed at calibration (see below) set the gain and offset of
the inverting amplifier.

[edit] Calibration and use

For very precise work the pH meter should be calibrated before each measurement. For
normal use calibration should be performed at the beginning of each day. The reason for
this is that the glass electrode does not give a reproducible e.m.f. over longer periods of
time.

Calibration should be performed with at least two standard buffer solutions that span the
range of pH values to be measured. For general purposes buffers at pH 4 and pH 10 are
acceptable. The pH meter has one control (calibrate) to set the meter reading equal to the
value of the first standard buffer and a second control (slope) which is used to adjust the
meter reading to the value of the second buffer. A third control allows the temperature to
be set. Standard buffer sachets, which can be obtained from a variety of suppliers, usually
state how the buffer value changes with temperature.

The calibration process correlates the voltage produced by the probe (approximately 0.06
volts per pH unit) with the pH scale. After each single measurement, the probe is rinsed
with distilled water or deionized water to remove any traces of the solution being
measured, blotted with a clean tissue to absorb any remaining water which could dilute
the sample and thus alter the reading, and then quickly immersed in another solution.
When not in use, the probe tip must be kept wet at all times. It is typically kept immersed
in an acidic solution of around pH 3.0. In an emergency, acidified tap water can be used,
but distilled or deionised water must never be used for longer-term probe storage as the
relatively ionless water "sucks" ions out of the probe through diffusion, which degrades
it.

Occasionally (about once a month), the probe may be cleaned using pH-electrode
cleaning solution; generally a 0.1 M solution of Hydrochloric Acid (HCl) is used [1],
having a pH of about one.

Types of pH meters

A simple pH meter

pH meters range from simple and inexpensive pen-like devices to complex and expensive
laboratory instruments with computer interfaces and several inputs for indicator (ion-
sensitive, redox), reference electrodes, and temperature sensors such as thermoresistors or
thermocouples. Cheaper models sometimes require that temperature measurements be
entered to adjust for the slight variation in pH caused by temperature. Specialty meters
and probes are available for use in special applications, harsh environments, etc. Pocket
pH meters are readily available today for a few tens of dollars that automatically
compensate for temperature (ATC, Automatic Temperature Compensation)

History
The first commercial pH meters were built around 1936 by Radiometer in Denmark and
by Arnold Orville Beckman in the United States. While Beckman was an assistant
professor of chemistry at the California Institute of Technology, he was asked to devise a
quick and accurate method for measuring the acidity of lemon juice for the California
Fruit Growers Exchange (Sunkist). Beckman's invention helped him to launch the
Beckman Instruments company (now Beckman Coulter). In 2004 the Beckman pH meter
was designated an ACS National Historical Chemical Landmark in recognition of its
significance as the first commercially successful electronic pH meter.[2]

In the 1970s Jenco Electronics of Taiwan designed and manufactured the first portable
digital pH meter. This meter was sold under Cole-Parmer's label.

Building a pH meter
Because the circuitry of a basic pH meter is quite simple, it is possible to build a
serviceable pH meter or pH controller with parts available at a neighborhood electronics
retailer. (pH probes, however, are not so easily acquired and must usually be ordered
from a scientific instrument supplier.) For a walkthrough of how to build the simplest
possible pH meter or a detailed description of how to build a pH meter/pH controller, see
The pH Pages. The application note for the LM6001 chip at the National Semiconductor
web site also has a very simple demonstration circuit. Although the application note is for
a specialty IC, serviceable pH meters can be built from any operational amplifier with a
high input impedance, such as the common and inexpensive National Semiconductor
TL082 or its equivalent.