Classifying Chemicals
1. Reagent Grade: Reagent grade chemical conform
Chemical Apparatus, and Unit
Operation of Analytical Chemistry
3. Special-Purpose Reagent: chemicals that have
been prepared for a specific application. Included
among these are solvents for spectrophotometry and
high-performance liquid chromatography.
Rules for Handling Reagents and
Solutions
5. Never insert spatulas, spoons, or knives into
a bottle that contains a solid chemicals.
6. Keep the reagent shelf and the laboratory
balance clean and neat.
7. Observe regulations concerning the disposal
of surplus reagents and solutions.
to the minimum standards set forth by the Reagent
Chemical committee of the American Chemical
Society and are used wherever possible in analytical
work.
2. Primary Standard Grade: Extraordinary purity is
required for a primary standard. Primary standard
reagent is carefully analyzed and the assay is printed
on the container label.
Rules for Handling Reagents and
Solutions
1. Select the best grade of chemical available
for analytical work.
2. Replace the top of every container
immediately after removal of the reagent.
3. Hold the stoppers of reagent bottles
between your fingers.
4. Never return any excess reagent to a bottle.
Cleaning and Marking Laboratory Ware
• Every beaker, flask, or crucible that will contain the
sample must be thoroughly cleaned before being
used.
• The apparatus should be washed with a hot detergent
solution and then rinsed, initially with tap water and
finally with several small portions of deionized
water.
• Organic solvents such as benzene or acetone may be
used to remove grease films.
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 Types of Analytical Balances Types of Analytical Balances

An analytical balance is a weighing Semimicroanalytical balances have a

instrument with a maximum capacity that maximum load of 10 to 30 g with a precision
ranges from 1 g to a few kilograms with a of 0.01mg.
precision of at least 1 part in 105 at Microanalytical balance has a capacity of 1
maximum capacity. to 3 g and a precision of 0.001mg.

Macrobalances have a maximum capacity

ranging between 160 and 200 g;
measurement can be made with a standard
deviation of 0.1mg.

Desiccators and Desiccants

• Oven drying is the most common way of

removing moisture from solids.

• This approach is not appropriate for

substances that decompose or for those from
which water is not removed at the
temperature of the oven.

Desiccators and Desiccants

• Dried material are stored in desiccator while

they cool so as to minimize the uptake of
moisture.

• The base section of the desiccator contains a

chemical drying agent (desiccants) such as
anhydrous calcium chloride, calcium sulfate,
magnesium perchlorate or phosphorus
pentoxide.
Desiccator

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 Weighing by Difference

Weighing by difference is a simple method for

determining a series of sample weights.
• First the bottle and its contents are weighed.
• Sample is then transferred from the bottle to a
container; gentle tapping of the bottle with its
top and slight rotation of the bottle control over
the amount of sample removed.
• Following transfer, and its residual contents are
weighed.
• The mass of the sample is the difference
Weighing bottles
between the two weighings.

Volume Measurement
 Pipets

 Burets

 Volumetric flask

 Measuring cylinder

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 Pipets
• Pipette Calibration and Pipetting Accuracy
Pipette - a high precision laboratory
instruments that requires proper maintenance
on a regular basis.
Ways to Stop Pipetting Errors
From Ruining Your Experiments
. Limitations:
Without accurate pipetting
• experiments would not be reproducible
• stock solutions would be inaccurate
• assays would have such large errors that comparing
them would be meaningless.
Know How Pipets Work!!!
Precision instruments they may be, but the accuracy of
your micropipettes depends on you
Pipette Storage – Where and How for Pipetting
Accuracy
Proper storage -very important factor in maintaining
an accurate pipetting result.
• Upright –prevent any liquids from corroding the
internal workings of your pipette.
• Environmental Factors – Store your pipettes away
from the window, heating sources and in an
environment with low moisture
Air Displacement Pipette
• works a bit like a syringe, except that there is an air-filled
cushion between the piston and the sample.
• Temperature and pressure affects the volume
of the air cushion, which affects pipetting
accuracy.
• volatile solvents can evaporate into the air
cushion, which leads to an inaccurate and
lower dispensed volume than what is
displayed on the pipette.
• barrel of air displacement pipettes is
vulnerable to contamination by the pipetted
solution, corrosives or bio-hazardous
material, this can be a problem.
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Positive displacement pipettes

• also work like a

syringe, but they
don’t have an air
cushion—unlike air
displacement
pipettes.
• more accurate for
pipetting volatile
solvents because
there is no place (air
cushion) for the
solvent to evaporate.

The lack of air cushion decreases the chance of contamination

when pipetting corrosives and bio-hazardous material, which
makes positive displacement pipettes more suitable to working
with those reagents.

5
 How do positive displacement
pipets work?
How do air displacement pipets
work?
Works like a syringe. There is no
air cushion between the
When the push button is pressed,
disposable piston and the sample.
the piston moves down to let the
With no elastic air cushion to
air out. Volume of air displaced is
expand or contract, the aspiration
equal to the volume of liquid
force remains constant,
aspirated.
unaffected by the physical
properties of the sample.

This allows to pipette very

viscous or high density samples
such as mercury or toothpaste.

Accuracy and precision: The most critical aspects

Reliable results affecting pipetting results are accuracy and precision.
Scientists rely on pipettes that are accurate and
reproducible to guarantee the success of an experiment.

Factors to improve pipetting results:

• Physical properties of liquid: aqueous, viscous and

volatile.
• Most liquids are of the aqueous type, making air
displacement pipettes the first choice.
• positive displacement pipettes in very viscous or
volatile liquids.

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 For maximum pipetting reliability: 4. High quality pipettes and tips

1. Volume transferred
• As a rule of thumb, always choose the smallest pipette
capable of handling the required volume
-accuracy decreases when the set volume is close to the
pipette’s minimum capacity.
2. Easy calibration
• Some electronic pipettes have useful features, such as
setting a calibration reminder or saving the calibration
history.
3. Volume adjustment locking
• Volumes set on traditional manual pipettes can change
while pipetting, due to unintentional plunger turns.
Do your pipette tips ever loosen, leak or fall off?
This is a common issue in laboratories, caused by the use
of universal pipette tips. Such tips require ‘hammering on’,
which stretches the pipette tip rim. This can cause leaking
or misaligned tips, or even cause the pipette tips to fall off
the pipette completely!
Choosing micropipettes which were designed together
with the tips ensures secure connections and tips that do
not leak or fall off.
Color coded pipettes and pipette tips: Color coding helps
you to choose the right tips for your pipette.

No matter how light and well balanced a pipette is,

Efficiency prolonged pipetting tasks remain physically and
mentally challenging.
Multichannel pipettes
allow you to transfer Switch between the manual and
multiple samples at once. automated pipetting tasks, relieving
of the routine pipetting.

Transfer from tubes to plates

using a VOYAGER pipette

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 Take Care of Your Pipette
Clean Your Pipettes Each Day Before Use
Have Your Pipette Serviced Every 6-12 months
• wipe pipette with 70% ethanol
• need to have it serviced more often depending on
Store Your Pipette Vertically, Using a Pipette Holder
your requirements for accuracy. The service should
include re-calibration, greasing of the moving parts,
• Storing your pipette this way prevents any liquids that are in the
and replacement of any worn out seals or other
pipette barrel from getting any further inside and causing
parts.
corrosion.
Check Your Pipettes Daily for Damage
Never Put Your Pipette on Its Side With Liquid in the Tip
Examine the nose of the barrel (where the tip is fitted)
for any obvious damage. If there is a problem, have it • There’s nothing preventing the liquid from rolling down and into
serviced—because it is unlikely to be fit for the job. the pipette barrel . Any liquid inside the pipette barrel results in
contamination at the least and can cause some serious corrosion
Clean Your Pipettes Each Day Before Use damage.

Wiping your pipette with some 70% ethanol should do it.

6. Store Your Pipette Vertically, Using a Pipette Holder

Storing your pipette this way prevents any liquids that are in the
pipette barrel from getting any further inside and causing corrosion.
7. Never Put Your Pipette on Its Side With Liquid in the Tip

Use Well-Fitting Tips

• Poorly fitting tips allow air to escape when drawing up and

dispensing liquid, leading to inaccurate results.

Typical pipets

Automatic pipet

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9
Correct Operation

10
11
QUICK DIAGNOSIS

Solutions:

1. Use a positive
displacement pipet
because it has no air
cushion.
2. Saturate the air-cushion
of your pipette with
solvent vapor by
aspirating and
distributing solvent
repeatedly. The leak will
stop when pressure
equilibrium is reached.

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Multichannel pipettes with tip spacing feature
How can you avoid discomfort or arm injuries caused by excessive pipetting?
How can you avoid discomfort or arm injuries caused by excessive pipetting?
(Multible answers possible)
Use heavy pipettes in order to have more force to securely load tips
Low tip loading and ejection forces
Only use pipettes that have their mass center at the lowest point
Having the pipette in your hand for the shortest amount of time possible

Calibration of Pipettors

Use of Gravimetric Method

Based on the determination of the weight of water samples

delivered by the pipette.

Implementation of this method requires the strict monitoring of

environmental conditions, and the systematic use of adequate
and controlled equipment.

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 Replicate Data on the Calibration of a 10ml Pipette
No. Vol, ml. No. Vol, ml. No. Vol, ml

1 9.988 18 9.975 35 9.976

2 9.973 19 9.980 36 9.990
3 9.986 20 9.994 37 9.988
4 9.980 21 9.992 38 9.971
5 9.975 22 9.984 39 9.986
6 9.982 23 9.981 40 9.978
7 9.986 24 9.987 41 9.986
8 9.982 25 9.978 42 9.982
9 9.981 26 9.983 43 9.977
10 9.990 27 9.982 44 9.977
11 9.980 28 9.991 45 9.986
12 9.989 29 9.981 46 9.978
13 9.978 30 9.969 47 9.983
14 9.971 31 9.985 48 9.980
15 9.982 32 9.977 49 9.983
16 9.983 33 9.976 50 9.979
17 9.988 34 9.983

Mean volume 9.982 ml Median volume 9.982 ml

Spread 0.025 ml Standard deviation 0.0056 ml

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GLOSSARY

Burets and volumetric flask

15
 Reading a buret
Using pipet

Simple Crucibles

Simple crucibles serve only as containers.

Porcelain, aluminum oxide, silica and platinum
crucibles maintain constant mass and are used
principally to convert a precipitate into a suitable
weighing form.
• The solid is first collected on filter paper.
• The filter and contents are then transferred to a
weighed crucible,
• the paper is ignited.

Titration

Filtering Crucibles

Filtering crucibles serve not only as containers but also

as filters.
• A vacuum is used to hasten the filtration
• a tight seal between crucible and filtering flask is
accomplished with any of the several types of rubber
adapters.

Filtering Crucible

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 Sintered-glass Crucibles
Sintered-glass crucibles are manufactured in fine,
medium, and coarse porosities. The upper temperature
limit for sintered glass crucible is ordinarily about
200oC. Filtering crucibles made entirely of quartz can
tolerate substantially higher temperatures.

Filter Paper
Paper is an important filtering medium. Ashless paper is
manufactured from cellulose fibers that have been
treated with HCl and HF acids to remove metallic
impurities and silica, NH3 is then used to neutralize the
acids. It is necessary to destroy the paper by ignition if
the precipitate collected on it is to be weighed.
Decantation and transferring precipitate

Folding and seating a filter paper Vacuum Filtration

Heating Equipment Heating Equipment

Many precipitate can be weighed directly after being Microwave laboratory ovens are currently appearing
brought to constant mass in a low temperature drying on the market. Where applicable, these greatly
oven. shorten drying cycles.

Such an oven is electrically heated and capable of
maintaining a constant temperature to within 1oC. The
maximum attainable temperature ranges from 140 to
260oC, depending on make and model, for many
precipitate 110oC is a satisfactory drying temperature.
Muffle furnace (a heavy duty electric furnace) is
capable of maintaining controlled temperatures of
1100oC or higher. Long handled tongs and heat
resistance gloves are needed for protection.

17
 Calibrating Glassware

Volumetric glassware is calibrated by measuring

the mass of a liquid (water) of known density
and temperature that is contained in the
volumetric ware.

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 Laboratory Notebook
1. Record all data and observations
2. Supply each entry with a heading
3. Date each page of the notebook
4. Never attempt to erase an incorrect entry, cross
it out with single horizontal line
5. Never remove a page from the notebook
6. Do not overcrowd entries
7. Keep first few pages for table of contents

Lab note book

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