Flow Diagram Showing the Steps in a Quantitative

Analysis

WHAT IS ANALYTICAL CHEMISTRY?

➢ Analytical chemistry is a measurement science
consisting of a set of powerful ideas and methods
that are useful in all fields of science and
medicine.

Qualitative Analysis vs. Quantitative Analysis

➢ Qualitative Analysis reveals the identity of the
elements and compounds in a sample.
➢ Quantitative Analysis indicates the amount of
each substance in a sample.
Picking a Method
The Role of Analytical Chemistry ➢ One of the first questions to be considered in the
➢ Analytical chemistry is applied throughout selection process is the level of accuracy
industry, medicine, and all the sciences. required.
➢ Quantitative analytical measurements also play a ➢ A second consideration related to economic
vital role in chemistry, biochemistry, biology, factors is the number of samples to be analyzed.
geology, physics, and the other sciences. ➢ The complexity of the sample and the number of
➢ Many scientists devote much time in the components in the sample always influence the
laboratory gathering quantitative information choice of method to some degree.
about systems that are important and interesting
to them. Acquiring the Sample
➢ Sampling involves obtaining a small mass of a
Classifying Quantitative Analytical Methods material whose composition accurately
✓ We classify analytical methods according to the represents the bulk of the material being
nature of this final measurement. sampled.
➢ Gravimetric Method – determines the ➢ Sampling is frequently the most difficult step in
mass of the analyte or some compound an analysis and the source of greatest error. The
chemically related to it. final results of an analysis will never be any more
➢ Volumetric Method / Titrimetric reliable than the reliability of the sampling step.
Method – determines the volume of a Processing the Sample
solution containing sufficient reagent to ➢ Under certain circumstances, no sample
react completely with the analyte. processing is required prior to the measurement
➢ Electroanalytical Method – Involves the step.
measurement of such electrical ➢ Under most circumstances, we must process the
properties as voltage, current, sample in any of a variety of different ways.
resistance, and quantity of electrical ➢ The first step in processing the sample is often
charge. the preparation of a laboratory sample.
➢ Spectroscopic Method – are based on ➢ Preparing a Laboratory Sample
measurement of the interaction between ✓ A solid sample is ground, mixed to
electromagnetic radiation and analyte ensure homogeneity, and stored for
atoms or molecules or on the production various lengths of time before analysis
of such radiation by analytes. begins.
✓ Because any loss or gain of water
changes the chemical composition of
solids, it is a good idea to dry samples
just before starting an analysis.

1
 ✓ Alternatively, the moisture content of the Remain Steps of a Typical Quantitative Analysis
sample can be determined at the time of ➢ Calibration and Measurement
the analysis in a separate analytical ▪ Ideally, the measurement of the property
procedure. is directly proportional to the
✓ Liquid samples are subject to solvent concentration.
evaporation ➢ Calculating Results
✓ If the analyte is a gas dissolved in a ▪ Computing analyte concentrations are
liquid, analyte must be kept inside a based on the raw experimental data
second sealed container to prevent collected in the measurement step, the
contamination by atmospheric gases. characteristics of the measurement
✓ Extraordinary measures, including instruments, and the stoichiometry of the
sample manipulation and measurement analytical reaction.
in an inert atmosphere, may be required ➢ Evaluating Results by Estimating Their
to preserve the integrity of the sample. Reliability
✓ Replicate samples, or replicates, are ▪ Analytical results are incomplete without
portions of a material of approximately an estimate of their reliability.
the same size that are carried through an
analytical procedure at the same time
and in the same way.
✓ Replication improves the quality of the
results and provides a measure of their LABORATORY SAFETY (LAB)
reliability. ➢ OSHA – Occupational Safety and Health Act
✓ Quantitative measurements on replicates ➢ OSHA PUBLIC LAW 91-596
are usually averaged, and various ✓ Enacted by the U.S. Congress in 1970
statistical tests are performed on the ✓ Authorized to conduct on-site inspections
results to establish their reliability. ✓ GOAL: to provide all employees (clinical
➢ Preparing Solutions: Physical and Chemical laboratory personnel included) with a
Changes safe work environment
✓ Ideally, the solvent should dissolve the
entire sample, including the analyte, OSHA STANDARDS
rapidly and completely. ➢ Bloodborne pathogen standard
✓ The sample may require heating with ➢ Formaldehyde standard
aqueous solutions of strong acids, strong ➢ Laboratory standard
bases, oxidizing agents, reducing ➢ Hazard communication standard
agents, or some combination of such ➢ Respiratory protection standard
reagents. ➢ Air contaminants standard
✓ It may be necessary to ignite the sample ➢ Personal protective equipment standard
in air or oxygen or perform a high-
temperature fusion of the sample in the BLOODBORNE PATHOGENS
presence of various fluxes. ➢ Applies to all exposure to blood or other
➢ Eliminating Interferences potentially infectious materials.
✓ Few chemical or physical properties of ➢ It defines terminology relevant to such exposures.
importance in chemical analysis are ➢ Mandates the development of an exposure
unique to a single chemical species. control plan.
✓ Species other than the analyte that affect ➢ UNIVERSAL PRECAUTIONS
the final measurement are called ▪ All human blood, tissue, and most fluids
interferences, or interferents. are known to be infectious
✓ An interference is a species that causes ✓ Human immunodeficiency virus
an error in an analysis by enhancing or (HIV)
attenuating (making smaller) the quantity ✓ Hepatitis B virus (HBV)
being measured. ✓ Other bloodborne pathogens
2
Bloodborne Pathogens Standard ✓ Sharps Injury Log: Must include
➢ History: Published in 1991. Revised in 2001 description & location of incident device
following passage of Needlestick Safety & involved. Employee privacy must be
Prevention Act to include stronger requirements protected.
for employers to evaluate & adopt safer medical
devices. HAZARD COMMUNICATION
➢ Purpose: To protect health-care workers from ➢ OSHA HazCom Standard
occupational exposure to bloodborne pathogens ➢ Ensure that the hazards of all chemicals used in
(BBP; e.g., HIV, HBV, HCV) the workplace have been evaluated.
➢ Primary Requirements: ➢ Defines hazardous substances and provides
✓ Exposure Control Plan: Determination guidance for evaluating and communicating
of employees’ risk of exposure and identified hazards.
implementation of method to control
exposure. Plan must be reviewed and SAMPLE CHEMICAL LABEL
updated annually to reflect new ✓ Statement of hazard
technologies. Documentation of ✓ Hazard class
evaluation and adoption of safer devices ✓ Safety precautions
is required. Nonmanagerial employees ✓ National Fire Protection Agency (NFPA) hazard
must be involved in evaluation and code
selection devices. ✓ Fire extinguisher type
✓ Universal Precautions: All blood and ✓ Safety instructions
certain body fluids are to be handled as if ✓ Formula weight; Lot number
known to be infectious or bloodborne
pathogens.
✓ Engineering Controls: Control
measures that isolate or remove a
hazard from workplace, e.g., sharps,
containers, self-sheathing needles,
plastic capillary tubes, Plexiglas shields.
✓ Work Practice Controls: e.g.,
handwashing, disposal of needles, with
safety device activated and holder
attached, ban on eating/drinking/smoking
in lab.
✓ Personal Protective Clothing & Hazard Communication
Equipment: e.g., lab coats, face shields. ✓ History: issued by OSHA in 1983. Written for
Employer must provide and must launder manufacturing industry, but courts expand
lab coats. jurisdiction to clinical labs.
✓ Housekeeping: e.g., proper disposal of ✓ Also known as “Right-to-Know Law”; HAZCOM
biohazardous waste, decontamination of ✓ Purpose: To inform employees about the
work surfaces. chemical hazards in workplace & protective
✓ Training: On assignment and annually measures.
thereafter. ✓ Primary Requirements:
✓ Medical Surveillance: Postexposure ▪ Written hazard communication plan
evaluation & follow-up at no cost to ▪ Inventory of hazardous chemicals on site
employee. ▪ Hazard labelling
✓ Hepatitis B Vaccine: Provided by ▪ Material Safety Data Sheet (MSDS) for
employer within 10 days of assignment at each chemically readily accessible to
no cost to employee. employees on each shift.
▪ Training on initial assignment & when
new hazard introduced.
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FIRE SAFETY MSDS: MATERIAL SAFETY DATA SHEET
➢ FIRE: a chemical reaction that involves the rapid ➢ Product name and identification
oxidation of a combustible material / fuel, with the ➢ Hazardous ingredients
subsequent liberation of heat and light. ➢ Permissible exposure limit
➢ FIRE TETRAHEDRON: ➢ Physical and chemical data
✓ (1) Fuel ➢ Health hazard data and carcinogenic potential
✓ (2) Heat / Ignition source ➢ Primary routes of entry
✓ (3) Oxygen ➢ Fire and explosion hazards
➢ ACT CASE OF A FIRE: ➢ Reactivity data Spill and disposal procedures
✓ Use RACE acronym ➢ PPE recommendations
✓ Rescue: Help anyone who is in ➢ Handling Emergency and first aid procedures
immediate danger get away from the fire. ➢ Storage and transportation precautions
✓ Alarm: pull the fire alarm to alert others. ➢ Chemical manufacturer’s name, address, and
✓ Contain: close doors to keep the fire telephone number
from spreading ➢ Special information section
✓ Extinguish/Evacuate: Use a fire
extinguisher if the fire is small or if the is Occupational Exposure to Hazardous Chemicals in
too big, start removing others from the Laboratories (OSHA Lab Standard)
facility immediately. ➢ History: issued by OSHA in 1990. Extension of
HCS written specifically for labs.
➢ Also known as “Laboratory Standard”
“Chemical Hygiene Standard”
➢ Purpose: to limit employee exposure to
hazardous chemical to levels at below
permissible exposure levels. (PELs)
➢ Primary Requirements:
✓ Written chemical hygiene plan outlining
standard operating procedures for use,
storage, exposure control and disposal of
hazardous chemicals.
✓ Designation of chemical hygiene officer.
✓ Hazard identification & labelling
✓ Material safety data sheet (MSDS) for
each chemically readily accessible to
employees on each shift.
✓ Use of personal protective equipment
✓ Proper maintenance of fume hoods &
other equipment
✓ Monitoring of employee exposure to
hazardous chemicals
✓ Medical exams at no cost in cases of
suspected overexposure
✓ Training on initial assignment & before
assignments involving new exposures.

4
 TYPES OF HAZARDS
➢ CHEMICAL HAZARDS
➢ BIOLOGICAL HAZARDS
➢ PHYSICAL HAZARDS
▪ ERGONOMIC HAZARDS
▪ IONIZING RADIATION
▪ NON-IONIZING RADIATION
▪ NOISES
➢ ELECTRICAL HAZARDS
➢ MECHANICAL HAZARDS
CHEMICAL HAZARDS
➢ Flammable/Combustible chemicals
✓ Flammable: Flash Point below 37.8C
(100F)
✓ Combustible: Flash Point above 37.8C
(100F)
➢ Corrosive chemicals
➢ Reactive chemicals
➢ Carcinogenic chemicals
➢ FLAMMABLE AND COMBUSTIBLE SOLVENTS:
✓ Acetone
✓ Benzene
✓ Ethanol
✓ Heptane
✓ Isopropanol
✓ Methanol
✓ Toluene
✓ Xylene
➢ Flammable Gases: Hydrogen
➢ Flammable Solids: Paraffin
➢ Corrosive Chemicals – Injurious to the skin or
eyes by direct contact or to the tissue of the
respiratory and gastrointestinal tracts if ingested.
▪ ACIDS: Acetic, Sulfuric, Nitric,
Hydrochloric Acid
▪ BASES: Ammonium Hydroxide,
Potassium Hydroxide, Sodium
Hydroxide.

5
 • Reactive Chemicals – Substances that under with, or contaminated needles or by aerosol
certain conditions can spontaneously explode or dispersion.
ignite or that evolve heat/flammable/explosive • Non-Ionizing Radiation – a type of
gases. electromagnetic radiation that does not carry
✓ Hydrogen is liberated of alkali metals enough energy to ionize atoms.
(sodium or potassium) are mixed with
water or acids, and spontaneous
combustion also may occur.
✓ Mixture of oxidizing agents, such as
peroxides, and reducing agents, such as
hydrogen, generates heat and may be
explosive.
✓ Carcinogenic Chemicals – Substances
that have been determined to be a
cancer-causing agent ▪ Ionizing Radiation – It is generated
• Benzidine is a common example of a through nuclear atoms, by very high
known carcinogen. temperature, via production of high
energy particles or due to acceleration of
charged particles by electromagnetic
fields.
▪ Types of Ionizing Radiation:
✓ Cosmic Rays
✓ X-Rays
✓ Gamma Rays
✓ Beta Particles
✓ Ultraviolet
PHYSICAL HAZARDS
➢ NOISE: Anything that has the potential to cause
hearing loss.
➢ Exposure to an equivalent sound pressure level
of an 85dB (decibels) over an 8-hour period
workday.
➢ Cumulative Trauma Disorders – Injuries
involving the musculoskeletal and/or nervous
system in response to long-term repetitive
twisting, bending, lifting or assuming static
postures for an extended period of time.
▪ Constant or excessive repetitive actions,
mechanical pressure, vibrations, or
compressive forces on the arms, hands,
wrists, neck or back.
▪ Human error by pushing beyond one’s
limits or when productivity limits are set
BIOLOGICAL HAZARDS too high.
• Expose an unprotected individual to bacteria, ✓ CARPAL TUNNEL SYNDROME – compression
viruses, parasites or other biological entities that and entrapment of nerve from wrist to hand.
can result in injury. ✓ TENDONITIS – inflammation of the tendon.
• Exposure occurs from ingestions, inoculation, ✓ TENOSYNOVITIS – inflammation or injury to
tactile contamination or inhalation of infectious synovial sheath that surrounds a tendon.
material from patients or their body fluids/tissues, ✓ BURSITIS – inflammation of one of the bursa of
supplies or materials they have been in contact synovial fluid.
6
 ➢ PRIMARY CONTRIBUTORS OF REPETITIVE SAFETY RULES INSIDE THE LABORATORY
STRAIN DISORDERS: ✓ Dress appropriately for the lab
✓ Position or posture ✓ Know what safety equipment is available and how
✓ Applied force to use it.
✓ Frequency of repetition ✓ Know the dangers of chemicals in use and read
ELECTRICAL HAZARDS labels carefully
➢ Associated Deaths: ✓ Dispose of chemicals according to instruction
✓ Death ✓ Always slowly add acid to water to avoid
✓ Shocks splattering.
✓ Burns ✓ Never point heating test tubes at yourself or
✓ Fire others.
✓ Explosion ✓ Do not pipette anything by mouth
MECHANICAL HAZARDS ✓ Use the fume hood when dealing with toxic fumes
➢ Sources of Hazards: ✓ Do not eat or drink in the lab
✓ Centrifuges ✓ Follow all direction(s)
✓ Autoclave
✓ Homogenizer
✓ Glasswares Tools of Analytical Chemistry
COMPRESSED GASES HAZARDS
➢ Associated Hazards: TOOLS OF ANALYTICAL CHEMISTRY
✓ Danger of Fire Selecting and Handling Reagents and Other
✓ Explosion Chemicals
✓ Asphyxiation
✓ Mechanical Injuries Classification of Chemicals
CRYOGENIC MATERIAL HAZARDS Reagent Grade
➢ Associated Hazards: ➢ Reagent-grade chemicals conform to the
✓ Fire minimum standards set forth by the Reagent
✓ Explosion Chemical Committee of the American Chemical
✓ Asphyxiation Society (ACS) and are used whenever possible in
✓ Pressure build-up analytical work. Some suppliers label their
✓ Embrittlement of Materials products with the maximum limits of impurity
✓ Tissue damage similar to that of thermal allowed by the ACS specifications while others
burns print actual concentrations for the various
WASTE MANAGEMENT impurities.
➢ Collection, transport, processing or disposal, Primary-standard grade
managing and monitoring of waste materials. ➢ The qualities required of a primary standard, in
FOUR BASIC WASTE DISPOSAL TECHNIQUES addition to extraordinary purity, primary-standard
✓ Landfill burial reagents have been carefully analyzed by the
✓ Incineration supplier, and the results are printed on the
✓ Flushing down the drain to the sewer system container label. The national institute of
✓ Recycling or Resource recovery standards and technology (NIST) is an excellent
source for primary standards. This agency also
prepares and sells reference standards, which
are complex substances that have been
exhaustively analyzed.
Special-purpose Reagent Chemicals
➢ Chemicals that have been prepared for a specific
application are also available. Included among
these are solvents for spectrophotometry and
high-performance Liquid chromatography.
Information pertinent to the intended use is
7
 supplied with these reagents. Data provided with RULES in HANDLING REAGENT and SOLUTION
a spectrophotometric solvent, for example, might Evaporating Liquids
include its absorbance at selected wavelengths ➢ Bumping is the sudden, often violent boiling that
and its ultraviolet cutoff wavelength. tends to spatter solution out of its container.
➢ Chemicals that have been prepared for a specific ➢ Wet washing is the oxidation of the organic
application are also available. constituents of a sample with Oxidizing reagents
➢ Included Among these are solvents for such as nitric acid, Sulfuric acid, hydrogen
spectrophotometry and high-performance Liquid peroxide, aqueous Bromine, or a combination of
chromatography. Information pertinent to the these reagents.
intended use is supplied Measuring Mass
➢ With these reagents. Data provided with a ➢ In most analyses, an analytical balance must be
spectrophotometric solvent, for example, might used to measure masses with high accuracy.
include its absorbance at selected wavelengths Less accurate laboratory balances are also used
and its ultraviolet cutoff Wavelength. for mass measurements when the demands for
reliability are not critical.
RULES in HANDLING REAGENT and SOLUTION Types of Balances
➢ We observe the following rules to prevent the ➢ Analytical Balance
accidental contamination of reagents and ➢ Electroanalytical balance
solutions: ➢ Single pan mechanical analytical balance
✓ Select the best grade of chemical available for
analytical work. Whenever possible, pick the Analytical Balance
smallest bottle that is sufficient to do the job. ➢ An analytical balance is an instrument for
✓ Replace the top of every container immediately determining mass with a maximum capacity that
after removing reagent. Do not rely on someone ranges from 1 g to a few kilograms with a
else to do so. precision of at least 1 part in 105 at maximum
✓ Hold the stoppers of reagent bottles between capacity. The precision and accuracy of many
your fingers. Never set a stopper on a desk top. modern analytical balances exceed 1 part in 106
✓ Unless specifically directed otherwise, never at full capacity.
return any excess reagent to a bottle. The money ✓ Macrobalance – most common
saved by returning excesses is seldom worth the analytical balance have a maximum
risk of contaminating the entire bottle. capacity ranging between 160 and 200 g.
✓ Unless directed otherwise, never insert spatulas, With these balances, measurements can
spoons, or knives into a bottle that contains a be made with a standard deviation of
solid chemical. Instead, shake the capped bottle 60.1 mg.
vigorously or tap it gently against a wooden table ✓ Semimicroanalytical balance have a
to break up an encrustation. Then pour out the maximum loading of 10 to 30 g with a
desired quantity. These measures are precision of 60.01 mg.
occasionally ineffective, and in such cases a ✓ Microanalytical balance has a capacity
clean porcelain spoon should be used. of 1 to 3 g and a precision of 60.001 mg
✓ Keep the reagent shelf and the laboratory (1 μg).
balance clean and neat. Clean up any spills Electron Analytical Balance
immediately. A modern electronic analytical balance provides
✓ Follow local regulations concerning the disposal unprecedented speed and ease of use. For example, one
of surplus reagents and solutions. instrument is controlled by touching a single bar at various
positions along its length
RULES in HANDLING REAGENT and SOLUTION
Cleaning Laboratory Ware
➢ An organic solvent, such as methyl ethyl ketone
or acetone, may be effective in removing grease
films. Chemical suppliers also market
preparations for eliminating such films.
8
The Single-Pan Mechanical Analytical Balance 5. Keep the balance and its case
scrupulously clean. A camel’s-hair brush
is useful for removing spilled material or
dust.
6. Always allow an object that has been
heated to return to room temperature
before weighing it.
7. Use tongs, finger pads, or a glass.
Sources of Error in Weighing
Correction for Buoyancy
➢ Buoyancy or upthrust, is an upward force
exerted by a fluid that opposes the weight of an
immersed object. In a column of fluid, pressure
Auxiliary Balances increases with depth as a result of the weight of
➢ Balances that are less precise than analytical the overlying fluid.
balances find extensive use in the analytical ➢ A buoyancy error will affect data if the density of
laboratory. These balances offer the advantages the object being weighed differs significantly from
of speed, ruggedness, large capacity, and that of the standard masses. The error has its
convenience. Low-precision balances should be origin in the difference in buoyant force exerted
used whenever high sensitivity is not required. by the medium (air) on the object and on the
➢ Top-loading auxiliary balances are particularly masses. Buoyancy corrections for electronic
convenient. A sensitive top-loading balance will balances may be accomplished with the
accommodate 150 to 200 g with a precision of equation.
about 1 mg—an order of magnitude less than a ➢ Attempts to weigh an object whose temperature
macroanalytical balance. is different from that of its surroundings will result
➢ Some balances of this type tolerate loads as great in a significant error. Failure to allow sufficient
as 25,000 g with a precision of 60.05 g. Most are time for a heated object to return to room
equipped with a taring device that brings the temperature is the most common source of this
balance reading to zero with an empty container problem. Errors due to a difference in
on the pan. Some are fully automatic, require no temperature have two sources.
manual dialing or mass handling, and provide a ▪ convection currents within the balance
digital readout of the mass. Modern top-loading case exert a buoyant effect on the pan
balances are electronic. and object.
➢ Use auxiliary laboratory balances for determining ▪ warm air trapped in a closed container
masses that do not require great accuracy. weighs less than the same volume at a
Precautions in Using an Analytical Balance LOWER TEMPERATURE.
➢ An analytical balance is a delicate instrument that Equipment and Manipulations Associated with
you must handle with care. Observe the following Weighing
general rules for working with an analytical WEIGHING BOTTLES
balance regardless of make or model: ➢ Weighing bottles are convenient for drying and
1. Center the load on the pan as well as storing solids. Two common varieties of these
possible. handy tools. The ground-glass portion of the cap-
2. Protect the balance from corrosion. style bottle shown on the left is on the outside and
Objects to be placed on the pan should does not come into contact with the contents. This
be limited to nonreactive metals, design eliminates the possibility of some of the
nonreactive plastics, and vitreous, or sample becoming trapped on the ground-glass
glasslike, materials. surface and subsequently being lost.
3. Observe special precautions (see Ruggedness is a principal advantage of using
Section 2E-6) for the weighing of liquids. plastic weighing bottles rather than glass, but
4. Consult your instructor if the balance plastic abrades easily and is not as easily cleaned
appears to need adjustment. as glass.
9
Manipulating Weighing Bottles previously weighed containers with snugly fitting
➢ Heating at 105°C to 110°C for 1 hour is sufficient covers (such as weighing bottles). The mass of
to remove the moisture from the surface of most the container is subtracted from the total mass.
solids. The weighing bottle is contained in a ➢ A volatile or corrosive liquid should be sealed in a
labeled beaker with a cover glass. weighed glass ampoule. The ampoule is heated,
Weighing by Difference and the neck is then immersed in the sample. As
➢ Weighing by difference is a simple method for cooling occurs, the liquid is drawn into the bulb.
determining a series of sample masses. ➢ The ampoule is then inverted and the neck sealed
➢ First, the bottle and its contents are weighed. One off with a small flame. The ampoule and its
sample is then transferred from the bottle to a contents, along with any glass removed during
container. Gentle tapping of the bottle with its top sealing, are cooled to room temperature and
and slight rotation of the bottle provide control weighed. The ampoule is then transferred to an
over the amount of sample removed. Following appropriate container and broken. A volume
transfer, the bottle and its residual contents are correction for the glass of the ampoule may be
weighed. needed if the receiving vessel is a volumetric
➢ The mass of the sample is the difference between flask.
the two masses. It is essential that all the solid Filtration and Ignition of Solids
removed from the weighing bottle be transferred ➢ Several techniques and experimental
without loss to the container. arrangements allow solids to be filtered and
ignited with minimal contamination and error.
Equipment and Manipulations Associated with ➢ Apparatus
Weighing ✓ Simple crucible
Desiccators and Desiccants ✓ Filtering crucible
➢ Oven drying is the most common way of removing ✓ Sintered-glass (also called fritted-glass)
moisture from solids. This approach is not crucibles
appropriate for substances that decompose or for ✓ Gooch crucible
those from which water is not removed at the ➢ Simple crucibles serve only as containers.
temperature of the oven. To minimize the uptake Porcelain, aluminum oxide, silica, and platinum
of moisture, dried materials are stored in crucibles maintain constant mass—within the
desiccators while they cool. limits of experimental error— and are used
Weighing Hygroscopic Solids principally to convert a precipitate into a suitable
➢ Hygroscopic substances rapidly absorb moisture weighing form. The solid is first collected on a
from the atmosphere and, therefore, require filter paper. Simple crucibles of nickel, iron, silver,
special handling. You need a weighing bottle for and gold are used as containers for the high-
each sample to be weighed. temperature fusion of samples that are not
➢ Place the approximate amount of sample needed soluble in aqueous reagents. Attack by both the
in the individual bottles and heat for an atmosphere and the contents may cause these
appropriate time. When heating is complete, crucibles to suffer mass changes.
quickly cap the bottles and cool in a desiccator. ➢ Filtering crucibles serve not only as containers
➢ Weigh one of the bottles after opening it but also as filters. A vacuum is used to hasten the
momentarily to relieve any vacuum. Quickly filtration. A tight seal between crucible and
empty the contents of the bottle into its receiving filtering flask is made with any of several types of
vessel, cap immediately, and weigh the bottle rubber adaptors.
again along with any solid that did not get ➢ Sintered-glass (also called fritted-glass)
transferred. crucibles are manufactured in fine, medium, and
➢ Repeat for each sample and determine the coarse porosities (marked f, m, and c). The upper
sample masses by difference. temperature limit for a sintered-glass crucible is
Weighing Liquids usually about 200°C. Filtering crucibles made
➢ The mass of a liquid is always obtained by entirely of quartz can tolerate substantially higher
difference. Liquids that are noncorrosive and temperatures without damage. The same is true
relatively nonvolatile can be transferred to
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 for crucibles with unglazed porcelain or aluminum 3. Allow a crucible that has been subjected
oxide frits. The latter are not as costly as quartz. to the full flame of a burner or to a muffle
furnace to cool momentarily (on a wire
➢ A Gooch crucible has a perforated bottom that gauze or ceramic plate) before
supports a fibrous mat. Asbestos was at one time transferring it to the desiccator.
the filtering medium of choice for a Gooch 4. Keep the tongs and forceps used to
crucible. handle heated objects scrupulously
Filter Paper clean. In particular, do not allow the tips
➢ Paper is an important filtering medium. Ashless to touch the benchtop.
paper is manufactured from cellulose fibers that
have been treated with hydrochloric and Measuring Volume
hydrofluoric acids to remove metallic impurities Units of Volume
and silica; ammonia is then used to neutralize the ➢ The unit of volume is the liter (L), defined as one
acids. cubic decimeter. The milliliter (mL) is one one-
thousandth of a liter (0.001 L) and is used when
the liter represents an inconveniently large
volume unit. The microliter (µL) is 10-6 L or 10-3
mL.
The Effect of Temperature on Volume
➢ Measurements The coefficient of expansion for
dilute aqueous solutions (approximately
0.025%/°C) is such that a 5°C change has a
measurable effect on the reliability of ordinary
volumetric measurements.

Heating Equipment PIPETS / PIPETTES

➢ The maximum attainable temperature ranges
from 140°C to 260°C, depending on make and
model.
➢ For many precipitates, 110°C is a satisfactory
drying temperature. The efficiency of a drying
oven is greatly increased by the forced circulation
of air.
➢ Microwave laboratory ovens are currently quite
popular, and where applicable, they greatly
shorten drying cycles.
➢ A heavy-duty electric furnace (muffle furnace) is
capable of maintaining controlled temperatures of
1100°C or higher. Long-handled tongs and heat-
resistant gloves are needed for protection when
transferring objects to or from such a furnace. Pipets
➢ Pipets permit the transfer of accurately known
Rules for Manipulating Heated Objects volumes from one container to another.
✓ Careful adherence to the following rules will Pipettes
minimize the possibility of accidental loss of a ➢ Another type of volumetric glassware used
precipitate: extensively in the laboratory.
1. Practice unfamiliar manipulations before ➢ Calibration are according to deliver or transfer a
putting them to use. specific volume from one vessel to another.
2. Never place a heated object on the
benchtop. Instead, place it on a wire Types of Pipettes
gauze or a heat-resistant ceramic plate. ➢ According to manner of calibration
11
 ➢ According to graduation Unopette
➢ Specialized pipettes ➢ A special disposable micropipette used in the
▪ Micropipettes hematology laboratory.
▪ Unopette ➢ It is self-filling pipette accompanied by
▪ Capillary pipette polyethylene reagent reservoir.
▪ Automatic pipettor ➢ A capillary pipette is fitted in a plastic holder and
According to calibration: fill automatically with blood by means of capillary
➢ To deliver (TD) – calibrated to deliver the amount action.
of fluid designated on the pipette; this volume will Capillary Pipette
flow out of the pipette by gravity. ➢ Inexpensive, disposable micropipette.
▪ Calibration is usually performed by ➢ It is filled up to the calibrated line by capillary
measuring the amount of water delivered action and measured liquid is delivered by
by the pipette. positive pressure as with a medicine dropper.
➢ To contain (TC) – calibrated by introducing exact Automated Pipettor
amount of volume or weight of mercury. ➢ Allows rapid, repetitive measurement and
▪ it contains exact amount however does delivery of predetermined volumes of reagents or
not deliver the exact volume. specimens 0.5-500 uL
➢ To blow out pipette – similar to TD except that the
volume is obtained when the last drop is being Pipetting Technique
blown out. An etched or frosted ring indicated this ➢ Check pipette before using – wet, chipped or
calibration. broken.
➢ Calibrated between the marks – exact volume is ➢ Hold properly between thumb and forefinger.
calibrated to fill the volume between 2 calibrates ➢ Wipe the pipette with a soft tissue or lint free cloth.
points on the pipette. ➢ Hold the pipette vertically.
According to graduations: ➢ Do not mouth pipette.
➢ Volumetric or transfer pipette – has a ➢ Read the meniscus
cylindrical bulb located midway the mouthpiece ✓ Bottom of the meniscus
and the tip. ✓ Upper meniscus
▪ Oswald – Folin pipette – similar to ✓ Eye level
volumetric pipette but has a larger bulb Cleaning
closer to delivery tip and it has an etched ➢ Draw detergent solution to a level 2 to 3 cm above
ring that indicates that it is a blow-out. the calibration mark of the pipet.
▪ Used to measure viscous substance ex. ➢ Drain this solution and then rinse the pipet with
Blood and serum. several portions of tap water. Inspect for film
➢ Measuring or graduated pipette – plain narrow breaks, and repeat this portion of the cleaning
tube draws out to a tip and graduated uniformly cycle if necessary.
along its length; calibrated to deliver fractional ➢ Finally, fill the pipet with distilled water to perhaps
quantity specifically reagents. one third of its capacity and carefully rotate it so
➢ Types that the entire interior surface is wetted. Repeat
▪ Mohr this rinsing step at least twice.
▪ Serologic Pipette Burets
Micropipettes ➢ Burets, like measuring pipets, make it possible to
➢ Calibrated to contain or to wash out the pipettes. deliver any volume up to the maximum capacity
➢ 1 lambda = 1uL = 0.001Ml of the device. The precision attainable with a
Example of Micropipettes buret is substantially greater than the precision
✓ Kirk transfer pipette – TC with a pipet.
✓ Self-filling transfer pipette – TC ➢ A buret consists of a calibrated tube to hold titrant
✓ Lang levy pipette – TD plus a valve arrangement by which the flow of
✓ Overflow pipette – TC titrant is controlled. This valve is the principal
source of difference among burets. The simplest
pinchcock valve consists of a close-fitting glass
12
 bead inside a short length of rubber tubing that Volumetric Flasks
connects the buret and its tip. Only when the ➢ Volumetric flasks are manufactured with
tubing is deformed does liquid flow past the bead. capacities ranging from 5 mL to 5 L and are
➢ A buret equipped with a glass stopcock for a valve usually calibrated to contain (TC) a specified
relies on a lubricant between the ground-glass volume when filled to a line etched on the neck.
surfaces of stopcock and barrel for a liquid-tight They are used for the preparation of standard
seal. Some solutions, notably bases, cause glass solutions and for the dilution of samples to a fixed
stopcocks to freeze when they are in contact with volume prior to taking aliquots with a pipet.
ground glass for long periods. Therefore, glass ➢ Some are also calibrated on a to-deliver (TD)
stopcocks must be thoroughly cleaned after each basis, and they are distinguished by two
use. Most burets made in the last several of reference lines on the neck. If delivery of the
decades have Teflon® valves, which are stated volume is desired, the flask is filled to the
unaffected by most common reagents and upper line.
require no lubricant. Using Volumetric Equipment
➢ A buret must be scrupulously clean before it is Cleaning
used, and its valve must be liquid-tight. ➢ A brief soaking in a warm detergent solution is
Cleaning usually sufficient to remove the grease and dirt
➢ Thoroughly clean the tube of the buret with responsible for water breaks. Prolonged soaking
detergent and a long brush. Rinse thoroughly with should be avoided because a rough area or ring
tap water and then with distilled water. Inspect for is likely to develop at a detergent/air interface.
water breaks. Repeat the treatment if necessary. This ring cannot be removed and causes a film
Lubricating a Glass Stopcock break that destroys the usefulness of the
➢ Carefully remove all old grease from a glass equipment.
stopcock and its barrel with a paper towel and dry ➢ After being cleaned, the apparatus must be
both parts completely. Lightly grease the thoroughly rinsed with tap water and then with
stopcock, taking care to avoid the area adjacent three or four portions of distilled water. It is
to the hole. Insert the stopcock into the barrel and seldom necessary to dry volumetric ware.
rotate it vigorously with slight inward pressure. A Avoiding Parallax
proper amount of lubricant has been used when ➢ The top surface of a liquid confined in a narrow
(1) the area of contact between stopcock and tube exhibits a marked curvature, or meniscus. It
barrel appears nearly transparent, (2) the seal is is common practice to use the bottom of the
liquid-tight, and (3) no grease has worked its way meniscus as the point of reference in calibrating
into the tip. and using volumetric equipment.
Notes: ➢ This minimum can be established more exactly
➢ Grease films that are unaffected by cleaning by holding an opaque card or piece of paper
solution may yield to such organic solvents as behind the graduations. In reading volumes, the
acetone or alcohols. Thorough washing with eye must be at the level of the liquid surface to
detergent should follow such treatment. Silicone avoid an error due to parallax. Parallax is a
lubricants are not recommended because condition that causes the volume to appear
contamination by such preparations is difficult—if smaller than its actual value if the meniscus is
not impossible—to remove. viewed from above and larger if the meniscus is
➢ So long as the flow of liquid is not impeded, viewed from below.
fouling of a buret tip with stopcock grease is not a
serious matter. Removal is best accomplished Directions for Using a Volumetric Flask
with organic solvents. A stoppage during a ✓ Before being put into use, volumetric flasks
titration can be freed by gentle warming of the tip should be washed with detergent and thoroughly
with a lighted match. rinsed. Only rarely do they need to be dried.
➢ Before a buret is returned to service after ✓ If required, however, drying is best accomplished
reassembly, it is advisable to test for leakage. by clamping the flask in an inverted position.
Simply fill the buret with water and establish that ✓ Insertion of a glass tube connected to a vacuum
the volume reading does not change with time. line hastens the process.
13
Direct Weighing into a Volumetric Flask ✓ At 20°C: V 5 24.976 g 3 1.0037 mL/g 5 25.07 mL
✓ The direct preparation of a standard solution
requires the introduction of a known mass of Calibrating a Volumetric Pipet
solute to a volumetric flask.
✓ Use of a powder funnel minimizes the possibility
of losing solid during the transfer.
✓ Rinse the funnel thoroughly, and collect the
washings in the flask.
✓ The foregoing procedure may be inappropriate if
heating is needed to dissolve the solute. Instead,
weigh the solid into a beaker or flask, add solvent,
heat to dissolve the solute, and allow the solution
to cool to room temperature.
✓ Transfer this solution quantitatively to the
volumetric flask, as described in the next section.

CALIBRATING VOLUMETRIC GLASSWARE

➢ Volumetric glassware is calibrated by measuring
the mass of a liquid (usually Distilled or deionized ✓ Determine the empty mass of the stoppered
water) of known density and temperature that is receiver to the nearest milligram.
contained in (or delivered by) the volumetric ✓ Transfer a portion of temperature-equilibrated
ware. water to the receiver with the pipet, weigh the
➢ In carrying out a calibration, a buoyancy receiver and its contents (again, to the nearest
correction must be made since the density of milligram), and calculate the mass of water
water is quite different from that of the masses. delivered from the difference in these masses.
➢ The calculations associated with calibration are a ✓ With the aid of Table 2-3, calculate the volume
bit time consuming if done manually, but they can delivered.
be automated in a spreadsheet so that they ✓ Repeat the calibration several times, and
require little more time than entering the data. calculate the mean volume delivered and its
➢ Next, the volume of the apparatus at the standard deviation.
temperature of calibration (T) is obtained by Calibrating a Buret
dividing the density of the liquid at that ✓ Fill the buret with temperature-equilibrated water
temperature into the corrected mass. and make sure that no air bubbles are trapped in
➢ Finally, this volume is corrected to the standard the tip.
temperature of 20°C. Corrections for buoyancy ✓ Allow about 1 minute for drainage, and then lower
with respect to stainless steel or brass mass (the the liquid level to bring the bottom of the meniscus
density difference between the two is small to the 0.00-mL mark.
enough to be neglected) and for the volume ✓ Touch the tip to the wall of a beaker to remove
change of water and of glass containers have any adhering drop. Wait 10 minutes and recheck
been incorporated into these data. Multiplication the volume.
by the appropriate factor from Table 2-3 converts ✓ If the stopcock is tight, there should be no
the mass of water at temperature T to (1) the perceptible change. During this interval, weigh (to
corresponding volume at that temperature or (2) the nearest milligram) a 125-mL conical flask
the volume at 20°C. fitted with a rubber stopper.
• A 25-mL pipet delivers 24.976 g of water weighed ✓ Once tightness of the stopcock has been
against stainless steel mass at 25°C. Use the data established, slowly transfer (at about 10 mL/min)
in Table 2-3 to calculate the volume delivered by this approximately 10 mL of water to the flask.
pipet at 25°C and the volume if the weighing were ✓ Touch the tip to the wall of the flask.
carried out at 20°C. ✓ Wait 1 minute, record the volume that was
Solution apparently delivered, and refill the buret. Weigh
✓ At 25°C: V 5 24.976 g 3 1.0040 mL/g 5 25.08 mL the flask and its contents to the nearest milligram.

14
 ✓ The difference between this mass and the initial over an experiment. Regular prescription glasses
value is the mass of water delivered. Use Table are not adequate substitutes for eye protection
2-3 to convert this mass to the true volume. approved by the Office of Safety and Health
Subtract the apparent volume from the true Administration (OSHA). Contact lenses should
volume. never be used in the laboratory because
✓ This difference is the correction that should be laboratory fumes may react with them and have a
applied to the apparent volume to give the true harmful effect on the eyes.
volume. Repeat the calibration until agreement • There is necessarily a degree of risk associated with
within 60.02 mL is achieved. any work in a chemical laboratory. Accidents can
✓ Starting again from the zero mark, repeat the and do happen. Strict adherence to the following
calibration, this time delivering about 20 mL to the rules will go far toward preventing (or minimizing the
receiver. Test the buret at 10-mL intervals over its effect of) accidents:
entire volume. ➢ 3. Most of the chemicals in a laboratory are toxic,
✓ Prepare a plot of the correction to be applied as a some are very toxic, and some— such as
function of volume delivered. The correction concentrated solutions of acids and bases—are
associated with any interval can be determined highly corrosive. Avoid contact between these
from this plot. liquids and the skin. In the event of such contact,
Calibrating a Volumetric Flask immediately flood the affected area with large
✓ Weigh the clean, dry flask to the nearest quantities of water. If a corrosive solution is
milligram. Then fill to the mark with equilibrated spilled on clothing, remove the garment
water and reweigh. With the aid of Table 2-3, immediately. Time is of the essence, so do not be
calculate the volume contained. concerned about modesty.
Calibrating a Volumetric Flask Relative to a Pipet ➢ 4. NEVER perform an unauthorized experiment.
✓ The calibration of a volumetric flask relative to a Unauthorized experiments are grounds for
pipet provides an excellent method for disqualification at many institutions.
partitioning a sample into aliquots. ➢ 5. Never work alone in the laboratory. Always be
✓ Carefully transfer ten 50-mL aliquots from the certain that someone is within earshot.
pipet to a dry 500-mL volumetric flask. Mark the ➢ 6. Never bring food or beverages into the
location of the meniscus with a gummed label. laboratory. NEVER drink from laboratory
✓ Cover with a label varnish to ensure permanence. glassware. NEVER smoke in the laboratory
• Dilution to the label mark permits the same pipet to ➢ 7. Always use a bulb or other device to draw
deliver precisely a one-tenth aliquot of the solution liquids into a pipet. NEVER pipet by mouth.
in the flask. Note that recalibration is necessary if ➢ 8. Wear adequate foot covering (no sandals).
another pipet is used. Confine long hair with a net. A laboratory coat or
SAFETY IN THE LABORATORY apron will provide some protection and may be
➢ 1. Before you begin work in any laboratory, learn required.
the location of the nearest eye fountain, fire ➢ 9. Be extremely tentative in touching objects that
blanket, shower, and fire extinguisher. Learn the have been heated because hot glass looks
proper use of each, and do not hesitate to use this exactly like cold glass.
equipment if the need arises. ➢ 10. Always fire-polish the ends of freshly cut-
➢ 2. Wear eye protection at all times. The potential glass tubing. NEVER attempt to force glass
for serious and perhaps permanent eye injury tubing through the hole of a stopper. Instead,
makes it mandatory that adequate eye protection make sure that both tubing and hole are wet with
be worn at all times by students, instructors, and soapy water. Protect your hands with several
visitors. Eye protection should be donned before layers of towel while inserting glass into a
entering the laboratory and should be used stopper.
continuously until it is time to leave. Serious eye ➢ 11. Use fume hoods whenever toxic or noxious
injuries have occurred to people performing such gases are likely to be evolved. Be cautious in
innocuous tasks as computing or writing in a testing for odors. Use your hand to waft vapors
laboratory notebook. Incidents such as these above containers toward your nose.
usually result from someone else’s loss of control
15
 ➢ 12. Notify your instructor immediately in the event ➢ The ångstrom unit, Å, is a non-SI unit of length
of an injury. that is widely used to express the wavelength of
➢ 13. Dispose of solutions and chemicals as very short radiation such as X-rays (1 Å = 0.1 nm
instructed. It is illegal to flush solutions containing = 10-10 m). Typical X-radiation lies in the range of
heavy metal ions or organic liquids down the drain 0.1 to 10 Å.
in most localities. Alternative arrangements are The Mole
required for the disposal of such liquids. ➢ The mole (mol) is the SI unit for the amount of a
chemical species.

Calculations in Analytical Chemistry ➢ It is always associated with a chemical formula

and represents Avogadro’s number (6.022 × 1023)
of particles represented by that formula.
SI Units ➢ The molar mass (M) of a substance is the mass
➢ Scientists throughout the world are adopting a in grams of one mole of the substance.
standardized system of units known as the ➢ Molar masses are calculated by summing the
International System of Units (SI). atomic masses of all elements appearing in a
➢ This system is based on the seven fundamental chemical formula.
base units. Numerous other useful units, such as MILLIMOLE
volts, hertz, coulombs, and joules, are derived ➢ Sometimes it is more convenient to make
from these base units.

calculation with millimoles (mmol) rather than

moles, where the mmol is 1/1000 of a mole. The
mass in grams of a millimole of a substance, the
millimolar mass, is likewise 1/1000 of the molar
mass.
➢ 1 mmol = 10-3 mol
➢ 1 mol = 1000 mmol
➢ Example:
➢ Determine the number of moles and millimoles of
benzoic acid (C7H6O2) in 2.00 grams of the pure
acid.
Calculating the amount of a Substance in Mole or
Millimoles
➢ Example:
➢ Determine the number of moles and millimoles of
benzoic acid (C7H6O2) in 2.00 grams of the pure
acid.

➢ SI is the acronym for the French “Système

International d’Unités.”
16
 Molarity
• Example: Calculate the molar concentration of
ethanol in an aqueous solution that contains 2.30
g of C2H5OH in 3.50 L of solution.
Solution:
1. Convert the mass of ethanol to corresponding number
of moles.

2B-1 Expressing Solution Concentrations

➢ Analytical Molarity – The analytical molarity of a
Distinguishing between Mass and Weight solution gives the total number of moles of a
➢ Mass is an invariant measure of the amount of solute in 1 L of the solution, or alternatively, the
matter in an object. total number of millimoles in 1 mL.
➢ Weight is the force of attraction between an object ➢ Equilibrium Molarity – The equilibrium molarity,
and its surroundings, principally Earth. or species molarity, expresses the molar
Gravitational attraction varies with geographic concentration of a particular species in a solution
location, the weight of the object depends on at equilibrium.
where you weight it. MOLALITY
➢ Example: weight of crucible is less in Denver than ➢ Molality (m) is the number of moles of solute per
in Atlantic City because of the altitude. While the kilogram of solvent
mass of the crucible is constant regardless where
you measure it.
➢ Weight and mass are related by the familiar
expression:
➢ W=mg ➢ Remember that the denominator is the mass of
➢ where: W= weight of the object solvent, not of the entire solution
➢ m= mass of the object
➢ g= acceleration due to gravity
Solutions and Their Concentrations
➢ Expressing Solution Concentrations
o Molar Concentration
▪ The molar concentration (cx) of a
solution of a chemical species X
is the number of moles of the
solute species that is contained
in one liter of the solution (not
one liter of the solvent).
▪ Molar concentration, or molarity
M, has the dimensions of mol L-
1.

17
NORMALITY OH- ions per molecule, so the valence is 2. Gram
➢ Normality (N) is a term of concentration based on equivalent weight is:
the ability of some substances to release
hydrogen (H+) or hydroxide (OH- ) ions in solution
➢ In biological systems, the number of H+ or OH-
ions that are released per molecule in solution is
equivalent to valence, which is the bonding ability
of a molecule
➢ The simplest definition of normality is based on
molarity and valence

➢ Example, Ca(OH)2 dissociates in solution to

release two OH- ions per molecule, so the valence
is 2.
➢ Therefore the normality of a 1-M solution of
Ca(OH)2 is 2 N

2B-1 Expressing Solution Concentrations

Percent Concentration
➢ Chemists frequently express concentrations in
terms of percent (parts per hundred).
➢ For substances with a valence of 1, normality ➢ Three common methods are
equals molarity
➢ Normality is also defined as equivalents per liter
and can be determined given the weight of solute
and the volume of solution using the following
steps:
➢ 1. Determine valence to calculate the gram
equivalent weight of the solute, which is the
weight of 1 mole of the substance (also known as
formula weight) divided by its valence
➢ 2. Calculate the number of equivalents in 2B-2 Density and Specific Gravity of Solutions
solution, which is the weight of the solute in the ➢ Density is the mass of a substance per unit
sample divided by the gram equivalent weight volume. In SI units, density is expressed in units
➢ 3. Solve for normality with the equation, of kg / L or alternatively g/mL.
➢ N= Eq/L ➢ Specific gravity is the ratio of the mass of a
substance to the mass of an equal volume of
water.

➢ EXAMPLE: What is the normality of a 1-L solution

containing 46.3 g Ca(OH)2? (MW of Ca(OH)2 =
74.1 g/mol)
➢ 1. Determine the valence to calculate gram
equivalent weight. Ca(OH)2 dissociates into two

18