HANDOUT FOR BEGINNERS

IN THE LABORATORY

Prepared By:

Mary Njeakor (MIPAN) [Link] Analytical

Chemistry
Laboratory Consultant

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INTRODUCTION:
For graduates who have had several technical knowledge of the
chemistry laboratory, its relevant role in the quality control unit of any
company especially pharmaceutical, there is crucial need to acquire
equivalent practical experience where knowledge are brought into reality
and applied to yield notable results.
Basic operations in a chemistry lab include, measuring mass & volume,
heating substances, mixing solutions, preparing reagents and separating
mixtures.

The personnels that work in the lab must be:

(a) Observant to changes, colour, temperature, gas, precipitate and
the environmental conditions of the lab.
(b) Apt in recording data generated from analysis and honestly
draw conclusions.
(c) Identify and use various glass wares such as:
i. Breakers/conical flasks for mixing and reactions
ii. Graduated cylinders for accurate volume.
iii. Pipettes and burettes for small volumetric amounts.

EXAMPLES OF MEASURING EQUIPMENTS:

i. Digital/analytical weighing balance for mass.
ii. Thermometer/hydrometer for temperature and humidity
respectively
iii. pH strips or meter for checking acidity or alkalinity.

EXAMPLES OF HEATING EQUIPMENT:

- Bunsen burner or hot plate

- Test tube/boiling tube holders
- Distiller, water bath
- Oven and furnace

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EXAMPLES OF SEPERATION TOOLS:
- Sohxlet extractor
- Separating funnel
- Filter paper
- Evaporating dish
-

SAFETY CONCIOUSNESS (NON NEGOTIABLE)

This is 90% of being a good beginner in the lab

a. You must wear PPE (Personal Protective Equipment) such as lab

coat, goggles, closed shoes, gloves when handling chemicals.
b. In case of accidents, you must identify an eyewash station, a
shower, fire extinguishers, waste bins.
c. Always read the labels: never taste or smell chemicals rather
directly waft vapors with your hands.
d. Clean as you go: for spill, broken glass and clutter. All these
causes most accidents in the lab.
e. Always ask before you do: when not sure about a step, check with
the instructor or senior analyst.

A TYPICAL FLOW CHAT OF A LABORATORY/SESSION:

(a) Pre lab:
First read the procedure, take note of hazards and what you are
trying to find out.
(b) Set up:
Get the equipments need for a particular analysis ready, label
everything before you start work. check measurements twice.
(c) Experiment:
Adhere to each step in its order. change only one variable at a
time.
(d) Observe and record:
Write down what observed and not what you expect, record
time, colour, temperature , precipitates etc.
(e) Clean up and make report:

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 Dispose chemicals properly, wash glass wares, clean the
equipments and write down your lab report with the raw rata,
calculations (if necessary) then conclusion.

MINDSET OF AN ANALYST:

As a beginner in the lab, you need to have a:

(1) Accuracy over speed: a wrong measurement ruins the whole
reaction.
(2) Do not multitask with [Link] focused to avoid accidents
(3) It is okay to be wrong: every failed experiments teaches you
the most important lessons.

Take note of what went wrong before you experiment

(4) Documentation matters: What you do not document means you
did not do it.
Your lab note book is your memory. Insert date, title, steps and
calculations in pen.

STARTER’S TIPS
- Start with simple reactions preparation of reagents,
standardization of volumetric solutions, acid – base
titrations,precipitation, crystallization etc.
- Learn to read a meniscus: read the button of the curve at eye
level.
- Do not return unused chemicals to the original bottle, store it in a
different bottle and label.
- Always convert units before you start any analysis not during the
experiment.

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REAGENTS:

Preparation & standardization:

Reagents: Defination

Reagents are substances used to cause or effect a chemical reaction

basically, whatever you add to make something happen in laboratory, in
manufacturing or in a test kit.

Lab/Analytical Reagents: these are acids, bases, solvents, indicators etc.

They are used for experiments, titrations, quality control. They are
of different grades HPLC grades, AR ie analytical reagents, etc.
- Reagents in chemical labs are prepared in molarity or in
percentages.

WHAT TO NOTE IN PREPRATION & STANDARDIZATION OF

REAGENTS:

These are key words, accuracy, treaceability and consistency.

(a) Use the right grade of reagent. Analytical reagents AR grade

are use mostly for quantitative analysis, while technical grade
are for cleaning.
(b) Check the label and MSDS (Material Safety Data Sheet of that
reagent) it contains the formula, percentage purity, molecular
weight, hazard, precautions and, storage conditions of that
reagent.
(c) In weighing and measuring,
- Use calibrated weighing balances and volumetric glasswares,
- Ensure to rinse glasswares at lease twice with distilled or deionized
water.
- For solids: weigh by difference, account for hygroscopicity, dry
primary standards in an oven first if required before use.
- For liquids: measure at eye level, at 20oc use pipettes/burettes not
graduated cylinders for precision.
-

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 (d) Solvent and water quality:
- Use distilled or deronized water.
- Carbon-dioxide free water for preparation of bases like NaOH.

(e) Order of addition:

- Add acid to water
NB: Never add water to acid also dissolve a salt completely before
making up to the desired volume.

(f) Mixing:

Invert volumetric flask 10-15 times to mix very well.

(g) Labelling:

Always place the name, concentration, molarity/normality of any

solvent prepared, who prepared it , expiry date and hazard symbol on
the container of the prepared solution.
(h) Storage:
- Use the appropriate container, temperature, light conditions to
store reagents.
NB: place volatile reagents in fridge, bases in plastic and acids in
glass.

Always check for degradation of the reagents over a period of time.

STANDADIZATION OF REAGENTS:
Standardization means to find out the exact concentration of a solution
after preparation, most reagents acts in different ways hence the need
to standardize them.

Example: sodium hydroxides absorbs carbon-dioxide(CO2) over a period

of time, hydrochloric acids are volatile, potassium permanganates
decomposes easily with time.

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TIPS:

(a) Pick a primary standard.

These primary standards should be high in purity, stable, non
hydroscopic, have large molar, mass, easy to weigh and reacts
stiochrometrically.

EXAMPLE: use
- Potassium hydrogen phthalate to standardize sodium hydroxide or
a standardized hydrochloric acid solution,
- Use a pure sodium bicarbonate (Anhydrous) salt to standardized
hydrochloric acid and use zinc metal to standardize EDTA i.e
disodiumedetate solution.

PROCEDURE:
- Weigh accurately within 3-4 decimal places.
- Use indicator with sharp end point and matching pH range.
- Do at least 3 replicate titrations. RSD should be 0.2%.
- Run blank to correct for indicator/reagent error.

CALCULATIONS:
- Use M,V, = M2 V2 for titrations
- Record temperature

NB: Molarity changes with temperature.

- Calculate the exact concentration, then the standardization factor
(also known as concentration factor) of = actual molarity
(observed) stated theoretical molarity.

Use this factor in all your work:

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- Documentation: date prepared, date standardized, calculated
concentration, who standardized and next due date for re-
standardization.
- Fix schedules for re-standadization either weekly, bi-weekly or
monthly.

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ANALYSIS OF WATER

INTRODUCTION:
Water is an essential requirement for human and industrial
development. It is used directly or indirectly in many industrial processes
in large quantities. It is a neutral compound containing hydrogen and
oxygen with a variety of dissolved & suspended solids.

MAJOR SOURCES OF WATER:

- Springs
- Ponds
- Streams
- Rivers, oceans, rain and underground water.

NB: the variety of water sources bring in a variety of impurities in the

water and the presence of these impurities reduces the use to which the
water may be deployed.

ANALYSIS OF WATER:

Basic criteria to check during analysis of water (grouped into)

(a) PHYSICAL PARAMETERS
- Colour
- Turbidity
- Taste
- Odour
- pH = (pH, - log10 (H+)
- Conductivity
- Appearance
- Temperature

(b) CHEMICAL
- Dissolved minerals
a. Calcium and magnesium which causes the hardness of water.
b. Sodium
c. Potassium

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 d. Iron
e. Chloride/fluoride
f. Alkalinity (carbonates/bicarbonates)
g. Total dissolve solid (TDS)
h. Nitrates
i. Heavy metals (mercury, cadmium, lead, arsenic
j. Carbon-dioxide

(c) BIOLOGICAL/MICROBIOLOGICAL TESTS

a. E – coli
b. Coliform bacteria
c. Other pathogens

COMMON TESTS USED IN ANALYSIS OF WATER:

(i) Titrimetric method: this is used to check for the hardness,
alkalinity, chlorine.
(j) TDS/Conductivity Meter: to check for the total dissolved solids
and conductivity of water.
(k) Spectrophotometric method: to check for the nitrates,
phosphates and heavy metals.
(l) Membrane filtration and incubation method: to check for the
bacteria count in water.

ADVANTAGES OF ANALYSIS OF WATER .

- Water plays a crucial role in the maintenance of human, animal&
plant life:
- Hence there is need to ensure that water for drinking,
pharmaceutical aid, agricultural industry and other purposes,
remain safe and clean i.e free from unwanted ions and bacteria
For example: water used for pharmaceutical aid must be ions and
bacteria free else they constitute in the degradation of drugs also,
there is need to ensure that water meets the standard quality set
by regulatory bodies e.g WHO, SON, NAFDAC etc

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RAW MATERIAL / FINISHED PRODUCT ANALYSIS:

Generally in testing for raw materials, food, drugs, chemicals, finished

products etc, ensure you get a representativeie: a sizeable quality that
will represent the whole sample you are to analyze, your analysis must
be accurate and repeatable.

WHAT TO KNOW:
(a) Sampling is important (avoid cross contamination): If
you do not sample well, you will have a bad result. Use the
suitable and clean sampling tools (gloves, scoops, sealed
containers etc.)
(b) Sample preparation: Batch date received, samples by who
label properly.
- Ensure to grind, mix and dissolve samples well to have a
homogenius mixture before analysis begins.
NB: take note of samples that are heat or moisture sensitive, also
ensure you store sample well under a good condition to avoid
degradation before you carry out the analysis.
(c) Identify the sample properly and use validated methods to do
your analysis and use Sop well.
(d) Do blank, use a certified reference standard and repeat your
analysis.
(e) Use equipments that are well calibrated and well maintained
e.g: weighing balance, pH meters, HPLC, GC and have a log
book for them.
(f) Practice good lab (GLP): document all observations and results
proper as it avoid manipulation of results to ensure data
integrity.
(g) Out-of-spec Protocol: for analysis that fail, do a retest,
investigate and find out why.

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OPERATION, CALIBRATION AND OF EQUIPMENTS USED IN
LABORATORY.

(a) Always read the SOP (Standard Operating Procedure) or the

work instruction of any equipment first before using it. It tells
you the warm up time, limits and what not to do on the
equipment.
(b) Check for drainage, leaks, loose cables, dirty optics, etc before
using the equipment.
(c) Ensure equipment’s are kept in an environment free of dust,
vibrations, direct sunlight and humidity, swings, keeps benches
clean, stable.
(d) Always give time for warm up of equipment after you switch it
on. Some equipment’s like weighing balance, spectrophoto
meter, HPLC use 15-30 minutes to stabilize before functioning.
(e) Use the appropriate accessories such as reagents, culverts,
columns and gases to avoid variation in results and ensure
accuracy and precision.
(f) Have a detailed log book, for each equipment it should
carrydate, user name, sample ID, calibrations, repair, setting
and any error or change that occurs in that equipment.

CALIBRATION:
It is the adjustment or fine tuning of an instrument with a known
standardor reference to ensure a reliable and accurate result or
measurement.

This should be done frequently.

Some equipment’s are calibrated:

(a) Daily before use such as weighing balances, pH meters.
(b) Weekly or monthly: such as HPLC (high performance liquid
chromatogram), centrifuge, fume chambers etc.

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- Always use traceable standards NIST – traceable weights for
balance certified buffers for pH meters, Class A glass wave for
pipettes.
- Calibrate equipmentsunder the same working conditions: use same
temperature, humidity and location for the laboratory and
equipment to avoid a drift (variation)
- Do a multipoint calibration for pH meter, spectrophotomer, use at
least 2-3 standards that covers your working rangee.g buffer 4.7,
7.0, and 10.0 solution to calibrate your pH meter. It covers the
acidic, neutral and basic conditions of your samples.
- Record properly: your date, standard lot number or ID number,
who calibrated, result before and after calibrations and next due
date for calibration.
- Place a calibration sticker on your equipment, it automatically
indicate the calibration status of your equipment as at when used.
- Use only trained personnelto operate your equipment. Train all
personnel same way to avoid inconsistent handling of equipment
which is number one cause for calibration failure.

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 GOOD LAB PRACTICES

Procedure
1. While performing any analysis always follow respective Standard
operating Procedure.
2. Use only pure (AR / GR grade) material for standardization.
3. Use calibrated volumetric glassware only, when accurate dilutions are
required.
4. Ensure that the temperature during dilution, at the time of volumetric
measurement and standardization is maintained I. e. at about 25°C.
5. Rinse the burette or other vessel used to collect the volumetric
solution with the solution to be standardized.
6. While dilution always add concentrated acid slowly to the water, and
never add water to acid.
7. Handle the chemicals and solvents in a way to avoid spillage on the
working bench and the floor.
8. In case of any spillage Immediately clean the affected area,
according the type of spillage.
9. Wear a full-seal goggles and safety mask while handling
concentrated acids and bases.
10. Never try and adjust the pH of a solution in a narrow-neck container.
Always use a beaker.
11. Never stick a spatula in to a container of a solid reagent. Always roll
the container to deliver the reagent in a controlled manner to the
receiving beaker.
12. Never return unused reagent to the reagent container.
Contamination of the whole reagent supply is more costly than the
small amount of reagent discarded.
13. Standardize the volumetric solution with that method only which is
used for end point determination.
14. Store the light sensitive solution in the light resistant (amber colored)
containers only.
15. Label all the reagents for Name, strength, prepared by & on and
shelf life.

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16. While performing titration in case of colorless solutions in the
burette, consider the lower meniscus as the reading. In case of
colored solutions consider upper meniscus.
17. Always fill burette taking it away from the stand in to the hand and
using a glass funnel. Adjust the reading to zero before starting
titration.
18. Use approximately 0.1ml of indicator, during titration or
standardization, unless otherwise specified.
19. Never use broken glass-wares and in case of iodometric titration use
iodine flask.
20. Store toxic and flammable chemicals in the separate area under lock
and key.
21. Store all the reference standards and stock of working standards in
their Specified area.
22. Working standards In use are to be stored in the desiccator provided.
And the bottles are sealed immediately after their use.
23. Never leave any chemical reaction taking place in the gaseous form
i.e. boiling, distillation, refluxing unattended to.
24. While boiling anything, keep the container opened, if not refluxing.
25. Check the LPG cylinder and pressure tubing for any damage or
leakage; change the tubing if damage or leakage is found.
26. Always close LPG valve first and then put off the compressor while
working with them, there is a chance of explosion when compressor
is closed first.
27. Always keep the burner OFF while mopping the LAF bench.
28. After completion of any microbiological work, put on the UV light for
sterilization.
29. Never look directly in to the UV light.
30. For analysis always use calibrated instruments
31. If instrument / equipment found damaged, do not use it and consult
Q. C. Manager / executive to get it repaired or changed.
32. While cleaning any electrode of either pH meter or conductivity
meter, clean them with water for injection and wipe off with the help
of cotton wool gently.
33. Never rub the electrode and keep all the electrodes dipped constantly
on the distilled water
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34. Conduct a regular training and re-training of the personnels in the
laboratory with equivalent evaluations to determine their competency
in the laboratory.

5.2 ABBREVIATIONS

 SOP - Standard Operating Procedure

 QC - Quality Control LAF - Laminar Air Flow
 LPG - Liquid Petroleum Gas
 UV - Ultra Violet
 AR - Analytical Grade Reagent
 GR - General Reagent
Thank you.

Prepared By:

Mary Njeakor (MIPAN) [Link] Analytical Chemistry

Laboratory Consultant

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