ENV460 Course File

FAKULTI SAINS KESIHATAN
UNIVERSITI TEKNOLOGI MARA

42300 BANDAR PUNCAK ALAM
SELANGOR DARUL EHSAN

FAIL KURSUS
FSK-ENV 460

ANALYTICAL TECHNIQUE AND
INSTRUMENTATION

Disediakan oleh: Diluluskan oleh:
Tandatangan : Tandatangan :

NAMA : AHMAD RAZALI BIN ISHAK NAMA : DR ABDUL MUJID ABDULLAH
JAWATAN : PENSYARAH KANAN
TARIKH : 30 SEPTEMBER 2020 JAWATAN : KOORDINATOR PROGRAM

TARIKH : 30 SEPTEMBER 2020

CENTRE OF ENVIRONMENTAL HEALTH
AND SAFETY

FACULTY OF HEALTH SCIENCES

ENV 460
ANALYTICAL TECHNIQUE AND INSTRUMENTATION

SYLLABUS

UNIVERSITI TEKNOLOGI MARA
COURSE INFORMATION
Confidential

Course Code : ENV460

Course Name (English) : ANALYTICAL TECHNIQUE AND INSTRUMENTATION APPROVED

Course Name (Malay) : TEKNIK ANALITIKAL DAN INSTRUMENTASI

Course Level : 6 - Bachelors Degree

SLT : 80 Hours

Equivalent to 2 Credits

Face to Face: 32 Hours

Non Face to Face: 38 Hours

Student Preparation Time: 10 Hours

Pre-Requisite Courses : No course recommendations

Co-requisite Courses : No co-requisite Courses listed

Equivalent Courses : No equivalent Courses listed

Teaching Period Duration : 17 Weeks

Resource Person : SHANTAKUMARI A/P RAJAN

National Education Code : Health and Welfare

Delivery Period Duration : 1 Semester

Faculty Name : FACULTY OF HEALTH SCIENCES Start Year : 2020
© Copyright Universiti Teknologi MARA Review Year : 2023

Course Learning Outcomes

At the end of the course, students should be able to:

1. Explain the principles and functions of analytical instruments used for environmental sample analysis. ( C2 )
2. Determine the appropriate techniques and requirements for environmental sample collection, extraction, preservation,

storage and handling. ( C5 )
3. Demonstrate autonomous learning through environmental sample preparation and analysis using standard analytical

methods and techniques. ( P5 )

Course Description

The course gives a description of instrumental chemical analytical techniques and sampling techniques for different types
of samples (including water, sediments, soil and air); the importance of appropriate sample collection, preservation,
storage techniques and handling requirements. On completion of this course, students will be able to explain the principles
of analytical techniques such as Atomic Absorption Spectrometry (AAS), spectrophotometry (UV/VIS), Gas
Chromatography (GC), High Performance Liquid Chromatography (HPLC), for the analysis of organic and inorganic
pollutants in environmental and biological samples. Laboratory activities include analysis of various important components
in biological samples, soil, air and water, and are especially dedicated to natural environmental chemistry and
environmental monitoring.

Syllabus Content

1. Basic analytical and sampling skills

laboratory hazards safety, standard precautions
Fundamentals of environmental sampling
Practical aspects of environmental sampling
Sample preservation, storage techniques, and handling requirements

2. Sample Preparation

Dry ashing
Acid digestion
Direct analysis of metals
Soxhlet extraction
Shake flask extraction
Ultrasonic extraction
Microwave assisted extraction (MAE)
Liquid Liquid extraction (LLE)
Solid phase extraction (SPE)

3. Principles of spectroscopy

Electromagnetic radiation
Atomic and molecular energy levels
Absorption and emission of radiation, spectra
Beer's law relationship between absorp

4. Principles of Chromatography

Principles of chromatography: column, separation modes; chromatogram
Gas chromatography (GC)
Liquid Chromatography (LC)

5. Validation of Analytical method

Principles of method validation
Validation tools
Method performance characteristics

Teaching Methodologies

Faculty Name : FACULTY OF HEALTH SCIENCES Start Year : 2020
© Copyright Universiti Teknologi MARA Review Year : 2023

Blended Learning
Lab Work
Lectures
Practical Classes

Proposed % Marks Approved by KPP

Question Difficulty Levels Range
C1 - C2 20 - 30
C3 - C4 40 - 60
C5 - C6 20 - 30

Assessment

Continuous 100.00%
Assessment: Assignment - 60% out of 0 on Week 14. Passing Mark(s): 0
n/a
CLO: 3

Final Test - 20% out of 0 on Week 14. Passing Mark(s): 0
Online Test 3
CLO: 2

Test - 10% out of 0 on Week 8. Passing Mark(s): 0
Online test 1
CLO: 1

Test - 10% out of 0 on Week 12. Passing Mark(s): 0
Online test 2
CLO: 2

Transferable Skills
data analysis, practical skills, teamwork, written communication
Special Regulation
This Module has no special regulation
Reassessment Requirement
Repeat the Course
Reassessment Description
The assessment of this Course is inextricably linked to the delivery. The student must reattend the Course in its entirety in
order to be reassessed.
Recommended Text

1. Maria Csuros,Csaba Csuros, Environmental Sampling and Analysis for Metals, CRC Press, 2016, ISBN: 1420032348
2. Bryan F.J. Manly,Jorge A. Navarro Alberto, Introduction to Ecological Sampling, CRC Press, 2014, ISBN: 1466555149
3. Cazes J. and Ewing G. W, Analytical Instrumentation Handbook, 3rd, Marcel Dekker Inc., 2005, ISBN: n/a

Faculty Name : FACULTY OF HEALTH SCIENCES Start Year : 2020
© Copyright Universiti Teknologi MARA Review Year : 2023

References

1. Koren H., et al., Handbook of Environmental Health. Vol. 2: Pollutant Interactions in Air, Water and Soil, 4th Ed, CRC
Press, 2002, ISBN: n/a

2. Khandpur R. S, Handbook of analytical instruments, 1st, McGraw-Hill., 2007, ISBN: n/a

Other References

1. Magnusson and U. Örnemark, Eurachem Guide: The Fitness for Purpose of Analytical Methods – A Laboratory Guide
to Method Validation and Related Topics, n/a, 2014, URL: www.eurachem.org

Course Extra Information
This Module has no extra information

Faculty Name : FACULTY OF HEALTH SCIENCES Start Year : 2020
© Copyright Universiti Teknologi MARA Review Year : 2023

UNIVERSITY TE

COURSE CODE COURSE LEARNING OUTCOMES - MOE LEARNING OUTCOMES
COURSE NAME (IN ENGLISH) ENV460
COURSE NAME (IN MALAY) ANALYTICAL TECHNIQUE AND INSTRUMENTATION
TEKNIK ANALITIKAL DAN INSTRUMENTASI

  Achievement of MOHE Learning Ou

Course Learning Outcomes (CLO) LO1 LO2 LO3 LO4 LO5 LO6

CLO1 Explain the principles and /
functions of analytical
instruments used for
environmental sample analysis.
( C2 )

CLO2 Determine the appropriate /
techniques and requirements for
environmental sample collection,
extraction, preservation, storage
and handling.
( C5 )

CLO3 Demonstrate autonomous /
learning through environmental
sample preparation and analysis
using standard analytical
methods and techniques.
( P5 )

Ministry of Education Learning Outcomes

i. Knowledge
ii. Practical Skills
iii. Thinking and Scientific Skills
iv. Communication Skills
v. Social skills, Teamwork and Responsibilities
vi. Values, Ethics, Moral and Professionalism
vii. Information Management and Life Long Learning
viii. Management and Entrepreneurship
ix. Leadership Skills

Faculty Name : FACULTY OF HEALTH SCIENCES
© Copyright Universiti Teknologi MARA

EKNOLOGI MARA

- SOFT SKILLS LEARNING OUTCOMES RELATIONSHIP MATRIX

FACULTY / ACADEMY FACULTY OF HEALTH SCIENCES

CREDIT UNIT 2

Resource Person SHANTAKUMARI A/P RAJAN

utcomes LO8 LO9 Achievement of Soft Skills Learning Outcomes
LO7 SS1 SS2 SS3 SS4 SS5 SS6 SS7

/

Soft Skills Learning Outcomes

i. Critical Thinking and Problem-solving Skills
ii. Communication Skills
iii. Team Skills
iv. Ethics & Moral Professionalism
v. Life-long Learning & Information Management
vi. Entrepreneurial Skills
vii. Leadership Skills

Start Year : 2020
Review Year : 2023

UNIVERSITY TE

COURSE CODE COURSE LEARNING OUTCO
COURSE NAME (IN ENGLISH) ENV460
COURSE NAME (IN MALAY) ANALYTICAL TECHNIQUE AND INSTRUMENTATION
TEKNIK ANALITIKAL DAN INSTRUMENTASI

COURSE OUTCOMES COGNITIVE
C1 C2 C3 C4 C5 C6

Explain the principles and functions of

1 analytical instruments used for /
environmental sample analysis.

( C2 )

Determine the appropriate techniques and /
requirements for environmental sample
2 collection, extraction, preservation, storage
and handling.
( C5 )

Demonstrate autonomous learning through
environmental sample preparation and
3 analysis using standard analytical methods
and techniques.
( P5 )

Faculty Name : FACULTY OF HEALTH SCIENCES
© Copyright Universiti Teknologi MARA

EKNOLOGI MARA

OMES - TAXONOMY LEVELS FACULTY OF HEALTH SCIENCES
FACULTY / ACADEMY 2
CREDIT UNIT SHANTAKUMARI A/P RAJAN
Resource Person

PSYCHOMOTOR AFFECTIVE

P1 P2 P3 P4 P5 P6 P7 A1 A2 A3 A4 A5

/

Start Year : 2020
Review Year : 2023

UNIVERSITY TE

COURSE O

COURSE CODE ENV460
COURSE NAME (IN ENGLISH) ANALYTICAL TECHNIQUE AND INSTRUMENTATION
COURSE NAME (IN MALAY) TEKNIK ANALITIKAL DAN INSTRUMENTASI

ACHIEVEMENT OF MOE LEA

COURSE LEARNING OUTCOMES

LO1 LO2 LO3 LO4 LO5

Explain the principles and functions of /
1 analytical instruments used for environmental

sample analysis.

Determine the appropriate techniques and

2 requirements for environmental sample /
collection, extraction, preservation, storage

and handling.

Demonstrate autonomous learning through

3 environmental sample preparation and /
analysis using standard analytical methods

and techniques.

Faculty Name : FACULTY OF HEALTH SCIENCES
© Copyright Universiti Teknologi MARA

EKNOLOGI MARA

OUTCOMES FACULTY OF HEALTH SCIENCES
FACULTY / ACADEMY 2
CREDIT UNIT SHANTAKUMARI A/P RAJAN
Resource Person

ARNING OUTCOMES

LO6 LO7 LO8 LO9 TEACHING METHODOLOGY ASSESSMENT
Test
Lecture
Laboratory Test
Blended Learning Final Test

Lecture Assignment
Laboratory
Blended Learning

Lecture
Laboratory
Blended Learning

Start Year : 2020
Review Year : 2023

CENTRE OF ENVIRONMENTAL HEALTH
AND SAFETY

FACULTY OF HEALTH SCIENCES

ENV 460
ANALYTICAL TECHNIQUE AND INSTRUMENTATION

TIME TABLE



CENTRE OF ENVIRONMENTAL HEALTH
AND SAFETY

FACULTY OF HEALTH SCIENCES

ENV 460
ANALYTICAL TECHNIQUE AND INSTRUMENTATION

LESSON PLAN

Lesson Plan

Course: Analytical Technique & Instrumentation

Venue : Open and Distance Learning (ODL)

Time : Lecture: 1000 – 1100

Date Title Week
Week 1
25 Feb Lesson 1 Introductory session: Analytical Technique & Week 2
Instrumentation Week 3
Week 4
3 Mac Lesson 2 Fundamental of Environmental Sampling
Week 5
10 Mac Lesson 3 Sample preparation for environmental analysis - Inorganic Week 6
17 Mac Lesson 4 compound from Aqueous & Solid liquid samples Week 7
Sample preparation for environmental analysis -Organic
compound from Solid samples Week 8
Week 9
18 March – 12 April 2020 :MCO due to Covid 19 Week 10
Week 11
1-12 April Semester Break
Week 12
14 April Lesson 5 Sample preparation for environmental analysis – Organic Week 13
compound from liquid sample Week 14

21 April Test 1 and Assignment Progress

28 April Lesson 6 Sample preparation for environmental analysis - Volatile
5 May Lesson 7 compounds and pre-concentration using solvent
evaporation
Instrumental Inorganic Analysis – Atomic Absorption
Spectroscopy (AAS)

12 May Lesson 8 Instrumental Organic Analysis - Introduction to
Chromatographic analysis

19 May Test 2 and Assignment progress

9 June Lesson 9 Instrumental Organic Analysis – High Performance Liquid
Chromatography (HPLC)

25 May – 3 Jun 2020: Cuti Sempena Hari Raya Aidil Fitri

16 June Lesson 10 Lesson 8 Instrumental Organic Analysis - Gas
Chromatography (GC)

23 June Lesson 11 Method Validation

30 June Test 3 and Assignment Submission and presentation

CENTRE OF ENVIRONMENTAL HEALTH
AND SAFETY

FACULTY OF HEALTH SCIENCES

ENV 460
ANALYTICAL TECHNIQUE AND INSTRUMENTATION

CLASS
ATTENDANCE













CENTRE OF ENVIRONMENTAL HEALTH
AND SAFETY

FACULTY OF HEALTH SCIENCES

ENV 460
ANALYTICAL TECHNIQUE AND INSTRUMENTATION

COURSE NOTES

15/12/2020

ENV460 ANALYTICAL Course Outcome
TECHNIQUE AND
Upon successful completion of this course, students
INSTRUMENTATION should be able to:
• Explain the principles of environmental analytical
LESSON 1: INTRODUCTORY SESSION
chemistry.
AHMAD RAZALI ISHAK • Illustrate with examples the principles of extraction

CENTRE OF ENVIRONMENTAL HEALTH & SAFETY methods.
FACULTY OF HEALTH SCIENCES • Describe the procedures and activities for basic

UiTM PUNCAK ALAM, 42300 KUALA SELANGOR laboratory skills needed for environmental sample
preparation.
• Discuss environmental pollution monitoring instruments

2

ACADEMIC CALENDAR 2020 ENV460 ANALYTICAL TECHNIQUE &
INTRUMENTATION – LESSON PLAN

34

ENV460 ANALYTICAL TECHNIQUE & ENV460 ANALYTICAL TECHNIQUE &
INTRUMENTATION ASSESSMENT INTRUMENTATION TEST

CONTINUOUS ASSESSMENT = 100% Test 1 (10%) – 21/03/2020 – 4 Short Essay
3 Test (50%) Test 2 (10%) – 19/05/2020 – 4 Short Essay
Assignment (50%) Test 3 (30%) – 30/06/2020 – 6 Short Essay
FINAL EXAMINATION = NO Final Exam
6
5

1

ENV460 ANALYTICAL TECHNIQUE & 15/12/2020
INTRUMENTATION - INDIVIDUAL ASSIGNMENT
Thank you.
Write a review paper in 3000 – 4000 words regarding the
following title:
Detection and analysis of pollutants in the environment;
PAHs, Endocrine Disrupting Chemicals, Heavy Metals,
Halogenated hydrocarbon, pesticides.

Introduction
Methodology
Results & Discussion
Conclusion
References
Using Prisma : http://www.prisma-statement.org/

7

2

15/12/2020

ENV460 ANALYTICAL TECHNIQUE & INSTRUMENTATION Learning outcomes

LESSON 2: FUNDAMENTAL OF At the end of this lesson, student should be able to;
ENVIRONMENTAL SAMPLING • Explain the purpose of Environmental Sampling
• Explain 4 criteria of sampling plan
AHMAD RAZALI ISHAK • Describe a few type of Environmental sampling

CENTRE OF ENVIRONMENTAL HEALTH & SAFETY strategies
FACULTY OF HEALTH SCIENCES • Explain general guidelines to all environmental

UiTM PUNCAK ALAM, 42300 KUALA SELANGOR sampling techniques

2

The purpose of environmental sampling Environmental Sampling design

 To determine how much pollutant enters into the Planning and sampling protocols
environment through stack emission, wastewater • Data quality objectives
discharge, and so forth in order to comply with • Sampling plan
regulatory requirement
Environmental sampling strategies
 To measure the ambient background concentration and • Where and when
assess the degree of pollution and to identify the short • How many samples
and long term trends
4
 The detect accidental release and evaluate risk and
toxicity to human and biota

 To study the fate and transport of contaminants and
evaluate the efficiency of remediation systems.

3

Planning and sampling protocols Data Quality Objectives

• Systematic planning to generate performance and acceptance criteria for
collecting environmental data.

• Seven-step planning approach to develop sampling designs for data
collection activities that support decision making.

56

1

Sampling Plan 15/12/2020

Environmental Sampling strategies

78

Judgemental Simple random

• Selection of sampling locations based on • Arbitrary collection of samples by a process that gives
professional judgment using prior information on the each sample unit in the population the same
sampling site, visual inspection and/or personal probability of being chosen
knowledge and experience
• Assumes variability of sampled medium is insignificant
• Schedule and budget - tight, early stage when – homogenous population
objective is just screen the area
• Applies for sites with little background information
• Primary representative sampling approach for • Not applicable for heterogeneous population
groundwater assessment • Ignoring prior information leads to more samples
• Statistical analysis of data - simple and straight
• No randomization and does not support any
statistical interpretation of sampling results forward

9 10

Stratified random Systematic sampling

• Sampling population is divided into several non • Systematic random - subdivides the area into grids
overlapping strata and collects samples using simple random sampling

• Each strata is more homogenous than whole • Systematic Grid - easy to implement
population • Uniform distribution over the space or time domain

• Strata could be temporal or spatial Critical part - choose right grid spacing
• Sample size can be adjusted

11 12

2

15/12/2020

Others Where and when: sampling
contaminants
Composite sampling
• Sampling cost much less than analytical cost • In space domain - contaminant variations can be in 3
• Average concentration rather than variability dimensions and hence sampling points can be
• e.g., Trace metal analysis designed by the coordinates in 1-D, 2-D or 3-D

Transect sampling • In time domain, there is only one dimension and hence
• Variation of systematic grid sampling - one or more sampling points can be designed in a time period such
as days, weeks, months or years
transect lines across a surface
• Regular intervals along the transect lines • Space and time are of interest – both spatial and
• Parallel or non parallel to one another temporal patterns of a contamination
• e.g. characterizing waste piles and water flow
• Obtain representative samples Solids - contaminants
13 accumulates at certain depth
• Air - wind velocity and direction
• Water – seasonal variations
• Biological – different species, size, sex

14

14

Cont.. 4. Sampling techniques

How many samples? General guidelines common to all environmental
• Largest sample number possible sampling
• Avoid taking too few samples 1. Sequence of sampling matrices
• No Universal formula 2. Sample amount
• Simple random sampling 3. Sample preservation and storage
• n= 4* variability2 / acceptable error2 4. Selection of sample containers
5. Selection of sampling equipments

15 16

Cont.. Cont..

1. Sequence of sampling matrices Water/waste water samples
• Least to most contaminated sampling locations • 100 ml for trace metals
• Sediment and water at same site -collect water first • 1 L for total organics
• Sampling at different depths - collect surface water • 20~40 L for an effluent acute toxicity test

samples first Soil/sediment/solid waste samples
• 200 g per sample
2. Sample amount
• Sufficient to perform all required laboratory analyses Air samples
• Trial and error method
and with an additional amount remaining for • 10 m3 may be required per sample
QA/QC analysis
• Representativeness factor 18

17

3

15/12/2020

Cont.. Cont..

3. Sample preservation and storage
Purpose to minimize any physical, chemical and
biological changes from time of sample collection to
the time of analysis

(Zhang, 2007. “Fundamentals of Environmental Sampling & Analysis” Wiley) 20
19

Cont.. Cont..

4. Selection of sample containers

• Glass vs. plastic
• Headspace vs. no headspace
• Special containers
• Biological samples -aluminium foil and closed glass

containers with inert seals or cap liners
• Aluminium foils should not be used if mercury is the

target

21 22

Cont.. 5. Environmental sampling

5. Selection of sampling equipments 24

• Made of plastic, glass, Teflon, stainless steel and other 4
materials for
• Surface water and waster water sampling
• Groundwater sampling
• Soil sampling
• Sediment sampling
• Hazardous waste sampling
• Biological sampling
• Air and stack emission sampling

23

Cont.. Cont.. 15/12/2020

2. Surface water and waste water sampling 26

• Pond sampler - near shore sampling 28
• weighted bottle sampler - collect samples in a
30
water body at a predetermined depth
• Kemmerer bottle – Teflon, acrylic or stainless steel 5

tube attached to a rope and best used when
access is from a boat or structure such as bridge or
pier

25

Cont.. Cont..

3. Ground water sampling
• Collected from a well by a bailer
• Bailer – an open pipe with an open top and a check

valve at the bottom.
• Peristaltic pump – rotor with ball bearing rollers
• Well – with a small diameter and has a depth limitation

of 25 ft

27

Cont..

4. Sediment sampling
• Scoops and trowels – for sample sediments around

shoreline and slow moving waters
• Ekman dredge – small and light weight (10 lbs) and

collects soft sediments
• Petersen or Ponar dredges

29

Cont.. 15/12/2020

5. Hazardous waste sampling 32
• Ponar or Ekman sampler – sludge sampling
• Composite liquid waste sampler – stratified liquid in

drums and other similar containers
• Thief – drum sampling device particularly useful for

grain like materials
• Trier – sampling sticky solids and loosened soils

31

6. Biological sampling Cont..
• Unique and diverse equipments
• Mammals – trapping 7. Air and stack emission sampling
• Fish – trawl nets gill nets
• Vegetation – harvested during growing season • Direct reading instruments and type of monitoring instruments
• Benthic macro invertebrate samples – Petersen and • Expensive and complex techniques
• Professional stack – testing firms
Ekman dredges can be used • High volume, total suspended particle (TSP) sampling system PM-10

sampling system

33 34

6. Quality Assurance/Quality Control

• Standard Methods -EPA Methods for Air, Water,
Wastewater and Hazardous waste

• Project point of view
• Objectives for accuracy, precision
• Sampling & custody
• Analytical procedures
• Calibration (when, how)
• Data analysis & management
• Corrective action plan

35 36

6

15/12/2020

TERIMA KASIH.

7

15/12/2020

ENV460 ANALYTICAL TECHNIQUE & INSTRUMENTATION Learning outcomes

LESSON 3: SAMPLE PREPARATION FOR At the end of this lesson, student should be able to;
ENVIRONMENTAL ANALYSIS – • Identify the common category of pollutants in the

INORGANIC FROM AQUEOUS AND environment
SOLID SAMPLE • Explain the classical and modern analytical techniques.
• Describe the purpose of sample preparation
AHMAD RAZALI ISHAK • Describe different type of sample preparation methods.
• Explain sample preparation techniques for trace metals
CENTRE OF ENVIRONMENTAL HEALTH & SAFETY
FACULTY OF HEALTH SCIENCES analysis.

UiTM PUNCAK ALAM, 42300 KUALA SELANGOR 2

Common pollutant in the environment Common pollutant in the environment

• Estimated 100K chemical present in the environment. 1. Elements: Metals (Cu, Zn, Pb, Cd, Ni, Hg, Cr) and
• A few hundred commonly considered as important metalloids (As, Se)

environmental pollutant 2. Inorganic compounds: Cyanide, CO, NOx, asbestos
• Priority chemicals has been established by various 3. Organo-metallic and metalloid compound: tetraethyl

organization based on selected factors such as lead and trybutyl tin.
quantity, persistence, bioaccumulation, toxicity and 4. Hydrocarbons: Saturated and unsaturated aliphatic
other adverse effect.
• USEPA published 129 priority pollutants (114 organic and and aromatic hydrocarbons including BTEX and
15 inorganic) in water. polycyclic aromatic hydrocarbons (PAHs)
• Important chemical pollutants can be divided into 9 5. Oxygenated compounds: Alcohol, aldehyde, ether,
categories based on their chemical characteristics. organic acid, ester, ketone, phenol.

3 4

Common pollutant in the environment Classical and Modern Analytical and
monitoring techniques
6. Nitrogen compound: Amine, amide, nitro aromatic
hydrocarbons, nitrosamines • Environmental analysis are achieved by various
classical & modern techniques.
7. Halogenated hydrocarbons: Aliphatic and aromatic
halogenated hydrocarbon, polychlorinated biphenyls • Classical - Volumetric and gravimetric methods.
(PCB) and dioxin. • Modern – Spectrometric, electrometric and

8. Organosulfur compound: Thiols, thiophenes, chromatographic methods – will be discussed in detail
mercaptans and many pesticides in Lesson 4 & 5.

9. Phosphorus compounds :Many pesticides to replace • Gravimetric methods is weighing with analytical
organo-chlorine pesticides. balance - Rarely used in environmental contaminant
analysis - due to trace concentration.
5
• E.g. Solid, moisture, oil and grease, particulate matter.

6

1

Classical and Modern Analytical and 15/12/2020
monitoring techniques
8
• Volumetric wet chemical methods is still widely used for a
variety of parameters.

• Titration is based on the simple stoichiometric relationship
of chemical reaction.

• Based on the four major reaction; acid base reaction,
redox reaction, complexation/chelation and
precipitation reaction

• Wet chemistry based method were developed that
altered the spectroscopic properties of chemicals and
made these chemical suitable for colorimetric
determination.

7

Sample preparation The purpose of sample preparation

• The step after samples have been collected and • To homogenize sample or remove moisture
preserved but before introduced into instrument for • To increase or decrease analyte concentration
further analysis. • To remove interfering chemicals
• To change the sample phase
• Very rarely can environmental samples be directly • To liberate analyte from sample matrix
injected into instrument without any pre-treatments • To modify chemical structure

• Often the most labour-intensive and time-consuming
fraction for the entire measurement process.

• 61% of total time spent in a typical chromatographic
analysis is devoted to sample preparation.

9 10

Types of sample preparation Types of sample preparation

• Different preparation technique exist between solid and Metals Liquid Sample Dissolved metal Organics
liquid samples • Ion exchange (VOCs)
• Chelation • Static head space
• For each matrix type, sample preparation differs extraction
fundamentally between organic and metal analysis. Solid Sample • Purge and trap
• Acid Digestion
• Sample preparation procedures for organic compounds • Microwave 12
in turn also vary with the volatility of the organic • Alkaline digestion
compounds. • Hydride generation

11 Organics Liquid Sample
(SVOCs, • Liquid-liquid extraction
NVOCs) • Solid Phase extraction
• Solid Phase micro extraction

Solid Sample
• Soxhlet, soxtec
• Ultrasonic extraction
• Pressurised fluid extraction
• Supercritical fluid extraction

2

15/12/2020

Common sample preparation Common Laboratory techniques
techniques
3. Concentration
1. Filtration • Achieved by Kuderna Danish evaporator or rotary
• Used to remove /collect material from a liquid/air matrix evaporator.
in which the materials are suspended. • To remove excess solvent from mixture of analyte and
• Filter media can filter papers under gravity or vacuum solvent, so that the concentration of analyte will be
filtration. sufficient enough to be detected.
• Quantitative analysis of solid – e.g. Whatman
• Small particle (0.025 -14 um) – membrane filter (Millipore 4. Digestion
and Gelman) • Commonly used for metals.
• The digestion procedure break down organically
2. Centrifugation bound substance and convert the substance to the
• Used to separate solids or liquid particles of different analysed form using oxidizing agent such as HNO3,
densities or size by rotating them in a centrifuges tubes H2SO4
• Low speed – 5000 RPM, high speed 100 000 RPM
14
13

Common Laboratory techniques Sample preparation (SP) for metal analysis

5. Extraction
• Chemical have different solubility in different solvents
• This can be used to selectively remove a solute from a
mixture.
• Often used as sample preparation to concentrate
trace organic compounds

15 16

SP for Metals determination SP for Total metals analysis in solid samples
in aqueous samples
Two common SP for total metal analysis from solid samples:
• Metal in aqueous samples are present in various form • Dry Ashing
• Can be defined operationally into Dissolved metals, • Wet digestion

Suspended metals and Total metals Dry Ashing
• Dissolved metals – unacidified sample pass through 0.45 • Simplest of all decomposition systems
• involves heating of samples in a porcelain crucible in a
μm membrane filter
• Suspended metals – metal in an acidified sample that muffle furnace in the presence of air at 400-500˚C
• After decomposition, the residue is dissolved in acid and
are retained by a 0.45 μm membrane filter.
• Total metals – unfiltered sample from vigorous acid transferred to a volumetric flask prior to analysis.
• Allow organic matter to be destroyed.
digestion (Dissolved + Suspended).
• USEPA method 18

• 3005A, 3010 - Digestion of water for FLAA or ICP
• 3015 – Microwave digestion for aqueous samples
• 3020A - Digestion of water for GFAA

17

3

15/12/2020

Dry ashing Wet digestion

Disadvantages of dry ashing method: • The use of mineral or oxidizing acids and an external heat
• Loss due to volatilization (Hg, Cd, Ca, As, Sb, Cr and Cu) sources to decompose the sample matrix.
• Resistance to ashing by some materials
• Difficult dissolution of ashed materials • The purpose of acid digestion is to dissolve metal from
• High risk of contamination sample matrix – metals can be in a measurable form.

19 • The choice of acid is dependent upon the nature of the
matrix to be decomposed and instrument analysis.
• Clean sample – HNO3 alone is adequate
• Readily oxidizable organic matter – HNO3-HCl or
HNO3/H2SO4
• Difficult to oxidize organic matter – HNO3/HClO4
• HCl is not preferred if using GFAA for analysis due to
chloride interference

20

Wet digestion Classical Hotplate digestion method

• 2 common wet digestion method procedures: • USEPA 1g soil sample

• Classical Hotplate digestion method methods Add 10 mL 1:1 HNO3, cover with watch glass Heat <BP for 15 min
• Microwave assisted acid digestion method (3050) –
Cool the sample, Add 5 mL of conc. HNO3. Cover with watch
• Both methods use mineral/oxidizing acid as an external Acid glass. Heat <BP for 30 min-Additional HNO3 until no brown fume
heat source to decompose the matrix and liberate the digestion
metal in an analysable form. given off
for
• Hotplate digestion is conducted under a ventilation hood Reduce volume to < 5mL and cool. Add 2 mL water and 3mL 30%
for minimal exposure to metals while allowing corrosive sediment, H2O2 and heat <BP. Additional H2O2 until effervescence
acids to be washed from hood after digestion is sludge
completed. ceased. Process continue for 2h at T <BP. Reduce the volume to
and soil. <5mL
21
Add 10mL HCL and heat Filter if necessary. Quantitatively
or 15 min transfer to 100mL VF. Analysis by

Filter and transfer to 100 GFAAS or ICPMS
mL VF for FLAAS or ICPMS

22

Classical Hotplate digestion method Classical Hotplate digestion method

• The digestion of 5g of samples 24

food samples can 10 mL conc. HNO3; Heat to 95ºC for 1h
be carried out in a
Cool; add 5 mL conc. H2SO4; Heat until
similar manner charring
• A typical procedure
Cool; add 5 mL conc. HNO3; Heat 180ºC
for digestion of the sample appear clear

foodstuffs (cereals, Cool; add 1 mL conc. H2O2 (per
meats, fish and 500g)heat until 200ºC;repeat until no

vegetable) for more brown fume
determination of Cool; add 10 mL water and 0.5 mL
HNO3, heat until 200ºC until white fume
total metal content
evolved
(Cd, Cu, Fe, Pb, Zn) Cool; transfer to volumetric flask for

analysis

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Microwave assisted acid Microwave assisted acid digestion systems
digestion systems
• USEPA method
• Microwave assisted acid digestion systems uses a • 3051-Microwave assisted acid digestion of sediments,
specially designed microwave that is acid proof along
with safety features for acid fume collection and sludges, soils and oils
programmable temperature and pressure control. • 3052 –Microwave assisted acid digestion of siliceous and

• Offer faster and more reproducible results than the organically based matrices.
conventional hotplate methods.

25 26

Thank you.

5

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ENV460 ANALYTICAL TECHNIQUE & INSTRUMENTATION Learning outcomes

LESSON 4: SAMPLE PREPARATION FOR At the end of this lesson, student should be able to:
ENVIRONMENTAL ANALYSIS
• To identify different approaches available for the
-ORGANIC COMPOUND FROM SOLID preparation of solid samples for organic analysis.
SAMPLES
• To explain typical procedure of Soxhlet extraction,
AHMAD RAZALI ISHAK shake flask, ultrasonic, supercritical fluid,
microwave-assisted extraction, pressurized fluid
CENTRE OF ENVIRONMENTAL HEALTH & SAFETY extraction and matrix solid phase dispersion in a
FACULTY OF HEALTH SCIENCES safe and controlled manner.

UiTM PUNCAK ALAM, 42300 KUALA SELANGOR 2

Introduction Extraction of Solid

• Extraction of organic pollutants from solid or semi-solid • Extraction of environmental pollutants from solid or
matrices has been widely investigated in recent years. semi-solid matrices can be divided into several
categories based on the method of extraction, mode of
• New instrumental approaches enhance the speed of heating and presence or not of some type of agitation.
extraction, reduction in organic solvent consumption
and the introduction of automation.

• However require higher price tag than the conventional
approaches

34

1. Soxhlet Extraction Cont..

• The basic Soxhlet • A Soxhlet uses a range of organic solvents to remove
extraction organic compounds
apparatus consists
of a solvent • The solid sample (10 g) and a similar mass of anhydrous
reservoir, an sodium sulfate are placed in the porous thimble
extraction body, a (cellulose) and inserted in the inner tube of the Soxhlet
heat source (e.g. an apparatus.
isomantle) and a
water-cooled reflux • The apparatus is then fitted to a round-bottomed flask
condenser of appropriate volume containing the organic solvent of
choice, and to a reflux condenser.

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Cont.. Cont..

• The solvent is then boiled gently using an isomantle – the • The whole process is repeated frequently until the pre-
solvent vapour passes up through the tube marked (A), set extraction time is reached.
is condensed by the reflux condenser, and the
condensed solvent falls into the thimble and slowly fills • As the extracted analyte will normally have a higher
the body of the Soxhlet apparatus. boiling point than the solvent, it is preferentially retained
in the flask and fresh solvent recirculates.
• When the solvent reaches the top of the tube (B), it
syphons over into the round-bottomed flask the organic • This ensures that only fresh solvent is used to extract the
solvent containing the analyte extracted from the analyte from the sample in the thimble.
sample in the thimble.

• The solvent is then said to have completed one cycle.

78

Cont.. Automatic Soxhlet extraction (Soxtex)

• Utilizes a 3-stage process to obtain rapid extractions.

• Stage 1- a thimble containing the sample is immersed in
the boiling solvent for approximately 60 min.

• Stage 2 - The thimble is elevated above the boiling
solvent and the sample extracted as in the standard
Soxhlet extraction approach. This is carried out for up to
60 min.

• Stage 3 - involves the evaporation of the solvent directly
in the Soxtec apparatus (10–15 min).

9 10

Cont.. 2. Shake-Flask Extraction

Advantages of Soxtec • Conventional liquid–solid extraction, in the form of
• It is faster than normal Soxhlet extraction, i.e. shake-flask extraction.

approximately 2 h per sample). • is carried out by placing a soil sample into a suitable
• it uses only 20% of the solvent volume of Soxhlet glass container, adding a suitable organic solvent, and
then agitating or shaking.
extraction.
• The sample can be concentrated directly in the • Agitating or shaking is carried out for a pre-specified
time-period.
Soxtec apparatus.
• After extraction, the solvent containing the analyte
11 needs to be separated from the matrix by means of
centrifugation and/or filtration.

• In some instances, it may be advisable to repeat the
process several times with fresh solvent and then
combine all

12

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Cont.. 15/12/2020
Cont..
Cont.. 3. Ultrasonic Extraction

• The use of sound waves to agitate a sample immersed
in an organic solvent.

• Use a sonic probe or a sonic bath.
• The sample is placed in a suitable glass container and

enough organic solvent is then added to cover the
sample.
• The system is then sonicated for a short time-period,
typically 3 min, using the sonic bath or probe.
• After extraction, the solvent containing the analyte is
separated by centrifugation and/or filtration and fresh
solvent added.
• The whole process is repeated three times and all of the
solvent extracts are then combined.

13 14

4. Supercritical Fluid Extraction

• SFE relies on the diversity of properties exhibited by a
supercritical fluid (CO2) to selectively extract analytes
from solid, semi-solid or liquid matrices.

• SFE properties - Good solvating power, high diffusivity and
low viscosity and minimal surface tension.

• Consists of six basic components;
• Supply of high-purity CO2
• Supply of high-purity organic modifier
• Two pumps
• The oven for the extraction cell
• The pressure outlet or restrictor
• The collection vessel.

15 16

Cont..

17 18

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5. Microwave-Assisted Extraction 5. Microwave-Assisted Extraction

• MAE utilizes organic solvent and heat to extract organic
pollutants from solid matrices.

• The major difference between this approach and others
is the use of a microwave oven as the heat source.

19 20

6. Pressurized Fluid Extraction 6. Pressurized Fluid Extraction

• Pressurized fluid extraction uses heat and pressure to
extract analytes rapidly and efficiently from solid
matrices.

• Also known as accelerated solvent extraction (ASE).
• A commercial PFE system currently available is a fully

automated consists of a solvent-supply system,
extraction cell, oven, collection system and purge
system.

21 22

6. Pressurized Fluid Extraction 7. Matrix Solid-Phase Dispersion

• Matrix solid-phase dispersion (MSPD) is analogous to
solid-phase extraction (SPE)

• The main difference being that MSPD is used for solid
samples.

• The sample is mixed with an SPE sorbent, e.g. C18
(octadecylsilane (ODS)) from the cartridge. After
returning the sorbent sample mixture to the cartridge, it
is eluted as in SPE.

• The main purpose of the C18 sorbent is to act as an
abrasive, thus disrupting the sample’s structure, and
hence promoting its dispersion within the sorbent and
creating a large surface area for solvent interaction.

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7. Matrix Solid-Phase Dispersion EPA method

25 26

Thank you.

5

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ENV460 ANALYTICAL TECHNIQUE & INSTRUMENTATION Learning outcomes

LESSON 5: SAMPLE PREPARATION OF At the end of this lesson, student should be able to:
LIQUID SAMPLES FOR ORGANIC
ANALYSIS • Describe the need of sample preparation for
organic compounds in liquid sample.
AHMAD RAZALI ISHAK
• Explain the principal of liquid-liquid extraction (LLE),
CENTRE OF ENVIRONMENTAL HEALTH & SAFETY solid phase extraction (SFE) and Solid-Phase Micro-
FACULTY OF HEALTH SCIENCES extraction (SPME).

UiTM PUNCAK ALAM, 42300 KUALA SELANGOR • Describe the typical procedure for LLE, SPE and
SPME.

2

Why sample prep for organic Liquid-Liquid Extraction (LLE)
compounds in solution is important?

• the analyte is present at a low concentration in a
water sample.

• used to pre-concentrate the analyte from a large
volume of water into a small sample volume.

• used to clean-up the analyte from its matrix.

3

Liquid-liquid extraction Liquid-Liquid Extraction
principal
• Selectivity and efficiency is critically governed by the
• The sample is distributed or partitioned between two choice of the two immiscible solvents.
immiscible solvents due to the analyte and matrix
have different solubilities. • Using aqueous and organic - the more hydrophobic
compounds prefer the organic solvent while the more
• The main advantage of this approach is the wide hydrophilic compounds prefer the aqueous phase.
availability of pure solvents and the use of low-cost
apparatus. • Two common approaches of LLE –discontinuous and
continuous method
5
• Discontinuous extraction - equilibrium is established
between two immiscible phases.

• Continuous - equilibrium may not be reached.

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Discontinuous method Discontinuous method

• The most common approach- separating • then the organic solvent is run off and
funnel retained in a collection flask.

• The sample is introduced into a separating • Fresh organic solvent is then added to the
funnel and a suitable organic solvent is separating funnel and the process is
added. repeated.

• The vessel is sealed with a stopper, and • This should be carried out at least 3 times
shaken vigorously for 1–2 min. • The 3 organic extracts should be

• This shaking process allows thorough combined for analysis
interspersion between the two immiscible
solvents

• periodically vent the excess pressure
generated during this shaking process.

7 8

Typical procedure of LLE Continuous method

• Continuous liquid–liquid extraction 10
can be used in dealing with large
volume of sample.

• fresh organic solvent is boiled,
condensed and allocated to
percolate repetitively through the
analyte-containing aqueous sample.

• Two type of continuous liquid
extractors - lighter-than or heavier-
than-water organic solvents.

• Extractions usually take several hours.

9

Continuous LLE

• 1 L sample is added to the continuous Solid Phase Extraction (SPE)

extractor. Then, 300 -500 mL of organic

solvent of volume is added.

• The solvent is boiled and the extraction

process allowed to occur for between

18–24 h. • Heavier than water

• After completion of the extraction

process and sufficient cooling time, the

boiling flask is detached and can be

proceed with solvent evaporation.

• Lighter than water
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Solid Phase Extraction (SPE) Introduction

• Referred to as Liquid-Solid Extraction • the sample is forced by
• involves bringing a liquid sample into pressure/vacuum through the
sorbent.
contact with a solid phase or sorbent
• the analyte is selectively adsorbed • the analyte should be retained
by the sorbent with other
onto the surface of the sorbent. extraneous material present in
• The solid-phase sorbent is packed into the sample.

small tubes or cartridges • The extraneous material can
• also available in round, flat sheets be washed from the sorbent
by the passing of an
which can be mounted in a filtration appropriate solvent.
apparatus.
• the analyte of interest can
13 then be eluted from the
sorbent by using a suitable
solvent.

14

Method of SPE operation Type of SPE media

• The SPE sorbents can be divided into three classes
method of • Normal phase
SPE can be • Reversed phase
divided into • Ion-exchange
four steps
• The most common sorbents are based on silica
particles to which functional groups are bonded to
surface silanol groups

• In addition to silica, some other common sorbents
are based on florisil, alumina and macro reticular
polymers.

15 16

Solvent selection Typical procedure of SPE

• The choice of solvent 18
directly influences the
retention of the 3
analyte on the sorbent
and its subsequent
elution

• the solvent polarity
determines the solvent
strength (or ability to
elute the analyte from
the sorbent in a
smaller volume than a
weaker solvent).

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Solid Phase Micro Extraction Solid Phase Micro Extraction
(SPME) (SPME)

• is a solvent-free sample prep
technology that is fast, economical,
and versatile.

• the process of an analyte is adsorbed
onto the surface of a coated-silica fibre

• followed by desorption of the analytes
into a suitable instrument for separation
and quantitation.

• The main principle of operation of SPME
is partitioning of analytes between an
aqueous sample and a stationary
phase

20

SPME Procedure SPME Procedure

• the fibre is inserted into the sample-containing vial 22
• the fibre is exposed to the analyte by pressing down

the plunger for a pre-specified time.
• the fibre is withdrawn back into its protective syringe

barrel and withdrawn from the sample vial.
• The SPME device is then inserted into a suitable

instrument for separation and quantitation.
• SPME can carried out either manually or by an auto

sampler.

21

Summary References

• This present chapter has summarized the main methods • Zhang, C. (2007). Fundamentals of environmental
available for such preconcentration procedures. sampling and analysis. John Wiley & Sons.

• The traditional approach has utilized liquid–liquid • Dean, J. R. (2003). Methods for environmental trace
extraction. analysis (Vol. 12). John Wiley and Sons.

• However, since the 1970s solid-phase extraction (SPE) 24
has become increasingly popular, particularly as it is
possible to automate the procedure.

• Most recently (the 1990s), the use of solid-phase
microextraction (SPME) has offered an alternative
approach to pre-concentration. However, it is not
foreseen that SPME will replace SPE, but rather offer an
alternative method which is ‘portable’ and hence can
be applied outside of the laboratory.

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Thank you.

5

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ENV460 ANALYTICAL TECHNIQUE & INSTRUMENTATION Learning outcomes

LESSON 6: SAMPLE INTRODUCTION TECHNIQUES At the end of this lesson, student should be able to:
FOR GAS CHROMATOGRAPHY
• To describe sample introduction techniques of
AND SAMPLE PRE-CONCENTRATION USING volatile and semi volatile compound prior GC
SOLVENT EVAPORATION analysis

AHMAD RAZALI ISHAK • To explain the concept of Headspace, Purge &
trap and Thermal desorption techniques
CENTRE OF ENVIRONMENTAL HEALTH & SAFETY
FACULTY OF HEALTH SCIENCES • to identify and discuss a few methods available for
solvent evaporation such as rotary evaporator,
UiTM PUNCAK ALAM, 42300 KUALA SELANGOR centrifugal evaporator, Kuderna-Danish Evaporator
and nitrogen blow evaporator.

2

Sample introduction techniques for GC Liquid Injection

• Liquid injection • Most samples for GC are introduced as liquid
• SPME • Micro syringe is normally used for injecting liquid
• Headspace
• Purge and Trap sample
• Thermal Desorption • Syringe capacity: usually 10 microliter

34

SPME Headspace

• Analytes are adsorbed onto the fiber coating materials • Indirect determination of volatile constituents in
then thermally desorbed into GC injector liquid/solid by analyzing the vapors phase that is in
thermodynamic equilibrium with the sample in a closed
system.

• Headspace provides an extract limited to volatile
components which are ideally suited to GC.

• The volatile nature of the headspace sample reduces
the need for cleaning and maintenance of the injector,
column and detector.

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Purge and Trap Purge and Trap

Purge • Widely used for the extraction of volatile organic
• Analytes are compounds from aqueous samples

extracted by the • The analytes are purged out from the matrix by using
purge gas from the inert gas, then trapped in a solid sorbent material,
sample transferred to e.g. Tenax, carbon
the U-shaped glass
tube. • Sorbent is heated to release target compounds into
Trap the GC for analysis
• The extracted
analytes are 78
collected to the
adsorbent contained Purge and Trap 10
in the trap tube.
Desorption drain
Thermal desorption
Desorption tube • The collected
analytes are
desorbed by heating
the trap tube.

• then back-flushed
with nitrogen to send
the sample to the GC
column.

9

Thermal desorption

• Sample is collected in a tube containing solid sorbent
(e.g. Tenax TA, activated carbon) by flowing inert gas
over the sample and through the tube.

• Tube is then heated to release (desorb) the target
compounds into the GC injector for subsequent GC
analysis

11 12

2