ANALYTICAL CHEMISTRY LEC analysis, accuracy, interpretation and

Topic 1 evaluation

Definition of Terms
Analytical Chemistry 1. Analyte - the specific components in a sample
- specific study that made Chemistry the central that is determined
science
- seek to know the composition of matter 2. Matrix - component of the sample other than
- highly relevant scientific field the analyte components that are not of interest
- enabled the other disciplines to dig further
- one of the avenues of delving deeper into 3. Interferrent - a substance that can potentially
discovering the unknown interfere with the analysis
- knowing more about the things that exists in
the present: relevant or detrimental to the 4. Signal - concerned about the analyte
society
- Branch of chemistry 5. Noise - unwanted information of the data
- deals with the chemical characterization lowers the accuracy and precision of analysis
(quanti or quali) of matter
 Quantitative 6. Limit of Detection (LOD) - smallest
- how many are present concentration or amount of analyte that can be
- determination of how much of reliably detected in a given analytical method
one or more constituent is
present
 Qualitative Environmental Science
- what is present
 increasing effect of pollution to the
- deals with the identification of
environment
elements, ions or compounds
 monitoring pollutants in the
present in a sample
environment
Analyst  effect to its inhabitants
- must know what information is really needed
Food and Feed Analysis
- obtain a representative sample
 ensuring that the products produced
Medical Laboratory Analyses are true to what is indicated in the
- used as bases to diagnose patients packaging (nutritional labelling, health
- founded on concepts of analytical chemistry claims)
 Blood gas analysis  FDA
- blood is examined - Food and Drug Administration
- monitor the levels of oxygen - tests products
and carbon dioxide - certify that the contents of the
- acid-base status products are safe for
- glucose levels consumption
 Ionized calcium in the blood - if it follows acceptable levels of
- quantitatively measured to chemical and microbiological
detect parathyroid disease substances
 Pregnancy kits
Functional Foods and Drug Discovery
- urine as a sample
 exploring the possible health benefit of
Analytical Process
food products against diseases:
o Define the problem
 cancer
what is needed to be identified? use of the
 diabetes
information obtained
 cardiovascular diseases
o Select a method
 further elucidation of its contents
sample type, size, preparation, conc.
 research institutions are initiating in
Tools/instruments availability
taking steps to harness the potential of
o Obtain a representative sample
the biodiversity
type, size and error
 examining the potential of plants for
o Prepare the sample for analysis
drug discovery and development
state of sample, interference
o Perform any necessary chemical separations Clinical Chemistry Tests
distillation, precipitation, extraction
 chemical test done to human biological
o Perform the measurement
specimens:
calibration, validation
 urine
o Calculate the results and reports
 blood
 tissues
  ultraviolet-visible (UV-
Vis) diode array
 qualitatively or quantitatively measure detector (DAD)
the levels of chemical components: - detection and early
 glucose identification of acetone:
 electrolytes  initial indicator of
 enzymes detection of all these
 hormones physio pathological
 lipids conditions and the
 proteins biological monitoring
 to anchor medical diagnosis of a test
patient’s condition  Limit Test
- specific substance is below a
Mettalurgical specific limit
 Ex. Carbon content of steel that - make a simple pass/fail
contributes to its quality evaluation of the sample
o Forensic - presence of trace elements - relies on the comparison of an
o Pharmaceutical Chemistry - purity of the drugs unknown sample to a standard
that influence their efficacy solution
- rely on a visual evaluation by
Laboratory Tests the technician
 Urinalysis Quantitative Analysis
- examine for presence of red
blood cells, white blood cells,  Sample composition will often be
microorganisms, or levels of known
proteins Quantitative Analytical Methods
- used to check for drugs and
other chemical substances  Classical Methods
 Blood Tests  Advantages:
- examine cell count  cheap initial cost
- examine presence of  cheap operating cost if
pathogenic microorganisms used in a small – scale
- measure quantitative amounts operation
of chemicals in blood  Disadvantages
 Fecalysis  high LOD
- examine presence of pathogens  less specific
 procedures are done manually
Analytical Methods
 more expensive operating cost
 Qualitative if used in a large-scale
- aims to know the chemical 1. Gravimetric
identity of the species in the - measured signal: weight
sample - Gravimetry is a sub-branch of analytical
- identify the presence of a chemistry: concept of conservation of mass
specific substance in a sample - measurement of the weight of a pure substance
- results: giving meaning, of known composition
confirming, verifying - a convenient method
- testing whether the hormone, - only requires an analytical balance and furnace
human chorionic gonadotropin
(HCG) is present 2. Volumetric
 Quantitative - measured signal: volume
- determine the relative - Titrimetry: a titrant is slowly added to the
amounts of species, or analyte until an equivalence point is reached
analytes, in numerical forms - equivalence point: the point where the analyte
- Derivatizing reagent: and the titrant are in stoichiometric amounts
 high-performance liquid - Uses indicator: substance that changes color at
chromatography (HPLC) the end point
 2,4- - ideally the equivalence point is equal to the end
dinitrophenylhydrazine point
(2,4-DNPH)
- quantitative determination of
acetone in human blood:  Types of Titration Based on Reactions
 carried out at 365 nm  Acid – Base Titration (CH3OOH
titrated with NaOH)
  Precipitimetric Titration (Cl titrated o can detect many cations in one
with AgNO3) reading - portable
 Compleximetrix Titration (Ca2+
titrated with EDTA) 4. Spectroscopic
 Redox Titration - measured signal: absorbance or emission
- energy levels in atoms are discrete
Qualitative Test
- energy difference between energy levels is the
 Performed by selective chemical property of each specific element
reactions/instrumentation - Absorbance Spectrometer
 Selective – rxn or test occur with other o most common instrument
substances but exhibits a degree of o cost effective
preference for the substance of interest o can be utilized for various types of
 Specific – occurs only with the analysis
substance of interest - UV-VIS Spectrometer
o for measuring concentrations of
Selective reactions
substances
 Strategies for achieving selectivity:
Analytical Method Validation
1. Sample preparation (extraction,
precipitation)  testing if an analytical method is suitable under
2. Instrumentation (selective detectors) actual conditions of use and for quality control
3. Target analyte derivatization (derivatize
Considerations Prior to Method Validation
specific functional groups)
4. Chromatography, separates the sample 1. Suitability of Equipment - equipment should be
constituents qualified based on its intended use
Instrumental Techniques o Check for the equipment’s accuracy,
linearity and precision
 measurement of a physical property of the
sample 2. Suitability of Materials - should also be
 Electrical property - absorption of qualified based on its use
electromagnetic radiation
 Spectrophotometry o use America Chemical Society (ACS)
 Fluorimetry Reagent Grade
 atomic spectroscopy 3. Suitability of Analyst - capability of the analyst
 mass spectroscopy to use the equipment and instruments
 NMR
 chromatography o laboratory techniques

Instrumental Methods  Specificity

- ability to assess unequivocally the analyte
 Advantage
against components which may be expected to
- very specific
be present
- low LOD
- matrix do not have a significant effect in the
- faster analysis time
reliability of the analysis
- less prone to human error
- cheaper operating cost
 example: Analysis of Vitamin C in Pineapple in
the Presence of Citric Acid
 Disadvantage
- high initial cost o Acid – Base Titration - presence of
- higher equipment maintenance cost citric acid may interfere with the
analysis and the result
3. Electroanalytic
- measured signal: volts and/or amperes o Redox Titration - can specifically detect
- Potentiometry the Vitamin C or ascorbic acid
 usually done when there is no suitable Specificity for Quantitative Methods
indicator
 uses Ion Selective Electrode  accuracy and precision at the specified range of
- Anodic Stripping Voltemetry concentration of the analyte in the matrix
 usually used to determine presence of  to check for accuracy and precision:
heavy metals  spiking
 Potentiostat  running the method concurrently with a
o electronic instrument that validated method and comparing the
controls the voltage of the results
electrode  Accuracy - the closeness of the test result
obtained by the method to the true value
measured by % Accuracy or % Error
  Precision - agreement of test results measured - store dehydrated samples
by standard deviation or relative standard
deviation
6. Burette
Specificity for Qualitative Methods
- used equipment in titration
 False Positives under substances that is - delivers known amounts of liquids
probably present - delivers the titrant during titration
 False Negatives under substances that is 7. Pipette
probably present - used to deliver liquids from one
container to another
Specificity for Limit Tests - more accurate than other glass wares
 Detection Limit
- lowest amount of analyte in a sample
which can be reliably detected
- based on:
 visual examination
 signal to noise ration
 relative standard deviation

Compedial Methods

 validated methods / established analytical

methods
 AOAC (Association of Official Agricultural
Chemists) International
 USP (US Pharmacopeia)
 BP (British Pharmacopeia)
 To qualify:
 Instruments: USP and AOAC manual
 Materials: USP and AOAC manual
 Adding Controls

Basic Tools of Analytical Chemistry

1. Analytical Balance
- most basic tool in a chemistry lab
- a precision instrument
- allows measurement of masses to a
limit of ± 0.0001 g
- highly relevant in every analytical
laboratory
- electromagnetic force balance or
electronic balance
- uses electric current to generate a
magnetic force
2. Furnace
- drying samples or removing moisture
- used during gravimetric analysis
- used in sterilizing laboratory equipment
- recording of time start after the furnace
reached the desired temperature
(preheat)
3. Desiccator
- an airtight container
- uses desiccant to maintain low moisture
- used in cooling hot objects
- maintaining low moisture of dried
samples
4. Filter Paper
- used in gravimetric analysis
- separates the solid components from
the liquid component during filtration
5. Crucible
- made of porcelain
- used in heating samples
- moisture removal