Instrumental methods of analysis

By

Dr Hisham Ezzat Abdellatef

Prof. of Analytical Chemistry

Background:
Analytical Chemistry: The Science of Chemical Measurements.
Analyte: The compound or chemical species to be measured, separated
or studied
Types of analytical methods:
1.) Classical Methods (Earliest Techniques)
a.) Separations: precipitation, extraction, distillation
b.) Qualitative: boiling points, melting points, refractive index,
color, odor, solubilities
c.) Quantitative: titrations, gravimetric analysis
2.) Instrumental Methods (~post-1930s)
a.) Separations: chromatography, electrophoresis, etc.
b.) Qualitative or Quantitative: spectroscopy, electrochemical
methods, mass spectrometry, NMR, radiochemical
methods, etc.
Qualitative instrumental analysis is that measured property indicates
presence of analyte in matrix
Quantitative instrumental analysis is that magnitude of measured
property is proportional to concentration of analyte in matrix
Introduction

CHOOSING AN ANALYTICAL METHOD

1. What are the advantages or disadvantages of the technique versus
other methods?
2. How reproducible and accurate is the technique? Page
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3. How much or how little sample is required?
4. How much or how little analyte can be detected?
5. What types of samples can the method be used with?
6. Will other components of the sample cause interference?
7. Other factors: speed, convenience, cost, availability, skill required.

Types of Instrumental Methods: Example methods

Radiation emission Emission spectroscopy, fluorescence,
phosphorescence, luminescence
Radiation absorption Absorption spectroscopy, photometry,
spectrophotometry, NMR
Electrical potential Potentiometry
Electrical charge Coulometry
Electrical current Voltammetry - amperometry, polarography
Electrical resistance Conductometry
Mass-to-charge ratio Mass spectrometry

Example: Spectrophotometry
Instrument: spectrophotometer
Stimulus: monochromatic light energy
Analytical response: light absorption

Instrumental analysis Dr. Hisham E Abdellatef

Introduction

Transducer: photocell
Data: electrical current
Data processor: current meter
Readout: meter scale
Page
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Detector (general): device that indicates change in environment

Transducer (specific): device that converts non-electrical to electrical data
Sensor (specific): device that converts chemical to electrical data

Performance Characteristics: Figures of Merit:

How to choose an analytical method? How good is measurement?
How reproducible? - Precision
How close to true value? - Accuracy
How small a difference can be measured? - Sensitivity
What range of amounts? - Dynamic Range
How much interference? Selectivity

1. Accuracy: The degree to which an experimental result approaches

the true or accepted answer.
Ways to Describe Accuracy:
Error: An experimental measure of accuracy. The difference between the
result obtained by a method and the true or accepted value.
Absolute Error = (X m)
Relative Error (%) = 100(X m)/m
where: X = the experimental result
m = the true result
All Methods, except counting, contain errors dont know true value

Instrumental analysis Dr. Hisham E Abdellatef

Introduction

2. Precision: The reproducibility of results. The degree to which an

experimental result varies from one determination to the next.

Accuracy indicates proximity of measurement results to the true value, Page

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precision to the repeatability or reproducibility of the measurement

Ways to Describe Precision:

Range: the high to low values measured in a repeat series of experiments.
Standard Deviation: describes the distribution of the measured results
about the mean or average value.

Instrumental analysis Dr. Hisham E Abdellatef

Introduction

i N

(X i X )2
Absolute Standard Deviation (SD): S
i 0
N 1
SD Page
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Relative Standard Deviation (RSD): RSD(%) ( )100
X
where: n = total number of measurements
Xi = measurement made for the trial
X = mean result for the data sample

3. Response: The way in which the result or signal of a method varies

with the amount of compound or property being measured.

Ways to Describe Response:

Calibration Curve: A plot of the result or signal vs. the known amount of a
known compound or property (standard) being measured.

A calibration curve plot showing limit of detection (LOD), limit of

quantification (LOQ), dynamic range, and limit of linearity (LOL).

Calibration expression is
Absorbance = slope [Analyte (ppm)] + intercept

Instrumental analysis Dr. Hisham E Abdellatef

Introduction

Parameters used to Describe a Calibration Curve:

S = mc + Sbl
S measured signal Page
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c analyte concentration
Sbl instrument signal for blank
m - slope

Sensitivity: ability to discriminate

between small
differences in analyte
concentration. Slope
and reproducibility of the calibration curve.
(Larger slope of calibration curve m, more sensitive measurement)

Selectivity: degree to which the method is free from interference by other

species the sample
No analytical method is completely
free from interference by
concomitants. Best method is more
sensitive to analyte than interfering
species (interferent).

Matrix with species A&B:

Signal = mAcA +mBcB + Signal blank
Selectivity coefficient:
kB,A = mB/mA
k's vary between 0 (no selectivity)
and large number (very Selective).

Instrumental analysis Dr. Hisham E Abdellatef

Introduction

Interested in detecting species A, but signal will be a combination of signal

from the presence of species A and species B.

Calibration methods Page

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Basis of quantitative analysis is magnitude of measured property is
proportional to concentration of analyte
Signal [ x ] or Signal = m[x]+ Signal blank

[ ]

Dynamic Range: linear region of calibration curve where the lower limit is
ten times the standard deviation of the blank.
LOQ - limit of quantitation
LOL - limit of linearity

Detection Limit: The smallest [analyte] that can be determined with

statistical confidence. Analyte must produce an analytical signal that is
statistically greater than the random noise of blank. (i.e. analytical signal =
2 or 3 times std. dev. of blank measurement (approx. equal to the peak-peak
noise level).
Calculation of detection limit
The minimum detectable analytical signal (Sm) is given by:
Sm = Sbl + k(stdbl); for detection use k =3

To Experimentally Determine
Perform 20 30 blank measurements over an extended period of time.
Treat the resulting data statistically to obtain Sbl (mean blank signal) and
stdbl (std. dev. of blank signals). Use these to obtain S m value.
Using slope (m) from calibration curve. Detection limit (Cm) is calculated
by: (Rearranged from Sm = mc + Sbl)

Instrumental analysis Dr. Hisham E Abdellatef