Instrumental Techniques for Environmental Analysis Chapter 1 : Introduction

Rezaul Karim
Environmental Science and Technology

Jessore Science and Technology University

Chapter outline
Analytical chemistry; its scope and application; instrumental analysis; instruments for analysis, representative sample; sample storage, its pre-treatment and preparation, sample pre-treatment, calibration of instrumental methods; selecting an analytical methods

Reference
Skoog, Holler & Crouch 2007, Instrumental Analysis, Brooks Cole Cengage Learning, USA. Gray, Cakvin & Bhatia, 2009, Instrumental Methods of Analysis, 1st edition, CBS, New Deli, India

Analytical chemistry

Analytical Chemistry deals with methods

for determining the chemical composition of samples of matter. A qualitative method yields information about the identity of atomic or molecular species or the functional groups in the sample. A quantitative method provides numerical information as to the relative amount of one or more of these components.

Classification of analytical methods

Classical methods sometimes called wetchemical method preceded by a

Instrumental methods
physical properties as conductivity, electrode potential, light absorption or emission, mass-tocharge ratio paralleled the development of the electronics and computer industries.
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century or more Gravimetric or by volumetric

measurements

Types of instrumental methods

Characteristic Properties INSTRUMENTAL METHODS

Radiation Spectrophotometer and e.g. Emission, photometry adsorption, scattering, refracting, rotating, Diffraction, refraction Electrical charge Electrical current Mass coulometry Amperomtry; palaeography Gravimetric

Instruments for analysis

An instrument for chemical analysis converts information about the physical or chemical characteristics of the analyte to information that can be manipulated and interpreted by a human. An analytical instrument can be viewed as a communication device between the system under study and the investigator.
Stimulus System under study response

To retrieve the desired information from the analyte, it is necessary to provide a stimulus, which is usually in the form of electromagnetic, electrical, mechanical, or nuclear energy.
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Data domain
The

measurement process is aided by a wide variety of devices that convert information from one form to another. It is important to understand how information can be encoded (represented) by physical and chemical characteristics and particularly by electrical signals, such as current. voltage, and charge. The various modes of encoding information are called data domains. types of domain nonelectrical domains electrical domains
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Nonelectrical domains

The measurement process begins and ends in nonelectrical domains. The physical and chemical information / characteristics are length, density, chemical composition, intensity of light. pressure, and etc. The information representing the mass of the object in standard units is encoded directly by the experimenter who provides information processing by summing the masses to arrive at a number. Home task: TABLE 1-2 Some Examples of Instrument Components

Data domain Map

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Electrical domains

The modes of encoding information as electrical quantities are : analog domains time domains, the digital domain For example, the measurement of the molecular fluorescence intensity of a sample of tonic water containing a trace of quinine The intensity of the fluorescence is significant in this context because it is proportional to the concentration to the quinine in the tonic water, which is ultimately the information that we desire.
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A block diagram of fluorometer

(a) a general diagram of the instrument, (b) a diagrammatic representation of the flow of information through various data domains in the instrument (c) the rules governing the data-domain transformations during the measurement process.
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The intensity of the fluorescence emission, which is nonelectrical information, is encoded into an electrical signal by a special type of device, called an input transducer.
In this example, the input transducer converts the fluorescence from the tonic water to an electrical current I, proportional to the intensity of the radiation. The current is then passed through a resistor R, which according to Ohm's law produces a voltage V that is proportional to I, which is in turn proportional to the intensity of the fluorescence.
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 The mathematical relationship between the electrical output and the input radiant power impinging on its surface is called the transfer function of the transducer

Finally, V is measured by the digital voltmeter to provide a readout proportional to the concentration of the quinine in the sample.
Devices that serve to convert data from electrical to non-clectrical domains are called output transducers e.g. Voltmeters, alphanumeric displays, electric motors, computer screens
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Analog-Domain Signals

is encoded as the

magnitude of one of the electrical quantities voltage, Current, charge,

or power. These quantities are continuous in both amplitude and time. Magnitudes of analog quantities can be measured continuously, they can be sampled at specific points in time dictated by the needs at a particular experiment or instrumental.

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Time-Domain Information
Information is stored in the time domain as the time relationship of signal fluctuations.

The time relationships between transitions of the signal from HI to LO or from LO to HI contain the information of interest.
For instruments that produce periodic signals, the number of cycles of the signal per unit time is the frequency and the time required for each cycle is its period.

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Digital information

Data are encoded in the digital domain in a twolevel scheme. The characteristic that these devices share is that each of them must be in one of only two states. For example, lights and switches may be only ON or OFF and logic-level signals may be only HI or LO. The measurement task is to count the pulses during a fixed period of time
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Detector

Detector refers to a mechanical, electrical or chemical device that identifies, records. or indicates a change in one of the variables in its environment, e.g.
pressure, temperature, electrical charge, electromagnetic radiation, nuclear radiation.

An example is the UV (ultraviolet) detector often used to indicate and record the presence of eluted analytes in liquid chromatography.
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Transducer

those devices that convert

information in nonelectrical domains to information in electrical domains and the converse.

photodiodes, photomultipliers, and other electronic photodetectors

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Sensor

analytical devices that are capable of monitoring specific chemical species continuously and reversibly.
the glass electrode ion-selective electrodes, the Clark oxygen electrode, and liber-optic sensors (optrodes)

Sensors consist of a transducer coupled with a chemically selective recognition phase.

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Readout device
A readout device is a transducer that converts information from an electrical domain to a form that is understandable by a human observer. Usually, the transduced signal takes the form of
the alphanumeric or graphical output of a cathode-ray tube, a series of numbers on a digital display, the position of a pointer on a meter scale, a tracing on a recorder paper.
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Computer in instruments

Most modern analytical instruments contain or are attached to one or more sophisticated electronic devices and data-domain converters.
operational amplifiers, integrated circuits, analog-to-digital digital-to-analog converters, microprocessors, and Computers.
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A complete analysis

five main steps:

Sampling; selecting a representative sample of the material to be analyzed Dissolution of the sample Conversion of the analyte into a form suitable for measurement Measurement and Calculation and interpretation of the measure.
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Representative sample

The aims of analysis are understood and an appropriate sampling procedure adopted. Heterogeneous material have to be homogenized prior to obtaining a laboratory sample if an average or bulk composition is required. Typical examples Surface water, rivers, seawaters Ores, minerals and alloys Laboratory , industrial or urban atmosphere
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Sample storage
After collecting sample, it may elapse several days or weeks before they are received by the laboratory. The working load Sample containers and storage conditions must be controlled

Temperature, humidity, light levels and exposure to the atmosphere

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Effects for consideration

Increases temperature leading to the loss of volatile analytes ii. Decrease in temperature that lead to the formation of deposits or the precipitation iii.Changes in humidity that affect the moisture content of hygroscopic solids and liquids iv.UV radiation that induces photochemical reactions, photo-decompostion, or polymerisation v. Air induced oxidation vi.Physical separation of the sample
i.
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Sample pre-treatment

Samples arriving in an analytical laboratory come in a very wide assortment of sizes, conditions, and physical forms They can contain analytes from major constituents down to ultra-trace levels. They have moisture content Examples of pre-treatment

Dry at 100-120C to remove moisture Weighing before and after drying Separating into groups e.g. Distillation, filtration Concentrating analytes

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Sample preparation

A laboratory sample generally needs to be prepared for anlytical measurement by treatment with reagents, that converts the analyte into an appropriate chemical form for the selected technique and method.
if the material are readily soluble in aqueous or organic solvents, a simple dissolution step may suffice. Some solution need to be decomposed to release the analyte and facilitate specific reactions in solution. Need to be diluted or concentrated Stabilizations of solutions
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Calibration determines the relationship between the analytical response and the analyte concentration. Usually this is determined by the use of chemical standards. Almost all analytical methods require some type of calibration with chemical standards. do not rely on calibration

Calibration of instrumental methods

Gravimetric methods and some coulometric methods

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Several types of calibration procedures

Comparison with Standards Direct comparison Titrations External standard calibration The least Squares methods Errors in external calibrations Multivariate calibration Standard addition methods The internal standard methods

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Direct comparison
Some analytical procedures involve comparing a property of the analyte (or the product of a reaction with the analyte) with standards such that the property being tested matches or nearly matches that of the standard. For example, colorimeters, the color produced as the result of a chemical reaction of the analyte was compared with the color produced by reaction of standards.

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Titrations

In a titration, the analyte reacts with a standardized reagent (the titrant) in a reaction. Usually the amount of titrant is varied until chemical equivalence is reached, as indicated by the color change of a chemical indicator or by the change in an instrument response. The titration is thus a type of chemical comparison.

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External standard calibration

An external standard is prepared separately from the sample. External standards are used to calibrate instruments and procedures when there are no interference effects from matrix components in the analyte solution. A series of such external standards containing the analyte in known concentrations is prepared. Ideally, three or more such solutions are used in the calibration process. However, in some routine analyses, two-point calibrations can be reliable.

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Calibration is accomplished by obtaining the response signal (absorbance, peak height, peak area) as a function of the known analyte concentration.
A calibration curve is prepared
by plotting the data or by fitting them to a suitable mathematical equation e.g. the method of linear least squares. obtained for the sample and used to predict the unknown analyte concentration, cx from the calibration curve or best-fit equation.

In the prediction step, the response signal is

The concentration of the analyte in the original bulk sample is then calculated from cx , by applying the appropriate dilution factors from the sample preparation steps.

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The Least-Squares Method: two assumption

i.

is the y intercept (the value of y when x is zero), m is the slope of the line. ii. We also assume that any deviation of
the individual points from the straight line arises from error in the

there is actually a linear relationship between the measured response and the standard analyte concentration: regression model, Y= mx + b; where b

measurement.

we assume there is no error in the x values of the points (concentrations).

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Basic least-squares analysis may not be

appropriate i. whenever there is significant uncertainty in the x data, a more complex correlation

analysis may be necessary.

ii.

when the uncertainties in the y values vary significantly with x. In this case, it may be necessary to apply a weighted least-squares

analysis

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the determination of isooctane in a hydrocarbon sample.

Here, a series of isooctane standards was injected into a gas chromatograph. the area of the isooctane peak was obtained as a function of concentration. The ordinate is the dependent variable, peak area, and the abscissa is the independent variable, mole percent (mol %) of isooctane. As is typical and usually desirable, the plot approximates a straight line, because of the indeterminate errors in the measurement process to draw the "hest straight line among the data points. Regression analysis provides the means for objectively obtaining such a line and also for specifying the uncertainties associated with its subsequent Line.
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Errors in External-Standard Calibration

When external standards are used, it is assumed that the same responses will be obtained when the same analyte concentration is present in the sample and in the standard. Thus, the calibration functional relationship between the response and the analyte concentration must apply to the sample as well. Usually, in a determination, the raw response from the instrument is not used. Instead, the raw analytical response is corrected by measuring a blank.
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An ideal blank is identical to the sample but without the analyte.

In practice, with complex samples, it is too time-consuming or impossible to prepare an ideal blank and a compromise must be made. Most often a real blank is used in sample preparation

a solvent blank , containing the same solvent in which the sample is dissolved, a reagent blank, containing the solvent plus all the reagents
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Matrix effects, due to extraneous species in the sample that are not present in the standards or blank, can cause the same analyte concentrations in the sample and standards to give different responses. systematic errors can occur during the calibration process. i. if the standards are prepared incorrectly, an error will occur. ii. errors can occur if the calibration function is obtained without using enough standards to obtain good statistical estimates of the parameters. Random errors can also influence the accuracy of results obtained from calibration curves.
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Standard-Addition Methods

Standard-addition methods are particularly useful for analyting complex samples in which the likelihood of matrix effects is substantial. A standard-addition common forms involves adding one or more increments of a standard solution to sample aliquots containing identical volumes. This process is often called spiking the sample. Each solution is then diluted to a fixed volume before measurement. Measurements are made on the original sample and on the sample plus the standard after each addition.
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The internal standard method

An internal standard is a substance that is added in a constant amount to all samples, blanks, and calibration standards in an analysis. Alternatively, it may be a major constituent of samples and standards that is present in a large enough amount that its concentration can be assumed to be the same in all cases. Calibration then involves plotting the ratio of the analyte signal to the internal-standard signal as a function of the analyte concentration of the standards. This ratio for the samples is then used to obtain their analyte concentrations from a calibration curve. An internal standard, if properly chosen and used, can compensate for several types of both random and systematic errors.
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selecting an analytical method

Defining the Problem To select an analytical method intelligently, it is essential to define clearly the nature of the analytical problem. Such a definition requires answers to the following questions: 1. What accuracy is required? 2. How much sample is available? 3. What is the concentration range of the analyte? 4. What components of the sample might cause interference? 5. What are the physical and chemical properties of the sample matrix? 6. How many samples are to be analyzed?

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Performance Characteristics of Instruments: Numerical Criteria

i.
ii. iii. iv. v.

vi.

Precision: Absolute standard deviation, relative standard deviation, coefficient of variation. variance Bias: Absolute systematic error, relative systematic error Sensitivity: Calibration sensitivity. analytical sensitivity Detection limit: Blank plus three times standard deviation of the blank Dynamic range: Concentration limit of quantitation (LOQ) to concentration limit of linearity (LOL) Selectivity: Coefficient of selectivity
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Other Characteristics to Be Considered in Method Choice

i. Speed ii. Ease and convenience iii. Skill required of operator iv. Cost and availability of equipment v. Per-sample cost

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Precision
Precision provides a measure of the random, or indeterminate, error of an analysis. Figures of merit or precision include

Absolute standard deviation (S= Relative standard deviation, RSD Standard error of the mean, Sm Coefficient of variation, CV and Variance, S2
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Bias
Bias provides a measure of the systematic, or determinate, error of an analytical method. Bias , is defined hy the equation: = Where is the population mean for the concentration of an analyte in a sample and is

the true value,

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Sensitivity

There is general agreement that the sensitivity of an instrument or a method is a measure of its ability to discriminate between small differences in analyte concentration. Two factors limit sensitivity:
the slope of the calibration curve and the reproducibility or precision of the measuring device.

A corollary to this statement is that if two methods have calibration curves with equal slopes, the one that exhibits the better precision will be the more sensitive.
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Detection Limit

Detection limit is that it is the minimum concentration or mass of analyte that can be detected at a known confidence level. This limit depends on
the ratio = the magnitude of the analytical signal the size of the statistical fluctuations in the blank signal.

The analytical signal is larger than the blank by some multiple k of the

variation in the blank due to random

errors, it is impossible to detect the analytical signal with certainty.

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Dynamic Range

the delinition of the

dynamic range of an analytical method It extends from the

lowest concentration at which quantitative measurements can be made (limit of quantitation, or LOQ) to the concentration at which the calibration curve departs from linearity by a specified amount (limit of linearity, / LOL).
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Dynamic Range

Usually, a deviation of 5 % from linearity is considered the upper limit. Deviations from linearity are common at high concentrations because of no ideal detector responses or chemical effects. The lower limit of quantitative measurements is generally taken to be equal to ten times the standard deviation of repetitive measurements on a blank.
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Selectivity
Selectivity of an analytical method refers to the degree to which the method is free from interference by other species contained in the sample matrix. Unfortunately, no analytical method is totally free from interference from other species Frequently steps must be taken to minimize the effects of these interferences.

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