Environmental Science and Engineering (3.0.

0) Water and wastewater analysis : Basic concepts and instrumental methods of analysis ; Determination of major parameter of water such as pH, acidity, alkalinity, hardness, BOD, COD, Solids, Fluoride, Nitrogen, Iron, manganese, sulphate, phosphate, volatile acids and trace contaminants. Water treatment: coagulation, Softening, reactors, mixing and flocculation, sedimentation,
filtration, disinfection, adsorption.

Wastewater treatment:

wastewater microbiology, domestic wastewater, municipal wastewater treatment systems, unit operations of pretreatment, primary and secondary treatment, sludge treatment and disposal.

Atmosphere, Composition & Behavior: Gaseous & particulate constituents of the

atmosphere, temperature and pressure profile of atmosphere, Atmospheric Photochemistry: Electromagnetic radiations, kinetics of thermal and photochemical processes, Reactions in the upper atmosphere, photo processes in the troposphere, photochemical smog, photosynthesis, Ozone chemistry.

Air pollution: Standards, effect of air pollutants, origin and fate of air pollutants, atmospheric
dispersion, air pollution control at stationary and mobile sources, Introduction to Hazardous waste management, Environmental impact statement and global pollution issues.Introduction to Environmental legislation, regulation, ethics and system overview

Instrumental Methods: Intro

q q q q Types of Instrumental Methods Fundamental Components of an Instrument Instruments Measure Voltages and Currents! Basics of Analytical Methods Some notes and figures in this course have been taken from Skoog, Holler and Neiman, Principles of Instrumental Analysis, 5th or 6th Edition, Saunders College Publishing.

Basic Instrument Components

Source: produces some form of energy or mass that is relevant to the measurement at hand Sample Holder or Cell: contains the sample with your analyte of interest Discriminator: selects the desired signal from the source or the sample Input Transducer: detects the signal from the sample, source or discriminator. Processor: manipulates the signal electronically or mechanically to produce some useful value. Readout: displays the signal in some useful form.

Some Basic Definitions (Review) A sample is collected or taken An aliquot is usually selected from the larger, bulk sample for preservation, preparation and/or analysis A technique implies the use of a specific type of instrument for analysis A method is the procedure followed when utilizing an instrumental technique A protocol is a regulatory or officially recognized method that must be adhered to

GLP stands for Good Laboratory Practice GMP stands for Good Manufacturing Practice

Instruments Measure 1 of 2 things.

VOLTAGE (V), volts, electrical potential across two electrodes. Current (A), amperes, the flow of electrons across some point. V = IR R= resistance in Ohms

Basic Questions Regarding All Analytical & Instrumental Methods o What accuracy and precision are required?

o How much sample do I have available, and how much money do we have available for the analysis? Time + Complexity = Money o What concentration is the analyte present at and can we pre-concentrate or dilute the sample?
o What interferences might be present and can we eliminate or mask them? o What are the properties of the sample matrix?

Relevant Analytical Parameters

These are new. You should be familiar with accuracy, precision, average, standard deviation, %relative standard deviation, etc. Analytical Sensitivity: The slope of the calibration curve (IUPAC Definition)
Analytical Sensitivity slope of calibration curve standarddeviation of the measuremen t

Thus, other factors being equal, the method with the steepest calibration curve will be more sensitive
Better ability to discriminate between numerically close concentrations.

Calibration Curves UV-VIS

1.2

AAS

Linear (AAS)

Linear (UV-VIS)

Absorbance (AU)

1 0.8 0.6 0.4 0.2 0 0 0.2 0.4

y = 1.0279x - 0.0055

y = 0.204x + 0.0018

0.6 [Y] ppb

0.8

Detection Limit (DL, LOD, MDL)

Most widely disputed term in instrumental methods. The minimum concentration of analyte that can be detected, based on the analytical signal. DETECTED, not necessarily known with any great confidence! LOD (C m)= [Mean Blank Signal] + 3 x Std. Dev. Blank Signal In general, 3 is chosen as the multiplier because at + 3 STDEV, you are 99% confident you are not measuring signal from noise, background, etc. Measurements at or near the limit of detection are not necessarily precise (high %RSD)! This is what instrument manufacturers will quote you, as measured under the most ideal, not regularly attainable, conditions! The STDEVBlank signal is often replaced with the standard deviation for some very, very low (near the DL) sample you have prepared. This signal is then used with the cal. curve to calculate a DL.

Limit of Quantitation (LOQ)

Another somewhat disputed term. The LOQ is generally considered the minimum concentration of analyte that can be accurately and precisely determined. Exact definitions vary, however..

LOQ Blank Signal 10 x STDEVVERY LOW CONC. SAMPLE

You measure a blank AND a VERY low concentration sample that is near the detection limit) numerous times, and then use that data. 10 times is the typical number of replicates This signal is used in the calibration curve to calculate the MDL.

Dynamic Range
Usually called the Linear Dynamic Range, this is the concentration range over which the calibration curve has a linear shape. You have probably seen an instrument exceed its linear dynamic range with the SPEC 20
Beers Law fails at increasing concentrations.

Sample matrix, analyte and method dependent. You usually want to work with linear calibration curves if at all possible (much less complex than quadratic, exponential or polynomial fits)

Determination of Metals by AAS : 1-3 orders of magnitude Determination of Metals by ICP-AES: 5-8 orders of magnitude

Calibration Curve for Absorbance Measurement 0.6 0.5

Absorbance (AU)

0.4 0.3 0.2 0.1 0 0 0.5 1 [Analyte] M 1.5 2

Beers Law Begins to Fail Here!

Standard Additions Curve for Determination of Se by Fluorescence 2 1.8

Absorbance (AU)

1.6 1.4 1.2 1 0.8 0.6 0.4 0.2 0 -1 -0.5 0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 5.5 [Added Se] uM

Selectivity
Also known as discrimination The ability to discern different, yet closely spaced analytical signals. The spectrometer on the SPEC 20 can discriminate wavelengths of light that are about 20 nm apart (even if you can set wavelengths only 5 nm different) The spectrometer on our Varian ICP can discriminate wavelengths of light that are 0.005 nm apart! Better selectivity means you can be sure which signal is which when you have more than one analyte in the sample! However, if all other conditions are equal, increasing selectivity will decrease the amount of signal you can measure (reduce the LOD)!

Bandwidth is closely related to selectivity in optical spectrometers. It is a measure of what range of light we allow to strike the detector at any given time.

There is a difference - you need both

Errors in Analytical Measurements

Determinant - unidirectional errors ascribable to a definite cause Indeterminate - uncertainties from unknown or uncontrollable factors - generally random - noise

Systematic errors - sources

Inhomogeneity - handling & storage Contamination - sampling to reagents Adsorption on surface or volatilization Unwanted or incomplete chemical reactions Matrix effects on generation of analytical signal Incorrect standards or calibration

Recognition of systematic errors

Reproducibility gives NO information on accuracy (high std. dev. hints at problems) Make comparisons with other methods Check standard reference materials Run blanks (be sure background is small and reproducible)

Gaussian Distribution
Random fluctuations Bell shaped curve Mean and standard deviation 1sigma 68.3%, 2sigma 95.5%, 3sigma 99.7% Absolute Vs Relative standard deviation Accuracy and its relationship to the measured mean

EVERYTHING YOU DO IN THIS CLASS WILL BE A BATTLE! THE BATTLE BETWEEN SIGNAL AND SELECTIVITY! There is no way to maximize both. You have to choose some happy medium, where you get enough signal to detect the analyte, but can also be selective enough so that you are sure of what you are detecting.

Questions to ask???
Why? Is sample representative What is host matrix? Impurities to be measured and approximate concentrations Range of quantities expected Precision & accuracy required

More things to ask.

Where is analysis to be conducted How many samples (per day & total) How soon are results needed Are there standards (analytical & QC) Long term reliability Form of answer required Special facilities available

Limit of detection
signal - output measured as difference between sample and blank (averages) noise - std dev of the fluctuations of the instrument output with a blank S/N = 3 for limit of detection S/N = 10 for limit of quantitation

Sources of Noise
Environmental - 60 Hz electrical, vibrational (shield) Johnson (thermal) noise - random fluctuations in charge carriers (cool) Shot noise - pulses 1/f (flicker) noise - important at low frequencies

Noise Reduction

Avoid (cool, shield, etc.) Electronically filter Average Mathematical smoothing Fourier transform

Single channel scanning

3 objects each measured 3 times (averaged to reduce noise) Balance requires 9 measurements Monochromator - broad band source to dispersive device and then wavelengths are selected one at a time Increase intensity by scanning slower or increasing bandpass

Multidetector Spectrometer
Get 3 balances and measure all 3 samples simultaneously on separate balances Can make measurements in 1/3 time or measure 3 times as much (noise is random and proportional to square root of number of measurements) Use of diode arrays instead of slits

Signal Transformation
Double pan balance - mesure multiple objects simultaneously & measure linear combinations y(1)=X(1) + X(2) y(2)=X(1) + X(3) y(3)=X(2) + X(3) 3 equations & 3 unknowns (each object measured twice in half the time)

Hadamard multiplexing (transform)

Use one detector and replace the slit with a mask of slits at certain locations (n)- some are open & others closed (2n1 slits in mask with just more than half open) For n=3 a mask of 11011 (1 is open) can be slid to give 110, 101, 011 Linear equations improve S/N

Fourier advantage
Put all weights on 2 pan balance at the same time Change what is measured (not weights but angle of pointer showing difference in the 2 pans) Z(1)=X(1) + X(2) - X(3) Z(2)=X(1) - X(2) + X(3) Z(3)= -X(1) + X(2) + X(3)

h(t) = a cos 2 pi freq. x time

sum = cos(2pi((f1+f2)/2)t beat or difference = cos(2pi((f1-f2)/2)t 5104-sine-wa

Fourier transform - beat frequency (time domain)

We can sample the time domain at N equally spaced time intervals Represent each measurement in terms of a series of frequencies Decoding procedure to decode N algebraic equations Fourier transform requires a computer

An analytical checklist
Have the analytical tasks and goals been defined? Have issues of sampling been defined?(eg. size, homogeneity, composites) Are there facilities for sample storage (custody) available and is there a means of identification and retreival)

Checklist 2
Is pretreatment (eg. extraction, dissolution) necessary? (facilities, equipment, reagents) Is the sample analyzed representative? (mixing, weighing, size) Are the instruments appropriate for the required measurements? (sensitivity, sample state)

Checklist 3
What is the time required for each analysis? What expertise is needed to prepare, analyze, and interpret? How is data captured, calculated, presented, and stored for future comparisons? Are there appropriate quality controls? Define time line for tasks and analysis and then calculate overall costs