InstrumentalAnalysis:SpectroscopyandSensors

Errorand Atomic Spectroscopy&

Chemistry Spectroscopy Chromatography
Review (Elements)
Sensors

WEEK7!
1. OverviewofIRanduvvis Electrochemical
2. Instrumentationfor Spectroscopy
Sensors
spectrophotometrs
3. Interprepting IRanduvvis
spectra

IR and UV-Vis Ion selective electrodes

Instrumentation Solid state fuel cells
Typical spectra

Week 7-8
 Infrared and ultraviolet-visible
spectroscopy: Overview
Week 7, Lecture 1

(A) The electromagnetic spectrum and the units of

light energy

(B) Electronic transitions in molecules versus

vibrational transitions in molecules towards
absorbance

(C) Typical IR and UV-VIS Spectra

 The goal of spectroscopy:
A Spectrum

Molar Absorptivity,

Dichloromethane, near-infrared, creative comments

X-axis units are wavelength, y-axis units are the amount of light absorbed or transmitted
Electromagnetic
Radiation: X-Axis
Converting EM
Radiation Units

h: plancks constant
c: speed of light
: wavenumber (1/)
 Examples of
converting units
You have light incident on a sample for absorption spectroscopy. Its wavelength
is 500 nm. What is the energy of the light in Joules? What kind of spectroscopy is
this? This is uv-vis (I know from the wavelength range)

6.63 10 3.00 10 /
E 3.98 10
500 10
An infrared spectrometer designed to measure ethanol in breath uses two filters at
3.400 microns and 9.500 microns to detect light. What do these filter wavelengths
correspond to in wavenumbers (cm-1)?
1 1 1
2941
3.400 10 100

1 1 1
1052
9.500 10 100
 Ultra-Violet Visible:
Electronic Transitions

Molecules have quantized When energy is absorbed a

energy levels, like atoms peak appears in the spectrum
 Ultra-Violet Visible:
Broad peaks in solution

Ultra-violet visible spectra are

often very broad in solution not
great for qualitative fingerprinting
Infrared: Vibrational
Transitions

Dancing Molecules https://en.wikipedia.org/wiki/Infrared_spectroscopy

Interpreting Infrared
Spectra
 KEY Differences
IR spectroscopy is more powerful qualitatively for molecules
to identify structural groups
UV-VIS spectroscopy is most powerful quantitatively to
figure out how much

Molar Absorptivity,

But both techniques require spectrometers to measure the spectra of samples

 The y-axis in a spectrum:
Beers Law and Absorbance
Week 7, Lecture 2

(A) Beers law for absorbance and units

(B) Quantitative analysis using beers law

(C) Absorbance versus transmittance

 Uv-vis absorption:
pretty colors

Solutions must be clear and if

absorbing at visible wavelengths
may have a color
What you see is not the
absorption wavelength!
590
590
570 450
 Quantitative Analysis:
Absorbance & Transmittance

Transmittance
Ranges 0 1
Near 1 not a lot of
absorption

Absorbance
Ranges 0 4
Near 4 a lot of absorption
If too small, hard to detect
P0 is the power incident If too big, not enough light
P is the power after passing through
 Beers Law for
Absorbance
One must run a blank!

Can lose light from many other

factors, not just absorption!
Written
as this for
rest of
lectures

molar absorptivity units M-1cm-1

b path length in cm
c concentration in moles/Liter (M)

(note: there is another less specific unit called

absorptivity has versatile units)
 Absorbance to
Transmittance
If you are measuring a substance with an
absorbance of .1 what is its transmittance?

10 .794

If you are measuring a substance with a

transmittance of .5 or 50% what is its
absorbance

.5 .303
Beers Law In Action

Potassium permanganate in solution the

numbers are the concentration of manganese
in ppm. The cell path length, b, is 1.00 cm.
Can you find the molar absorptivity?

PathL 1 cm
Manga 54.938
PotPerm 158.034
ppm moles/L DeltaA Epsilon
4 7.28E05 0.18 2.47E+03
8 1.46E04 0.33 2.27E+03
12 2.18E04 0.5 2.29E+03
16 2.91E04 0.68 2.33E+03
20 3.64E04 0.85 2.33E+03
 Summary of
Quantitative Terms

molar absorptivity units M-1cm-1

b path length in cm
c concentration in moles/Liter (M)
 Spectrophotometers: Sources
and Detectors
Week 7, Lecture 3

(A) Typical light sources for IR and optical

spectroscopy

(B) Sample sources for IR and uv-vis

(C) The ever important block diagram

 Understanding the
Magic Box

Spectrometers for optical and infrared spectroscopy

Place sample in
this compartment

Diagram the instrument (major components)

List options for each major component know details!
Demonstrate choosing options based on measurement need
 A Pretty Typical
Research Grade Uv-Vis

Wikimedia Commons
 The Block Diagram:
Absorption Specto(photometer)

Second path for the blank

The Materials Matter

Finding containers for

samples and windows for
the instrument can be
challenging! Always think
about it
 Vials for uv-vis
(liquids)

Must remember that the

solutions are liquids and
that the vials have to be
made of the appropriate
materials (e.g. quartz if
have true uv peaks)
 For infrared samples: Salt
plates or pellet presses

Biggest challenge in IR is
typically will not use
solvents need solids
that dont absorb or use
ATR sampling
 Light Sources

(a) Is a continuum source while (b) has

discrete wavelengths
 Covering the Full
Range of Wavelengths

Tungsten lamps are

cheap, easy BUT they
peter out in the all
important ultraviolet

Deuterium lamps give

great ultraviolet signal so
the system switches
over during operation
 Spectrophotometers II:
Splitting the Light
Week 7, Lecture 4

(A) Monochromators: the basic language and issues

(B) Slit widths and resolution needs

(C) Fourier transforms and the special ways that

infrared spectrometers have to split light
 Double vs.
Single Beam
 Multichannel
Detection

From www.chem.agilent.com
 Constructive Interference:
Key to a Grating

Constructive Destructive

The groves per millimeter govern HOW SPREAD OUT and hence the resolution:
Better resolution in wavelength means fewer photons to detect!
 Gratings Help
Disperse the Light

Wikipedia on monochromators/commons license

 Slit Widths on
Spectrometers

The slit controls the range of angles

of light that enter the spectrometer

5 microns wide 200 microns wide

1 mm high 1 mm high
http://sensing.konicaminolta.us/learning-center/spectroscopy/the-slit/
 Why slit width and
ruling density matter!

Spectra for cytochrome 1, 5, 10

and 20 micron slit widths
Interference Filters:
Cheap and Easy
 Fourier Transform IR
Spectroscopy

In a FT spectrometer, data is
collected in the time domain and
fourier transformed into frequency:

No issue with background light

Better resolution
Higher light gathering capability
 Spectrophotometers II:
Splitting the Light
Week 7, Lecture 4

(A) Monochromators: the basic language and issues

(B) Slit widths and resolution needs

(C) Fourier transforms and the special ways that

infrared spectrometers have to split light
 Spectrophotometers III:
Detecting the Light
Week 7, Lecture 5

(A) Detector basics

(B) Thinking through detector selection

 Overview of
Detectors
This table leaves out SENSITIVITY:
Can you see single photons?
Also leaves out the hassle factor :>

You want a sensitive detector that works in your wavelength range!

Ideally for low cost and low maintenance (secondary)
Photomultiplier
Tubes (PMTs)

Photoelectric effect light makes

current

Every photon makes 100,000 to

10,000,000 electrons

Can do photon counting for very low

incident light
 Photodiode Arrays

Create biased junctions in silicon (pn)

When light hits, it changes the balance

of electrons and/or holes creating current
From wikimeida commons
Can make them 20 microns in diameter!
2 cm by 2 cm chip, 200 diodes!
Charge-Coupled
Devices (CCDs)

Light forms carriers (holes)

They migrate to negative bias

When the well is full (a million e-)

can sweep the voltages and
collect at amplifiers

Intensified CCDs can see single

photons!
 Key Questions for
Detector Selection

What is your wavelength range?

Do you need multiwavelength detection?
How sensitive does your measurement
need to be?
What is your budget?
 UV-VIS Spectra: Interpreting
What You Measure
Week 7, Lecture 5

(A) Anatomy of a uv-vis spectrum

(B) Quantitative analysis with beers law

(C) Identifying compounds from their uv-vis (not a

common use)
 Interpreting optical and IR
spectra
Week 7, Lecture 6

(A) Anatomy of a uv-vis or infrared spectrum

(B) Breaking down molecules into their parts

(C) Gleaning structural information from an IR

spectrum
Ultraviolet Absorbance:
Most Common
The ONLY molecular
functional groups likely to
absorb light in the 200 to 800
nm range are pi-electron
functions or hetero-atoms.
Called Chromophores
 Molecular Orbital
Picture

These are the only

ones with low
enough energies to
be in uv-vis

Pi to pi-star will have the largest molar absorptivities!

 Examples: Size
Matters

More aromatic character

shifts uv-vis to RED

https://www2.chemistry.msu.edu/faculty/reusch/VirtTxtJml/Spectrpy/UV-Vis/spectrum.htm
Infrared: Vibrational
Transitions
Interpreting Infrared
Spectra
 Some Important
KEY IR Peaks
frequency, cm1 bond functional group
36403610 (s,
OH stretch, free hydroxyl alcohols, phenols
sh)
35003200 (s,b) OH stretch, Hbonded alcohols, phenols
34003250 (m) NH stretch primary, secondary amines, amides
33002500 (m) OH stretch carboxylic acids
C(triple bond)CH: CH
33303270 (n, s) alkynes (terminal)
stretch
31003000 (s) CH stretch aromatics
31003000 (m) =CH stretch alkenes
30002850 (m) CH stretch alkanes
28302695 (m) HC=O: CH stretch aldehydes
The only two Ill 22602210 (v) C(triple bond)N stretch
22602100 (w) C(triple bond)C stretch
nitriles
alkynes
want you to know 17601665 (s C=O stretch
17601690 (s) C=O stretch
carbonyls (general)
carboxylic acids

for this class are 17501735 (s) C=O stretch

17401720 (s) C=O stretch
esters, saturated aliphatic
aldehydes, saturated aliphatic
17301715 (s) C=O stretch alpha,betaunsaturated esters
the carbonyl and 1715 (s) C=O stretch ketones, saturated aliphatic
17101665 (s) C=O stretch alpha,betaunsaturated aldehydes, ketones
the OH stretch! 16801640 (m) C=C stretch alkenes
16501580 (m) NH bend primary amines
16001585 (m) CC stretch (inring) aromatics
15501475 (s) NO asymmetric stretch nitro compounds
15001400 (m) CC stretch (inring) aromatics
14701450 (m) CH bend alkanes
13701350 (m) CH rock alkanes
13601290 (m) NO symmetric stretch nitro compounds
13351250 (s) CN stretch aromatic amines
13201000 (s) CO stretch alcohols, carboxylic acids, esters, ethers
13001150 (m) CH wag (CH2X) alkyl halides
13001150 (m) CH wag (CH2X) alkyl halides
12501020 (m) CN stretch aliphatic amines
1000650 (s) =CH bend alkenes
950910 (m) OH bend carboxylic acids
910665 (s, b) NH wag primary, secondary amines
900675 (s) CH "oop" aromatics
850550 (m) CCl stretch alkyl halides
725720 (m) CH rock alkanes
C(triple bond)CH: CH
700610 (b, s) alkynes
bend
690515 (m) CBr stretch alkyl halides
Take the challenge: Do the unknowns at m=medium, w=weak, s=strong, n=narrow, b=broad, sh=sharp
http://wwwchem.csustan.edu/Tutorials/INFRARED.HTM
 Quick IR Analysis
Look for absorption bands in decreasing order of importance:

1.the C-H absorption(s) between 3100 and 2850 cm-1. An absorption above 3000 cm-1 indicates C=C, either alkene or aromatic.
Confirm the aromatic ring by finding peaks at 1600 and 1500 cm-1 and C-H out-of-plane bending to give substitution patterns
below 900 cm-1. Confirm alkenes with an absorption at 1640-1680 cm-1. C-H absorption between 3000 and 2850 cm-1 is due to
aliphatic hydrogens.

2.the carbonyl (C=O) absorption between 1690-1760cm-1; this strong band indicates either an aldehyde, ketone, carboxylic acid,
ester, amide, anhydride or acyl halide. The an aldehyde may be confirmed with C-H absorption from 2840 to 2720 cm-1.

3. the O-H or N-H absorption between 3200 and 3600 cm-1. This indicates either an alcohol, N-H containing amine or amide, or
carboxylic acid. For -NH2 a doublet will be observed.
4. the C-O absorption between 1080 and 1300 cm-1. These peaks are normally rounded like the O-H and N-H peak in 3. and are
prominent. Carboxylic acids, esters, ethers, alcohols and anhydrides all containing this peak.
5. the CC and CN triple bond absorptions at 2100-2260 cm-1 are small but exposed.
6. a methyl group may be identified with C-H absorption at 1380 cm-1. This band is split into a doublet for isopropyl(gem-
dimethyl) groups.
7. structure of aromatic compounds may also be confirmed from the pattern of the weak overtone and combination tone bands
found from 2000 to 1600 cm-1.

http://wwwchem.csustan.edu/Tutorials/quickir.htm
 Infrared Spectra:
A Fingerprint

http://www.chemguide.co.uk/analysis/ir/interpret.html
 More IR Practice

Know: A broad absorption around

3000 to 3400 cm-1 is a OH Bond

Know: A strong and narrow peak

from 1680 to 1800 cm-1 is a C=O

http://www.chemguide.co.uk/analysis/ir/interpret.html
 Case Study 3: Breathalyzers
for Fighting Crime
Week 7, Lecture 7

(A) The problem: understanding someones medical

state through measurements of their breath

(B) The specifics of blood alcohol measurement

(C) Options for your police officers to consider in

order to better serve their community
 The Setting:
WeRFamous U.

3000 students, 9 dormitories, 80% of students live on campus

Wet campus with many traditions surrounding alcohol consumption
Independent and irreverent student body, strong student governance
 The Problem:
Student Drinking

Under-age drinking on campus is commonplace

Last academic year, twenty emergency room visits for alcohol poisoning,
students arrested for drunk driving (state police), one tragic accident

Campus police have not been equipped with breathalyzers

 Alcohol in the Human
Body

A small percentage of
alcohol is exhaled as gas
through the lungs.

The assumption is that

the % alcohol content in
the lungs is 2100 times
lower than the % alcohol
content in the blood.

Chemistry Today %BAC=gEthanol x2100mLBreath x100%

52.5mLBreath1mLBlood

When alcohol is consumed, 90% is metabolized (liver)

For an adult male (58, 160 lbs), drinking > 4 beers/hour can lead to legal intoxication.
Blood alcohol content: 0.08 grams alcohol per 100 ml of blood is considered intoxicated
 The Solution: Train Up Campus Police Force
1: (Police) Need an evidentiary standard method for BAC for drivers stopped
on the campus premises: two policeman on duty most days

2: (Campus Lawyer) Need a secondary evidentiary method for drivers with high BAC

3: (Parents) Suggested permanently installed breathalyzers on parking gates

4: (Students) Suggested alcohol breath strips as part of alcohol awareness

5: (Students) Suggested fixed breathalyzers in some dormitories

6: (Police) Want handheld patrol devices (5 campus officers on party nights)

Budget for bringing campus police up to date in

measurement of blood alcohol levels: $250,000
Cost of a medical technician to handle human blood:
$200,000/year
Cost of sending out blood samples to city police:
$1,000/sample
 The Solutions:
Breathalyzers!

Keychain
Breathalyzers! Car-GPS
Breathalyzers

IPOD
Breathalyzers!

Coin Operated
Blow
In Bars
breathalyzers
Different Approaches to Breathalyzers

An Intoximeters Alcosensor IV breathalyzer

Old technology colorimetric changes due to alcohol

Handheld breath analyzers electrochemical fuel cell technology
Tguchi cell based sensors (not as stable as fuel cells or IR)
State of the art: benchtop infrared spectrometers
 Colormetric breathalyzer
(student interest)

8H+ + Cr2O72- + 3C2H5OH 2Cr3+ + 3C2H4O + 7H2O

Chromium (VI) is orange and is reduced to the green chromium (III) ion
(Requires sulfuric acid (100 ml) to make the reaction go)
Slow process on the order of about 30 minutes

Use of alcohol dehydrogenase plus various

markers (MTT) to cause color change.
Headspace GC-MS
 Fuel Cell Technology

Apparatus consists of two platinum electrodes

with acidic electrolyte material between them
Ethanol in breath oxidized at surface of anode
to give acetic acid, protons, and electrons
Atmospheric oxygen reduced at cathode to
give two oxygen atoms
Protons and electrons from anode travel to
the cathode and combine with oxygen to
form water
Movement of electrons produces a current
that is proportional to the amount of alcohol in the breath sample
Microprocessor measures the current and calculates BAC

Labianca, Dominick A. The Chemical Basis of the Breathalyzer: A Critical Analysis, Journal of Chemical
Education. (1990). 67(3). 259-261.
State of the art in breathalyzers
Most commercial handheld devices use
electrochemical fuel cell technology
Fuel cell has two platinum electrodes with a
porous acid electrolyte
As exhaled air goes past, Pt oxidizes any
alcohol into acetic acid, 2 protons and 2
electrons.
Electrons flow through a wire hooked to a
current meter a microprocessor measures the
electrical current which is calibrated to the BAC
 Infrared Spectrometer
Uses Infrared spectroscopy to detect ethanol (C-O, O-
H, C-H, C-C bonds)
IR energy passes through sample chamber containing
breath sample and then through narrowband IR filter
Filtered energy focused on photocell detector which
converts it to electrical pulses
Microprocessor interprets pulses and calculates BAC

Breath Sample In Breath Sample Out

Microprocessor
Sample Chamber
Photocell

Quartz Lamp Lenses

Filter Wheel

http://www.youtube.com/watch?v=YwlA1ejfC8E
 Checking at 3.4 and 9.5
microns

3.4 microns = 3333 cm-1

9.5 microns = 1052 cm-1

 Questions to consider
1. What are the various interests and needs of the stakeholders
(A) The police
(B) The campus lawyers
(C) The parents
(D) The students

2. Of the various measurement devices what are the most appropriate for each group?

3. How does the budgetary constraint limit the solutions?

4. How would you propose WeRFamous handle their problem?