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Analytic Chem
Analytic Chem
Analytic Chem
Chemistry 155
Introduction to Instrumental
Analytical Chemistry
Unit 1
Spring 2010
San Jose State University
Roger Terrill
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Chem 155 Unit 1 Page 2 of 316
1 Overview and Review ........................................................................................ 7
2 Propagation of Error......................................................................................... 56
3 Introduction to Spectrometric Methods ............................................................ 65
4 Photometric Methods and Spectroscopic Instrumentation ............................... 86
5 Radiation Transducers (Light Detectors): ...................................................... 103
6 Monochromators for Atomic Spectroscopy: ................................................... 117
7 Photometric Issues in Atomic Spectroscopy .................................................. 138
8 Practical aspects of atomic spectroscopy: ..................................................... 152
9 Atomic Emission Spectroscopy ...................................................................... 163
10 Ultraviolet-Visible and Near Infrared Absorption .......................................... 178
11 UV-Visible Spectroscopy of Molecules ........................................................ 194
12 Intro to Fourier Transform Infrared Spectroscopy ........................................ 211
13 Infrared Spectrometry: ................................................................................. 234
14 Infrared Spectrometry - Applications ............................................................ 247
15 Raman Spectroscopy: .................................................................................. 259
16 Mass Spectrometry (MS) overview: ............................................................. 279
17 Chromatography .......................................................................................... 294
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1 Overview and Review ................................................................................ 7
1.1 Tools of Instrumental Analytical Chem. ............................................. 8
1.2 Instrumental vs. Classical Methods. ................................................ 12
1.3 Vocabulary: Basic Instrumental ....................................................... 13
1.4 Vocabulary: Basic Statistics Review................................................ 14
1.5 Statistics Review ............................................................................. 15
1.6 Calibration Curves and Sensitivity ................................................... 23
1.7 Vocabulary: Properties of Measurements ....................................... 24
1.8 Detection Limit ................................................................................ 25
1.9 Linear Regression ........................................................................... 31
1.10 Experimental Design: ................................................................... 35
1.11 Validation Assurance of Accuracy: ............................................ 43
1.12 Spike Recovery Validates Sample Prep. ..................................... 45
1.13 Reagent Blanks for High Accuracy: ............................................. 46
1.14 Standard additions fix matrix effects: ........................................... 47
1.15 Internal Standards ........................................................................ 52
2 Propagation of Error ................................................................................. 56
3 Introduction to Spectrometric Methods ..................................................... 65
3.1 Electromagnetic Radiation: ............................................................. 66
3.2 Energy Nomogram .......................................................................... 67
3.3 Diffraction ........................................................................................ 68
3.4 Properties of Electromagnetic Radiation: ........................................ 71
4 Photometric Methods and Spectroscopic Instrumentation ....................... 86
4.1 General Photometric Designs for the Quantitation of Chemical
Species .................................................................................................... 87
4.2 Block Diagrams ............................................................................... 88
4.3 Optical Materials ............................................................................. 89
4.4 Optical Sources ............................................................................... 90
4.5 Continuum Sources of Light: ........................................................... 91
4.6 Line Sources of Light:...................................................................... 92
4.7 Laser Sources of Light: ................................................................... 93
5 Radiation Transducers (Light Detectors): ............................................... 103
5.1 Desired Properties of a Detector: .................................................. 103
5.2 Photoelectric effect photometers ................................................... 104
5.3 Limitations to photoelectric detectors: ........................................... 106
5.4 Operation of the PMT detector: ..................................................... 107
5.5 PMT Gain Equation: ...................................................................... 108
5.6 Noise in PMTs and Single Photon Counting: ................................ 110
5.7 Semiconductor-Based Light Detectors: ......................................... 112
5.8 Charge Coupled Device Array Detectors: ..................................... 115
6 Monochromators for Atomic Spectroscopy:............................................ 117
6.1 Adjustable Wavelength Selectors .................................................. 118
6.2 Monochromator Designs: .............................................................. 119
6.3 The Grating Equation: ................................................................... 120
6.4 Dispersion ..................................................................................... 123
6.5 Angular dispersion:........................................................................ 124
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6.6 Effective bandwidth ....................................................................... 126
6.7 Bandwith and Atomic Spectroscopy .............................................. 127
6.8 Factors That Control EFF ............................................................ 128
6.9 Resolution Defined ........................................................................ 129
6.10 Grating Resolution ..................................................................... 130
6.11 Grating Resolution Exercise:...................................................... 131
6.12 High Resolution and Echelle Monochromators .......................... 133
7 Photometric Issues in Atomic Spectroscopy .......................................... 138
8 Practical aspects of atomic spectroscopy:.............................................. 152
8.1 Nebulization (sample introduction): ............................................... 153
8.2 Atomization ................................................................................... 157
8.3 Flame Chemistry and Matrix Effects.............................................. 158
8.4 Flame as sample holder: ............................................................. 159
8.5 Optimal observation height: ........................................................... 160
8.6 Flame Chemistry and Interferences: ............................................. 161
8.7 Matrix adjustments in atomic spectroscopy: .................................. 162
9 Atomic Emission Spectroscopy .............................................................. 163
9.1 AAS / AES Review: ....................................................................... 164
9.2 Types of AES: ............................................................................... 165
9.3 Inert-Gas Plasma Properties (ICP,DCP) ....................................... 166
9.4 Predominant Species are Ar, Ar+, and electrons ........................... 166
9.5 Inductively Coupled Plasma AES: ICP-AES .................................. 167
9.6 ICP Torches .................................................................................. 168
9.7 Atomization in Ar-ICP .................................................................... 169
9.8 Direct Current Plasma AES: DCP-AES ........................................ 170
9.9 Advantages of Emission Methods ................................................. 171
9.10 Accuracy and Precision in AES .................................................. 173
10 Ultraviolet-Visible and Near Infrared Absorption .................................... 178
10.1 Overview .................................................................................... 178
10.2 ........................................................................................................ 178
10.3 The Blank ................................................................................... 179
10.4 Theory of light absorbance:........................................................ 180
10.5 Extinction Cross Section Exercise: ............................................ 181
10.6 Limitations to Beers Law: .......................................................... 182
10.7 Noise in Absorbance Calculations: ............................................ 184
10.8 Deviations due to Shifting Equilibria: .......................................... 185
10.9 Monochromator Slit Convolution in UV-Vis: ............................... 188
10.10 UV-Vis Instrumentation: .......................................................... 190
10.11 Single vs. double-beam instruments: ...................................... 191
11 UV-Visible Spectroscopy of Molecules ................................................... 194
11.1 Spectral Assignments ................................................................ 195
11.2 Classification of Electronic Transitions ....................................... 196
11.3 Spectral Peak Broadening ......................................................... 197
11.4 Aromatic UV-Visible absorptions: ............................................... 201
11.5 UV-Visible Bands of Aqeuous Transition Metal Ions .................. 202
11.6 Charge-Transfer Complexes ...................................................... 205
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11.7 Lanthanide and Actinide Ions: .................................................... 206
11.8 Photometric Titration .................................................................. 207
11.9 Multi-component Analyses: ........................................................ 208
12 Intro to Fourier Transform Infrared Spectroscopy .................................. 211
12.1 Overview: ................................................................................... 212
1 molecular vibrations ............................................................................... 212
12.2 IR Spectroscopy is Difficult! ....................................................... 215
12.3 Monochromators Are Rarely Used in IR..................................... 216
12.4 Interferometers measure light field vs. time ............................... 217
12.5 The Michelson interferometer: ................................................... 218
12.6 How is interferometry performed? .............................................. 219
12.7 Signal Fluctuations for a Moving Mirror ...................................... 220
12.8 Mono and polychromatic response ............................................ 222
12.9 Interferograms are not informative: ............................................ 223
12.10 Transforming time frequency domain signals: .................... 224
12.11 The Centerburst: ..................................................................... 225
12.12 Time vs. frequency domain signals:........................................ 226
12.13 Advantages of Interferometry. ................................................ 227
12.14 Resolution in Interferometry .................................................... 228
12.15 Conclusions and Questions: ................................................... 232
12.16 Answers: ................................................................................. 233
13 Infrared Spectrometry: ........................................................................... 234
13.1 Absorbance Bands Seen in the Infrared: ................................... 235
13.2 IR Selection Rules ..................................................................... 236
13.3 Rotational Activity ...................................................................... 238
13.4 Normal Modes of Vibration:........................................................ 239
13.5 Group frequencies: a pleasant fiction! ........................................ 242
13.6 Summary:................................................................................... 246
14 Infrared Spectrometry - Applications ...................................................... 247
14.1 Strategies used to make IR spectrometry work - ....................... 248
14.2 Solvents for IR spectroscopy: .................................................... 249
14.3 Handling of neat (pure no solvent) liquids: .............................. 249
14.4 Handling of solids: pelletizing: .................................................... 250
14.5 Handling of Solids: mulling: ........................................................ 250
14.6 A general problem with pellets and mulls: .................................. 251
14.7 Group Frequencies Examples.................................................... 252
14.8 Fingerprint Examples ................................................................. 253
14.9 Diffuse Reflectance Methods: .................................................... 254
14.10 Quantitation of Diffuse Reflectance Spectra: .......................... 255
14.11 Attenuated Total Reflection Spectra: ...................................... 256
15 Raman Spectroscopy: ............................................................................ 259
15.1 What a Raman Spectrum Looks Like ......................................... 261
15.2 Quantum View of Raman Scattering. ......................................... 262
15.3 Classical View of Raman Scattering .......................................... 263
15.4 The classical model of Raman: .................................................. 265
15.5 The classical model: catastrophe! .............................................. 266
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15.6 Raman Activity: .......................................................................... 267
15.7 Some general points regarding Raman: .................................... 269
15.8 Resonance Raman .................................................................... 271
15.9 Raman Exercises ....................................................................... 272
16 Mass Spectrometry (MS) overview: ....................................................... 279
16.1 Example: of a GCMS instrument: ............................................... 279
16.2 Block diagram of MS instrument. ............................................... 280
16.3 Information from ion mass.......................................................... 281
16.4 Ionization Sources ..................................................................... 282
16.5 Mass Analyzers:......................................................................... 287
16.6 Mass Spec Questions: ............................................................... 292
17 Chromatography Chapter 26 ............................................................... 294
17.1 General Elution Problem / Gradient Elution ............................... 307
17.2 T-gradient example in GC of a complex mixture. ....................... 309
17.3 High Performance Liquid Chromatography ................................ 310
17.4 Types of Liquid Chromatography ............................................... 311
17.5 Normal Phase: ........................................................................... 311
17.6 HPLC System overview: ............................................................ 314
17.7 Example of Reverse-phase HPLC stationary phase: ................. 315
17.8 Ideal qualities of HPLC stationary phase: .................................. 316
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QUANTITATIVE ANALYSIS
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Gas Chromatography GC
Powerful but Suitable for Volatile chemicals only
Chromatogram
absorbance
time / s
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1.1.4 Electrochemistry
Simple, sensitive, limited to certain chemicals
Dynamic electrochemistry
measure current (i) resulting from redox
reactions at an driven by a controlled voltage
at an electrode surface
i(E,t) [concentration]
1.1.5 Gravimetry
Precipitate and weigh products
very precise, very limited
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Relax, you dont need
1.2 Instrumental vs. Classical Methods. to memorize this table
just humor Dr. Terrill
while he talks about it.
Methods of Analytical Chemistry
# Chemicals # Chemicals
Isolated / hr Isolated / hr
Classical Instrumental
and amount and amount
(g) (g)
High Performance
Extraction 1-2 g Liquid 10 ng
Chromatrography
Gas
Distillation 1-2 g 100 ng
Chromatography
Separation
Precipitation 1-2 g
Electrophoresis 50 pg
Crystallization 1-2 g
0.1%
Mass
Quantitation Gravimetry 0.01 amount
1 ppm Spectrometry 10-13 M
Precision % -4
10 %
mass
Colorimetry NMR 100
10% 1 ppm
Spectroscopy 1% pppm
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2 n
squares of the random
measurement errors. n 1
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6 2.94
a 73.278
84.096
Chem 155 Unit 1 Page 15 of 316
93.404
103.493
113.431
1.5 Statistics Review 123.458
133.337
142.478
1.5.1 Precision and Accuracy 153.035
80
Mean
Number of times it was observed
60
Mean -
one Mean +
40 standard one
deviation standard
20
deviation
0
0 1 2 3 4 5 6
Value observed
2
xN
Population
lim xN Standard N
N x lim
N Deviation: N N
Mean :
N
2
xN xN xavg
Sample
N Standard s N
Average: xavg N Deviation: x N 1
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Smaller is better
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This leaves only z and t what are they? These numbers represent
the multiple of one standard deviation ( or s) that correspond to the
confidence interval. In the second case, s is only an estimate of , so
the error in s needs to be taken into account, so t is a function of the
number of degrees of freedom. For our purposes, i.e. averaging
multiple identical measurements, the number of degrees of freedom
is simply N-1.
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Chem 155 Unit 1 Page 18 of 316
An example:
Assume that we do our best to measure the concentration of basic
amines in a fish tank. Our answers are 4.2, 4.6, 4.0.
N= 3
s= (4.2-4.27)2+(4.6-4.27)2+(4.0-4.27)2 = 0.31
3-1
Number of degrees of freedom for confidence interval = 3-1=2
4.5
3.5
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ANSWER:
Realize this fact: Sample standard deviations are
only estimates.
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1.5.5 CI PROBLEM:
This imperfect camera thermometer is going to be used to screen
passengers boarding an airliner. Passengers with a high temperature
may have avian flu.
So, there is a 90% probability that the true mean lies within or above
the 80% CI.
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% confidence interval
freedom 50 80 95 99
xav N
1 1.00 3.08 12.71 63.66 texp
2 0.82 1.89 4.30 9.92 s
3 0.76 1.64 3.18 5.84 = tes t value
( 103.76 102) 3
t exp
1.34
t exp 2.275
Can you state with the given confidence that this persons
temperature differs from the expected value of 102?
99%? No
95%? No
80%? Yes
50%? Yes
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Chem 155 Unit 1 Page 23 of 316
sensitivity C
10
S S
i
S
5 But, can we
really
distinguish
C between small
changes in
0 0
0 20 40 60 80 100concentration?
0 C 100
i
Analyte Concentration (C)
Bad
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SM 3 b = mCM
By convention, m=3.
Derive a formula for CM based on B and m:
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Dynamic Range
1.8.3
Between LOQ and LOL your instrument is most
useful!
This is called:
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Selectivity
So:
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and
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Regression
Standard Additions A nearly matrix-effect free form of analysis. A
standard additions plot is a linear plot of
Plot instrument signal versus quantity of a standard
analyte solution spiked or added to the unknown
analyte sample. The unknown analyte
concentration is derived from the concentration
axis-intercept of this plot.
Sample Matrix / The matrix is the solution, including solvent(s)
and all other solutes in which an analyte is
Standards Matrix dissolved or mixed
Matrix Effect A matrix effect refers to the case where the
instrumental sensitivity is different for the sample
and standards because of differences in the
matrix.
Internal Standards A calibration method in which fluctuation in the
instrument signals due to matrix effects are,
ideally, cancelled out by monitoring the
fluctuations in the instrument sensitivity to
chemicals, internal standards, that are chemically
similar to the analyte.
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But, one can not just draw the line between any two
points because all the points have some error. So,
one mathematically attempts to minimize the
residuals.
2
= (yi (a + bxi))2
1.5
1.5
Instrument Signal (S)
S
i 1
S
i
F
i
F 0.5
i
0 0
0 2 4 6 8 10
0 C 10
i
Analyte Concentration (C)
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2
The values of a and b for which is a minimum are
the following:
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1.00 g
10 mL => sucrose
100 10.0 g
mL => sucrose
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C1V1 = C2V2
C1 = 100 ppth
C2 = 1 ppth
V2 = 10.0 mL
V1 = ? = volume to pipet over
V1 = C2V2/C1
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Acids etc.
Dilute to Analyze
100 mL 140
digest filter volume ppm 14.0 mg
found
Sample
Dilute to Analyze
100 mL 187
digest filter volume ppm 18.7 mg
found
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No Sample sample.
Ultrapure
water blank
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EQUAL TO
the sensitivity of the instrument to the analyte in the sample matrix
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1. Add sample
2. Add standard
3. Dilute to volume,
and mix mix mix!
a'
b'
a
a/b= a/b
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VXCX/VT
Dilution Calc 2:
Contribution to concentration of analyte from spike:
VSCS/VT
Slope = m = kCS/VT
intercept = b = kVXCX/VT
we can get b and m from linear regression
kVXCX/VT
and we want CX so b/m = = VXCX/CS
kCS/VT
bCS
so : CX = mVX
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On the other hand, if all the samples were spiked with the same
amount of acetonitrile then the sharp nitrile stretch at 2250 cm-1 could
be used as an internal standard.
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Often the volume of the injection will vary from injection to injection,
so the peaks will vary in size!
Peak 1 is the Peaks 2, 3 and 4 are other
internal standard. ingredients in the sample.
100 ppm NaNO3,
mAu.s
480All Peak 5 is the
1
analyte, a caffeine
1 standard, 25 ppm,
530 mAu.s
absorbance
0.5
Chromatogram
0
1 is a standard.
0 400 800 1200 1600 2000 2400 2800 3200 3600 4000
time / s
Did the caffeine
1
concentration
increase from
2 chromatogram
absorbance
1 to 2?
0.5
3 Unknown
caffeine conc.
absorbance
0
0 400 800 1200 1600 2000 2400 2800 3200 3600 4000
time / s
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Standard
S' IS STD 480 mAu.s
CIS STD 100 ppm
S ' A STD C IS STD m AN 530 100
k 4.417 unitles s
S ' IS STD C A STD mIS 480 25
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2 Propagation of Error
Skoog Chapters Covered:
Appendix a1B-4 a1B-5 and eqn. a1-28 table a1-5
Lets consider the sum of two measurements a and b that both have
some fluctuation sa = 0.5 lb and sb = 0.5 lb.
Lets say that we are weighing a and b and putting them into a box for
shipment. We need to know the total weight. Our scale is really bad
(poor precision, lots of fluctuation), and it cant weigh both a and b at
the same time because it has a limited capacity. So, we have to first
weigh a, then b and then calculate the total weight. But we know that
there is a problem with fluctuations, so we repeatedly weigh the same
items a and b and do the following experiment:
This is somewhat artificial and is not quite right, but it gives you the
general idea. Errors in a and b propagate into c.
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From this example above, one would conclude that the fluctuation in
the sum is equal to the fluctuation in the numbers summed but this
is not right.
Which is bigger?
a. the fluctuations in the individual numbers summed
b. the fluctuations in the sum
a+sa+b+sb
theoretical
a+sa+b-sb
c
a-sa+b-sb
theoretical
a+sa a-sa+b+sb
a
distribution
a-sa
Real
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error propagation through sums :
0 0 0
0 2.781 0 3.554 0 6.334
1 2.66 1 3.331 1 5.991
a b c
2 2.763 2 1.98 2 4.744
3 2.524 3 3.482 3 6.006
4 2.157 4 3.848 4 6.005
a_dist histogram ( 40 a)
2 2
0.5 0.5 0.707
b_dist histogram ( 40 b )
c_dist histogram ( 40 c)
1000
a_dist j 1
b_dist j 1 500
c_dist j 1
0
0 2 4 6 8 10
a_dist j 0 b_dist j 0 c_dist j 0
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c a b
i i i mean ( c) 9.003
stdev ( c) 2.156
0
0 9.881 a_dist hist ogram ( 40 a)
1 8.861 b_dist hist ogram ( 40 b )
2 5.473
c_dist hist ogram ( 40 c)
3 8.789
4 8.301
1000
5 8.057
6 7.561
c 7 9.909 a_dist j 1
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Absorbance
A = -log (P/Po)
1.5
A 1
i
0.5
0
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1
P
i
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Obviously, the answer has to do with the way that the dependent
variable (A) in this case changes as a function of the independent
variable (P).
S = f(a,b,c)
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2 2 2 2
S a b c
2 2 2 2
S a b c
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Chem 155 Unit 3 Page 65 of 316
distance
=c MEDIUM = cMEDIUM
= wavelength
MEDIUM = VAC /
= frequency
c = light speed cMEDIUM = cVAC /
Note: When the refractive index ( ) changes
Stays same
___________
___________
changes
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Energy Nomogram
Energy nomogram -
Hard UV 100 nm to Far IR 1mm
102 105 103
101 105
1015 102
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3.2 Diffraction
Diffraction is the basis of wavelength selection in most
spectrometers:
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=n
n = -2,-1,0,1,2 i.e. any integer
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frequencies phases
Coherence:
EM Radiation is coherent if all rays have identical:
frequency phase
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3.3.1 Polarization:
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= cVAC / cMEDIUM
=c/ 1 2.5
Refractive Wavelength Frequency s-1 Energy
J
index,
1.00 500 nm 3x108 ms-1 / 6x1015 s-1 x
500x10-9 m = 6.626x10-34 Js
6x1015 s-1 =
4x10-20 J
1.50 500/1.5 = 6x1015 s-1 4x10-20 J
333 nm
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Index = f(wavelength)
Blue
Blue Red Red
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3.3.4 Reflection:
Transmitted beam
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3.3.5 Scattering:
Incident Transmitted
Scattered
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Light
current
electrons
Metal Slope =
Free e-
Electron Energy
h
(KE)
EF
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Quantum
electronic electronic electronic
States
vibrational
ctronic
vibrational phonon
rotational
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Light Frequency
Energy
Absorbance
Emission or
Light Frequency
Energy
Heated Mat.
Emission or
Light Frequency
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Energy
Chem 155 Unit 3 Page 85 of 316
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Chem. 155 Unit 4 Page 86 of 316
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Chem. 155 Unit 4 Page 87 of 316
4.1.5 Quantitation
P = PO + mC P = Light Power at Detector
PO = Light Power for Blank
c = concentration in any unit
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2.1 Fluorescence
2.2 Raman
Scattering
3. Chemiluminescence
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$$ Water Soluble!
LiF
Sapphire Al2O3
Crystal $$$$ Hardness!
$$ UV-Vis
$ vis only
$ Water Soluble!
$$$$ IR Only
$$ non-linear dispersion
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Chem. 155 Unit 4 Page 90 of 316
$$ VUV Only?
$$ Short Lifetimes
$ IR only
$ IR Only
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H2 + e- H2* H + H + h
UV-Vis-Near IR
b. Ar, Xe, Hg
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Chem. 155 Unit 4 Page 92 of 316
+
+ Ne + Fe* Fe +h
Ne
Atomic Emission
Fe Metal Cathode
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An Introduction to Lasers:
L Light
LASER is an acronym for A Amplification by
a light amplification S Stimulated
process:
E Emission of
Partially
transmitting R Radiation
mirror
Fully
reflective
mirror
1.
relatively
long lived
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Chem. 155 Unit 4 Page 94 of 316
2.
3.
4.
Monochromatic
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Chem. 155 Unit 4 Page 95 of 316
beam out
3
Fast Decay
2
Lasing!
Pump 1
0
Fast Decay 4-state (or more)
Page 95 of 316
Chem. 155 Unit 4 Page 96 of 316
absorption
population in state Ex
lasing amplification
population in state Ey
Page 96 of 316
Chem. 155 Unit 4 Page 97 of 316
Page 97 of 316
Chem. 155 Unit 4 Page 98 of 316
Page 98 of 316
Chem. 155 Unit 4 Page 99 of 316
Page 99 of 316
Chem. 155 Unit 4 Page 100 of 316
Kr* + F
KrF* Excimer!
This favors:
population inversion!
LS = lower state
US = upper state
GS = ground state
stimulated
h + US 2h + LS gain
emission
h + LS US absorption loss
spontaneous
US LS + h loss
emisson
Laser Questions:
Indicate all that apply:
Stimulated
Emission
High Gain:
For each photon that strikes the
detector, a large signal is generated.
Low Noise:
+ -
Readout
Amplifier
Slope of
Photoelectron energy / j
this line =
Planks Constant:
h = 6.626x10-34 Js
or 1 photoelectron
About 10%
10 photons
Faceplate material:
UV-transparent
Silica
Photocathode potential:
From www.hamamatsu.com pmtconstruct.pdf -100 to 1000V
Question:
1 Photon Photocathode
Quantuum
Efficiency -
How many photoelectrons
per incident photon are
emitted? Secondary
electron yield -
How many secondary Dynode 1
electrons?
Dynode 2
Dynode 12
n
PMT Gain = G =
= Quantum efficiency
= Secondary electron yield
n= Number of dynodes
Ultimate sensitivity =
h e- at e- strikes n- n
electrons
photocathode dynodes strike anode
3x10-9s
F = 96485 coulombs/mole e-
Mol = 6.023x1023
Current (amperes) = # coulombs / s
410 = 1048576
Reject: cosmic
current / microamperes
ray e- cascade.
Accept: photon-
electron from
photocathode
Reject:
time / s Thermal e- from
dynodes
Muon: created by very high energy cosmic ray interaction with upper atmosphere, similar to
electron (-1 charge, spin 1/2) but about 200 times heavier.
About 10,000 muons reach every square meter of the earth's surface a minute; these charged
particles form as by-products of cosmic rays colliding with molecules in the upper atmosphere.
Traveling at relativistic speeds, muons can penetrate tens of meters into rocks and other matter
before attenuating as a result of absorption or deflection by other atoms.
Mark Wolvertron, science writer, Scientific American magazine, September 2007, page 26 "Muons for
Peace"
Retain signal
noise
Reject
2. Intensity limitations:
In general, the PMT is used in
LOW LIGHT ONLY ambient
light will destroy the tube.
+
+ P
-
B
- +
B P
+ -
Holes! Electrons!
B Mobile P Mobile
(p-dopant) holes. (n-dopant) electrons
p-n
junction
s
+
-
+ +
- -
+ -
-
+
+ -
Photodiode:
+
+ -
-
+
+ -
Steps in Photoconduction:
h current transduction
Al contact
SiO2
- - -
- - -
n-Si - - -
- - - - - -
1. e- are 2. h strikes 3. h+
repelled depletion accumulate
from Al region at Al
contact contact
-10V
+ + + + + +
-10V
-10V
+ + + + + +
-10V
+ + + + + +
Monochromatic
Why do we care about monochromators? light is useful!
Wavelength / nm
455.3
Czerney-Turner
blue
red
Prism
Wavelength
Dispersion:
Linear
Nonlinear with
UV-Absorption
Nonlinear
Grooves or blazes:
Lines or rulings etched into grating that scatter or
diffract the light.
Light emitted from the grating surface is called:
Diffracted light
i. b+i=? 90
ii. b=? 90 - i
a + (90 i) + 90 = 180
a i +180 = 180
a=i
n = DB + BC
6.4 Dispersion
Dispersion characterizes the extent to which a
monochromator separates (disperses) the different
wavelengths of light.
Angular dispersion:
d ( sin(i) + sin(r) )
n =
d ( sin(i) + sin(r) )/ n
(r) =
d/dr[dsin(i)] /n + d/dr[dsin(r)] / n
d / dr =
d / dr d cos(r) / n
Angle of
r=
diffraction
wavelength
=
sin(r) r
sin(r) y/F
d d
dy Fdr
D-1 =
d cos(r) / nF
D-1 =
D-1 = d / dy = d cos(r) / nF
What is dispersion?
Consider a typical monochromator:
d = 1000 lines / mm r = 45 n = 1 F = 1m
F= F = 1m = 1000 mm
-1 W = slit width in mm
EFF = wD D-1 = dispersion in nm / mm
W = slit width in mm
D-1 = dispersion in nm / mm
EFF 15 nm
EFF 2 nm
Resolution is defined as R /
Low resolution
(1 nm)
Light throughput
R / = nN
n= Diffraction order
R= / = nN
A Czerny-Turner monochromator is set to 300.00 nm and the slits are set so that
the effective bandwidth is 1.0 nm. If a broadband UV light source is directed into
the monochromator, what wavelengths of light will exit?
How big must the grating be in order to achieve this resolution (R).
1. narrow 2. numerous
EFF R
w should be small -
d should be small small
F should be large large
n should be large large
What about n?
n is the key
n = 2dsin( ) n1 = n2 2
1
but
62x??? = 2dsin( ) also!
n / nm
61 309.83
62 304.84
63 300
64 295.31
65 290.77
#3 Understand how narrow atomic bands, that are narrower than the
attainable effective bandwidth, yield a difficult stray light problem.
#6 Understand how Zeeman splitting samples the flame background but not
the analyte absorbance.
a. inefficient
b. problematic
c. hard to do reproducibly
Nebulizer types:
1. concentric
2. cross flow
3. fritted disk pneumatic
4. Babbington
5. ultrasonic
Concentric:
Nebulizers, cont:
Cross flow:
Babbington:
Fritted disk:
Ultrasonic:
8.2 Atomization
Atoms and
ions diluted
Atoms
Optimal AES
region
Optimal AAS
region
Molecules
Particles
Droplets
Optimize:
Observation region
Flame stoichiometry
-
Fe+ + e Fe
Releasing agents:
Compete with analyte for interferant:
Adding Sr+2 to react with PO4 and release
Ca+2 and Mg+2 for analysis.
Protecting agents:
Form stable but volatile complex with
analyte e.g. EDTA
Standard Additions:
Match matrix of sample and standards
Overview
Nomenclature
Plasma Types
Pros and Cons
Spectrometers
Bandwidth Considerations
Multi-channel Designs
M+ - ions
-e- +e-
Emission
M*h [M] = k(P-Po)
h
Absorption
HCL Atomic gas [M] = -klog(P/Po)
M
Molecular
gas
Solid
particles
Nebulize
9.2.1 PLASMA
9.2.2 FLAME
Water-cooled induction
coils with 27 or 41 MHz
large AC currents
flowing through them.
Quartz envelope
Tangential Ar
flow pushes
Sample plasma away
aerosol from quartz
housing
Excitation =
Optimal
observation
region.
Desolvation /
Atomization
Properties:
Inert
Hot
Electron rich
Inert = lowAtomization
Analyte oxide formation, chemical interference low
/ Ionization:
Hot = efficient atomization
Excellent atomizer
Ar flow:
Relative to ICP:
P0
Atomization is High
Chemical Interference is Low
Oxidation is Low
E
Excited Excited
N g kT
Recall: e
No go
The Solution: (when very high accuracy or precision
is needed)
Internal Standards
EA
g kT
SA QA NA e
g0
EIS
g kT
SIS QIS NIS e
g0
EA E IS
The
answer:
SA QANA kT
e
S IS Q IS N IS
So:
10.1 Overview
Ultraviolet 190-400 nm
Visible 400-750 nm
Near Infrared 750-2500 nm
We group these wavelengths together for two
reasons.
Analytes:
Optics:
P(x) = Po exp(-aNx)
a = the cross section of the absorber in cm2
N = number of absorbers per cm3 (# concentration)
x = distance
a is about 1 nm
theoretical cross section is about
the same size as the molecule!
Pi Pi P stray
P stray 0.05 A true log A app log
i Po i P o P stray
Pi A true A app
i i 4
0
1 0
1 3
0.1 0.845
2
0.01 1.243
3 A true 2
110 -3 1.314 i
4
110 -4 1.321
1
0
0 1 2 3 4
C
i
Po P Po Po
A= A= A=
Case 1. Case 2.
Hard to distinguish P and Po Hard to distinguish P and zero
For example:
For example: P=0.010.01
P=0.990.01 Po=1.000.01
Possible : Possible: Possible :
P = 0.00
0.99
P/Po 1.00
A = -Log(P/Po)
1.01
A=-Log(P/Po)
Infinite!
NEGATIVE
2
a=1
2 b b 4ac
ax bx c 0 x
2a b = KEQ
c = KEQ[HA0]
2
- K K 4 K HAo
[A ] = x( K HAo )
2
4
4 10
4
3 10
x K HAo
4
2 10
HAo
4
1 10
0
5 4 4 4 4 4
0 5 10 1 10 1.5 10 2 10 2.5 10 3 10
HAo
Fraction ionized:
x K HAo z
HAo 0.1
z
0.01
7 6 5 4 3
1 10 1 10 1 10 1 10 1 10 0.01 0.1
HAo
z
Light Sources
a. D2 Lamps 190-400nm,
excellent UV sources
Electrical Discharge in D2 gas in a sealed, SiO2
bulb.
D2 D2* (dissociative molecular state)
D2* Da + Db + h
E = KE(Da) + KE(Db) + h
Wavelength
Kinetic Energy Continuum continuum
b. Tungsten(W)-Halogen 350-2500nm ,
excellent Vis-NIR (visible-near Infrared) sources
blackbody radiators (e.g. 2500K)
halogen helps to re-deposit W onto filament and
improves lifetime for very hot filaments
drift
i.e. slow change in:
o source intensity
o detector sensitivity
gives
absorbance error.
H
C O Lewis Structure of
Formaldehyde
H
Orbital geometries
Energy
6
5 10
4
5
Absorbance
4 n
2 3 3
0
200 250 300 350 400
2
Wavelength / nm 1
0
UV spectrum & energy levels of a fictitious
carbonyl:
If: * * n * n * are allowed,
assign peaks 1, 2 and 3 to specific transitions:
Transition Energy
From Diagram From Spectrum
Possible Energy / Peak # / nm / cm-1
Transitions arbitrary units
* 10-0 = 10 1 200 50,000
n * 10-4 = 6 2 250 40,000
* 9-1 = 8 3 330 30,000
n * 9-4 = 5
E (spectrum) E (digram) / nm Assignment
50,000 10 200 *
40,000 8 250 n *
30,000 6 330 *
n *
1. *
1.1. Highest energy!
1.2. Shortest wavelength < 200 nm
1.3. O2, N2, all molecules have -bonds so: this
radiation is absorbed by air and SiO2 so
spectroscopy must be done in vacuum and is
very difficult called VUV or vacuum
ultraviolet spectroscopy
2. n *
2.1. 150-250nm most all transitions are found
in the ultraviolet
2.2. Molecules with lone (nonbonding) pairs of
electrons participate such as: O, N, Halogens
2.3. 100 to 1000 medium strength absorbers
3. n *, *
3.1. Longest wavelength transitions 200-700 nm
3.2. Strongest absorbers 100 to
-1 -1
100,000 M cm
3.3. Limited to molecules with -bonds e.g.
alkenes, alkynes, aromatics, carbonyls, azides
etc.
They are
molecules, not
atoms so
electronic
transitions,
vibrational
transitions,
rotational
transitions all
happen
simultaneously
Solution phase
interactions
perturb the energy
levels of each
molecule
differently. The
spectrum
averages over
many molecules,
creating a
continuum of
energy levels for
the transition
11.3.3 Solvatochromism:
The shift in wavelength of an absorbance band with
changes in polarity (dielectric constant) of the solvent.
Primary E2 B
(nm) 184 204 256
-1 -1
MAX (M cm ) 60,000 7,900 200
V. Strong Strong Intermed
I- < Br- < Cl- < F- < OH- < C2O4-2 < H2O < SCN < NH3
< Ethylenediamine < o-phenanthroline < NO2- < CN-
Example:
Fe(III)3+-SCN- + h Fe(II)2+SCN0
electron transfer from SCN- to Fe(III)
5,000
Fe(II)2+(o-phenanthroline)3 + hv
Fe(III)3+(o-phenanthroline)-1(o-phenanthroline)2
electron transfer from Fe(II) to (o-phenanthroline)
12,000
Electronic
configurations differ in
energy due primarily to
orbital angular
momentum, i.e.
magnetic interactions
described by Russell-
Saunders and other
coupling schemes.
S+TP
S = Analyte metal ion e.g. Fe3+
T = Titrant e.g. SCN-
P = Product e.g. FeSCN2+
Why might titration be a more accurate and precise way to compute C than
using C=A/ b (i.e. from an absorbance calibration curve).
Bi
( 0.25A) i ( 0.75B) i 4
( 0.5A) i ( 0.5B) i
( 0.75A) i ( 0.25B) i
2
Ai
0
200 250 300 350 400
wi
Absorbance
2
A2 CA B2 CB A2
0.72
III II
5.3
0.72 0.72 0.72
III 0.72 CA 5.3 CB 3.47
5.3 5.3 5.3
III 0.098 CA 0.72 CB 0.471 Subs titute CA into I and II (to check):
Matrix approach:
A1 CA B1 CB A
1 A
1
A1 B1
A2 CA B2 CB A2 C A
A2 B2 A 2
.
A3 B3 C A
A3 CA B3 CB A
3 B 3
A4 B4 A
A4 CA B4 CB A 4
4
12.1 Overview:
Infrared (IR) spectroscopy 1 molecular
deals mainly with: vibrations
so
IR radiation has a lower frequency: IR = c/ IR
so
Because:
IR sources are somewhat weak and
IR radiation is somewhat hard to detect, and
IR radiation is hard to focus and
IR spectroscopy must be done at very low effective
bandwidth,
Grating or prism-monochromator based IR
spectrometers are usually:
slow noisy
Interferometry
time!
1
c c
c
Signal / Volts
0 5 10 15 20 25 30
Time / femtoseconds
This can be done with an optical device called an:
Interferometer
Take this on faith for now, and lets explore what that
might look like
10 Light Electric
cos( ) 6
Fields Sum
5
cos( ) 4
cos( ) cos( )
Constructive
0
Interference
5
0 0.5 1 1.5 2 2.5 3
2
10
Destructive
cos( ) 6
5 Interference
cos( ) 4
cos( ) cos( ) 0
5
0 0.5 1 1.5 2 2.5 3
Constructive interference:
n = f( sin(i)+sin(r) ) for a grating
n = f( time ) for an interferometer
AC(0) = AB
AC(t) = AB + vt Constant Velocity Mirror
= 2v = / 2v
monochromatic-
one frequency
only.
Two frequencies
(lines) only.
Polychromatic
many
frequencies.
1000
x
0
1000
0 20 40 60 80 100
T ime
1 .5
Amp li tud e1 0
Amp li tud e 1
0 .5
1
0 0 .5 1 1 .5 0
ti me_s econ ds 0 1 2 3 4
Freq uency_ Hz
1 2
1 .5
+ 1
0 0 .5 1 1 .5
0 .5
0
0 1 2 3 4
ti me_s econ ds Freq uency_ Hz
------------------------------------------------------------------------------------------
2
2
1 .5
Amp li tud e3 0
Amp li tud e 1
0 .5
2
0 0 .5 1 1 .5 0
0 1 2 3 4
ti me_s econ ds
Freq uency_ Hz
signals:
= 0!
100 20
Amplitude
Amplitude
10
50
0
0 10
0 0.5 1 1.5 0 20 40
Frequency Frequency
2000
Interferogram
Amplitude
2000
0 20 40 60 80 100
T ime
10
Amplitude
10
20 25 30 35 40
T ime
Page 226 of 316
Chem 155 Unit 12 227 of 316
cos( ) 6
4
cos( 1.02 ) 3
0.999013 2
0 10 20 30 40 50 60 70 80 90 100
0 100
2
P( ) = Pi cos(2 1) + Pi cos(2 2)
For identical B
P( ) = P ( cos(2 1) + cos(2 2) )
What is P( ) when = 0?
B( cos(0)+cos(0) ) = 2B
1 = some integer
2 = some integer 1
1 = 1 1
1 - 2 = 1
12.18 Answers:
cm 1 -1
f 2v 21 3000 cm 6000s
s
1 1 1 1 1
effective bandwidth incm : EFF 0.5cm
2d 2 1 cm
7 nm 7 nm
10 10
cm cm
0.555nm
effective bandwidth in nm
: 1 1
3000 cm 3000.5cm
13 Vibrational Spectroscopy:
1,000,000 10 14
Vibrations Rotations
E( )
+ H H H
- Cl Cl Cl
In Out In Force
E( )
N N N
N N N
E( )
Cl H H Cl
J= J=
v= v=
Energy
Page 238 of 316
Chem 155 Unit 13 Page 239 of 316
translations
rotations
vibration!
Modes of vibration:
For example CO2: Linear or Non-linear?
# of Modes: 3( ) - ( ) =
Overtones: v = 2, 3
Combinations: C = A + B and A - B
Characteristic group
modes and show up
in IR spectra.
Examples of groups:
C-H
C=O
C N
NH3
OH etc.
is somewhat
variable because:
environment
other bonds
other modes
influence the group
vibration.
F= ky
m1 m2
= reduced mass
m1 + m2
E = (v + )h m
v = vibrational quantuum number = 0,1,2
v = 0,1 = the selection rule for vibrational transitions
1 k
again: m =
2
so:
h k
E= vh m = 1 =h
2
=c
13.6 Summary:
3. Sample Handling:
use solvents transparent in selected spectral
regions
5 10 tons of pressure
a few minutes
2-5 mg
14.5 Handling of Solids: mulling:
sample 2 m
Nujol =
mineral
(aliphatic ) oil
or
1. Grind sample finely Fluorolube =
fluorocarbon oil
2. Mix with mulling oil
3. Sandwich w/ KBr/NaCl
plates
A = -log[P/Po]
qualitative
The two pieces of information that are useful:
Spectral fingerprinting
O-H or C-Cl
Why?
Reduced mass of C-Cl much
larger so (k/ )1/2 for C-Cl smaller
Penetration depth
15 Raman Spectroscopy:
vibrational spectroscopy with
Rayleigh Raman
elastic scattering
inelastic scattering
High resolution
monochromator /
Scattered Beam interferometer +
sensitive detector
Laser
Transmitted Beam
Incident Beam
If the laser is, e.g. a HeNe laser ( =632.8 nm), most of the scattered
radiation will be:
Rayleigh Line
400 425 450 475 500 525 550 575 600 625 650
Wavelength / nm
Energy S1
So
Vibrational coordinate
Hypothetical Raman Scattering Spectrum
Light Power
400 425 450 475 500 525 550 575 600 625 650
Wavelength / nm
time
E E0 cos 2 ex t
time
m E E0 cos 2
ex t
d d
r re r cos 2
vib t
0 dr 0 dr A
Incident E-field
Oscillating
polarizability
Amplitude
modulation in
induced dipole
Recall that:
1 1
cos ( x) cos ( y) 2
cos ( x y) 2
cos ( x y)
So m becomes:
m E0 0 cos 2 ex t
1 d
E0 r cos 2 cos 2
2 dr A ex vib ex vib
4 4 4 4 4 4 4 4 4 4 4
1.6 10 1.68 101.76 101.84 101.92 10 2 10 2.08 102.16 102.24 102.32 10 2.4 10
Wavelength / nm
but
4 4 4 4 4 4 4 4 4 4 4
1.6 10 1.68 101.76 101.84 101.92 10 2 10 2.08 102.16 102.24 102.32 10 2.4 10
Frequency / cm-1
C NO YES YES NO
Asymmetric
O stretch
YES NO NO YES
C
Bend
O
YES YES YES YES
C
1 k
vib
2
So
Pros
1. Intensity greatly enhanced (102 106)
2. Selective for vibrations of chromophore
Cons
1. Sample degradation because of absorption.
2. Fluorescence can swamp Raman signals.
400 425 450 475 500 525 550 575 600 625 650
Wavelength / nm
7
10 1
las er frequency: 20000 cm
500
7
10 1
Stokes frequency: 17857 cm
560
7 7
10 10 1
2143 cm Effective bandwidth: Must res olve 20000 and 20001 cm-1.
500 560
7 7
7 7 10 10
10 10 1 0.025 nm high res olution needed!
2759 cm 20000 20001
500 580
7 7
10 1 10
1520 17916 cm 558.2 nm
514.5 17916
E E
N1 kT kT 7
e e 8.784 10
N0
b. Assuming
that the bond
m1 m2
strengths are 1 k
VIB
equal, 2 m1 m2
calculate the
vibrational 1 kDCl
frequency DCl 2 DCl DCl HCl
that you since kHCl kDCl
HCl 1 kHCl HCl DCl
expect for the
2
isotope DCl35. HCl
35 1 35 2 HCl
HCl DCl 0.717
35 1 35 2 DCl
S1
Virtual state
S0
Fluorescence Raman-Stokes Raman-AntiStokes
7 7 7 7
10 10 10 10
3504 3483
250 274 300 335
Sample
Introduction
Ionization
Source
Mass
Analyzer
Pattern recognition to ID
molecule
C2H5+ + M M C2H5+
ethyl transfer M+15
Electron impact
ionization
chemical
ionization (CH4)
o Soft
o Pulsed introduction of solids
o Large molecules proteins / biomolecules
o Gives predominance of M+
16.5.1.1 Resolution: R M/ M
m/ M = 28 / 0.012 = 2300
m/ M = 149,190 / 1 = 150,000
R attainable: 100,000
Resolution up to:
10,000
DC field
M/z
A given AC amplitude + DC field
yields a stable trajectory for one:
mass filter
Like E and B sectors this is a:
TOF analyzers:
Element Iso %
12 13
Carbon C 98.90% C 1.10%
1 2
Hydrogen H 99.99% H 0.01%
16 17 18
Oxygen O 99.76 O 0.038% O 0.2%
17 Chromatography Chapter 26
Electrophoresis.
Some definitions:
Mobile Phase is the gas, liquid or supercritical fluid
that passes through separation column or slab.
Stationary Phase is the solid or immobilized liquid
into which the solutes or analytes partition, adsorb or
bind.
Band or Zone describes the region (stripe, plug) of
the separation column or slab that contains the solute
or analyte.
Retention is the word used to refer to the delay of the
solute or analyte in reaching the detector relative to
the mobile phase.
x
Page 302 of 316
Chem 155 Unit 17 Chromatography Page 303 of 316
2 2
xi i 100 25
10 0.625
1000 1000
1 1 2 2 2
S x exp x 50 12.5
2 2 2.5 0.156
2 1000 1000
x
2 2 2
1000 1000
N: 100 1600 N
100 25
2 2
1000 1000
400 6400
50 12.5
0.0 3
6400 plates
0.0 25
0.0 2
signal
1600 plates
0.0 15
0.0 05
100 plates
0
0 20 0 40 0 60 0 80 0 10 00 12 00 14 00 16 00 18 00 20 00
ret ent ion time or column lengt h
x
2 2 2
400 1200
N: 1024 9216 N
12.5 12.5
2 2
800 1600
4096 16384
12.5 12.5
0.0 3
0.0 25
0.0 2
signal
0.0 15
0.0 1
0.0 05
0
0 20 0 40 0 60 0 80 0 10 00 12 00 14 00 16 00 18 00 20 00
ret ent ion time or column lengt h
12
10
H ( 1 2 0.5 u)
2
0 2 4 6 8 10 12
u
A 2 dp
B 2 DM
B
H A Cu
u
C = com plex function of particle
s ize, coating, diffus ion coefficient,
favored in general by large
diffus ivity and s m all particle s ize
a.
b.
x
A mixture of molecules with strongly differing k values
is hard to separate because:
x
o temperature gradient elution (gas
chromatography)
x
Page 309 of 316
Chem 155 Unit 17 Chromatography Page 310 of 316
x
Skoog Holler and Nieman Principles of Instrumental Analysis 5th ed.
x
17.5 Normal Phase:
stationary phase: hydrophilic (SiO2 particles)
mobile phase: hydrophobic mobile phase
(hexane, methanol).
weak solvent: hexane
strong solvent: methanol
retained: polar compounds
eluted first: non-polar compounds
GPC
Gel permeation chromatography: porous polymer
gel particles small molecules penetrate and are
retained, large molecules elute more quickly.
x
PAGE polyacrlamide gel electrophoresis:
Aqueous gel: -CH2-CONH- used for
electrophoresis, large molecules experience more
drag and are retained.
Ion Exchange:
Stationary phase is an ionic polymer e.g.
polystyrenesulfonic acid:
x
SO3-
*
*
n
x
Injection valve
x
Page 315 of 316
Chem 155 Unit 17 Chromatography Page 316 of 316
x
strong adsorption.
Chemically inert.