Announcements

Office hours tomorrow 9:05 am at the Mandeville coffee cart

Salinity lab report grades available (review comments, feedback)

Nitrite lab report due Thursday before midnight

(See Lecture Oct 25 for details)
Each team will submit ONE written report and ONE Excel file
Use “SUBMIT WORK” folder on TritonED
Duplicate submissions will be penalized

Do not forget to make comparisons! Consider the nitrite concentration of the

contaminated and local seawater samples. Put your results into perspective.
Are your results unusual, surprising, or as expected?
Are further tests needed? What improvements would you make?
What does the EPA consider to be the maximum allowable limit in
drinking water or seawater? WHY.
 Monochromator
Flame atomizer Iref Detector
Electronics

Lamp Readout

Detector output Electronics includes

Chopper current is Current-to-voltage
Beam proportional to converters;
alternately
reflects &
recombiner photon flux (Iref, I) Amplifiers;
Bandpass filters;
passes Analog-to-digital
beam at steady
frequency.
Analyte converters.

in solution

Design principles
Matrix effects / Chemical Interference
 Light Source

4 1 P1 (4s, 4p)

422.7 nm

41S0 (4s2)
What would create a
photon with the EXACT
energy required for Ca
absorption by calcium?

3
 Light Source

4 1 P1 (4s, 4p) 4 1 P1 (4s, 4p)

422.7 nm 422.7 nm

41S0 (4s2) 41S0 (4s2)

Ca* Ca

A calcium lamp emitting at 422.7 nm can be used to investigate the

transparency (or absorption) of samples at 422.7 nm
Hollow cathode lamp
cathode coated with
element that you want to
analyze with AAS

anode (W)
transparent window

glass shielding
high voltage leads He or Ar gas (~1-5 torr

5
 +
500 Volts Ar
- Ar
Ar

High voltage strips electrons from the gas in the tube

6
 +
Ar+
500 Volts Ar+
- Ar+

Positively charged gas plasma accelerates towards the cathode

7
 +
Ar+
500 Volts Ar+
- Ar+

Positively charged argon ions strike calcium-coated cathode

with sufficient impact to send Ca atoms into the gas phase,
and promote the metal to an excited state.
Ca Ca*
8
 +
500 Volts
Ca*
-

The excited-state Ca atom relaxes back to ground state,

and releases a photon.

Ca* Ca
9
Instrument has a full turret of hollow cathode lamps– can
analyze Pb, Mg, Hg, Zn, Ca, K
 Source: https://www.acs.org/content/acs/en/education/resources/highschool/chemmatters/past-issues/2016-2017/december-
2016/flint-water-crisis.html

Source: http://www.npr.org/sections/thetwo-way/2016/04/20/465545378/lead-laced-water-in-flint-a-step-by-step-look-
at-the-makings-of-a-crisis
 Atomic Absorption Spectrophotometers: schematics
Single-beam AA spectrophotometer

Monochromator Electronics
Flame atomizer Detector
Lamp Readout

Detector output Electronics includes

Chopper current is Current-to-voltage
alternately proportional to converters;
reflects & photon flux (Iref, I) Amplifiers;
passes Bandpass filters;
beam at steady Analog-to-digital
frequency. Analyte converters.

in solution

Question: Why “chop” the lamp (light source) at a fixed frequency, before it reaches the flame?
Answer: The electronics are TUNED to select signals at the chopping frequency. This tuning is
like the tuning of a radio receiver. Therefore, the electronics can REJECT much of the steady or
flickering background signal from the flame that happens to pass through the monochromator
and reach the detector.

Question: How do you “zero” or null a single-beam spectrophotometer?

Answer: Lamp output must be steady. Measure Io before analyte introduced.
A = -log(Io/Io) = 0.000
 Atomic Absorption Spectrophotometers: schematics
Double-beam AA spectrophotometer

Monochromator
Flame atomizer Iref Detector
Electronics

Lamp Readout

Detector output Electronics includes

Chopper current is Current-to-voltage
Beam proportional to converters;
alternately
reflects &
recombiner photon flux (Iref, I) Amplifiers;
Bandpass filters;
passes Analog-to-digital
beam at steady
frequency.
Analyte converters.

in solution

Question: What is the advantage of the double-beam system, over the single beam?
Answer: The reflecting chopper alternately sends the beam through the flame or around the
flame. The detector reads the light intensity via both paths. The rapid alternation at a fixed
frequency compensates for source fluctuation and other noise sources.

Question: How do you “zero” or balance a double-beam spectrophotometer?

Answer: Lamp stability is not so important– I and Iref are rapidly alternated. When no analyte,
A = -log (I / Iref) = 0.000
Optical schematic
From Skoog, Principles of Instrument Analysis
 Flame atomizer I Monochromator Electronics
ref Detector
Lamp Readout

Detector output Electronics includes

Chopper current is Current-to-voltage
Beam proportional to converters;
alternately
reflects &
recombiner photon flux (Iref, I) Amplifiers;
Bandpass filters;
passes Analog-to-digital
beam at steady
frequency.
Analyte converters.

in solution
 Flame atomizer Schematic of a pneumatic nebulizer

http://www.analyticalspectroscopy.net/ap5_html_m470afebb.jpg

Pneumatic Nebulizer:
• Oxidant (air) forced at high pressure past
the liquid at the capillary exit. Creates
aerosol (suspension of fine liquid particles)

• Large particles blocked by flow spoiler

(baffles); excess liquid sent to waste.
Spray chamber
• Small particles reach mixing chamber.
Sample mixes thoroughly with oxidant and
Graphic ,modified from:
Skoog Fundamentals of Analytical Chem 9th ed. fuel before reaching flame.
Figure 28-10, reprinted from Perkin-Elmer Corp.
From Skoog, Principles of Instrument Analysis
 Flame atomizer
(from Harris)

Advantages of Flame atomization Disadvantages of Flame atomization

-High reproducibility • Poor efficiency
-Low cost -(~ 5% sample in the form of small
particles; most are too large
-dilution with oxidizer and fuel gases

• Atoms in the optical path for only a short time.

 Complexities with flames
Position matters

Interconal region

from Harvey

(A classic picture of CH4 –air

combustion) Figure/text from Analytical Atomic Absorption Spectroscopy J. C.
Van Loon 1980

Figure from Skoog Principles of Instrumental Analysis,

(and in turn from Lewis and van Elbe J. Chem. Phys. 1943

Fuel-air ratio: Fuel rich (reducing)? Or fuel-lean (oxidizing)? The ratio acetylene to air is important.
 Important processes in a flame atomizer1

1. Nebulization (creation of small droplets of sample)

e.g. Ca2+(aq) + 2Cl- (aq) + H2O (bulk solution)  Ca2+(aq) + 2Cl- (aq)
+ nH2O (aerosol)

2. Desolvation (remove solvent with heat)

Ca2+ (aq) + 2Cl- (aq) + nH2O (aerosol)  CaCl2 (solid) + nH2O (g)

3. Atomization (atoms created from solid particles

CaCl2 (solid)  Ca (g) + 2Cl (g)

4. Spectroscopy (light/matter interaction– either absorption or emission)

5. Condensation of products (removal via exhaust)

1Adapted from: Spectrochemical Analysis by Atomic Absorption and Emission Spectroscopy (1st edition,1992) by Lajunen;
Spectrochemical Analysis by Atomic Absorption and Emission Spectroscopy (2nd edition, 2004) by Lajunen and Perämäki
 From Skoog, Principles of Instrument Analysis

Figure from Analytical Atomic Absorption Spectroscopy J. C. Van Loon 1980

22
 In the previous slide, the claim is made that the gas-phase solid particles (e.g. NaCl,
KCl, CaCl2 , etc.. tend to form neutral atoms.

Why don’t we form charged particles, e.g. K+ and Cl- , or Ca2+ and 2Cl- for your lab?
(actually, some ions are in fact formed, and that is a error in the technique that can be
addressed)

Fundamental question: What is the energy difference for homolytic and heterolytic
cleavage of KCl, CaCl2?

KCl(g)  K(g) + Cl(g) ΔH°homolytic CaCl2(g)  Ca(g) + 2Cl(g) ΔH°homolytic

KCl(g)  K+(g) + Cl-(g) ΔH°heterolytic CaCl2(g)  Ca2+(g) + 2Cl-(g) ΔH°heterolytic

K(g) + Cl(g)  K+(g) + Cl-(g) ΔH°heterolytic – ΔH°homolytic Ca(g) + 2Cl(g)  Ca2+(g) + 2Cl-(g) ΔH°heterolytic – ΔH°homolytic

The magnitude of ionization energies exceeds electron affinities for

elements. For example:
K(g)  K+(g) + e-(g) 418.8 kJ/mol Ca(g)  Ca2+(g) + 2e-(g) 1145.4 kJ/mol
Cl(g) + e-(g)  Cl-(g) -348.6 kJ/mol Cl(g) + e-(g)  Cl-(g) -348.6 kJ/mol

K(g) + Cl(g)  K+(g) + Cl-(g) +70.2 kJ/mol Ca(g) + 2Cl(g)  Ca2+(g) + 2Cl-(g) +448.2 kJ/mol

It takes considerably more energy to form the ions relative to the neutral atoms
**in the gas phase**.

Our intuitions can be biased by thinking of these atomic species as solvated. If water is
around, then ions are strongly stabilized.
 One alternative to flame atomizer– graphite furnace

Quantitative Chemical Analysis 8th ed - Harris

 Monochromator
Flame atomizer Iref Detector
Electronics

Lamp Readout

Detector output Electronics includes

Chopper current is Current-to-voltage
Beam proportional to converters;
alternately
reflects &
recombiner photon flux (Iref, I) Amplifiers;
Bandpass filters;
passes Analog-to-digital
beam at steady
frequency.
Analyte converters.

in solution
 Photomultiplier Tube (PMT)
Light/Photon Detection

http://www.olympusmicro.com/primer/digitalimaging/concepts/photomultipliers.html

Photodiode

Charge-coupled device

http://www.thorlabs.us/thorproduct.cfm?partnumber=FGAP71

http://machinedesign.com/archive/forgot-film-no-problem-digital-cameras
 Photomultiplier Tube (PMT)

Advantages
Extraordinary amplification: converts incoming
photons into a cascade of electrons (e.g. 106
electrons per photon)

High sensitivity – capable of detecting single

photons

Fast response time (can be sub-nanosecond)

Spectral window can cover from UV to near IR.

Disadvantages:

Relatively large detector

Expensive

http://www.olympusmicro.com/primer/digitalimaging/concepts/photomultipliers.html
Requires high voltage source (typically 1000 –
2000 Volts)
PMT (inside)

Hollow cathode Lamps

Flame atomizer
What if we lose our hollow cathode lamps – is all lost?
Flame atomic emission spectroscopy
Think sodium street lights
 Linear vs non-linear: LINEST does both, but clunky.

𝟐
1.000
𝒚
Absorbance

0.800

0.600

0.400
b~m slope
0.200

0.000
0.0 5.0 10.0 15.0 20.0 𝒚
Concentration Ca, ppm
1.000 𝟐

0.800
𝒚
Absorbance

0.600
=LINEST(C2:C6,A2:A6,TRUE,TRUE)
0.400
=LINEST(C2:C6,A2:A6^{1,2},TRUE,TRUE)
0.200
𝟐
0.000
0.0 5.0 10.0 15.0 20.0
Concentration Ca, ppm
 Linear vs non-linear: Our friend Analysis toolpak
SUMMARY OUTPUT

Regression Statistics
1.000 Multiple R 0.999965218
R Square 0.999930438
Adjusted R Square 0.999860876
Absorbance

0.800 Standard Error

𝒚 0.004199155
Observations

0.600 ANOVA
df SS
Regression 2 0.506933934
0.400 Residual 2 3.52657E-05
Total 4 0.5069692

0.200 Coefficients Standard Error

Intercept 0.013410494 0.004671837
x 0.06178785 0.001094238
0.000 x^2 -0.000719351 5.04793E-05
0.0 5.0 10.0 15.0 20.0
Concentration Ca, ppm b~m slope for eq 4-27
RESIDUAL OUTPUT
1.000
Observation Predicted y Residuals
1 0.074478993 -0.000478993
0.800 2 0.304365963 -0.000365963
Absorbance

3 0.559353866 0.003646134
4 0.778374203 -0.004374203
0.600 5 0.961426974 0.001573026

0.400 0.006
0.004
Residuals

0.200 0.002
𝟐 0
0.000 -0.002 0.0 5.0 10.0 15.0 20.0
0.0 5.0 10.0 15.0 20.0 -0.004
Concentration Ca, ppm -0.006
 WEEK 2
MATRIX EFFECTS
What about matrix effects?
Recall these are the effects of other species on the analyte
Standard (S=35) seawater has 10.78 g sodium per kg, or 10.78 ‰

Suppose you dilute your

seawater by a factor of 40.
What is the ppm level of sodium?

Should your calibration solutions

have sodium as well ?

And why am I focusing on

sodium in the first place, rather
than Cl-, or K+ or…..

Table 6.2 from: A.G. Dickson & C. Goyet. Handbook of methods for the analysis of the various parameters of the carbon dioxide system in sea water (1997)
From Agilent “Flame Atomic Absorption
Spectrometry Analytical Methods ” 2015
Sodium is relatively easily ionized, and it is a known “ionization suppressor”

Harris pg 545.

If you find in a couple of “spot checks” that you get significantly different signals for
calcium with and without added sodium, one way to solve that problem is to add 1000
ppm or 2000 ppm potassium to all solutions– standards and unknowns.

In that case, ionization of Ca is suppressed uniformly for all solutions.

 What about chemical interferents?
• 3PO43- + 2Ca2+  Ca3(PO4)2 impedes atomization of Ca
• SO42- + Ca2+  CaSO4 impedes atomization of Ca
• Al3+ can also have a detrimental effect (form Ca-Al oxides)

• Are these species in seawater? How much, and to what

should we compare?

• Does dilution help, as you have done in your lab?

• What do you do if there is a problem? Higher flame

temperature can help reduce or eliminate problem with
phosphates, and in general can overcome loss of signal
caused by oxide formation.

• Can add substances such as lanthanum ions (La3+) to trap

the PO43- ions. Called a RELEASING AGENT.

• Or can add a PROTECTING AGENT that binds Ca2+ and

allows it to atomize without a problem (e.g. EDTA,
ethylenediaminetetraacetic acid) http://en.wikipedia.org/wiki/File:Sea_salt-e-dp_hg.svg
 How much do interferents or matrix
effects matter in your analysis of
Ca2+ in seawater?

“Physical and thermodynamic data” Chapter 5 of Handbook of methods for

the analysis of the various parameters of the carbon dioxide system in sea
water A.D. Dickson, C. Goyet (Editors)
 From Agilent “Flame Atomic Absorption
Spectrometry Analytical Methods ” 2015

From Perkin Elmer “Analytical Methods for Atomic

Absorption Spectroscopy” 1996
The method of standard addition

In the method of standard addition, known quantity of analyte is added or “spiked”

to sample to create a set of calibration standards
The method of standard addition
Example from Harris,
Quantitative Chemical Analysis

Illustration with strontium (Sr):

Interferences affect the signal of Sr

that exists at the outset (i.e. no
added Sr2+) and also the sensitivity
(slope) upon addition of Sr2+

The use of external

standards

i.e. Sr2+ in pure H2O would

be a poor way to assess the
Sr2+ in aquarium water.
4.6 ppm is far off the value
of 7.41 ppm which is found
by the method of standard
addition.
Text from
Harris
 Slope is key to understanding standard addition and matrix effects.

(A)
𝟐

Ca2+ FROM STANDARD ADDITION (ppm after diluted)

(~ ) 𝑑𝑖𝑙𝑢𝑡𝑖𝑜𝑛 𝑓𝑎𝑐𝑡𝑜𝑟
𝐶 ., 𝑝𝑝𝑚 = −(𝑥 − 𝑖𝑛𝑡𝑒𝑟𝑐𝑒𝑝𝑡)
(~ ) 𝑓𝑜𝑟 𝑆𝑊
Harris pg. 108-109
A worthwhile problem to study:
From Harvey, pgs 631-633: “Representative Method
10.4-Determination of Sodium in a Salt substitute.”
The method employs atomic emission rather than
absorption, but similar concepts apply.

45
See Harris Chapter 5 regarding uncertainty in the x-axis determination
Also a few words in Appendix B. One point is that one should determine the
uncertainty in the x-intercept in a naïve way from x = -b/m.

Why not?
Compare: