About Instrumental Analysis (1)

Lecture 1

(1) About the class

(2) Instrumental analysis – what and why?

(3) Who you are and prerequisites

(4) How grading will work

 What is instrumental
analysis?

Instrumental analysis is the

study of how best to use
instruments to analyze the
molecular composition of stuff.
 Instrumental Analysis:
Why?

To keep To understand To ensure laws

children safe important things are obeyed
 Topics in Instrumental Analysis
Topic IA Part 1 IA Part 2
Error, calibration, QA/QC Yes
Spectroscopy: Atomic Yes
Mass Spectrometry I Yes
Chromatography Yes
Spectroscopy: Infrared Yes
Electrochemical Sensors Yes
Spectroscopy: Optical Yes
Spectroscopy: Fluorescence Yes
Electroanalytical methods Yes
Mass Spectrometry II Yes
Nuclear Magnetic Resonance (NMR) Yes
 Instrumental Analysis:
What’s ‘in’ or ‘out’

• ACS exams in analytical chemistry and

instrumental analysis provide topics

• This class is 80% of an instrumental

analysis class and 20% of a quantitative
analysis class
 Who are you?

• A student who needs to get this material for

degree purposes or exam purposes

• An adult learner with a keen interest in analysis

and chemistry

• Not an expert in analytical chemistry or

instrumental analysis (!)
 Background Needed for
Instrumental Analysis

• Take the course pre-quiz! (need chemistry)

What is sulfate? How do you find its molecular weight? What is more
polar, methanol or benzene?

• Have access to excel or equivalent

• Be willing to work up to 10 hours/week

 Homework and Exam Calendar
Certificate in the class: complete 70% of the assignments, participate all weeks
Certificate with distinction: complete 85% of the assignments, participate all weeks

Early May Late May Early June Late June

Case Study 1 Case Study 2 Case Study 3

Lead in toys Chocolate Breathalyzers
Error bars
Calibration Chromatography
Infrared spectroscopy
Managing Error
Electrochemical sensors
Atomic Spectroscopy

3 problem sets 3 problem sets 2 problem sets

6-9 quizzes 6-9 quizzes 6-9 quizzes
Case 1 Case 2 Final Exam
Case 3
 About Instrumental Analysis
Lecture 1

(1) The class is about instrumental analysis

(2) Most modern chemical analysis is done with

instruments- huge importance, multiple areas

(3) You need some chemistry background

(4) Weekly quizzes, problems sets, case studies and a

final exam all are part of the final grade.
 Chemistry
About Instrumental Review
Analysis (1)
Lecture Lecture 2Analysis
ME 2: Dimensional

Dimensional Concentration
Analysis and Solutions

(1) Units in calculations

(2) The concept of dimensional analysis

(3) Examples of applying dimensional analysis

 Dimensional Analysis:
Going from one unit to another

You have 72 apples, how many dozen?

 Dimensional Analysis:
Going from one unit to another

• Mass units: kilogram, gram, pound (rare)

• Volume units: liter, milliliter, microliter

• Concentration units: parts per million, parts per

billion, molarity, molality, weight %, volume %

http://chemistry.about.com/od/lecturenotesl3/a/concentration.html (about concentration units)

http://lamar.colostate.edu/~hillger/common.html (about the metric system
 Conversions: Essential
Information for Analysis

• Mass units

• Volume units

• Concentration units
Dimensional Analysis:
Multiply by 1
 Dimensional Analysis:
Examples

You have 34.5 kg of sodium sulfate, how many grams do you have?

You have 1.0 quart of milk, how many milliliters do you have?
 Hi Students: So in the original I had
grams in the problem on the bottom, and
of course it should have been kilograms. Dimensional Analysis:
Thanks for the catch and its fine now!
More Examples
Dr. Colvin

You have 34.5 kg of sodium sulfate, how many moles do you have?

You have .100 moles of aluminum nitrate, how much is that in kilograms?
About
Dimensional
Instrumental
Analysis
Analysis (1)
Lecture
Lecture 22: Dimensional Analysis

(1) Units in calculations

(2) The concept of dimensional analysis

(3) Examples of applying dimensional analysis

 Concentration in
Analysis
Lecture 3

Dimensional Concentration
Analysis and Solutions

(1) Typical units of concentration

(2) Calculating concentration (examples)

(3) Calculating dilutions (examples)

Concentrations of
Solutions

“Concentration”: Characterizes ‘how much’

of something (X) is in solution:

Molarity: moles of X in one Liter solution

ppm: milligrams of X in one Liter solution

ppb: micrograms of X in one Liter solution

 Approach to Concentration
Calculations

Moles Moles
(molecule) (atom) Molarity

Weight
(grams) = ppb (ug/L)

Volume
ppm (mg/L)
(liters)
 Examples of simple
concentration calculations

You have .345 mg of sodium sulfate in 1L of water, what is the molarity of the solution?

You have .100 moles of aluminum nitrate in .500 L of water, what is the ppm of Al?
 Hi Students: When I did the first problem, I typed in sodium phosphate into
wikipedia to get the molecular weight. A common short-cut and confusing
in this case! It gave me the molecular weight for monosodium phosphate
(120), not trisodium phosphate which is what I show below. Always best to
More
calculate from the formula which is what I used to do, and will now do
again! It did not affect the numeric answer in this example, but best idea is
to calculate MW from formula weights.
concentration
Had a transcription error – right answer is .03069 as shown not .0369 as
originally written. This is corrected.
Dr. C
examples
You have 1.023 molar tri-sodium phosphate, how many moles sodium are in 10.0 ml?
164 gm/mole

.03069

How many atoms of aluminum are in a 1 microliter of a 1 ppb solution of Al(NO3)3?

 Practice with Dilutions
You need to make 50 ml of a .050 M solution of aluminum from a 1 M stock of
aluminum nitrate: how will you do this?

Way 1 – more conceptual, less efficient

Way 2 – less conceptual, more efficient

 Practice with Dilutions

You diluted 2.00 ml of an unknown lead solution into .500 L and found a 10.3 ppb
concentration in the diluted sample. What was the molarity of the unknown?
 Units of Concentration in Analysis
Lecture 3A

(1) Typical units of concentration

(2) Calculating concentration (examples)

(3) Calculating dilutions (examples)

 Analysis Error:
Significant Figures
Lecture 4

Significant Replicate Confidence Error

Figures measurements limits propagation

(1) Precision and accuracy in measurement

(2) Instrument tolerances

(3) Significant figures

How good does your
number have to be?
The US government now says, less than
100 ppm lead in toys.

But is that

100.00? 100.0000? ~ 100? ppm

There are two main
types of error

• Random or indeterminate – precision

• Systematic or determinate – accuracy

ME Not precise OR not
accurate?
Are these
precise?
accurate?
 Precision: Central Issue to
Communicate

Beginner: significant figures

Answer = 50.2 ml
and estimating error
Advanced beginner: averages
Answer = 50.2 ± .3 ml
and standard deviation
Intermediate: combining errors Answer = 8.2 ± .3 gm/ml
and linear regressions
Advanced Intermediate: using Answer = 8.2 ± .5 gm/ml
confidence limits with 90% confidence
 Expected error:
Instrument Tolerances

Every instrument has an instrument tolerance – the accuracy one can expect
 Expected error:
Instrument Tolerances
Graduated cylinder ± 2 ml

Volumetric flask ± .01

Pan balance ± 0.1 gm

Analytical balance ± 0.0001 gm

Manufacturer’s will report tolerances as ± .0001 on primary unit of measurement

 Don’t Lie!
(or convey false precision)

The metal toy sample weighed 5.1235 grams

The metal toy sample weighed 5 grams

Significant figures are the minimum number of digits needed to

write a given value in scientific notation without loss of precision.

http://en.wikipedia.org/wiki/Significant_figures
 Analysis Error:
Significant Figures
Lecture 4B- Examples

Significant Replicate Confidence Error

Figures measurements limits propagation

(1) Precision and accuracy in measurement

(2) Instrument tolerances

(3) Significant figures

http://www.ruf.rice.edu/~bioslabs/tools/data_analysis/errors_sigfigs.html
 Some dense rules

• How many sig figs?

OPTION A: You the user set your personal tolerance in ability to use an instrument
OPTION B: OR you look up the manufacturer’s reported tolerance

TOLERANCE a ± value in units of measured value which defines expected uncertainty

RULES for combining significant figures

-- Multiplication/division: Answer rounded so it has same number of
sig figs as smallest number in calculation (weakest link)
-- Addition/subtraction: Number of sig figs in answer given by the
number with smallest number of places after decimal place

When rounding – look at all digits beyond last place desired. If exactly halfway, round
to nearest even digit (why do we do this?)
 Independent Exercise:
Significant Figures

Ex. 1: Round
(A) 1.2367 to 4 SF:
(B) 0.1352 to 3 SF:
(C ) 2.051 to 2 SF:
(D) 2.0050 to 3 SF:
(E) 3.0150 to 3 SF:

Ex 2: Write each answer with the correct SF:

(A) 1.021 + 2.69 =
(B) 12.3 – 1.63 =
(C) 4.34 * 9.2 =
(D) .0602 ÷ 2.113·104 =
 Independent Exercise:
Significant Figures

Ex 3: Find the molecular weight of KrF2. We know Fluorine’s atomic weight to very high
precision (18.9984032 ± .0000005 g/mol). Krypton is less well defined (83.90 ± .01
g/mol).

Ex. 4: Rewrite the number 3.12356 ± 0.16789% in the forms (a) the absolute uncertainty and (b)
the relative uncertainty. Use the correct number of digits.
Analysis Error: Significant Figures
Lecture 4

(1) Precision and accuracy in measurement

(2) Instrument tolerances

(3) Significant figures

 Error Bars from Replicate
Measurements Lecture 5

Significant Replicate Confidence Error

Figures measurements limits propagation

(1) How taking replicate datasets improves data

(2) Getting the error bar from replicate data

(3) Confidence limits and the error bar

 Uncertainty and
‘Doubt’

uncertainty (of measurement)*

parameter, associated with the result of a measurement, that characterizes the
dispersion of the values that could reasonably be attributed to the measurand

Does the paint have 90 ppm lead?

or 110 ppm lead?

* September 2008 “Evaluation of Measurement Data- Guide to the Expression of Uncertainty”

http://www.bipm.org/utils/common/documents/jcgm/JCGM_100_2008_E.pdf
 Do it Over and Over

Analytical balance ± 0.0001 gm

Manufacturer’s instrument tolerances should be < replicate measurement error!

 The “ERROR BAR”:
Communicating Uncertainty

Measurement = Average ± uncertainty

The result is only 20% of the game; the

real challenge is in defining the error.

Really good book if you want to learn more:

Statistics for Analytical Chemistry, J. C. Miller and J. N. Miller (Chichester, 1992)
 Replicate Measurements:
N
Resolving Precision
x
Answer  x  s
i
x i 1
N

The standard deviation, s, means

that ___________ % of
measurements will fall between
average –  and average plus .

 (x i  x) 2
s i 1
N 1
 Example: Average and
Standard Deviation
Hi Students: Note, when I did this the first time I used Excel’s stdev function and somehow got
a stdev which is a divide by N standard deviation – excel now has a whole variety of stdev –
stdev.p, stdev.s for example. So, I redid it with the stdev.s function which is the sample
standard deviation and the answer has been updated –note it hardly changed the answer
(before it was s=1.85 with N=6) but I wanted to be as clear as possible.
Thanks! Dr. C
You are measuring the lead in teething toys, and find in 6 samples lead measurements
of 102.3, 100.1, 98.56, 104.3, 99.89 and 101.0 ppm. What is the average and
uncertainty (both absolute and relative) of this dataset? How do you report the data?

The average of these six measurements is 101.0 ppm

The standard deviation is 2.028 ppm

You would report either 101.0 ± 2.0 ppm Or you would report 101.0 ± 2%
You should only seldom keep more than one significant figure in the standard deviation
because it has error too – possibly two significant figures are OK but never more.
 Error Bars from Replicate
Measurements Lecture 5

(1) How taking replicate datasets improves data

(2) Getting the error bar from replicate data

(3) Confidence limits and the error bar

 Error Bars with Confidence
ME Lecture 6

Significant Replicate Confidence Error

Figures measurements limits propagation

(1) The concept of a ‘confidence’ limit

(2) Expanding or contracting error bars for confidence

(3) Defining an error bar with confidence

 Different Approaches to
Conveying Uncertainty

Beginner: significant figures

Answer = 50.2 ml
and estimating error
Advanced beginner: averages
Answer = 50.2 ± .3 ml
and standard deviation
Intermediate: combining errors Answer = 8.2 ± .3 gm/ml
and linear regressions
Advanced Intermediate: using Answer = 8.2 ± .5 gm/ml
confidence limits with 90% confidence

Measurement = Average ± uncertainty (CL=90%)

 How certain is your uncertainty?
N

x i
x i 1
N

The standard deviation, s, means that 68.3 % of

measurements will fall between average –  and
average plus .

But 2s means that 95 % of measurements will fall

between average – 2 and average plus 2.
 Defining error more precisely:
confidence intervals

If factor < 1 then we will have LESS

confidence that measurements will
fall in this range

If factor = 1 then its just the

Answer  x  (factor )  s standard deviation and we have a
68% confidence that new
measurements are in this range

If factor >1 then we will have MORE

than 68% confidence that new
measurements are in this range
 A t-table gives the exact
factors
Less confident More confident

Larger range Smaller range Larger range

One
measurement Factor >> 1 Factor ~ 1 Factor >> 1
N: Degrees of freedom

Answer  x  (factor )  s

Many Smaller range

measurements Factor ~1
 Degree of confidence (%)
Degrees of Freedom
90 95 99 99.9
1 6.31 12.71 63.66 636.62
2 2.92 4.30 9.93 31.60
T-table 3 2.35 3.18 5.84 12.92
4 2.13 2.78 4.60 8.61
5 2.02 2.57 4.03 6.87
6 1.94 2.45 3.71 5.96
7 1.89 2.37 3.50 5.41
8 1.86 2.31 3.36 5.04
9 1.83 2.26 3.25 4.78
t s
Answer  x 
10 1.81 2.23 3.17 4.59
11 1.80 2.20 3.11 4.44
N
12 1.78 2.18 3.06 4.32
13 1.77 2.16 3.01 4.22
14 1.76 2.14 2.98 4.14
15 1.75 2.13 2.95 4.07
16 1.75 2.12 2.92 4.02
17 1.74 2.11 2.90 3.97
18 1.73 2.10 2.88 3.92
19 1.73 2.09 2.86 3.88
20 1.72 2.09 2.85 3.85
21 1.72 2.08 2.83 3.82
22 1.72 2.07 2.82 3.79
23 1.71 2.07 2.82 3.77
24 1.71 2.06 2.80 3.75
Infinity 1.65 1.96 2.58 3.29
 Example: Using Confidence
Limits
two different kinds of problems

• Type 1: you have a set of experimental data and can find ‘s’ from
calculating the standard deviation. You wish to report the error bars
and specify a confidence limit:

t s
Answer  x 
Specify degrees of freedom (N-1), confidence limit
Then use t-table to look up the “t” parameter
N
• Type 2: You know the s for a method (past history/experience/ error
validations) that it is the REAL . You collect experimental data with a
given N and wish to report the error bars and confidence limit:

t s t 
Some books call this t the

Answer  x   Answer  x 
parameter ‘z’ and pulls out the
N=infinity t-table in a separate
N N table
Specify an infinite degree of freedom and desired confidence to look up ‘t’
Remember for ‘N’ to use the number relevant in specific experimental case
 Hi Students: Note, when I did this the first time I used Excel’s stdev function which is a divide
by N standard deviation – even though it claims it isn’t! So, I redid it with the stdev.s function
which is the sample standard deviation and the answer has been updated –note it hardly
changed the answer (before it was s=.47) but I wanted to be as clear as possible.

Note that a similar issue was in lecture 5 as well. Check my posts to see more information.
Thanks! Dr. C

EXAMPLE: Using a method you developed yourself to measure zinc in tablets,

you find 14.1, 15.2, 14.8, 15.5, 15.3, 14.6 and 14.9 mg Zinc per capsule. Find the
error bars on the average value of zinc in the tablets at a 95% confidence limit.

ts ts
Answer  x   Answer 14.9 mg Zn 
N N

s: calculate the standard deviation using a calculator to be s = .52

t: we find ‘t’ from the t-test table. We use a DF = 7 – 1 = 6 and a 95% CL: t = 2.45
N: We use N= 7 as that is the actual measurements

2.45  .52
Answer 14.91 mg Zn   14.9 mg Zn  0.5 mg Zn
7
 Example: You measure the zinc level in vitamin tablets
using a NIST method that reports the standard error to
be ± .30 mg Zinc. How many replicate measurements
would you need to make to get the 95% confidence limits
to under ± .42 mg Zinc?

t s t  You get t from the t-table

Answer  x   error   .42mg at 95%  
N N BUT you use DF as infinite since you know 

 t    1.96  .30 
2 2

N      1.96 round to 2
  error   .42 
Error Bars with Confidence
Lecture 6

(1) The concept of a ‘confidence’ limit

(2) Expanding or contracting error bars for confidence

(3) Defining an error bar with confidence

 Combining Error Sources
Lecture 7

Significant Replicate Confidence Error

Figures measurements limits propagation

(1) The importance of error propagation

(2) Rules for error propagation

(3) Examples of error propagation

 How can we figure out what
these cylinders are made of?

Tolerance is _______
Tolerance +/- .1 grams

credit: PRHaney

ANSWER: 310 or 310.1 or 310.05 grams?

ANSWER: 38.7 or 38.75 or 38 ml?
 Error Propagation

If you are adding or subtracting different values then you

combine their absolute error by:

Answer = a + b - c

If you are multiplying or dividing then you combine their

relative errors as shown:

Answer = a·b/c
 Example: Error Propagation

You measure a metal rod of unknown composition and find using a graduated cylinder that it has a volume of
50.3 ml and you weigh it on an analytical balance and find a weight of 361 grams. (A) What are the standard
errors for volume and weight? (B) What is the density with error? (C) What is the rod made of?
Remember, we use instrument tolerance if we did not do replicate measurements
(A) For graduated cylinders it is ± .2 ml and for analytical balances it is ± .0005 gm (see first page)

Density is defined as mass over volume, so this means we need relative errors
.2 .0005
.00397 .0040 1.39 6
50.3 361

Now you just divide mass by volume, and propagate the error
361
7.18 /
50.3

So the answer would be 7.18 gm/ml ± .4% or if you convert back to absolute 7.18 ± .03 gm/ml
 Example: A 218.44 ± 0.01 gram cylinder has a diameter of 2.50
± 0.01 cm and is 5.00 ± 0.01 cm long. What is the density of
the metal?

Note: There
was a really
interesting
thread on
whether you
could lower the
error by writing
the volume in
terms of
diameter! An
interesting
conversation,
with no exact
resolution. My
take is that I
had envisioned
the error as
due to calipers
– so the error
on 1.25 would
be the same
as on 2.50
 .2

247

1.9% or 2%
Combining Error from
Multiple Sources
 Using Error Bars to Make Decisions
Lecture 8

(1) Characterizing bias in measurement

(2) Can I throw out a datapoint?

 Precision: Central Issue to
Communicate

Beginner: significant figures

Answer = 50.2 ml
and estimating error
Advanced beginner: averages
Answer = 50.2 ± .3 ml
and standard deviation
Intermediate: combining errors Answer = 8.2 ± .3 gm/ml
and linear regressions
Advanced Intermediate: using Answer = 8.2 ± .5 gm/ml
confidence limits with 90% confidence

THIS LECTURE: How do we use error bars?

 Decision 1: Is there
systematic error ?

• Before running an ‘unknown’ run a standard

• A known standard would have been verified by an outside company,
ideally using another method
• Many companies sell standard reference materials, and these should
be incorporated into experimental design

Systematic error? Run a known standard (ideally several) and troubleshoot

 Answer = Average ± uncertainty

The true value The true value

- uncertainty Average + uncertainty

1: Systematic error check. You are comparing an experimental result against a known
precise and accurate value. If the precise and accurate value is within your error bounds
then there is no systematic error.

x  known
t N if t  t X , N then with X% confidence they are the same
sx

• Compute the ‘t’ value for dataset and known standard

• Compare this t value to the t-table for desired confidence, N
• If experimental t is less than the desired t: difference is not significant
• If experimental t is larger than desired t: difference is significant

* By convention, x% is generally 95% confidence limit if not specified

 Is there systematic error? Example
You have bought a standard for chloride analysis that has a chloride content of
20.00%. Six repeated measurements of this standard result in an average value
of 20.06% ± 0.025%. Is there evidence of systematic error in your
measurements?
x  known
t N if t  t 95, N then with 95% confidence they are the same
sx

Now you simply plug in the values from the problem to get an actual t value

20.06  20.00
t 6  5.9  2.571  t95,5
0.025

The computed t-value is 5.9 which is greater than the t(95,5) or 2.571

We thus conclude that the known value and experimental values are not the same and
that there is a source of systematic error
 The Q-Test: Tossing out a Datapoint
• Q: Divide the ‘gap’ between outlier and nearest neighbor by
range of all measurements – call this Qexp

• If calculated Qexp is larger than Qrej from the table below,
you can neglect the datapoint in question
• This is a subjective decision about how confident you want or
need your measurement to be

Example: You have six measurements: 102, 97, 90, 110, 108, 65. Can you throw one out?
First put in descending order – outliers have to be biggest or smallest: 110, 108,102,97,90,60

The top is easy then, just the outlier – nearest value: 90-60 = 30 (note take the absolute values, sign won’t matter)

The bottom is the range of values including outlier: 110 -60 = 50

Qexp is this 30/50 = .6 and the number of measurements is six

You can throw out 60 with 90% but not 95% confidence!

Number 3 4 5 6 7 8 9 10
of values:
Q90%: 0.941 0.765 0.642 0.560 0.507 0.468 0.437 0.412

Q95%: 0.970 0.829 0.710 0.625 0.568 0.526 0.493 0.466

Q99%: 0.994 0.926 0.821 0.740 0.680 0.634 0.598 0.568

http://www.chem.uoa.gr/applets/AppletQtest/Text_Qtest2.htm and wikipedia.org

Using Error Bars to Make Decisions
Lecture 8

(1) Characterizing bias in measurement

(2) Can I throw out a datapoint?

 Summary of Error Bars
Beginner: significant figures
Answer = 50.2 ml
and instrument tolerances
Advanced beginner: averages
Answer = 50.2 ± .3 ml
and standard deviation
A) Communicating
the precision in a Intermediate: combining errors Answer = 8.2 ± .3 gm/ml
measurement and linear regressions
Advanced Intermediate: using Answer = 8.2 ± .595% gm/ml
confidence limits (e.g. with 95% CL)

Answer = Average ± uncertainty

Decisions

B) Using error to make decisions about your data

Evaluating your Deciding if two Deciding whether

data for datasets to
systematic error are different toss ‘bad’ data
Instrumental Analysis: Error Module Summary

(1) Accuracy and precision

(2) Instrument tolerances and significant figures

(3) Replicate measurements and confidence limits

(4) Propagating error

(5) Using error bars to make decisions