Noise and S/N,

Calibration methods,
Digital and analog signals

Lecture 2
Systematic errors
Present in all measurements made in the same way
and introduce bias.
Instrumental errors
Wacky instrument behavior, bad calibrations, poor
conditions for use
Electronic drift, temperature effects, 60Hz line noise,
batteries dying, problems with calibration equipment.
Personal errors
Originate from judgment calls
Reading a scale or graduated pipette, titration end points

Method errors
Non-ideal chemical or physical behavior
Evaporation, adsorption to surfaces, reagent degradation,
chemical interferences
 Noise
A signal is only of analytical value if it can be
definitively attributed to the species/system of interest
in the presence of noise.

2.5 2.5

2 2
signal

signal
1.5 1.5

1 1

0.5 0.5
0 20 40 60 80 100 0 20 40 60 80 100
data point data point
What is signal and noise?
Signal-to-noise ratio
Signal-to-noise ratio is a measure of the quality of a
method or of instrument performance
Ratio of the mean and the standard deviation of the signal
Higher value of S/N means that its easier to resolve
signal.
x
S
N s
1
S
N RSD

Rev. Sci. Inst., 1966, 37, 93-102.

Where does noise come from?
Chemical noise
Temperature, pressure, humidity, fumes, etc.
Instrumental noise
Detector and post-detector
noise
Thermal (Johnson) noise
Shot noise
Flicker (1/f) noise
Environmental noise
Popcorn (burst) noise
Microphonic noise
Thermal (Johnson) noise
Random motions of charge carriers
(electrons or holes) that accompany thermal
motions of solid lattice of atoms.
Lead to thermal current fluctuations that
create voltage fluctuations in the presence of
a resistive element
Resistor, capacitor, etc.

rms 4kTRf

rms = root-mean-square noise voltage R = resistance of element (W)

k = Boltzmans constant f = bandwith (Hz) = 1/3tr
T = temperature tr = rise time
Thermal (Johnson) noise
continued
Dependent upon bandwidth (f) but not f itself
white noise
Can be reduced by narrowing bandwidth
Slows instrument response time
More time required for measurement
Reduced by lowering T
Common to cool detectors
298K77K lowers thermal noise by factor of ~2
rms 4kTRf
rms = root-mean-square noise voltage R = resistance of element (W)
k = Boltzmans constant f = bandwith (Hz) = 1/3tr
T = temperature tr = rise time
Shot noise
Arises from statistical fluctuations in
quantized behaviors
Electrons crossing junctions or surfaces
Independent of frequency
Example: current
11 e-/s
10.5 e-/s
10 e-/s
irms = root-mean-square noise current
irms 2Ief I = average direct current
e = electron charge
f = bandwidth (Hz)
Flicker (1/f) noise
Magnitude is inversely proportional to the
frequency of the signal
Significant at frequencies lower than 100 Hz
Long-term drift
Origin is not well understood
Dependent upon materials and device shape
Metallic resistors have 10-fold less flicker noise than
carbon-based resistors.
Referred to as pink noisemore red (low
frequency) components
Environmental noise
Comes from the surroundings
Biggest source is antenna effect of
instrument cabling

J. Chem. Educ., 1968, 45, A533-542.

Noise contributions in
different frequency regimes
Frequency
independent

Supposedly 1/f
mostly at low
frequencies

Occurs at discrete
frequencies
Enhancing signal-to-noise
Hardware methods Software methods
Grounding and shielding Ensemble averaging
Difference and Boxcar averaging
Instrumentation Digital filtering
Amplifiers Correlation methods
Analog Filtering
Lock-In Amplifiers
Modulation and
Synchronous
Demodulation
Ensemble averaging
Averaging multiple
data sets taken in
succession
Divide sum of data
sets by number of
data sets
n

S
i 1
i
Sx
n
J. Chem. Educ., 1979, 56, 148-153.
Ensemble averaging
continued
Signal-to-noise improves with increasing
number of data sets

S N n
N
n S
i
 Instrument calibration
Determine the relationship between response
and concentration
Dose response curve
Calibration methods typically involve standards
Comparison techniques
External standard*
Standard addition*
Internal standard*
* calibration curve is required
Comparison techniques
Titrations
Does not require standards
Direct comparisons
Null measurements

C.G. Enke, The Art and Science of Chemical Analysis, 2001.

External standard
Generate calibration curve by measuring
signals and plotting versus known standard
concentrations
Linear regression via least squares analysis
Compare response of sample and solve for
unknown concentration
Subject to interferences due to matrix effects
Concomitant species in sample lead to different
responses for standards at same concentration
Example of external standard
calibration
Determine the amount of atrazine in a drinking water
sample given the following standard and unknown
integrated peak areas from a liquid chromatogram.
Standard Integrated peak area
concentration (ppm) (V)
0 0.02
2 0.64
4 1.21
6 1.74
8 2.38
10 2.98
unknown 1.44
Example of external standard
calibration
Example of external standard
calibration
Determine the amount of atrazine in a drinking water
sample given the following standard and unknown
integrated peak areas from a liquid chromatogram.
Standard
y 0.293 0.004 x 0.03 0.02
concentration Integrated peak area
(ppm) (V) x 4.81 0.09
0 0.02 But how do I calculate the error
2 0.64 in the slope and intercept? And
4 1.21 how do I determine the error in
6 1.74 my concentration?
8 2.38 Ans: use the linear regression
10 2.98 function on my calculator?
unknown 1.44 NO! (at least not for now)
Appendix a1D
y mx b How do I determine the errors in m and b?

Defined terms Sxx, Syy, Sxy (eqns a1-31 to a1-33).

Sxx is the sum of the square of the deviations of individual values
of x from the mean

Syy m 2 Sxx SSresid

sr
N 2 N 2
sr2
sm
Sxx

sb sr
x 2
i

N x x
2
2
i i
Appendix a1D
Now what about the unknown concentration
determined from this calibration curve?
If the mean value is <y> from M replicate analyses,
and the calibration curve contains N data points,
then the error in the determined concentration is

1 1 y y mean
2
sr
sc
m M N m 2 Sxx

ymean is the mean value of y for all N data points

used to generate the calibration curve.
Standard addition
Method for reducing matrix effects
Spikingadding known amounts of
standard solution to sample volume
Assumes that matrix is nearly identical after
standard addition
How to use standard additions
To multiple sample volumes of an unknown,
different volumes of a standard is added and
diluted to the same volume.
S mVs b
b Vx c x
kcs
kVs cs kVx cx m m cs
S Vt
Vt Vt bcs
kVx cx cx
b mVx
Vt
C1V1 C2V2 sy 1 0 y 2
sv 2
m N m S xx
 Determination via standard
curve extrapolation
kVs cs kVx cx
S
Vt Vt
Extrapolating to the x-intercept, where S = 0

kVs cs kVx cx

Vt Vt
Vs cs Vx cx
Vs cs
cx
Vx cs
sc sv
Vx
Internal standard
A substance added in a constant amount to all
samples, blanks, and standards or a major
component of a sample at sufficiently high
concentration so that it can be assumed to be
constant.
Plotting the ratio of analyte to internal-standard as a
function of analyte concentration give the calibration
curve.
Accounts for random and systematic errors.
Difficult to apply because of challenges associated
with identifying and introducing an appropriate
internal standard substance.
Similar but not too similar; cant be present in sample