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Instrumental Analysis

Sang Man Koo

Hybrid Nanoparticles Lab.


Dept. of Chemical Engineering
Hanyang Univ.
Seoul 133-791, Korea

Tel. +82-2-2220-0527
hp: http://hnp.hanyang.ac.kr
E-mail: sangman@hanyang.ac.kr
Instrumental Analysis

• Contents

1. Basics

2. Spectroscopy

3. Instrumentation

4. Spectroscopic Instruments (IR, UV/Vis, and etc.)

5. Classical Analytic Methods (Titrations)


1. Basics on Chemical Analysis
Chemical Analysis

• Chemical Analysis
separation, identification, and the determination of the relative
amount of the components in a sample (analyte) of matter

Classification of Chemical Analysis


1) qualitative analysis : identification of analyte
2) quantitative analysis : relative amount of analyte

Classification of Chemical Analysis


1) classical analysis : the measurement of the weight or
volume of sample to be analyzed
2) instrumental analysis : the measurement of some quantity
that is proportional to the amount of analyte in that sample
Quantitative Chemical Analysis

• Quantitative Chemical Analysis

- determination of relative amount of analyte


1) weight or volume (g, ml)
2) number (moles)
3) concentration: M(molar), N(normal), F(formal), ppm, %

- relationship between the expressions


1) no. of moles = wt/fw
2) no. of moles = C x V

- stoichiometric relationship
aA + bB  cC + dD
Classical vs. Instrumental Methods

• Classical
– Qualitative : identification by color, indicators,
boiling points, odors
– Quantitative : mass or volume
e.g. gravimetric, volumetric

• Instrumental
– Qualitative : identification by measuring physical property
e.g. spectroscopy, electrode potential,
chromatography, electrophoresis
– Quantitative : measuring property and determining
relationship to concentration
e.g. spectrophotometry, mass spectrometry
– Often, same instrumental method used for qualitative and
quantitative analysis
Types of Instrumental Methods

• Types of instrumental methods

1) Emission
–Emission spectroscopy : fluorescence, phosphorescence,
luminescence

2) Absorption
–Absorption spectroscopy : spectrophotometry, photometry,
nuclear magnetic resonance, electron spin resonance

3) Scattering
–Turbidity, Raman

4) Refraction
– Refractometry, interferometry

5) Diffraction X-ray, electron


Types of Instrumental Methods

6) Radiation rotation
– Polarimetry, circular dichroism

7) Electrical potential
– Potentiometry

8) Electrical charge
– Coulometry

9) Electrical current
– Voltammetry - amperometry, polarography

10) Electrical resistance


– Conductometry
Types of Instrumental Methods

11) Mass
– Gravimetry

12) Mass-to-charge ratio


– Mass spectrometry

13) Rate of reaction


– Stopped flow, flow injection analysis

14) Thermal
– Thermal gravimetry, calorimetry

15) Radioactivity
– Activation, isotope dilution
– (Often combined with chromatographic or electrophoretic
methods)
Steps in a Chemical Analysis
• Steps in a chemical analysis

1) selecting a method of analysis


2) sampling : representative of the bulk
3) preparing laboratory sample: small, homogeneous
4) defining replicate samples: improve the quality of results
and liability
- replicate samples: portions of a material of approximately
the same size that are carried through an analytical
procedure at the same time and in the same way
5) preparing solutions of sample
6) eliminating interferences (species other than the analyte that
affect the final measurement)
7) calibration and measurement: C(A) = kX
8) calculating results
9) evaluating results and estimating their reliability
Selecting a Method

• Defining the problem: general, selective or specific

– Precision/Accuracy
– Amount of sample
– Range of analyte concentration
– Interferences
– Physical and chemical properties of sample matrices
– Number of sample
– Time required for analysis
– Professionalism
– Availability of instrument
– Cost
Characteristics of Instrument

• Precision/Bias

• Sensitivity

• Detection limit

• Concentration range

• Selectivity
Precision and Bias

• Precision : Deviation
– Measure of random, or indeterminate error
– Reproducibility
– Standard deviation, Coefficient of Variation
– Figure of merit

• Bias : Error
– Measure of systematic or determinate error
– Bias = (population mean) – (true value)
Sensitivity

• Sensitivity

– Measure of its ability to discriminate between small


differences in analyte concentration
– Tow factors
• Slope of calibration curve - steeper slope
• Precision – better precision
Data Domains
• Data Domains

1) Way of encoding analytical response in electrical or non-


electrical signals : 정보를 코드화하는 방법들을 영역(도메인)이라고
한다

2) Interdomain conversions transform information from one


domain to another.

- Light Intensity => Photocell Current => Current Meter Scale

- Detector (general) : device that indicates change in


environment

- Transducer (specific) : device that converts non-electrical to


electrical data

- Sensor (specific): device that converts chemical to electrical


data
Time Domains

• In time domain, information are stored, as not the magnitude


of a signal, but the function of signal fluctuation with time

- Time
– vary with time
– frequency, phase, pulse width

- Analog
– continuously variable magnitude
– current, voltage, charge

- Digital
– discrete values
– count, serial, parallel, number
Sensitivity

• Sensitivity
– Calibration sensitivity: S
– larger slope of calibration curve means
more sensitive measurement

• Analytical Sensitivity
– Mandel and Stiehler
– AS = slope of calibration curve/std.
deviation
– AS is unchanged with the increase in the
signal amplification of instrument, because
std. deviation also increases with the
increase in the slope of calibration curve
Detection Limit

• Signal must be bigger than random noise of blank

• Minimum signal :
– Signal (min) = Average Signal of blank + ks (blank)
– From statistics k=3 or more
• (at 95% confidence level)
• Suggested by Long and Winefordner
Dynamic Range

• At detection limit we can say confidently analyte is present


but cannot perform reliable quantitation

• Level (Limit) of quantitation (LOQ): (k=10)s(blank)


– the lowest conc. limit for the measurement

• Limit of linearity (LOL)


– when signal is no longer proportional to concentration

• Dynamic range : possible conc. range for the measurement


Selectivity

• No analytical method is completely free from interference by


concomitants. Best method is more sensitive to analyte than
interfering species (interferent).

• Matrix with species A and B


– Signal = m(A)c(A) +m(B)c(B) + Signal blank

• Selectivity coefficient
– kB,A = m(B)/m(A)

• k's vary between 0 (no selectivity) and large number (very


selective)
Errors and Standard Deviations
• Definition

- Mean (arithmetic mean = average = x)


xm = Σ xi/N

- Median: the middle result when data are arranged by size

- spread (range): the difference between the largest value in


the set and the smallest

- Precision: the agreement (or closeness) between two or more


measurements that have been made in exactly the same way

- Accuracy: the closeness of a measurement to its true or


accepted value
Errors and Standard Deviations

• Precision

- Standard Deviation (s): same units as in data


s = [ Σ (xi-xm)2/(N-1)]2

- Relative Standard Deviation: RSD = (s/x) x 1000 (ppt)

- Variance: s2

- Coefficient of variation: CV = (s/x) x 100 (%)


Standard Deviations

• Standard Deviation

- Sums and differences: y = a(±sa) + b(±sb) – c(±sc)


sy = (sa2 + sb2 + sc2)1/2

- products and quotients: y = [a(±sa) x b(±sb)]/c(±sc)


sy/y = {(sa/a)2 + (sb/b)2 + (sc/c)2}11/2

- exponentials: y = a(±sa)x , sy/y = x x (sa/a)


cf. y = a(±sa) x a(±sa) x a(±sa),
sy/y = {(sa/a)2 + (sa/a)2 + (sa/a)2}11/2

- logarithms and antilogarithms:


y = log a(±sa), sy = 0.434 x (sa/a)

y = antilog a(±sa), sy/y = 2.303 sa


Significant Figures
• Significant Figures: all of the certain digits + the 1st uncertain digit
1) disregard all initial zeros
2) disregard all final zeros unless they follow a decimal point
3) all remaining digits including zeros between non-zero digits
are significant

- Sums and differences: smallest figures

- products and quotients: relative value

- logarithms and antilogarithms: to the right of the decimal point


in the original number
(mantissa; right digits of the decimal pomit,
characteristic; left digits of decimal point)

- rounding data
1) choose even or odd number
2) do not round until calculations are complete (guard digit)
Errors and Standard Deviations
• Accuracy

- Absolute Error (E with sign)


E = x(i) – x(t), x(t); true or accepted value

- Relative Error (Er)


percent relative error (Er) = [x(i) – x(t)]/x(t) x 100 (%)

• Types of Errors

- Indeterminate (random) Error: random scattered

- Determinate (systematic) Error: unidirectional (calibration curve)

- Gross Error (outlyer): large, occasional occurrence


affect only a single result
Determinate Errors
• Sources of Determinate Errors

- Instrumental Error: detectable and correctable

- Method Error: difficult to detect


(non ideal physical or chemical behavior)

- Personal Error : prejudice or personal bias

• Effects of Determinate Errors :

- Constant Error: independent on the size of the quantity measured

- Proportional Error: the presence of interfering contaminants in the


sample
Determinate Errors

• Detection of determinate instrumental and personal errors


- periodic calibration, care, and self-discipline

• Detection of determinate method errors


1) analysis of standard samples
standard reference materials (SRM)
2) independent analysis
3) blank determinations (for detecting constant errors)
4) variation in sample size

- bias: the determinate error associated with an analytical method

• Detection of gross errors


- eliminate through self-discipline
Statistical Treatments for Indeterminate Errors
• Statistical treatment of indeterminate errors
- population (universe): data exist in principle (N -> infinity )
- sample: a small set of data which represent a population

• Gaussian Distribution:
- Gaussian Curve: frequency vs. deviation from mean
Normal Error Curve: frequency vs. z {= (x- μ)/σ}

- Properties of normal error curve


1) the mean occurs at the central point of maximum frequency
2) the symmetric distribution
3) an exponential decrease {= exp(-2z2)/(2π)1/2σ}
±1σ ; 68.3%, ±2σ; 95.5%, ±3σ; 99.7%
- standard deviation; sm = s/N1/2, σm = σ/N1/2 (for N>20, s ~ σ)
pooled standard deviation;
spooled = [(s12 + s22 + s32….)/(N1 +N2 + N3 + …. - Ns)]1/2
Statistical Treatments for Indeterminate Errors
• Gaussian Curve vs. Normal Error Curve (NEC)

0.4
Relative Frequency

0.3
0.2
0.1
0.0
-3 -2 -1 0 1 2 3
Statistical Treatments for Indeterminate Errors

• Confidence limits : define an interval (confidence interval)


around xm that probably contains μ

- standard deviation ~ σ
CL for μ = x ± z σ
for N replicate measurement, CL for μ = xm ± z σ/N1/2

- Standard deviation ~ s
CL for μ = x ± ts,
for N replicate measurement, CL for μ = xm ± ts/N1/2
Statistical Treatments for Indeterminate Errors

• Rejection of Outlyers

- T-test:
Tn = |xq – xm|/s,
for entire set including the questionable result

- Q-test:
Qexp = |xq – xn|/w, xn; the nearest neighbor of xq
Statistical Treatments for Indeterminate Errors

• Comparison of experimental results: null hypothesis

- comparison of an experimental mean with a true value


xm – μ = ± ts/N1/2

for N>20, t z, s  σ; xm – μ = ± z σ /N1/2

- comparison between two experimental means


xm1 – xm2 = ± tspooled x [(N1 + N2)/(N1xN2)]1/2

- estimation of detection limits


Δxm = xm1 – xmb > tsb [(N1 + N2)/(N1xN2)]1/2

- comparison between the precision of measurements (F-test)


F = sA2/sB2 > 1