Analytical Chemistry Laboratory Techniques and Instrumentation

I. Basic Laboratory Tools and Operations: Terms to remember

Exercise 1. Match the following terms with the items below.
a. Reagent grade chemicals
h. Analytical balance
b.Primary-standard grade chemicals i. macrobalance
c.Special purpose reagent chemicals j. Semimicroanalytical
d. Tare
k. Microanalytical
e. Weighing by difference
l. Transfer pipets
f. Parallax
m. Measuring pipets
g. Meniscus
n. Micropipets

o. TC 20oC
p. Desiccator
q. Drierite
r. Decantation
s. MSDS

Items
1. mass of the empty vessel
2. the error that occurs when your eye is not at the same height as the top of the liquid
3. type of balance whose maximum capacity is 1g-1kg precision at max.cap.of at least 1/10 5
4. type of balance whose maximum load is 10-30g precision of 0.01 mg.
5. type of balance whose max. load is 1-3 g precision of 0.001 mg.
6. type of balance whose max. load is 160-200g precision of 0.l mg.
7. weighing technique used routinely and is necessary for hygroscopic reagents.
8. concave formed at the surface of most liquids.
9. means "to contain at 20C"
10. also known as Mohr pipet and is calibrated like a buret, used to deliver a variable volume.
11. type of pipet which is calibrated to deliver one fixed volume.
12. a closed chamber containing a drying agent called a dessicant.
13. A type of dessicant
14. Conform to the minimum standards set forth by the Reagent Chemical Committee of the
American Chemical Society (ACS) and are used wherever possible in analytical work.
15. Reagents that have been carefully analyzed by the supplier and the assay is printed on the
container label.

16. Chemicals prepared for a specific application

17. A listing of hazards and safety precautions for a chemical sold in the US. It also gives first aid
procedures and instructions for handling spills.
18. process of pouring a liquid gently so as to not disturb a solid in the bottom of the container.

II. Steps in a typical Quantitative Analysis

1. Select method -level of accuracy required, no. of samples to be analyzed, complexity and no of
components in the sample.
2. Sampling- involves obtaining a small mass of a material whose composition accurately
represents the bulk of the material being sampled; source of greatest error
3. Prepare a laboratory sample- decrease particle size to assure homogeneity, storage, drying
/moisture determination.
4. Define 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. Dissolve the samples- water, acids, bases, oxidizing agents, reducing agents, combination of
reagents, ignition, and high temperature fusion of sample in presence of fluxes

6. Eliminating interferences- an interference is a species that causes an error by enhancing or

attenuating (making smaller) the quantity being measured in an analysis. Specific techniqueswork for only one analyte; selective techniques- work only for only a few analytes.
7. Calibration and Measurement- Calibration- empirical determination of the relationship between the
quantity measured (X) in an analysis and the analyte concentration CA. Ideally CA = kX where k is a
proportionality.
8. Calculate results
9. Estimate the reliability of results: An analytical result without an estimate of reliability is of no
value.
III. Experimental Error and Basic Statistics
Terms to remember: mean, median, systematic error, random error, precision, accuracy, absolute
error, relative error, standard reference materials, blank, outlier, bias, gross error
Exercise 2: Identify the terms described.
1. contains the solvent and all of the reagents in an analysis but none of the sample.
2. a substance prepared and sold by the National Institute of Standards and Technology and certified
to contain specified concentrations of one or more analytes.
3. measures the systematic error associated with an analysis. It has a negative sign if it causes the results
to be low and a positive sign otherwise.
4. occasional result in replicate measurements that obviously differs significantly from the rest of
the results.
5. the absolute error divided by the true value.
6. difference between the measured value and the true value. It bears a sign.
7. closeness of a result to its true or accepted value.
8. closeness of data to other data that have been obtained in exactly the same way.
9. middle value in a set of data that has been arranged in order of size. Middle value for odd number
of results, mean of the middle pair for even number of results.
10. average value
Exercise 3. Identify types of errors
1. Instrument, method, personal. Originates from afixed cause; affects accuracy of results.
2. Originates from indeterminate processes, affects precision of results
3. Occur only occasionally and are often large; lead to outlier results
Basic Statistic Equations:
Standard deviation,

Variance = s2
Student's t test: used to compare one set of measurements with another to decide whether or not they are
the same. This is often used to decide whether two sets of replicate measurements obtained using two
different methods each with its own standard deviation, give "the same or "different" results within a
stated confidence level.

tcalculated compared with t from statistical table for n1 + n2 -2 degrees of freedom. If tcalculated is greater than
t t a b l e at the 95% confidence level, the two results are considered to be different.
Q-test - this is used to help decide whether to retain or discard a questionable datum. To apply the
Q test arrange the data in order of increasing value and calculate Q, defined as

where Xq = questionable result

Xn = nearest value
w = spread or range
If Q calculated > Q table, the questionable point should be discarded
Confidence interval: an expression stating that the true mean, , is likely to lie within a certain
distance from the measured mean, x . The confidence interval is given by

where s is the measured standard deviation, n is the number of observations, t is the

student's t taken from table.

Exercise 4: Traces of toxic, man-made hexachlorohexanes in North Sea sediments were extracted by a
conventional method and by a new procedure and measured by chromatography. Is the mean concentration
found by new procedure significantly different from that of the conventional procedure? t95% = 2.228 for 10
degrees of freedom.
.
Method
Conventional
New Procedure

Concentration found (pg/g)

34.4 3.6 (n=6)
42.9 1.2 (n =6)

Exercise 5: Using Q test, decide whether the value 216 should be rejected from the set of
results192, 216, 202, 195 and 204. For 5 measurements, the Q table at the 95% confidence level is 0.710.

IV. Uncertainties in Experimental Results

Expressed as standard deviation or confidence interval
Based on series of replicate measurements
Applies only to random error

Summary of rules for propagation of uncertainty

Function

Uncertainty

Function

Uncertainty

Retain one or more insignificant figures to avoid introduction of round-off errors

into later calculations.

Exercise 6: Calculate the absolute uncertainty and % RSD of the results in the following
arithmetic operations. Express final answer with the correct number of significant figures.
A. 1.76 ( 0.03) + 1.39 ( 0.02) - 0.59 ( 0.02) = 3.06 ?
B.

1.76(0.03) x 1.89(0.02) = 5.6

?
0.59(0.02)
C. 1.76(0.03)-0.59(0.02) = 0.6190 ?
1.89(0.02)

D. Find [H+] and its uncertainty for pH= 5.21 0.03

V. Calibration Methods
Calibration curve: shows the response of an analytical method to known quantities of analyte.
External calibration method.
Solutions containing known concentrations of analyte (standard solutions) are prepared covering
a convenient range of concentrations.
Measure response of the analytical procedure to these standards.
Subtract average response of the blank samples from each measured response to obtain corrected
response. The blank measures the response of the procedure when no analyte is present
Make a graph of corrected response vs. concentration of analyte standards.
Use least-squares procedure to find the best straight line through the linear portion of the data.
The equation of the linear calibration line is y = mx +b, where m= slope, b=y-intercept,
y=response, x=concentration.
Analysis of unknown. Read response of unknown. Obtain corrected absorbance.
Terms to remember:
Linear range of an analytical method is the analyte concentration range over which response is
proportional to concentration.
Dynamic range: concentration range over which there is a measurable response to analyte, even if the
response is not linear.

Standard Addition Method- Known quantities of analyte are added to the unknown. From the increase in signal we
deduce how much analyte was in the original unknown.
Terms to remember:
Matrix- everything in the unknown other than analyte.
Matrix effect- change in the analytical signal caused by anything in the sample other than analyte.
Spike- analyte deliberately added.
1. Single standard addition: When signal is directly proportional to analyte concentration.
Case a. Analytical signal is measured before and after addition of standard to a solution containing
analyte.

Rearranging gives
where

Cx = Concentration of analyte solution

Vx
Cs = concentration of standard solution
Vs = volume of standard solution add4d
Ax= Analytical signal of solution with analyte
Ax+s = Analytical signal of solution with analyte + standard
VT =total volume= Vx + Vs

Exercise 7: Serum containing Na+ gave a signal of 4.27 mV in an atomic emission analysis. Then 5.00 mL of 2.08
M NaCl were added to 95.0 mL of serum. This spiked serum gave a signal of 7.98 mV. Find the original
concentration of Na+ in the serum.
Case b. Two solutions are prepared; one solution contains analyte only and the other contains analyte
and standard. Both solutions are diluted to .the same volume before measurement of analytical signal.

Rearranging gives

Exercise 8: A 4.97 -g petroleum specimen was decomposed by wet-ashing and subsequently diluted to
500 mL in a volumetric flask. Cobalt was determined by treating 25.00 mL aliquots of this diluted
solution as follows:
Sample
25.00mL
' 25.00mL

Co(II), 3.00 ppm

0.00 mL
5.00 mL

Ligand
20.00mL
20.00mL

H2 O
5.00mL
0.00 mL

Total Volume Absorbance

50.00mL
0.398
50.00mL
0.510

Assume that the Co(II)/ligand chelate obeys Beers Law. Calculate the % of cobalt in the original sample.
2. Multiple Standard Addition
A Graphic procedure for standard addition with constant volume of sample. Technique used if
chemical analysis consumes solution.
Pipet equal volumes of unknown into several volumetric flasks
Add increasing volumes of standard to each flask and dilute to mark.
Analyze solutions and construct graph of analytical signal (dependent y-variable) v s . concentration
of added analyte, after it has been mixed with sample, [S] (independent x-variable)
The x-intercept of the extrapoted line is the concentration of unknown, [X]r after it has been
diluted to the final volume.
Therefore [X] i = [X]f x Vf /Vo
Or fit points with a least-square line y = mx + b. The x-intercept is obtained by setting y = 0:
0 = mx + b; x= -b/m = [X]f ; [X]i = [X]f x Vf/Vo
B. Successive standard addition to one solution.
Technique used when chemical analysis does not consume solution. (Ex. In electrical potential
measurement)
Measure signal at 0 addition.
Make a standard addition which increases total volume of sample, measure signal again.
Repeat process several times until original signal has increased by a factor of 1.5 to 3.
( )
( )
is the corrected response. x-intercept = [X]o = original concentration of unknown.
Internal Standards
A known amount of a compound, different from analyte is added to the unknown
Signal from analyte is compared with signal from the internal standard to find out how much
analyte is present.
Use in chromatographic analysis. Chromatographic separation of unknown X and internal
standard S. Relative areas of the signals from X and Sallow us to find out how much X is in the
mixture.
It is necessary first to measure the relative response of the detector to each compound.
To use an internal standard, we prepare a known mixture of standard and analyte to measure the
relative response of the detector to the two species.

[X]f and [S]f are concentration of analyte and standard after they have been mixed together.
Exercise 9: In a preliminary experiment a solution containing 0.0837 M X and 0.0666 M S gave peak areas of
Ax = 423 and As= 347. (Areas are measured in arbitrary units by the instruments computer.). To analyze the
unknown, 10.0 mL of 0.146 M S were added to 10.0 mL of unknown and the mixture was diluted to 25.0 mL
in a volumetric flask. From the chromatogram, Ax = 553 and As = 582. Find the concentration of X in the
unknown

VI. Titrimetric Methods

Titration

Titrant

Primary Standard

NaOH
HCl

KHP
Na2CO3

2. Redox
Iodimetry
Iodometry

Iodin
soln.
Na2S2O3

Permanganimetry

KMnO4

Anhydrous
Na2S2O3
KIO3
(strongly acidic)
Cu, CuSO4
K2C2O4

Dichromate
Titration

K2Cr2O7

FAS(ferrous
ammonium sulfate)

3. Complexometry
EDTA Titration

Na2EDTA

4. Precipitation
Mohr Titration
Direct Chloride
determination
Volhard Titration
(Indirect
determination of
halide: Ag(I) is
added in excess
Fajans Titration
(Adsorption
Indicator Method)

AgNO3

CaCO3
MgCO3
Zn or Zn salt
Cu or Cu salt
NaCl

K2CrO4

KSCN

NaCl

Fe(III)

AgNO3

NaCl

Fluorescein

1. Acid-base

Indicator (pH
range)
H2Ph(8.2-9.8)
Methyl
Orange(3.1-4.4)
Bromocresol
green(3.8-5.4)
starch
starch

KMnO4 self
Indicator
Diphenylaminesulfonic acid
EBT

Color
change/condition
Colorless to pink
Red to yellow
orange
Yellow to blue
Colorless to deep
blue
Deep blue to
colorless
From _ to purple
From reduced form
colorless to oxidized
form-violet.
Wine red to blue
(at pH 10)

Formation of redorange AgCrO4

precipitate (pH 7 to
10)
Formation of red
FeSCN2+ complex

The silver
fluoresceinate
adsorbed on the
surface of the silver
chloride precipitate.
The solution
surrounding the solid
turns red.

Exercise 10: Predict the effect of the given condition on the indicated (parameter/calculated result).
1. The coin sample which was weighed has a higher temperature than the balance
(Mass of coin).
2. The coins were weighed by difference. (average weight of coins)
3. Calcium oxalate was precipitated at pH 3.0 instead of pH 4.0. (% CaO)
4. The permanganate solution was not filtered prior to its standardization and was used as is a week
later for the analysis of the unknown sample. (Volume of permanganate for the unknown
sample).
5. Bromthymol blue (pH range 6.0-8.0) was used as indicator for the first endpoint in the titration of carbonate
mixtures rather than H2Ph. (Volume of HCl, first endpt.).
6. Zn metal was used to standardize the EDTA solution for total hardness determination of water samples.
(Total hardness)
7. In the iodometric determination of Cu(II), starch was added at the start of the titration of the standard
Cu . (N of standard thiosulfate solution).
VII. Solvent Extraction: separation method based on difference in solubility of solute in two immiscible solvents
Nernst Distribution law for solute species A:
Distribution constant or partition coefficient:
where CA = total concn of A

Distribution coefficient or Distribution ratio:

Fraction of solute transferred to organic phase, p:
Fraction of solute remaining in aqueous phase, q:
Fraction of solute in aqueous phase after n transfers , qn:
% solute remaining in aqueous solution after n transfers:
Concentration of solute A in aqueous phase after n transfers, [A]n:

%A = qn x 100
[A]n

x [A]o

Exercise 11: The distribution coefficient for Z between n-hexane and water is 6.25. Calculate % Z
remaining in 25.0 mL of water that was originally 0.0600 M in Z after extraction with five 5.00-mL portions.

VIII. Instrumental Methods of Analysis

Excitation Signal
Source

Sample
Cell

Detector/Sensor

Qualitative
Parameter

Quantitative
Parameter
Measured

UV Absorption
Spectrophotometry

Deuterium/Hydrogen
lamp

Quartz

max
(wavelength
of optimum
absorption)

Absorbance
(Beer's Law)

Vis Absorption
Spectrophotometry

Tungsten-halogen
lamp

Quartz,
glass or
plastic cell

max
(wavelength
of optimum
absorption)

Absorbance
(Beer's Law)

Flame-AAS
Atomic Absorption
Spectrophotometry

Hollow Cathode
Lamp

ElectrothermalAAS
Atomic Emission

Hollow cathode
lamp

Flame
(e.g.
acetyleneair)
Graphite
furnace
Flame or
ICP

Phototubes,
PMT, silicon
photodiodes,
diode-array,
photovoltaic
cells
Phototubes,
PMT, silicon
photodiodes,
diode-array,
photovoltaic
cells
Photomultiplier
tube(PMT)

Spectrofluorometry

Xenon lamp

Analytical Method

l.Spectro

Quartz cell

2. Chrom
GC:Gas Chrom
(Isothermal and
Temperature
Programming)

HPLC (Isocratic
and Gradient)

Appropriate
HPLC flow
cell for a
given
detector

Absorbance
(Beer's Law)

PMT
PMT

PMT

max
(wavelength
of optimum
emission)
max
(wavelength
of optimum
emission)

Absorbance
(Beer's Law)
Emission
Intensity, I

Fluorescence
Intensity, F
(F=kC)

Flame
ionization
(FID)
Thermal
Conductivity
(TCD)
Electron
Capture (ECD)

Retention
time

Peak area or
peak height
(PA or PH=
kC)

UV-Vis,
Refractive
index (RI)
Electrochemical
detector

Retention
time

Peak area or peak

height (PA or
PH=
kC)

3.
Electroanalytical
Ion-selective
electrode (ISE)
Method (Direct
Potentiometry)

Concentration
gradient at the
electrodesolution
interlace

Any
appropriate
sample
container

ISEs

Potential/pX
(Nernst
equation)

Indicator
electrodes such
As Pt, Ag and
Au
DME

ASV

Applied potential

Any
appropriate
sample
container
Polarographic
cell
Voltammetric
cell

Volume of
titrant at
equivalence
point

Polarography

Addition of titrant
causing a change in
the potential of the
system
Applied potential

Potentiometric
Titration(Indirect
Potentiometry)

GCE, Pt TFME

Diffusion
current
Anodic peak Anodic peak
potential
current or
E1/2

Performance Characteristics of Instrumental Method; Figures of Merit.

Criterion
1. Precision
2. Bias
3.Sensitivity
4. Detection limit
5.Concentration range
6. selectivity

Figure of merit
Absolute standard deviation; RSD, CV, variance
Absolute systematic error, relative systematic error
Calibration sensitivity, analytical sensitivity
3 blank/m
LOQ to LOL
Selectivity coefficient

UV-Vis- Spectroscopy- interaction of matter with the uv-vis region of the electromagnetic spectrum.
Visible wavelength region: 380-750 nm
Ultraviolet region: 200-380 nm

Types of Instruments used for absorption measurements:

1. Spectrophotometer- employ grating or a prism monochromator to provide a narrow band of
radiation for measurements. (scanning)
2. Photometers- use an absorption filter or an interference filter for wavelength selection
Designs:
1. Single beam- employs a fixed beam of radiation that irradiates first the solvent and then the analyte
solution.
2. Double beam- the solvent and solution are irradiated simultaneously
3. Multichannel or diode array spectrophotometers- detect the entire spectral range simultaneously.
Can produce a spectrum in < 1 second.
Quantitative analysis based on Beer's Law
Quantitative Approaches
1. Two component analysis
2. Standard addition Method

10

Exercise 12. The absorption spectra of two colored substances HMR and HMO are determined and the
following data obtained in a 1.00-cm cell:
Solution
HMR alone
HMO alone
HMR+HMO

Concentration
4.75 X 10-4M
1.68 X 10-4 M
unknown

A at 375nm
0.726
0.112
0.595

A at 650nm
0.0950
0.625
0.925

Calculate concentration of HMR and HMO in the unknown solution.

Exercise 13. Draw a schematic diagram of an instrument used for UV-vis measurements. Give a short description
for each instrument component.
Molecular Photoluminescence

Involves emission of certain wavelengths of light by some substances after electronic excitation by either UV
or vis light.
Types:
1. fluorescence- emission of a photon during a transition between states with the same spin quantum numbers.
2. phosphorescence- emission of a photon during a transition between states with different spin quantum
numbers.
Quantitative analysis based on
I = KPoC
Exercise 14: Draw a schematic diagram of an instrument used to measure a sample luminescence. Give a short
description for each component.
Atomic Absorption Spectroscopy
Involves interaction of monoatomic particles with UV-vis light
Involves electronic transitions in which one or more of the electrons of the atoms is raised to a higher
energy level.
Instrument Components
Hollow Cathode lamps
Flame or electrothermal atomizer
Monochromator
Detector- radiation detectors.
Quantitative analysis based on Beer's Law. Atomic absorption cannot provide qualitative information
sinceit does not provide complete absorption spectra because of the discontinuous nature of radiation
sources that must be used
Exercise 15: Draw a schematic diagram of an instrument used to measure atomic absorption. Give a short
description for each component.
Atomic Emission Spectrometry
Provide both qualitative and quantitative information about an analyte.
Identification of the elements present is based upon the peak wavelengths which are unique for each
element.
ICP (Inductively Coupled Plasma) is the most popular source for emission spectrometry. Flame emission is
also used.

Plasma- a hot partially ionized gas. It contains relatively high concentrations of ions and
electrons.
Atomic emission and atomic absorption instruments are similar except that no hollow cathode
lamp is required for emission measurement.

Exercise 16: Draw a schematic diagram of a typical atomic emission spectrometer. Give a short
description for each component.

Potentiometry
Based on the measurements of a potential difference between two electrodes immersed in an analyte solution.
1. Direct Potentiometry (ISE) - measure activities of ions
2. Potentiometric Titration-determines volume at equivalence point without use of an indicator.
Commonly used indicator electrodes 1.
1. Ion selective electrodes
2. Inert electrodes (Redox electrodes) - the only role of this type of electrode is to provide or accept electrons
Commonly used reference electrodes
1. Saturated Calomel electrode
2. Silver/silver chloride electrode
Quantitative analysis based on Nernst Equation.
Exercise 17: An aqueous solution is to be analyzed for its free fluoride ion concentration by direct potentiometry. A
100.0-mL aliquot of this solution was measured with a fluoride ISE electrode and gave a reading of -120.0 mV vs. a
suitable reference electrode. Exactly 1.00 mL of a 0.100 M solution of KF is added to the test solution with stirring
and the potential changed to -132 mV. Calculate the fluoride ion concentration in the sample.

Exercise 18.The benzoic acid (C6H5COOH) extracted from a 100.0 g banana catsup was titrated
potentiometrically with 0.05555 M NaOH. Given below is a portion of the potentiometric data for the
determination of % Na benzoate in the catsup sample. The extracted benzoic acid was diluted to 100.0 mL and
was then titrated with std. NaOH.
NaOH, mL
14.40
14.60
14.80

pH
5.25
5.32
' 5.89

NaOH, mL
15.00
15.20
15.40

pH
6.75
9.16
10.04

A Tabulate the 1st and 2nd derivative point

B. Determine the equivalence point from the 2 nd derivative graph or points.
C. Calculate % sodium benzoate (NaC 6H5COO) of the sample.
D. Determine Ka of benzoic acid.
Voltammetry
An electroanalytical technique in which a varying potential is applied to an indicator electrode and the current that is
generated is monitored as a function of the applied potential. The graph that results is called a voltammogram.
Typical indicator electrodes used

1. Dropping Mercury Electrode (DME) -most common. Voltammetry at DME is termed

polarography.
2. Pt wire
3. Other noble metal electrodes (Au)
Voltammetric equipment consists of:
1. Potentiostat - which applies a varying potential to the indicator electrode
2. Voltammetric cell- consisting of the indicator electrode, reference electrode and an auxiliary
electrode
Terms to remember: half-wave potential, limiting current, diffusion current, residual current
Quantitative analysis is based on Ilkovic Equation, where m in mg/s, D in cm2/s, id in A, c in mM.
(id)max = 708 n D1/2 m2/3 t1/6 c
(id)ave = 607 n D1/2 m2/3 t1/6 c
Exercise 19: A 5 x 10-3 M solution of CaCl2 in 0.1 M KCl shows a diffusion current at -0.8 V versus SCE of
50.0 A. The mercury is dropping at a rate of 18.0 drops per minute. Ten drops are collected and found to
weigh 3.82 x 10-2 g.
a) Calculate the diffusion coefficient D.
b) If the capillary were replaced by another, for which the drop-time is 3.0 sec., and 10 drops weigh
4.20 x 10-2 g, what will be the new value of the diffusion current?
Chromatography
Refers to any separation method in which the components are distributed between a stationary
phase and a moving (mobile) phase.
Base on types of mobile and stationary phases, classified as
1. liquid chromatography
2. gas chromatography
3. supercritical-fluid chromatography

Resolution Equation: R =
Optimize each term to increase resolution
k' = capacity factor = t'R /to (also known as retention factor, k)
= selectivity = t'R(B) / t'R (A)
N =theoretical plate number= 16 (tR/W)2
R = Resolution = t/ 1/2(W(A) + W(B) )
Gas-liquid Chromatography
Components:
l. Carrier gas- includes He, Ar, N2 and H2
2. flow controller or pressure regulator
3. Injection port
4. Column-conventional "packed" and Capillary column
5. Detector- BCD, FID and TCD
Liquid Chromatography
Components:
1. Eluent Reservoir
2. Pump
3. Sample Injector
4. Column
5. Detector- Refractive index, electrochemical, UV/vis

13

Separation Modes
1. Normal phase
2. Reversed phase
3. Adsorption
4. Size exclusion
5. Ion-exchange
Quantitation techniques:
1. Internal standard calibration
2. Area normalization
Supercritical fluid chromatography
a hybrid of gas and liquid chromatography, MP is a supercritical fluid (physical state of a substance
that is held above its critical temperature) - usually CO2
density of a supercritical fluid is 0.2 to 0.5 g/cm3 200 to 400 times greater than that of the corresponding
gas, and approaches that of the substance in its liquid state will dissolve large nonvolatile molecules
GC: temperature programming I HPLC: gradient elution or solvent programming I SFC: pressure
programming
Exercise 20. Standard Addition: An unknown sample of Ni 2+ gave a current of 2.36A in an electrochemical
analysis. When 0.500 mL of solution containing 0.0287 M Ni 2+ was added to 25.0 mL of unknown, the
current increased to 3.79 A. Find the concentration of Ni2+ in the unknown.
Calculate the ppm Cr m the sample.
Exercise 21. To prepare a solution of NaCl, you weigh out 2.634 ( 0.002)g and dissolve it in a
volumetric flask whose volume is 100.00( 0.08) mL. Express the molarity of solution, along with its
uncertainty with correct no. of significant figures. MW NaCl = 58.4425 ( 0.0009) g/mol.
Exercise 22. The chromium in an aqueous sample was determined by pipetting 10.0 mL of the unknown
into each of five 50.0 mL volumetric flasks. Various volumes of a standard containing 12.2 ppm Cr were
added to the flasks, and the solutions were then diluted to volume.
Unknown
10.0
10.0
10.0
10.0
10.0

Standard, mL
0.0
10.0
20.0
30.0
40.0

Absorbance
0.201
0.292
0.378
0.467
0.554

Calculate the ppm Cr in the sample.

Exercise 23: A mixture of methyl esters of fatty acids was chromatographed on a Carbowax 20M giving
The following peak areas and detector response factors:
ester
Methyl-n-butyrate
Methyl-iso-valerate
Methyl-n-octonoate
Methyl-n-decanoate

Peak, area, cm2

2.95
0.86
1.66
4.52

Calculate the percentage composition of the mixture by area normalization.

14

Response factor
0.81
0.88
0.98
1.00

Exercise 24. The following calibration data were obtained by an instrumental method for the
determination of the species X in aqueous solution.
Conc. X, ppm

No.Replications, N

Mean Analytical
Signal, S
0.031
0.173
0.422
0.702
0.956
1.248

0.00
25
2.00
5
6.00
5
10.00
5
14.00
5
18.00
5
A Calculate the calibration sensitivity.
B. Calculate the analytical sensitivity at each concentration.
C. What is the detection limit for the method

Standard
Deviation, ppm
0.0079
0.0094
0.0084
0.0084
0.0085
0.0110

Exercise 25. Internal Standard: A solution was prepared by mixing 5.00 mL of unknown (element X) with 2.00
mL of solution containing 4.13 g of standard (element S) per milliliter and diluting to 10.0 mL. The measured
signal ratio in an atomic absorption experiment was (signal due to X)/(signal due to S) = 0.808. In a separate
experiment, it was found that for equal concentrations of X and S, the signal due to X was 1.31 times more
intense than the signal due to S. Find the concentration of X in the unknown.
IX. Other Techniques
1. Nuclear Magnetic Resonance (NMR) spectroscopy is based on the measurement of absorption of
electromagnetic radiation in the radio frequency region. Nuclei absorb electromagnetic radiation in a strong
magnetic field as a result of the energy splitting that is induced by the magnetic field.
2. Mass Spectrometry (MS) is a technique in which gaseous molecules are ionized, accelerated by an electric
field, separated according to their mass-to-charge ratio, and the amount of each species is detected.
3. Nuclear and Related Techniques
a. X-ray fluorescence (XRF) spectroscopy is based on the emission of characteristic "secondary" (or
fluorescent) X-rays from a material that has been excited by bombarding with high-energy X-rays or gamma
rays. The technique is used for elemental analysis and chemical analysis.
b. Neutron activation analysis (NAA) is based on the detection and measurement of characteristic
gamma rays emitted from radioactive isotopes produced in the sample upon irradiation with neutrons. The
emitted radiation is a 'fingerprint' of the element, and the amount of radiation given off at a certain energy is
indicative of the amount of the element present in the sample.
4. Polarimetry measures the extent to which a substance interacts with plane polarized light (light which
consists of waves that vibrate only in one plane); whether it rotates plane polarized light to the left or to the
right (optically active), or not at all.
5. Refractometry measures how light is refracted when it passes through a given substance. The amount by
which the light is refracted determines the refractive index. Refractive index can be used to identify an
unknown liquid compound, or it can be used as a means of measuring the purity (if a liquid compound by
comparing it to literature values. Refractive index is defined as the ratio of the velocity of light in air to the
velocity of light in the medium being measured.
6. Turbidimetry is a method for determining the concentration of a substance in a solution by measuring the
loss in intensity of a light beam through a solution that contains suspended particulate matter.

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