1104 CHAPTER 37 Selected Methods of Analysis

point at 1.0 V should be the easier of the two to locate exactly. Removal of oxy-
gen is unnecessary, however, for the titration at 0 V.

PREPARATION OF SOLUTIONS

1. Supporting electrolyte. Dissolve 10 g of KNO3 and 8.2 g of sodium acetate in

about 500 mL of distilled water. Add glacial acetic acid to bring the pH to 4.2
(pH meter); about 20 mL of the acid will be required (sufficient for about 20
titrations).
2. Gelatin, 0.1%. Add 0.1 g of gelatin to 100 mL of boiling water.
3. Potassium dichromate, 0.01 M. Use a powder funnel to weigh about 0.75 g (to
the nearest 0.5 mg) of primary-standard K2Cr2O7 into a 250-mL volumetric
flask. Dilute to the mark with distilled water, and mix well.
4. Potassium nitrate salt bridge. See Section 37J-2.

PROCEDURE

Obtain the unknown (Note) in a clean 100-mL volumetric flask; dilute to the mark
with distilled water, and mix well. Perform the titration in a 100-mL beaker. Locate
a saturated calomel electrode in a second 100-mL beaker, and provide contact
between the solutions in the two containers with the KNO3 salt bridge.
 The corrected current is given by Transfer a 10.00-mL aliquot of the unknown to the titration beaker. Add 25 mL
of the supporting electrolyte and 5 mL of gelatin. Insert a dropping mercury
(id)corr  id a b
Vv electrode into the sample solution. Connect both electrodes to the polarograph.
V Measure the current at an applied potential of 0 V. Add 0.01 M K2Cr2O7 in 1-mL
increments; record the current and volume after each addition. Continue the
where V is the original volume, v is the titration to about 5 mL beyond the end point. Correct the currents for volume
volume of reagent added, and id is the change and plot the data. Evaluate the end-point volume.
uncorrected current. Repeat the titrations at 1.0 V. Here, you must bubble nitrogen through the
solution for 10 to 15 min before the titration and after each addition of reagent. The
flow of nitrogen must, of course, be interrupted during the current measurements.
Again, correct the currents for volume change, plot the data, and determine the
end-point volume.
Report the mass of Pb in the unknown in milligrams.

Note
A stock 0.0400 M solution can be prepared by dissolving 13.5 g of reagent-grade
Pb(NO3)2 in 10 mL of 6 M HNO3 and diluting to 1 L with water. Unknowns should
contain 15 to 25 mL of this solution.

METHODS BASED ON THE ABSORPTION

37N OF RADIATION
Molecular absorption methods are discussed in Chapter 26. Directions follow for
(1) the use of a calibration curve for the determination of iron in water, (2) the use
of a standard-addition procedure for the determination of manganese in steel, and
(3) a spectrophotometric determination of the pH of a buffer solution.
 37N Methods Based on the Absorption of Radiation 1105

37N-1 The Cleaning and Handling of Cells

The accuracy of spectrophotometric measurements is critically dependent on the
availability of good-quality matched cells. These should be calibrated against one
another at regular intervals to detect differences resulting from scratches, etching,
and wear. Equally important is the proper cleaning of the exterior sides (the win-
dows) just before the cells are inserted into a photometer or spectrophotometer. The
preferred method is to wipe the windows with a lens paper soaked in methanol; the
methanol is then allowed to evaporate, leaving the windows free of contaminants. It
has been shown that this method is far superior to the usual procedure of wiping the
windows with a dry lens paper, which tends to leave a residue of lint and a film on
the window.13

37N-2 The Determination of Iron in a Natural Water

Discussion
The red-orange complex that forms between iron(II) and 1,10-phenanthroline
(orthophenanthroline) is useful in determining iron in water supplies. The reagent
is a weak base that reacts to form phenanthrolinium ion, phenH, in acidic media.
Complex formation with iron is thus best described by the equation

Fe2  3phenH 8 Fe(phen)2

3  3H


The structure of the complex is shown in Section 19E-1. The formation constant for
this equilibrium is 2.5  106 at 25°C. Iron(II) is quantitatively complexed in the pH
range between 3 and 9. A pH of about 3.5 is ordinarily recommended to prevent
precipitation of iron salts, such as phosphates.
An excess of a reducing reagent, such as hydroxylamine or hydroquinone, is
needed to maintain iron in the 2 oxidation state. The complex, once formed, is
very stable.
This determination can be performed with a spectrophotometer set at 508 nm or
with a photometer equipped with a green filter.

PREPARATION OF SOLUTIONS

1. Standard iron solution, 0.01 mg/mL. Weigh (to the nearest 0.2 mg) 0.0702 g of
reagent-grade Fe(NH4)2(SO4)2 # 6H2O into a 1-L volumetric flask. Dissolve in
50 mL of water that contains 1 to 2 mL of concentrated sulfuric acid; dilute to
the mark and mix well.
2. Hydroxylamine hydrochloride (sufficient for 80 to 90 measurements). Dissolve
10 g of H2NOH # HCl in about 100 mL of distilled water.
3. Orthophenanthroline solution (sufficient for 80 to 90 measurements). Dissolve
1.0 g of orthophenanthroline monohydrate in about 1 L of water. Warm slightly
if necessary. Each milliliter is sufficient for no more than about 0.09 mg of Fe.
Prepare no more reagent than needed; it darkens on standing and must then be
discarded.
4. Sodium acetate, 1.2 M (sufficient for 80 to 90 measurements). Dissolve 166 g of
NaOAc # 3H2O in 1 L of distilled water.

13For further information, see J. O. Erickson and T. Surles, Amer. Lab., 1976, 8(6), 50.
1106 CHAPTER 37 Selected Methods of Analysis

PROCEDURE

Preparation of the Calibration Curve

Transfer 25.00 mL of the standard iron solution to a 100-mL volumetric flask and
25 mL of distilled water to a second 100-mL volumetric flask. Add 1 mL of
hydroxylamine, 10 mL of sodium acetate, and 10 mL of orthophenanthroline to
each flask. Allow the mixtures to stand for 5 min; dilute to the mark and mix well.
Clean a pair of matched cells for the instrument. Rinse each cell with at least
three portions of the solution it is to contain. Determine the absorbance of the
standard with respect to the blank.
Repeat this procedure with at least three other volumes of the standard iron
solution; attempt to encompass an absorbance range between 0.1 and 1.0. Plot a
calibration curve.
Determination of Iron
Transfer 10.00 mL of the unknown to a 100-mL volumetric flask; treat in exactly
the same way as the standards, measuring the absorbance with respect to the blank.
Alter the volume of unknown taken to obtain absorbance measurements for
replicate samples that are within the range of the calibration curve.
Report the concentration of iron in the unknown in parts per million.

37N-3 The Determination of Manganese in Steel

Discussion
Small quantities of manganese are readily determined photometrically by the oxi-
dation of Mn(II) to the intensely colored permanganate ion. Potassium periodate is
an effective oxidizing reagent for this purpose. The reaction is
5IO4  2Mn2  3H2O S 5IO3  2MnO4  6H
Permanganate solutions that contain an excess of periodate are quite stable.
There are few interferences to the method. The presence of most colored ions
can be compensated for with a blank. Cerium(III) and chromium(III) are excep-
tions; these yield oxidation products with periodate that absorb to some extent at
the wavelength used for the measurement of permanganate.
The method given here is applicable to steels that do not contain large amounts
of chromium. The sample is dissolved in nitric acid. Any carbon in the steel is oxi-
dized with peroxodisulfate. Iron(III) is eliminated as a source of interference by
complexation with phosphoric acid. The standard-addition method (see Section
8C-3) is used to establish the relationship between absorbance and amount of man-
ganese in the sample.
A spectrophotometer set at 525 nm or a photometer with a green filter can be
used for the absorbance measurements.

PREPARATION OF SOLUTIONS

Standard manganese(II) solution (sufficient for several hundred analyses). Weigh

0.1 g (to the nearest 0.1 mg) of manganese into a 50-mL beaker, and dissolve in
about 10 mL of 6 M HNO3 (use the hood). Boil gently to eliminate oxides of
 37N Methods Based on the Absorption of Radiation 1107

nitrogen. Cool; then transfer the solution quantitatively to a 1-L volumetric flask.
Dilute to the mark with water, and mix thoroughly. The manganese in 1 mL of the
standard solution, after being converted to permanganate, causes a volume of 50
mL to increase in absorbance by about 0.09.

PROCEDURE

The unknown does not require drying. If there is evidence of oil, rinse the sample
with acetone and dry briefly. Weigh (to the nearest 0.1 mg) duplicate samples (Note
1) into 150-mL beakers. Add about 50 mL of 6 M HNO3 and boil gently (use the
hood); heating for 5 to 10 min should suffice. Cautiously add about 1 g of ammonium
peroxydisulfate, and boil gently for an additional 10 to 15 min. If the solution is
pink or has a deposit of MnO2, add 1 mL of NH4HSO3 (or 0.1 g of NaHSO3) and
heat for 5 min. Cool; transfer quantitatively (Note 2) to 250.0-mL volumetric
flasks. Dilute to the mark with water, and mix well. Use a 20.00-mL pipet to
transfer three aliquots of each sample to individual beakers. Treat as follows:

Aliquot Volume of 85% H3PO4, mL Volume of Standard Mn, mL Mass of KIO4, g

1 5 0.00 0.4
2 5 5.00 (Note 3) 0.4
3 5 0.00 0.0

Boil each solution gently for 5 min, cool, and transfer it quantitatively to a 50-mL
volumetric flask. Mix well. Measure the absorbance of aliquots 1 and 2 using
aliquot 3 as the blank (Note 4).
Report the percentage of manganese in the unknown.

Notes
1. The sample size depends on the manganese content of the unknown; consult
with the instructor.
2. If there is evidence of turbidity, filter the solutions as they are transferred to the
volumetric flasks.
3. The volume of the standard addition may be dictated by the absorbance of the
sample. It is useful to obtain a rough estimate by generating permanganate in
about 20 mL of sample, diluting to about 50 mL, and measuring the
absorbance.
4. A single blank can be used for all measurements, provided that the samples
weigh within 50 mg of one another.

37N-4 The Spectrophotometric Determination of pH

Discussion
The pH of an unknown buffer is determined by addition of an acid/base indicator
and spectrophotometric measurement of the absorbance of the resulting solution.
Because there is overlap between the spectra for the acid and base forms of the
indicator, it is necessary to evaluate individual molar absorptivities for each form at
two wavelengths. See page 796 for further discussion.
1108 CHAPTER 37 Selected Methods of Analysis

The relationship between the two forms of bromocresol green in an aqueous

solution is described by the equilibrium

HIn  H2O 8 H3O  In

for which

[H3O ] [In ]
Ka   1.6  105
[HIn]

The spectrophotometric evaluation of [In] and [HIn] permits the calculation of

[H3O] and thus pH.

PREPARATION OF SOLUTIONS

1. Bromocresol green, 1.0  104 M (sufficient for about five determinations).

Dissolve 40.0 mg (to the nearest 0.1 mg) of the sodium salt of bromocresol
green (720 g/mol) in water, and dilute to 500 mL in a volumetric flask.
2. HCl, 0.5 M. Dilute about 4 mL of concentrated HCl to approximately 100 mL
with water.
3. NaOH, 0.4 M. Dilute about 7 mL of 6 M NaOH to about 100 mL with water.

PROCEDURE

Determination of Individual Absorption Spectra

Transfer 25.00-mL aliquots of the bromocresol green indicator solution to two 100-
mL volumetric flasks. To one add 25 mL of 0.5 M HCl; to the other add 25 mL of
0.4 M NaOH. Dilute to the mark and mix well.
Obtain the absorption spectra for the acid and conjugate-base forms of the
indicator between 400 and 600 nm, using water as a blank. Record absorbance
values at 10-nm intervals routinely and at closer intervals as needed to define
maxima and minima. Evaluate the molar absorptivities for HIn and In at
wavelengths corresponding to their absorption maxima.

Determination of the pH of an Unknown Buffer

Transfer 25.00 mL of the stock bromocresol green indicator to a 100-mL
volumetric flask. Add 50.0 mL of the unknown buffer, dilute to the mark, and mix
well. Measure the absorbance of the diluted solution at the wavelengths for which
absorptivity data were calculated.
Report the pH of the buffer.

37O MOLECULAR FLUORESCENCE

The phenomenon of fluorescence and its analytical applications are discussed in
Chapter 27. Directions follow for the determination of quinine in beverages, in
which the quinine concentration is typically between 25 and 60 ppm.