Colorimetry and Turbidimetry: Indicated in
Colorimetry and Turbidimetry: Indicated in
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ABSTRACT
Conditions and quantities for colorimetric or turbidimetric tests have been chosen to produce colors or turbidities that
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can be observed easily. These conditions and quantities approach but do not reach the minimum that can be observed.
Five minutes, unless some other time is specified, must be allowed for the development of the colors or turbidities
before the comparisons are made. If solutions of samples contain any turbidity or insoluble matter that might interfere
with the later observation of colors or turbidities, they must be filtered before the addition of the reagent used to
produce the color or turbidity.
Note: Conditions of the tests will vary from one reagent to another, but most of the tests fall close to the limits
indicated in Table 2-3.
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ARSENIC
This colorimetric comparative procedure is based upon the reaction between silver diethyldithiocarbamate and arsine.
Interferences
Metals or salts of metals, such as chromium, cobalt, copper, mercury, molybdenum, nickel, palladium, and silver, may
interfere with the evolution of arsine.
Antimony, which forms stibine, may produce a positive interference in color development with silver diethyldithiocarba-
mate solution. Although potassium iodide and stannous chloride in the generator tend to repress the evolution of stibine at
low levels of antimony, at higher levels repression is incomplete, a situation that can lead to erroneously high results for
arsenic. Interference from antimony, however, can be essentially eliminated by adding ferric ion to the generator. An
arsenic test employing this addition is included in the monograph for phosphoric acid.
Antimony can be determined, if desired, by atomic absorption or by differential pulse polarography. An atomic absorption
procedure for antimony is also included in the monograph for phosphoric acid.
Apparatus
The apparatus (Figure 2-1) consists of an arsine generator (a) fitted with a scrubber unit (c) and an absorber tube (e), with
standard taper or ground glass ball-and-socket joints (b and d) between the units. Alternatively, any apparatus embodying
the principle of the assembly described and illustrated may be used.
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Sample Solution
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Dissolve the specified weight of the sample in water and dilute with water to 35 mL or use the solution prepared as directed
in the individual reagent standard.
Note: The color comparison may be made in a photometer at the wavelength of maximum absorbance occurring
between 535 and 540 nm. If a photometer is used, the silver diethyldithiocarbamate solution should be free from any
trace of impurity (before the test is started). The use of capped cells is recommended.
CHLORIDE
The general test procedure cited here applies to the majority of reagent chemicals having a chloride specification.
Perform the test for the sample and the standard in glass tubes or cylinders of the same diameter, matched as closely as
practicable. Use identical quantities of the same reagents for the sample and the standard solutions. If after dissolution the
solution is not clear, filter it through a chloride-free filter paper. Prepare the filter paper by washing with water until the
filtrate gives a negative test for chloride. The filter may also be prewashed with dilute nitric acid (1:3).
Experience has shown that visual turbidimetric comparisons are best made between solutions containing 0.01 mg of chloride
ion in a volume of 20 mL.
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For samples yielding colored solutions, dissolve the sample, prepare the standard, and treat the two solutions as directed in
the specification. The comparison is best made by viewing both turbidities through the same depth and color of solution. Do
this as follows: Prepare a test tube with the specified quantity of sample in a volume of water equal to the total volume of
the test, and superimpose it over the test tube that contains the standard turbidity. Place a third tube containing the same
volume of water below the tube containing the sample and added reagents. The comparison tubes may be machine-made
vials, long style, of about 20 mL capacity.
DITHIZONE
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Because dithizone is extremely sensitive to many metal ions, all glassware used must be specially cleaned. After the usual
cleaning, it should be thoroughly rinsed with warm dilute nitric acid (1:1) and finally rinsed with reagent water. All glassware
used in the preparation and storage of reagents and in performing the tests must be made of lead- and zinc-free glass.
Alternatively, for solution storage, polyethylene containers may be used.
Two general test procedures are provided. Method 1 is to be used unless otherwise specified in the individual standard. This
method is used, generally, for those substances that yield clear, colorless solutions under the specified test conditions. Method
2 is used, generally, only for certain organic compounds. An alternate procedure ([Part 2: Colorimetry and Turbidimetry; Heavy
Metals (as Lead); Procedure for Heavy Metals, Method 2; Alternate Sulfide Reagent Solution and Use]) is provided for cases
where hydrogen sulfide is not suitable.
Note: Heavy metals may also be analyzed for some reagents by ICP–OES ([Part 2: Trace and Ultratrace Elemental
Analysis; Inductively Coupled Plasma−Optical Emission Spectroscopy (ICP–OES); Calculation of Heavy Metals (by
ICP–OES) Results]).
Unless otherwise directed in the individual specification, the test shall be carried out as follows.
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Te s t S o l u t i o n . Using the solution prepared as directed in the individual reagent monograph, adjust the pH to between 3
and 4 (by using a pH meter) with 1 N acetic acid reagent solution or 10% ammonium hydroxide reagent solution. Dilute with
pH 3.5 ammonium acetate dilution buffer solution to 40 mL, if necessary, and mix.
To each of the tubes containing the test solution and the standard/control solution, add 5 mL of freshly prepared hydrogen
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sulfide water and mix. Any brown color produced within 5 min in the test solution should not be darker than that produced
in the standard/control solution.
Te s t S o l u t i o n . Transfer the quantity of substance specified in the individual reagent monograph to a suitable crucible, add
sufficient sulfuric acid to wet the sample, and carefully heat at a low temperature until thoroughly charred. (The crucible
may be loosely covered with a suitable lid during the charring.) Add to the carbonized mass 2 mL of nitric acid and 0.25 mL
of sulfuric acid, and heat cautiously until white fumes of sulfur trioxide are no longer evolved. Ignite, preferably in a well-
ventilated muffle furnace, at 500–600 °C until the carbon is burned off completely. Cool, add 4 mL of 6 N hydrochloric acid,
and cover. Digest on a hot plate (~100 °C) for 15 min, uncover, and slowly evaporate on the hot plate to dryness. Moisten the
residue with 0.05 mL of hydrochloric acid, add 10 mL of hot water, and digest for 2 min. Add 10% ammonium hydroxide
reagent solution dropwise until the solution is just alkaline, and dilute with water to 25 mL. Adjust the pH of the solution to
between 3 and 4 (by using a pH meter) with 1 N acetic acid reagent solution, dilute with pH 3.5 ammonium acetate dilution
buffer solution (described in Method 1) to 40 mL, and mix.
S t a n d a r d / C o n t r o l S o l u t i o n . To 20 mL of water, add 0.02 mg of lead ion (Pb), and dilute with water to 25 mL. Adjust the pH
(within 0.1 unit) to the value established for the test solution (by using a pH meter) with 1 N acetic acid reagent solution or
10% ammonium hydroxide reagent solution, dilute with pH 3.5 ammonium acetate dilution buffer solution (described in
Method 1) to 40 mL, and mix.
To each of the tubes containing the test solution and the standard/control solution, add 5 mL of freshly prepared hydrogen
sulfide water and mix. Any brown color produced in the test solution within 5 min should not be darker than that produced
in the standard/control solution.
S o d i u m S u l fi
fidde S o l u t i o n . (Note: Before sodium sulfide is weighed, any excessive liquid on the reagent should be removed
on filter paper.) Dissolve 2.5 ± 0.05 g of sodium sulfide, nonahydrate, in a mixture of 5 mL of water and 15 mL of glycerol.
Store the solution in a small, tightly closed, light-resistant bottle. Use within 3 months. Make fresh when in doubt.
Add 0.05 mL or 1 drop of sodium sulfide solution to the test solution and the standard/ control solution, mix well, and allow
to stand for 5 min. Compare both solutions against a white background. Any brown color produced in the test solution
within 5 min should not be darker than that produced in the standard/control solution.
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Note: The final volume of the sample and control solutions must be the same in both Method 1 and Method 2 when the
pH 3.5 ammonium acetate dilution buffer solution is used.
IRON
Two procedures are provided for the determination of iron. Method 1, which uses thiocyanate, is usually performed in a 50 mL
solution containing 2 mL of hydrochloric acid, to which 30–50 mg of ammonium peroxydisulfate crystals and 3 mL of 30%
ammonium thiocyanate reagent solution are added. These quantities of acid and thiocyanate were selected so that the
production of the red color would not be affected significantly by anything except the iron. The use of peroxydisulfate to
oxidize the iron prevents fading of the color and eliminates the need for immediate comparison with the standard. For certain
alkaline earth salts, permanganate is used to oxidize the iron.
Method 2, which uses 1,10-phenanthroline, is used generally for phosphate reagents. It is usually performed by adjusting the
pH (when necessary) to between 4 and 6, adding 6 mL of hydroxylamine hydrochloride reagent solution and 4 mL of 1,10-
phenanthroline reagent solution, and diluting with water to 25 mL. Because development of the color may be slow, comparison
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of the color produced is not made until 60 min after addition of the 1,10-phenanthroline reagent solution. The hydroxylamine
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hydrochloride reagent solution is the unacidified reagent solution and not the acidified solution used in the dithizone test for
lead and other metals.
The two general methods, which are referred to in the individual reagent monographs, are described here. Where a difference
appears between the general test method and the directions set forth in the individual reagent monograph, the directions in
the monograph are to be followed.
NITRATE
Four different tests are applied to the determination of nitrate: brucine sulfate, diphenylamine, indigo carmine, and
phenoldisulfonic acid. General procedures are provided below for brucine sulfate and diphenylamine.
S a m p l e S o l u t i o n A . Dissolve the specified quantity or volume of the sample in 3 mL of water by heating in a boiling water
bath. Dilute with brucine sulfate reagent solution to 50 mL.
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C o n t r o l S o l u t i o n B . To the specified volume of nitrate ion (NO3) standard solution, add the same quantity of sample as in
sample solution A and sufficient water to make a volume of 3 mL. Dissolve the mixture by heating in a boiling water bath.
Dilute with brucine sulfate reagent solution to 50 mL.
B l a n k S o l u t i o n C . Use 50 mL of brucine sulfate reagent solution. Heat the three solutions in a preheated (boiling) water
bath for 15 min with periodic gentle swirling. Cool rapidly in an ice water bath to room temperature. Set a
spectrophotometer at 410 nm, and using 1 cm quartz cells, adjust the instrument to read zero absorbance with blank
solution C in the light path, and then determine the absorbance of sample solution A and of control solution B. Calculate the
nitrate content as the following:
The prescribed tests must be followed carefully to ensure the development of a reproducible and stable color. Under
slightly different conditions, the oxidation reaction may produce a somewhat different color, which is not stable.
Place the specified quantity of the sample in a dry beaker. Cool the beaker thoroughly in an ice bath, and add 22 mL of
sulfuric acid that has been cooled to ice-bath temperature. Allow the mixture to warm to room temperature, and swirl the
beaker at intervals to produce gentle dissolution with slow evolution of the acid vapor. Prepare a standard by evaporating to
dryness a solution containing the specified quantity of nitrate ion (NO3) standard solution and 0.01 g of anhydrous sodium
carbonate. Treat the residue exactly as the sample. Add 3 mL of diphenylamine reagent solution to each solution, and
digest on a hot plate (~100 °C) for 90 min. Compare the color of the solutions visually. Any blue color produced in the
solution of the sample should not exceed that in the standard.
PHOSPHATE
Three general procedures are provided to test for phosphate: direct molybdenum blue (Method 1), extracted molybdenum
blue (Method 2), and precipitation (Method 3). In Method 2, conditions are established that allow only the phosphomolybdate
to be extracted into ether, thus eliminating any interference from arsenate and silicate.
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conical flask. Add 13 mL of ammonium hydroxide, warm the solution to 40 °C, and add 50 mL of freshly filtered ammonium
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molybdate–nitric acid reagent solution. Shake vigorously for 5 min, and allow to stand for 2 h at 40 ± 5 °C. Any yellow
precipitate obtained from the sample should not exceed that obtained from the standard.
SILICATE
Small amounts of silica or silicate are determined by a molybdenum blue method. Conditions have been established that allow
the extraction of the silicomolybdate and thus eliminate any interference from arsenate or phosphate.
SULFATE
Note: Use 12% barium chloride reagent solution unless stated otherwise; for reagent monographs for phosphates, use
40% barium chloride reagent solution.
Dilute to 10 mL and add 1 mL of barium chloride reagent solution. Any turbidity should not exceed that produced by 0.05 mg
of sulfate ion (SO4) in an equal volume of solution containing the quantities of reagents used in the test. Unless otherwise
specified, compare turbidity 10 min after adding the barium chloride to the sample and standard solutions.
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