Accessed from 128.83.63.

20 by nEwp0rt1 on Sat Dec 03 02:26:06 EST 2011

USP 35 Reagents / Reagents 993

Reagents, Indicators and

Solutions
This section deals with the reagents and solutions re- Where a particular brand or source of a material or piece
quired in conducting the Pharmacopeial and the National of equipment, or the name and address of a manufacturer,
Formulary tests and assays. is mentioned, this identification is furnished solely for infor-
As is stated in the General Notices, listing of reagents, in- mational purposes as a matter of convenience, without im-
dicators, and solutions in the Pharmacopeia in no way im- plication of approval, endorsement, or certification.
plies that they have therapeutic utility; thus, any reference Atomic absorption and flame photometry require the use
to the USP in their labeling is to include the term “reagent” of a number of metal-ion standard solutions. While the indi-
or “reagent grade.” vidual monographs usually provide directions for prepara-
Reagents required in the tests and assays for the Pharma- tion of these solutions, use of commercially prepared stan-
copeial and National Formulary articles are listed in this sec- dardized solutions of the appropriate ions is permissible,
tion, generally with specifications appropriate to their in- provided that the analyst confirms the suitability of the solu-
tended uses. Exceptions to the latter include those reagents tions and has data to support their use.
for which corresponding specifications are presented in the Reagents are substances used either as such or as constit-
current edition of Reagent Chemicals, published by the uents of solutions.
American Chemical Society, and reagents for which specifi- Indicators are reagents used to determine the specified
cations could not be drafted in time for inclusion here. endpoint in a chemical reaction, to measure hydrogen-ion
Thus, where it is directed to “Use ACS reagent grade,” it is concentration (pH), or to indicate that a desired change in
intended that a grade meeting the corresponding specifica- pH has been effected. They are listed together with indica-
tions of the current edition of ACS Reagent Chemicals shall tor test papers.
be used. Where no such specifications exist, and where it is Buffer Solutions are referred to separately.
directed to “Use a suitable grade,” the intent is that a suita- Colorimetric Solutions, abbreviated “CS,” are solutions
ble reagent grade available commercially shall be used. Oc- used in the preparation of colorimetric standards for com-
casionally, additional test(s) augment the designation “suita- parison purposes.
ble grade,” as indicated in the text. Listed also are some, Test Solutions, abbreviated “TS,” are solutions of re-
but not all, reagents that are required only in determining agents in such solvents and of such definite concentrations
the quality of other reagents. For those reagents that are as to be suitable for the specified purposes.
not listed, satisfactory specifications are available in standard Volumetric Solutions, abbreviated “VS” and known also
reference works. as Standard Solutions, are solutions of reagents of known
In those instances in which a reagent required in a Phar- concentration intended primarily for use in quantitative de-
macopeial or National Formulary test or assay need not be terminations. Concentrations are usually expressed in terms
of analytical reagent quality, it suffices to refer to the mono- of normality.
graph for that article appearing in this Pharmacopeia or the Water—As elsewhere in the Pharmacopeia, where
National Formulary or the current edition of the Food “water”, without qualification, is mentioned in the tests for
Chemicals Codex (FCC). In such cases it is to be understood reagents or in directions for preparing test solutions, etc.,
that the specifications are minimum requirements and that Purified Water (USP monograph) is always to be used. Car-
any substance meeting more rigid specifications for chemi- bon dioxide-free water is Purified Water that has been boiled
cal purity is suitable. vigorously for 5 minutes or more and allowed to cool while
Where the name of a reagent specified in a test or assay is protected from absorption of carbon dioxide from the at-
the same as the title of a USP or NF article, and it does not mosphere, or Purified Water that has a resistivity of not less
appear among the following Reagent Specifications, a sub- than 18 Mohm-cm. Deaerated water, for purposes other
stance meeting the requirements of the USP or NF mono- than dissolution and drug release testing, is Purified Water
graph is to be used (e.g., Benzocaine, USP; or Propylparaben, that has been treated to reduce the content of dissolved air
NF). However, reference is specifically made, under Reagent by suitable means, such as by boiling vigorously for 5 min-
Specifications, to a reagent bearing the name of a USP or NF utes and cooling or by the application of ultrasonic vibra-
article: (1) where there are requirements for a reagent in tion. Particle-free water is water that has been passed
addition to the USP or NF monograph requirements (e.g., through a 0.22-µm filter.
Sodium Salicylate, USP; or Isopropyl Myristate, NF), (2) where Organic-free water is Purified Water that produces no sig-
a source other than the USP or NF monograph is specified nificantly interfering peaks when chromatographed as it is
(e.g., Lactose, ACS reagent; or Hydrochloric Acid, ACS rea- indicated in Identification, Control, and Quantification of Re-
gent), (3) where complete reagent specifications differ from sidual Solvents under Residual Solvents 〈467〉
the USP or NF monograph standards (e.g., Calcium Lactate; Chromatographic Solvents and Carrier Gases—The
or Thymol), or (4) where a standard material is included chromatographic procedures set forth in the Pharmacopeia
among the reagent specifications (e.g., Calcium Carbonate, may require use of solvents and gases that have been espe-
primary standard; or Sodium Carbonate, primary standard). cially purified for such use. The purpose may be (a) to ex-
Reagents and solutions should be preserved in tight con- clude certain impurities that interfere with the proper con-
tainers made of resistant glass or other suitable material. duct of the test procedure, or (b) to extend the life of a
Directions for storage in light-resistant containers should be column by reducing the build-up of impurities on the col-
carefully observed. umn. Where solvents and gases are called for in chromato-
Stoppers and stopcocks brought into contact with sub- graphic procedures, it is the responsibility of the analyst to
stances capable of attacking or penetrating their surfaces ensure the suitability of the solvent or gas for the specific
may be given a protective coating of a thin film of a suita- use. Solvents and gases suitable for specific high-pressure or
ble lubricant unless specifically interdicted. other chromatographic uses are available as specialty prod-
ucts from various reagent supply houses, although there is

Official from May 1, 2012

Copyright (c) 2011 The United States Pharmacopeial Convention. All rights reserved.
 Accessed from 128.83.63.20 by nEwp0rt1 on Sat Dec 03 02:26:06 EST 2011

994 Reagents / Reagents USP 35

no assurance that similar products from different suppliers uses of the solvents and gases and not for chromatographic
are of equivalent suitability in any given procedure. The rea- uses for which the especially purified specialty products may
gent specifications provided herein are for general analytical be required.

Reagents
For the purposes of the following specifications, these def- Arsenic in Reagents
initions apply: A blank consists of the same quantities of the
same reagents treated in the same manner as the specimen Select reagents for this test for a low arsenic content, so
under test. A control is a blank to which has been added the that a blank test results in either no stain or one that is
limiting quantity of the substance being tested for, or is a barely perceptible.
specified comparison solution prepared as directed in the APPARATUS—Prepare a generator by fitting a 1-hole rubber
particular test. stopper into a wide-mouth bottle of about 60-mL capacity.
The values given in boldface type following chemical sym- Through the perforation insert a vertical exit tube about 12
bols and formulas represent, respectively, atomic and molec- cm in total length and 1 cm in diameter along the entire
ular weights of the substances concerned. upper portion (for about 8 cm) and constricted at its lower
Color and turbidity comparisons are to be made in color- extremity to a tube about 4 cm in length and about 5 mm
comparison tubes that are matched as closely as possible in in diameter. The smaller portion of the tube should extend
internal diameter and in all other respects, as directed for to just slightly below the stopper. Place washed sand or a
Visual Comparison under Spectrophotometry and Light-scatter- pledget of purified cotton in the upper portion to about 3
ing 〈851〉. Such tubes frequently are called “Nessler tubes.” cm from the top of the tube. Moisten the sand or cotton
In making visual comparisons of the densities of turbid uniformly with lead acetate TS, and remove any excess or
fluids, compensate for differences in color, if necessary, by adhering droplets of the latter from the walls of the tube.
viewing the turbidity through a column of water, the depth Into the upper end of this tube fit a second glass tube 12
of which is determined by the volume specified in the indi- cm in length, having an internal diameter of 2.5 to 3 mm,
vidual reagent specification. Place the water in color-com- by means of a rubber stopper. Just before running the test,
parison tubes, and hold one of the tubes above the control place a strip of mercuric bromide test paper (see under Indi-
tube and the other below the specimen tube. cator and Test Papers) in this tube, crimping the upper end
Where an expression such as “Retain the filtrate” appears of the strip so that it will remain in position about 2 cm
it is to be understood, unless otherwise indicated, that the above the rubber stopper. Clean and dry the tube thor-
washings of the residue are not to be added to the filtrate oughly each time it is used.
obtained. In the test heading, Calcium, magnesium, and R2O3
precipitate, the expression R2O3 is intended to indicate the STANDARD ARSENIC SOLUTION—Use Standard Preparation pre-
residue on ignition from compounds precipitated upon the pared as directed under Arsenic 〈211〉.
addition of ammonium hydroxide, such as Fe2O3 and Al2O3. TEST PREPARATION—Add 1 mL of sulfuric acid to 5 mL of a
solution of the chemical substance (1 in 25), unless another
quantity is directed in the individual reagent specification.
GENERAL TESTS FOR REAGENTS Omit its addition entirely in the case of inorganic acids. Un-
less especially directed otherwise, add 10 mL of sulfurous
The following general test methods are provided for the acid. Evaporate the liquid in a small beaker, on a steam
examination of reagents to determine their compliance with bath, until it is free from sulfurous acid and has been re-
the specifications of the individual reagents and are to be duced to about 2 mL in volume. Dilute with water to 5 mL
used unless it is otherwise directed in such specifications. to obtain the Test Preparation. Substances subjected to spe-
cial treatments specified in the individual reagent specifica-
tion may be used directly as the Test Preparation.
Boiling or Distilling Range for Reagents [NOTE—Solutions prepared by the dissolving of the chem-
ical substances in dilute acids are not considered to have
Use the following procedure for determining the boiling undergone special treatment.]
or distilling range of reagents, unless otherwise directed in STANDARD STAIN—Place in the generator bottle 5 mL of po-
the individual specifications: tassium iodide TS, 2.0 mL of Standard Arsenic Solution, 5 mL
APPARATUS—Use apparatus similar to that specified for Dis- of acid stannous chloride TS, and 28 mL of water. Add 1.5
tilling Range—Method I 〈721〉, except that the distilling flask g of granulated zinc (in No. 20 powder), and immediately
is to be of 250-mL capacity, to have a short neck, and to be insert the stopper containing the exit tube. Keep the gener-
connected to the condenser by means of a three-way con- ator bottle immersed in water at 25° during the period of
necting tube fitted with standard-taper ground joints. the test to moderate the reaction so that the stain will take
PROCEDURE—Place the distilling flask in an upright position the form of a distinctive band to facilitate the comparison of
in the perforation in the asbestos board, and connect it to color intensity. When evolution of hydrogen has continued
the condenser. for 1 hour, remove the mercuric bromide test paper for
Measure 100 mL of the liquid to be tested in a graduated comparison. This stain represents 2 µg of arsenic.
cylinder, and transfer to the boiling flask together with PROCEDURE—Pipet into the generator bottle 5 mL of potas-
some device to prevent bumping. Use the cylinder as the sium iodide TS and 5 mL of the Test Preparation, and add 5
receiver for the distillate. Insert the thermometer, and heat mL of acid stannous chloride TS. Set the apparatus aside at
so as to distill at the rate of 3 mL to 5 mL per minute. Make room temperature for a period of 10 minutes, then add 25
a preliminary trial, if necessary, to determine the adjustment mL of water and 1.5 g of granulated zinc (in No. 20 pow-
for the proper rate of heating. Read the thermometer when der), and proceed as directed under Standard Stain. Remove
about 20 drops have distilled and thereafter at volumes of the mercuric bromide test paper, and compare the stain
distillate of 5, 10, 40, 50, 60, 90, and 95 mL. Continue the upon it with the Standard Stain: the stain produced by the
distillation until the dry point is reached. chemical tested does not exceed the standard stain in
The Boiling or Distilling Range is the interval between the length or in intensity of color, indicating not more than 10
temperatures when 1 mL and 95 mL, respectively, have parts of arsenic per million parts of the substance being
distilled. tested. Since light, heat, and moisture cause the stain to

Official from May 1, 2012

Copyright (c) 2011 The United States Pharmacopeial Convention. All rights reserved.