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1886 Colorimetric Solutions / Solutions

with 100 mL of water, and titrate the liberated iodine with
0.1 N sodium thiosulfate VS, adding 3 mL of starch TS as
the indicator. Perform a blank determination with the same
quantities of the same reagents, and make any necessary
correction. Each mL of 0.1 N sodium thiosulfate is equivalent to 27.03 mg of FeCl3 6H2O. Adjust the final volume of
the solution by the addition of enough of the mixture of
hydrochloric acid and water so that each mL contains
45.0 mg of FeCl3 6H2O.

INDICATOR SOLUTIONS
See TEST SOLUTIONS.

TEST SOLUTIONS (TS)

1. USE AS INDICATORS
Certain test solutions are intended for use as acid-base
indicators in volumetric analyses. Such solutions should be
so adjusted that when 0.15 mL of the indicator solution is
added to 25 mL of carbon dioxide-free water, 0.25 mL of
0.02 N acid or alkali will produce the characteristic color
change. Similar solutions are intended for use in pH measurement. Where no special directions for their preparation
are given, the same solution is suitable for both purposes.

2. VOLUMETRIC SOLUTIONS USED AS TEST

SOLUTIONS
Where it is directed that a volumetric solution be used as
the test solution in a qualitative procedure, standardization
of the solution used as the test solution is not required.

Reagents

3. SOLUTIONS PREPARED FRESH

In general, the directive to prepare a solution fresh indicates that the solution is of limited stability and must be
prepared on the day of use.
Acetaldehyde TSMix 4 mL of acetaldehyde, 3 mL of alcohol, and 1 mL of water. Prepare this solution fresh.
Acetate Buffer TSDissolve 320 g of ammonium acetate in
500 mL of water, add 5 mL of glacial acetic acid, dilute with
water to 1000.0 mL, and mix. This solution has a pH between 5.9 and 6.0.
Acetic Acid, Glacial, TSDetermine the water content of a
specimen of glacial acetic acid by the Titrimetric Method (see
Water Determination 921). If the acid contains more than
0.4% of water, add a few mL of acetic anhydride, mix, allow to stand overnight, and again determine the water content. If the acid contains less than 0.02% of water, add
sufficient water to make the final concentration between
0.02% and 0.4%, mix, allow to stand overnight, and again
determine the water content. Repeat the adjustment with
acetic anhydride or water, as necessary, until the resulting
solution shows a water content of not more than 0.4%.
Acetic Acid, Strong, TSAdd 300.0 mL of glacial acetic
acid, and dilute with water to 1000 mL. This solution contains about 30% (v/v) of CH3COOH and has a concentration
of about 5 N.
Acetic AcidAmmonium Acetate Buffer TSDissolve
77.1 g of ammonium acetate in water, add 57 mL of glacial
acetic acid, and dilute with water to 1000 mL.
Acetone, Buffered, TSDissolve 8.15 g of sodium acetate
and 42 g of sodium chloride in about 100 mL of water, and
add 68 mL of 0.1 N hydrochloric acid and 150 mL of acetone. Mix, and dilute with water to 500 mL.
Acid Ferric Chloride TSMix 60 mL of glacial acetic acid
with 5 mL of sulfuric acid, add 1 mL of ferric chloride TS,
mix, and cool.
Acid Ferrous Sulfate TSSee Ferrous Sulfate, Acid, TS.

USP 38
Acid Stannous Chloride TSSee Stannous Chloride, Acid,
TS.
Acid Stannous Chloride TS, StrongerSee Stannous Chloride, Acid, Stronger, TS.
Albumen TSCarefully separate the white from the yolk of
a strictly fresh hens egg. Shake the white with 100 mL of
water until mixed and all but the chalaza has undergone
solution; then filter. Prepare the solution fresh.
AlcoholPhenol TSDissolve 780 mg of phenol in alcohol
to make 100 mL.
Alcoholic TSIt contains 95 parts of specially denaturated
alcohol 3A with 5 parts of isopropyl alcohol. The final concentrations are approximately 90% alcohol, 5% methanol,
and 5% isopropanol.
[NOTEA suitable grade is available as Reagent alcohol,
catalog number R8382, available at www.sigma-aldrich.
com.]
Alcoholic Ammonia TSSee Ammonia TS, Alcoholic.
Alcoholic Mercuric Bromide TSSee Mercuric Bromide TS,
Alcoholic.
Alcoholic Potassium Hydroxide TSSee Potassium Hydroxide TS, Alcoholic.
Alkaline Cupric Citrate TSSee Cupric Citrate TS, Alkaline.
Alkaline Cupric Citrate TS 2See Cupric Citrate TS 2, Alkaline.
Alkaline Cupric Iodide TSSee Cupric Iodide TS, Alkaline.
Alkaline Cupric Tartrate TS (Fehlings Solution)See Cupric
Tartrate TS, Alkaline.
Alkaline MercuricPotassium Iodide TSSee
MercuricPotassium Iodide TS, Alkaline.
Alkaline Picrate TSSee Picrate TS, Alkaline.
Alkaline Sodium Hydrosulfite TSSee Sodium Hydrosulfite
TS, Alkaline.
Amaranth TSDissolve 20 mg of amaranth in 10 mL of
water.
Aminonaphtholsulfonic Acid TSAccurately weigh 5 g of
sodium sulfite, 94.3 g of sodium bisulfite, and 700 mg of
1,2,4-aminonaphtholsulfonic acid, and mix. Prepare aminonaphtholsulfonic acid TS fresh on the day of use by dissolving 1.5 g of the dry mixture in 10 mL of water.
Ammonia TSIt contains between 9.5% and 10.5% of
NH3. Prepare by diluting 350 mL of Ammonia Water,
Stronger (see in the section, Reagents) with water to make
1000 mL.
Ammonia TS 2Prepare by diluting 13.5 mL of Ammonia
Water, Stronger (see Reagent Specifications in the section Reagents) with water to make 100 mL.
Ammonia TS, AlcoholicA solution of ammonia gas in alcohol. Clear, colorless liquid having a strong odor of ammonia. Specific gravity: about 0.80. It contains between 9%
and 11% of NH3. Store it in alkali-resistant containers, in a
cold place.
Ammonia TS, StrongerUse Ammonia Water, Stronger (see
in the section Reagents).
AmmoniaAmmonium Chloride Buffer TSDissolve
67.5 g of ammonium chloride in water, add 570 mL of ammonium hydroxide, and dilute with water to 1000 mL.
AmmoniaCyanide TSDissolve 2 g of potassium cyanide
in 15 mL of ammonium hydroxide, and dilute with water to
100 mL.
Ammoniacal Potassium Ferricyanide TSDissolve 2 g of
potassium ferricyanide in 75 mL of water, add 25 mL of ammonium hydroxide, and mix.
Ammoniated Cupric Oxide TSSee Cupric Oxide, Ammoniated, TS.
Ammonium Acetate TSDissolve 10 g of ammonium acetate in water to make 100 mL.
Ammonium Carbonate TSDissolve 20 g of ammonium
carbonate and 20 mL of ammonia TS in water to make
100 mL.
Ammonium Carbonate TS 2Prepare a 158-mg/mL solution of ammonium carbonate in water.
Ammonium Chloride TSDissolve 10.5 g of ammonium
chloride in water to make 100 mL.

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USP 38

Solutions / Test Solutions 1887

phate solution (1 in 10) in a low-actinic glass bottle. Mix,
and allow to stand overnight before using. Protect from
light, and use within 7 days.
Bromocresol Blue TSUse Bromocresol Green TS.
Bromocresol Green TSDissolve 50 mg of bromocresol
green in 100 mL of alcohol, and filter if necessary.
Bromocresol GreenMethyl Red TSDissolve 0.15 g of
bromocresol green and 0.1 g of methyl red in 180 mL of
alcohol, and dilute with water to 200 mL.
Bromocresol Purple TSDissolve 250 mg of bromocresol
purple in 20 mL of 0.05 N sodium hydroxide, and dilute
with water to 250 mL.
Bromophenol Blue TSDissolve 100 mg of bromophenol
blue in 100 mL of diluted alcohol, and filter if necessary.
Bromothymol Blue TSDissolve 100 mg of bromothymol
blue in 100 mL of diluted alcohol, and filter if necessary.
Buffered Acetone TSSee Acetone, Buffered, TS.
Calcium Chloride TSDissolve 7.5 g of calcium chloride in
water to make 100 mL.
Calcium Hydroxide TSUse Calcium Hydroxide Topical Solution (USP monograph).
Calcium Sulfate TSA saturated solution of calcium sulfate
in water.
Ceric Ammonium Nitrate TSDissolve 6.25 g of ceric ammonium nitrate in 10 mL of 0.25 N nitric acid. Use within 3
days.
Chloral Hydrate TSDissolve 50 g of chloral hydrate in a
mixture of 15 mL of water and 10 mL of glycerin.
Chlorine TS (Chlorine Water)A saturated solution of chlorine in water. Place the solution in small, completely filled,
light-resistant containers. Chlorine TS, even when kept from
light and air, is apt to deteriorate. Store it in a cold, dark
place. For full strength, prepare this solution fresh.
Chromotropic Acid TSDissolve 50 mg of chromotropic
acid or its disodium salt in 100 mL of 75% sulfuric acid,
which may be made by cautiously adding 75 mL of sulfuric
acid to 33.3 mL of water.
CobaltUranyl Acetate TSDissolve, with warming, 40 g
of uranyl acetate in a mixture of 30 g of glacial acetic acid
and sufficient water to make 500 mL. Similarly, prepare a
solution containing 200 g of cobaltous acetate in a mixture
of 30 g of glacial acetic acid and sufficient water to make
500 mL. Mix the two solutions while still warm, and cool to
20. Maintain the temperature at 20 for about 2 hours to
separate the excess salts from solution, and then pass
through a dry filter.
Cobaltous Chloride TSDissolve 2 g of cobaltous chloride
in 1 mL of hydrochloric acid and sufficient water to make
100 mL.
Congo Red TSDissolve 500 mg of congo red in a mixture
of 10 mL of alcohol and 90 mL of water.
m-Cresol Purple TSDissolve 0.10 g of metacresol purple
in 13 mL of 0.01 N sodium hydroxide, dilute with water to
100 mL, and mix.
Cresol Red TSTriturate 100 mg of cresol red in a mortar
with 26.2 mL of 0.01 N sodium hydroxide until solution is
complete, then dilute the solution with water to 250 mL.
Cresol RedThymol Blue TSAdd 15 mL of thymol blue TS
to 5 mL of cresol red TS, and mix.
Crystal Violet TSDissolve 100 mg of crystal violet in
10 mL of glacial acetic acid.
Cupric Acetate TSDissolve 100 mg of cupric acetate in
about 5 mL of water to which a few drops of acetic acid
have been added. Dilute to 100 mL, and filter, if necessary.
Cupric Acetate TS, Stronger (Barfoeds Reagent)Dissolve
13.3 g of cupric acetate in a mixture of 195 mL of water
and 5 mL of acetic acid.
Cupric-Ammonium Sulfate TSTo cupric sulfate TS add
ammonia TS, dropwise, until the precipitate initially formed
is nearly but not completely dissolved. Allow to settle, and
decant the clear solution. Prepare this solution fresh.
Cupric Citrate TSDissolve 25 g of cupric sulfate, 50 g of
citric acid, and 144 g of anhydrous sodium carbonate in
water, and dilute with water to 1000 mL.

Official from August 1, 2015

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Reagents

Ammonium ChlorideAmmonium Hydroxide TSMix

equal volumes of water and ammonium hydroxide, and saturate with ammonium chloride.
Ammonium Molybdate TSDissolve 6.5 g of finely powdered molybdic acid in a mixture of 14 mL of water and
14.5 mL of ammonium hydroxide. Cool the solution, and
add it slowly, with stirring, to a well-cooled mixture of
32 mL of nitric acid and 40 mL of water. Allow to stand for
48 hours, and filter through a fine-porosity, sintered-glass
crucible. This solution deteriorates upon standing and is unsuitable for use if, upon the addition of 2 mL of dibasic sodium phosphate TS to 5 mL of the solution, an abundant
yellow precipitate does not form at once or after slight
warming. Store it in the dark. If a precipitate forms during
storage, use only the clear supernatant.
Ammonium Oxalate TSDissolve 3.5 g of ammonium oxalate in water to make 100 mL.
Ammonium Phosphate, Dibasic, TS (Ammonium Phosphate
TS)Dissolve 13 g of dibasic ammonium phosphate in
water to make 100 mL.
Ammonium Polysulfide TSYellow liquid, made by saturating ammonium sulfide TS with sulfur.
Ammonium Pyrrolidinedithiocarbamate, Saturated, TS
Add about 10 g of ammonium pyrrolidinedithiocarbamate
to a 1000-mL volumetric flask, and dilute with water to volume.
Ammonium Reineckate TSShake about 500 mg of ammonium reineckate with 20 mL of water frequently during
1 hour, and filter. Use within 2 days.
Ammonium Sulfide TSUse ACS reagent-grade Ammonium Sulfide Solution.
Ammonium Thiocyanate TSDissolve 8 g of ammonium
thiocyanate in water to make 100 mL.
Ammonium Vanadate TSDissolve 2.5 g of ammonium
vanadate in 500 mL of boiling water, cool, and add 20 mL
of nitric acid. Mix, cool, and add water to make 1 L. Store
in polyethylene containers.
Anthrone TSWithin 12 hours of use, rapidly dissolve
35 mg of anthrone in a hot mixture of 35 mL of water and
65 mL of sulfuric acid. Immediately cool in an ice bath to
room temperature, and filter through glass wool. Allow the
solution to stand at room temperature for 30 minutes before
use.
Antimony Trichloride TSDissolve 20 g of antimony
trichloride in chloroform to make 100 mL. Filter if necessary.
Barium Chloride TSDissolve 12 g of barium chloride in
water to make 100 mL.
Barium Hydroxide TSA saturated solution of barium hydroxide in recently boiled water. Prepare the solution fresh.
Barium Nitrate TSDissolve 6.5 g of barium nitrate in
water to make 100 mL.
Betanaphthol TSSee 2-Naphthol TS.
Biuret Reagent TSDissolve 1.5 g of cupric sulfate and
6.0 g of potassium sodium tartrate in 500 mL of water in a
1000-mL volumetric flask. Add 300 mL of carbonate-free sodium hydroxide solution (1 in 10), dilute with carbonatefree sodium hydroxide solution (1 in 10) to 1000 mL, and
mix.
Blue Tetrazolium TSDissolve 500 mg of blue tetrazolium in alcohol to make 100 mL.
Brilliant Blue G TSTransfer 25 mg of brilliant blue G to a
100-mL volumetric flask, add 12.5 mL of alcohol and 25 mL
of phosphoric acid, dilute with water to volume, and mix.
Bromine TS (Bromine Water)A saturated solution of bromine, prepared by agitating 2 to 3 mL of bromine with
100 mL of cold water in a glass-stoppered bottle, the stopper of which should be lubricated with petrolatum. Store it
in a cold place, protected from light.
BromineSodium Acetate TSDissolve 100 g of sodium
acetate in 1000 mL of glacial acetic acid, add 50 mL of bromine, and mix.
p-Bromoaniline TSAdd 8 g of p-bromoaniline to a mixture of 380 mL of thiourea-saturated glacial acetic acid,
10 mL of sodium chloride solution (1 in 5), 5 mL of oxalic
acid solution (1 in 20), and 5 mL of dibasic sodium phos-

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Reagents

1888 Test Solutions / Solutions

Cupric Citrate TS, AlkalineWith the aid of heat, dissolve
173 g of dihydrated sodium citrate and 117 g of monohydrated sodium carbonate in about 700 mL of water, and
filter through paper, if necessary, to obtain a clear solution.
In a separate container dissolve 17.3 g of cupric sulfate in
about 100 mL of water, and slowly add this solution, with
constant stirring, to the first solution. Cool the mixture, add
water to make 1000 mL, and mix.
Cupric Citrate TS 2, AlkalineWith the aid of heat, dissolve about 173 g of sodium citrate dihydrate and 117 g of
sodium carbonate monohydrate in about 700 mL of water,
and filter. In a second flask, dissolve about 27.06 g of cupric
sulfate (CuSO4 5H2O) in about 100 mL of water. Slowly
combine the two solutions while stirring, and dilute with
water to 1000 mL.
Cupric Iodide TS, AlkalineDissolve 7.5 g of cupric sulfate
(CuSO4 5H2O) in about 100 mL of water. In a separate
container dissolve 25 g of anhydrous sodium carbonate,
20 g of sodium bicarbonate, and 25 g of potassium sodium
tartrate in about 600 mL of water. With constant stirring,
add the cupric sulfate solution to the bottom of the alkaline
tartrate solution by means of a funnel that touches the bottom of the container. Add 1.5 g of potassium iodide, 200 g
of anhydrous sodium sulfate, 50 to 150 mL of 0.02 M potassium iodate, and sufficient water to make 1000 mL.
Cupric Oxide, Ammoniated, TS (Schweitzers Reagent)
Dissolve 10 g of cupric sulfate in 100 mL of water, add sufficient sodium hydroxide solution (1 in 5) to precipitate the
copper hydroxide, collect the latter on a filter, and wash
free from sulfate with cold water. Dissolve the precipitate,
which must be kept wet during the entire process, in the
minimum quantity of ammonia TS necessary for complete
solution.
Cupric Sulfate TSDissolve 12.5 g of cupric sulfate in
water to make 100 mL.
Cupric Tartrate TS, Alkaline (Fehlings Solution)
THE COPPER SOLUTION (A)Dissolve 34.66 g of carefully selected, small crystals of cupric sulfate, showing no trace of
efflorescence of adhering moisture, in water to make
500 mL. Store this solution in small, tight containers.
THE ALKALINE TARTRATE SOLUTION (B)Dissolve 173 g of
crystallized potassium sodium tartrate and 50 g of sodium
hydroxide in water to make 500 mL. Store this solution in
small, alkali-resistant containers.
For use, mix exactly equal volumes of Solutions A and B
at the time required.
Delafields Hematoxylin TSPrepare 400 mL of a saturated
solution of ammonium alum (Solution A). Dissolve 4 g of
hematoxylin in 25 mL of alcohol, mix it with Solution A, and
allow it to stand for 4 days in a flask closed with a pledget
of purified cotton and exposed to light and air (Solution B).
Then filter Solution B, and add to it a Solution C consisting of
a mixture of 100 mL of glycerin and 100 mL of methanol.
Mix, and allow the mixture to stand in a warm place, exposed to light, for 6 weeks until it becomes dark-colored.
Store in tightly stoppered bottles.
For use in staining endocrine tissue, dilute this test solution with an equal volume of water.
Denaturated Alcoholic TSA specially denaturated alcohol
containing either rubber hydrocarbon solvent of heptane or
toluene. [NOTEA suitable grade is available from www.lyondell.com or from www.sasol.com, as Ethanol SDA 2B HEP
200, or Ethanol SDA 2B TOL 200, or Ethanol SDA 2B TOL
190, or Alcohol SDA 2B-2.]
Denig`es ReagentSee Mercuric Sulfate TS.
Diazobenzenesulfonic Acid TSPlace in a beaker 1.57 g of
sulfanilic acid, previously dried at 105 for 3 hours, add
80 mL of water and 10 mL of diluted hydrochloric acid, and
warm on a steam bath until dissolved. Cool to 15 (some of
the sulfanilic acid may separate but will dissolve later), and
add slowly, with constant stirring, 6.5 mL of sodium nitrite
solution (1 in 10). Then dilute with water to 100 mL.

USP 38
Dichlorofluorescein TSDissolve 100 mg of dichlorofluorescein in 60 mL of alcohol, add 2.5 mL of 0.1 N sodium
hydroxide, mix, and dilute with water to 100 mL.
2,7-Dihydroxynaphthalene TSDissolve 100 mg of 2,7dihydroxynaphthalene in 1000 mL of sulfuric acid, and allow
the solution to stand until the yellow color disappears. If the
solution is very dark, discard it and prepare a new solution
from a different supply of sulfuric acid. This solution is stable
for approximately 1 month if stored in a dark bottle.
Diiodofluorescein TSDissolve 500 mg of diiodofluorescein
in a mixture of 75 mL of alcohol and 30 mL of water.
Diluted Lead Subacetate TSSee Lead Subacetate TS, Diluted.
p-Dimethylaminobenzaldehyde TSDissolve 125 mg of pdimethylaminobenzaldehyde in a cooled mixture of 65 mL
of sulfuric acid and 35 mL of water, and add 0.05 mL of
ferric chloride TS. Use within 7 days.
Dinitrophenylhydrazine TSCarefully mix 10 mL of water
and 10 mL of sulfuric acid, and cool. To the mixture, contained in a glass-stoppered flask, add 2 g of 2,4-dinitrophenylhydrazine, and shake until dissolved. To the solution add
35 mL of water, mix, cool, and filter.
Diphenylamine TSDissolve 1.0 g of diphenylamine in
100 mL of sulfuric acid. The solution should be colorless.
Diphenylcarbazone TSDissolve 1 g of crystalline
diphenylcarbazone in 75 mL of alcohol, then add alcohol to
make 100 mL. Store in a brown bottle.
Dithizone TSDissolve 25.6 mg of dithizone in 100 mL of
alcohol. Store in a cold place, and use within 2 months.
Dragendorffs TSMix 850 mg of bismuth subnitrate with
40 mL of water and 10 mL of glacial acetic acid (Solution A).
Dissolve 8 g of potassium iodide in 20 mL of water (Solution
B). Mix equal portions of Solution A and Solution B to obtain
a stock solution, which can be stored for several months in
a dark bottle. Mix 10 mL of the stock solution with 20 mL of
glacial acetic acid, and dilute with water to make 100 mL.
Edetate Disodium TSDissolve 1 g of edetate disodium in
950 mL of water, add 50 mL of alcohol, and mix.
Eosin Y TS (adsorption indicator)Dissolve 50 mg of eosin
Y in 10 mL of water.
Eriochrome Black TSDissolve 200 mg of eriochrome black
T and 2 g of hydroxylamine hydrochloride in methanol to
make 50 mL.
Eriochrome Cyanine TSDissolve 750 mg of eriochrome
cyanine R in 200 mL of water, add 25 g of sodium chloride,
25 g of ammonium nitrate, and 2 mL of nitric acid, and dilute with water to 1000 mL.
Fehlings SolutionSee Cupric Tartrate TS, Alkaline.
Ferric Ammonium Sulfate TSDissolve 8 g of ferric ammonium sulfate in water to make 100 mL.
Ferric Chloride TSDissolve 9 g of ferric chloride in water
to make 100 mL.
Ferroin TSDissolve 0.7 g of ferrous sulfate and 1.76 g of
o-phenanthroline monohydrochloride monohydrate in
water, and dilute with water to 100 mL.
Ferrous Sulfate TSDissolve 8 g of clear crystals of ferrous
sulfate in about 100 mL of recently boiled and thoroughly
cooled water. Prepare this solution fresh.
Ferrous Sulfate, Acid, TSDissolve 7 g of ferrous sulfate
crystals in 90 mL of recently boiled and thoroughly cooled
water, and add sulfuric acid to make 100 mL. Prepare this
solution immediately prior to use.
Folin-Ciocalteu Phenol TSInto a 1500-mL flask introduce
100 g of sodium tungstate, 25 g of sodium molybdate,
700 mL of water, 50 mL of phosphoric acid, and 100 mL of
hydrochloric acid. Gently reflux the mixture for about
10 hours, and add 150 g of lithium sulfate, 50 mL of water,
and a few drops of bromine. Boil the mixture, without the
condenser, for about 15 minutes, or until the excess bromine is expelled. Cool, dilute with water to 1 L, and filter:
the filtrate has no greenish tint. Before use, dilute 1 part of
the filtrate with 1 part of water. When used for protein determination (i.e., Lowry assay), this reagent must be further
diluted (1:5) with water. See Method 2 in Total Protein Assay

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USP 38

Solutions / Test Solutions 1889

make 250 mL. Add to the resulting solution an equal volume of sodium acetate solution freshly prepared by dissolving 13.66 g of anhydrous sodium acetate in water to make
500 mL and adjusting with 0.5 N acetic acid to a pH of 7.
Store in a refrigerator, and use within 2 weeks.
Intestinal Fluid, Simulated, TSDissolve 6.8 g of monobasic potassium phosphate in 250 mL of water, mix, and add
77 mL of 0.2 N sodium hydroxide and 500 mL of water.
Add 10.0 g of pancreatin, mix, and adjust the resulting solution with either 0.2 N sodium hydroxide or 0.2 N hydrochloric acid to a pH of 6.8 0.1. Dilute with water to
1000 mL.
Iodine TSUse 0.1 N Iodine (see in the section Volumetric
Solutions).
Iodine, Diluted TSTransfer 10.0 mL of 0.1 N iodine VS to
a 100-mL volumetric flask, dilute with water to volume, and
mix.
Iodine Monochloride TSDissolve 10 g of potassium iodide and 6.44 g of potassium iodate in 75 mL of water in a
glass-stoppered container. Add 75 mL of hydrochloric acid
and 5 mL of chloroform, and adjust to a faint iodine color
(in the chloroform) by adding dilute potassium iodide or
potassium iodate solution. If much iodine is liberated, use a
stronger solution of potassium iodate than 0.01 M at first,
making the final adjustment with the 0.01 M potassium iodate. Store in a dark place, and readjust to a faint iodine
color as necessary.
Iodine and Potassium Iodide TS 1Dissolve 500 mg of iodine and 1.5 g of potassium iodide in 25 mL of water.
Iodine and Potassium Iodide TS 2Dissolve 12.7 g of iodine and 20 g of potassium iodide in water, and dilute with
water to 1000.0 mL. To 10.0 mL of this solution, add 0.6 g
of potassium iodide, and dilute with water to 100.0 mL. Prepare immediately before use.
Iodine and Potassium Iodide TS 3Dissolve 0.127 g of iodine and 0.20 g of potassium iodide in water, and dilute
with water to 10.0 mL.
Iodobromide TSDissolve 20 g of iodine monobromide in
glacial acetic acid to make 1000 mL. Store in glass containers, protected from light.
Iodochloride TSDissolve 16.5 g of iodine monochloride in
1000 mL of glacial acetic acid.
Iodoplatinate TSDissolve 300 mg of platinic chloride in
97 mL of water. Immediately prior to use, add 3.5 mL of
potassium iodide TS, and mix.
IronPhenol TS (Iron-Kober Reagent)Dissolve 1.054 g of
ferrous ammonium sulfate in 20 mL of water, and add 1 mL
of sulfuric acid and 1 mL of 30 percent hydrogen peroxide.
Mix, heat until effervescence ceases, and dilute with water
to 50 mL. To 3 volumes of this solution contained in a volumetric flask add sulfuric acid, with cooling, to make 100
volumes. Purify phenol by distillation, discarding the first
10% and the last 5%, collecting the distillate, with exclusion
of moisture, in a dry, tared glass-stoppered flask of about
twice the volume of the phenol. Solidify the phenol in an
ice bath, breaking the top crust with a glass rod to ensure
complete crystallization. Weigh the flask and its contents,
add to the phenol 1.13 times its weight of the ironsulfuric
acid solution prepared as directed, insert the stopper in the
flask, and allow to stand, without cooling but with occasional mixing, until the phenol is liquefied. Shake the mixture vigorously until mixed, allow to stand in the dark for
16 to 24 hours, and again weigh the flask and its contents.
To the mixture add 23.5% of its weight of a solution of 100
volumes of sulfuric acid in 110 volumes of water, mix, transfer to dry glass-stoppered bottles, and store in the dark,
protected from atmospheric moisture. Use within 6 months.
Dispense the reagent from a small-bore buret, arranged to
exclude moisture, capable of delivering 1 mL in 30 seconds
or less, and having no lubricant, other than reagent, on its
stopcock. Wipe the buret tip with tissue before each addition.
Iron Salicylate TSDissolve 500 mg of ferric ammonium
sulfate in 250 mL of water containing 10 mL of diluted sulfuric acid, and add water to make 500 mL. To 100 mL of

Official from August 1, 2015

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Reagents

under Biotechnology-Derived ArticlesTotal Protein Assay

1057.
Formaldehyde TSUse Formaldehyde Solution (see in the
section Reagents).
FuchsinPyrogallol TSDissolve 100 mg of basic fuchsin in
50 mL of water that previously has been boiled for 15 minutes and allowed to cool slightly. Cool, add 2 mL of a saturated solution of sodium bisulfite, mix, and allow to stand
for not less than 3 hours. Add 0.9 mL of hydrochloric acid,
mix, and allow to stand overnight. Add 100 mg of pyrogallol, shake until solution is effected, and dilute with water to
100 mL. Store in an amber-colored glass bottle in a refrigerator.
FuchsinSulfurous Acid TSDissolve 200 mg of basic fuchsin in 120 mL of hot water, and allow the solution to cool.
Add a solution of 2 g of anhydrous sodium sulfite in 20 mL
of water, then add 2 mL of hydrochloric acid. Dilute the
solution with water to 200 mL, and allow to stand for at
least 1 hour. Prepare this solution fresh.
Gastric Fluid, Simulated, TSDissolve 2.0 g of sodium
chloride and 3.2 g of purified pepsin, that is derived from
porcine stomach mucosa, with an activity of 800 to 2500
units per mg of protein, in 7.0 mL of hydrochloric acid and
sufficient water to make 1000 mL. [NOTEPepsin activity is
described in the Food Chemicals Codex specifications under
General Tests and Assays.] This test solution has a pH of
about 1.2.
Gelatin TS (for the assay of Corticotropin Injection)Dissolve
340 g of acid-treated precursor gelatin (Type A) in water to
make 1000 mL. Heat the solution in an autoclave at 115
for 30 minutes after the exhaust line temperature has
reached 115. Cool the solution, and add 10 g of phenol
and 1000 mL of water. Store in tight containers in a refrigerator.
Glacial Acetic Acid TSSee Acetic Acid, Glacial, TS.
Glucose OxidaseChromogen TSA solution containing,
in each mL, 0.5 mol of 4-aminoantipyrine, 22.0 mol of
sodium p-hydroxybenzoate, not less than 7.0 units of glucose oxidase, and not less than 0.5 units of peroxidase, and
buffered to a pH of 7.0 0.1.
SUITABILITYWhen used for determining glucose in Inulin,
ascertain that no significant color results by reaction with
fructose, and that a suitable absorbance-versus-concentration slope is obtained with glucose.
[NOTEGlucose oxidase can be from Aspergillus niger.]
Glycerin Base TSTo 200 g of glycerin add water to bring
the total weight to 235 g. Add 140 mL of 1 N sodium hydroxide and 50 mL of water.
Gold Chloride TSDissolve 1 g of gold chloride in 35 mL
of water.
Hydrogen Peroxide TSUse Hydrogen Peroxide Topical Solution (USP monograph).
Hydrogen Sulfide TSA saturated solution of hydrogen
sulfide, made by passing H2S into cold water. Store it in
small, dark amber-colored bottles, filled nearly to the top. It
is unsuitable unless it possesses a strong odor of H2S, and
unless it produces at once a copious precipitate of sulfur
when added to an equal volume of ferric chloride TS. Store
in a cold, dark place.
Hydroxylamine Hydrochloride TSDissolve 3.5 g of hydroxylamine hydrochloride in 95 mL of 60% alcohol, and
add 0.5 mL of bromophenol blue solution (1 in 1000 of
alcohol) and 0.5 N alcoholic potassium hydroxide until a
greenish tint develops in the solution. Then add 60% alcohol to make 100 mL.
8-Hydroxyquinoline TSDissolve 5 g of 8-hydroxyquinoline in alcohol to make 100 mL.
Indigo Carmine TS (Sodium Indigotindisulfonate TS)Dissolve a quantity of sodium indigotindisulfonate, equivalent
to 180 mg of C16H8N2O2(SO3Na)2, in water to make 100 mL.
Use within 60 days.
IndophenolAcetate TS (for the assay of Corticotropin Injection)To 60 mL of standard dichlorophenol-indophenol solution (see in the section Volumetric Solutions) add water to

Accessed from 10.6.1.1 by gsk on Fri Oct 30 04:44:36 EDT 2015

1890 Test Solutions / Solutions

USP 38

the resulting solution add 50 mL of a 1.15% solution of sodium salicylate, 20 mL of diluted acetic acid, and 80 mL of a
13.6% solution of sodium acetate, then add water to make
500 mL. Store in a well-closed container. Protect from light.
Use within 2 weeks.
Lanthanum Nitrate TSDissolve 5.0 g of lanthanum nitrate
hexahydrate in 100 mL of water.
Lead Acetate TSDissolve 9.5 g of clear, transparent crystals of lead acetate in recently boiled water to make
100 mL. Store in well-stoppered bottles.
Lead Acetate TS, AlcoholicDissolve 2 g of clear, transparent crystals of lead acetate in alcohol to make 100 mL. Store
in tight containers.
Add the following:

Lead Nitrate Stock Solution TSDissolve 159.8 mg of

lead nitrate in 100 mL of water to which 1 mL of nitric acid
has been added, then dilute with water to 1000 mL. Prepare
and store this solution in glass containers free from soluble
lead salts.USP38
Lead Subacetate TSDissolve 40.0 g of lead acetate in
90 mL of carbon dioxide-free water. Adjust with 10 M sodium hydroxide to a pH of 7.5, centrifuge, and use the clear
supernatant. It contains NLT 16.7% (w/w) and NMT 17.4%
(w/w) of Pb in a form corresponding to the formula
C8H14O10Pb3. The solution remains clear when stored in a
well-closed container.
Lead Subacetate TS, DilutedDilute 3.25 mL of lead
subacetate TS with water, recently boiled and cooled, to
make 100 mL. Store in small, well-filled, tight containers.
Litmus TSDigest 25 g of powdered litmus with three successive 100-mL portions of boiling alcohol, continuing each
extraction for about 1 hour. Filter, wash with alcohol, and
discard the alcohol filtrate. Macerate the residue with about
25 mL of cold water for 4 hours, filter, and discard the filtrate. Finally digest the residue with 125 mL of boiling water
for 1 hour, cool, and filter.
Locke-Ringers SolutionSee Locke-Ringers TS.
Locke-Ringers TS (Locke-Ringers Solution)

Reagents

Sodium Chloride
Potassium Chloride
Calcium Chloride
Magnesium Chloride
Sodium Bicarbonate
Dextrose
Water, recently distilled from a hard-glass
flask, a sufficient quantity to make

9.0
0.42
0.24
0.2
0.5
0.5

g
g
g
g
g
g

1000 mL

Prepare fresh each day. The constituents (except the dextrose and the sodium bicarbonate) may be made up in
stock solutions and diluted as needed.
Magnesia Mixture TSDissolve 5.5 g of magnesium chloride and 7 g of ammonium chloride in 65 mL of water, add
35 mL of ammonia TS, set the mixture aside for a few days
in a well-stoppered bottle, and filter. If the solution is not
perfectly clear, filter it before using.
Magnesium Sulfate TSDissolve 12 g of crystals of magnesium sulfate, selected for freedom from efflorescence, in
water to make 100 mL.
Malachite Green TSDissolve 1 g of malachite green oxalate in 100 mL of glacial acetic acid.
Mallorys StainDissolve 500 mg of water-soluble aniline
blue, 2 g of orange G, and 2 g of oxalic acid in 100 mL of
water.
Mayers ReagentSee MercuricPotassium Iodide TS.
Mercuric Acetate TSDissolve 6.0 g of mercuric acetate in
glacial acetic acid to make 100 mL. Store in tight containers,
protected from direct sunlight.

MercuricAmmonium Thiocyanate TSDissolve 30 g of

ammonium thiocyanate and 27 g of mercuric chloride in
water to make 1000 mL.
Mercuric Bromide TS, AlcoholicDissolve 5 g of mercuric
bromide in 100 mL of alcohol, employing gentle heat to
facilitate solution. Store in glass containers, protected from
light.
Mercuric Chloride TSDissolve 6.5 g of mercuric chloride
in water to make 100 mL.
Mercuric Iodide TS (Valsers Reagent)Slowly add potassium iodide solution (1 in 10) to red mercuric iodide until
almost all of the latter is dissolved, and filter off the excess.
A solution containing 10 g of potassium iodide in 100 mL
dissolves approximately 14 g of HgI2 at 20.
Mercuric Nitrate TSDissolve 40 g of mercuric oxide (red
or yellow) in a mixture of 32 mL of nitric acid and 15 mL of
water. Store in glass containers, protected from light.
MercuricPotassium Iodide TS (Mayers Reagent)Dissolve
1.358 g of mercuric chloride in 60 mL of water. Dissolve 5 g
of potassium iodide in 10 mL of water. Mix the two solutions, and dilute with water to 100 mL.
MercuricPotassium Iodide TS, Alkaline (Nesslers Reagent)Dissolve 143 g of sodium hydroxide in 700 mL of
water. Dissolve 50 g of red mercuric iodide and 40 g of potassium iodide in 200 mL of water. Pour the iodide solution
into the hydroxide solution, and dilute with water to
1000 mL. Allow to settle, and use the clear supernatant.
Mercuric Sulfate TS (Denig`es Reagent)Mix 5 g of yellow
mercuric oxide with 40 mL of water, and while stirring
slowly add 20 mL of sulfuric acid, then add another 40 mL
of water, and stir until completely dissolved.
Mercurous Nitrate TSDissolve 15 g of mercurous nitrate
in a mixture of 90 mL of water and 10 mL of diluted nitric
acid. Store in dark, amber-colored bottles in which a small
globule of mercury has been placed.
Metaphenylenediamine Hydrochloride TSDissolve 1 g of
metaphenylenediamine hydrochloride in 200 mL of water.
The solution must be colorless when used. If necessary, decolorize by heating with activated charcoal.
MetaphosphoricAcetic Acids TSDissolve 15 g of metaphosphoric acid in 40 mL of glacial acetic acid and sufficient
water to make 500 mL. Store in a cold place, and use within
2 days.
Methoxyphenylacetic TSDissolve 2.7 g of methoxyphenylacetic acid in 6 mL of Tetramethylammonium Hydroxide TS,
and add 20 mL of dehydrated alcohol. Store in a polyethylene container.
Methyl Orange TSDissolve 100 mg of methyl orange in
100 mL of water, and filter if necessary.
Methyl Purple TSUse Methyl RedMethylene Blue TS.
Methyl Red TSDissolve 100 mg of methyl red in 100 mL
of alcohol, and filter if necessary.
Methyl Red TS 2To 1.86 mL of 0.1 M sodium hydroxide
and 50 mL of alcohol, add 50 mg of methyl red, and dilute
with water to 100 mL.
Methyl Red TS, MethanolicDissolve 1 g of methyl red in
100 mL of methanol, and filter, if necessary. Store protected
from light, and use within 21 days.
Methyl RedMethylene Blue TSAdd 10 mL of methyl red
TS to 10 mL of methylene blue TS, and mix.
Methyl Violet TSUse Crystal Violet TS.
Methyl Yellow TSPrepare a solution containing 0.10 mg
per mL in alcohol.
Methyl YellowMethylene Blue TSDissolve 1 g of methyl
yellow and 100 mg of methylene blue in 125 mL of methanol.
3-Methyl-2-benzothiazolinone Hydrazone Hydrochloride
TSDissolve 0.1 g of 3-methyl-2-benzothiazolinone
hydrazone hydrochloride monohydrate in 10 mL of water,
dilute the resulting solution with methanol to 100 mL, and
mix.
Methylene Blue TSDissolve 125 mg of methylene blue in
100 mL of alcohol, and dilute with alcohol to 250 mL.
Methylthionine Perchlorate TSTo 500 mL of potassium
perchlorate solution (1 in 1000) add dropwise, with con-

Official from August 1, 2015

Copyright (c) 2015 The United States Pharmacopeial Convention. All rights reserved.

Accessed from 10.6.1.1 by gsk on Fri Oct 30 04:44:36 EDT 2015

USP 38

Solutions / Test Solutions 1891

Perchloric Acid TSDilute 8.5 mL of perchloric acid with
water to 100 mL.
Phenol TSDissolve 1.2 g of phenol in alcohol to make
10 mL. Prepare weekly.
Phenol Red TS (Phenolsulfonphthalein TS)Dissolve 100 mg
of phenolsulfonphthalein in 100 mL of alcohol, and filter if
necessary.
pH 4.7 Phenol Red TSDissolve 33 mg of phenolsulfonphthalein in 1.5 mL of 2 N sodium hydroxide solution, dilute with water to 100 mL, and mix (Solution A). Dissolve
25 mg of ammonium sulfate in 235 mL of water, add
105 mL of 2 N sodium hydroxide solution and 135 mL of
2 N acetic acid, and mix (Solution B). Add 25 mL of Solution
A to Solution B, and mix. If necessary, adjust the pH of this
solution to 4.7.
Phenoldisulfonic Acid TSDissolve 2.5 g of phenol in
15 mL of sulfuric acid in a flask of suitable capacity. Add
7.5 mL of fuming sulfuric acid, stir well, and heat at 100
for 2 hours. Transfer the product, while still fluid, to a glassstoppered bottle, and, when desired for use, warm in a
water bath until liquefied.
Phenolphthalein TSDissolve 1 g of phenolphthalein in
100 mL of alcohol.
Phenylhydrazine Acetate TSDissolve 10 mL of phenylhydrazine and 5 mL of glacial acetic acid in water to make
100 mL.
PhenylhydrazineSulfuric Acid TSDissolve 65 mg of phenylhydrazine hydrochloride in 100 mL of a cooled mixture
of equal volumes of sulfuric acid and water.
Phloroglucinol TSDissolve 500 mg of phloroglucinol in
25 mL of alcohol. Store in tight containers, protected from
light.
Phosphatic Enzyme TSDissolve 5 g of phosphatic enzyme
in water to make 50 mL. Prepare this solution fresh.
Phosphomolybdic Acid TSDissolve 20 g of phosphomolybdic acid in alcohol to make 100 mL. Filter the solution,
and use only the clear filtrate.
Phosphotungstic Acid TSDissolve 1 g of phosphotungstic
acid in water to make 100 mL.
Picrate TS, AlkalineMix 20 mL of trinitrophenol solution
(1 in 100) with 10 mL of sodium hydroxide solution (1 in
20), dilute with water to 100 mL, and mix. Use within 2
days.
Picric Acid TSSee Trinitrophenol TS.
Platinic Chloride TSDissolve 2.6 g of platinic chloride in
water to make 20 mL.
PlatinumCobalt TSDissolve 1.246 g of potassium
chloroplatinate (K2PtCl6) and 1.000 g of cobalt chloride
(CoCl2 6H2O) in water, add 100 mL of hydrochloric acid,
and dilute with water to 1 L.
Potassium Acetate TSDissolve 10 g of potassium acetate
in water to make 100 mL.
PotassiumBismuth Iodide TSDissolve 12.5 g of tartaric
acid in 25 mL of water, then dissolve 1.06 g of bismuth subnitrate in this mixture (Solution A). Dissolve 20 g of potassium iodide in 25 mL of water (Solution B). Dissolve 100 g of
tartaric acid in 450 mL of water (Solution C). Add Solutions A
and B to Solution C, and mix.
Potassium Carbonate TSDissolve 7 g of anhydrous potassium carbonate in water to make 100 mL.
Potassium Chromate TSDissolve 10 g of potassium chromate in water to make 100 mL.
Potassium Dichromate TSDissolve 7.5 g of potassium
dichromate in water to make 100 mL.
Potassium Ferricyanide TSDissolve 1 g of potassium ferricyanide in 10 mL of water. Prepare this solution fresh.
Potassium Ferrocyanide TSDissolve 1 g of potassium ferrocyanide in 10 mL of water. Prepare this solution fresh.
Potassium Hydroxide TSDissolve 6.5 g of potassium hydroxide in water to make 100 mL.
Potassium Hydroxide TS, AlcoholicUse 0.5 N Potassium
Hydroxide, Alcoholic (see in the section Volumetric Solutions).
Potassium Hydroxide TS 2, AlcoholicDissolve 130 g of
potassium hydroxide, with cooling, in 200 mL of water. Add

Official from August 1, 2015

Copyright (c) 2015 The United States Pharmacopeial Convention. All rights reserved.

Reagents

stant shaking, methylene blue solution (1 in 100) until a

slight, permanent turbidity results. Allow the precipitate to
settle, decant the supernatant through paper, and use only
the clear solution.
Millons ReagentTo 2 mL of mercury in a conical flask
add 20 mL of nitric acid. Shake the flask under a hood to
break up the mercury into small globules. After about
10 minutes, add 35 mL of water, and, if a precipitate or
crystals appear, add sufficient dilute nitric acid (1 in 5, prepared from nitric acid from which the oxides have been
removed by blowing air through it until it is colorless) to
dissolve the separated solid. Add sodium hydroxide solution
(1 in 10) dropwise, with thorough mixing, until the curdy
precipitate that forms after the addition of each drop no
longer redissolves but is dispersed to form a suspension.
Add 5 mL more of the dilute nitric acid, and mix. Prepare
this solution fresh.
Molybdo-phosphotungstate TS (Folin-Denis Reagent)To
about 350 mL of water contained in a round-bottom flask
add 50 g of sodium tungstate, 12 g of phosphomolybdic
acid, and 25 mL of phosphoric acid. Boil the mixture under
a reflux condenser for 2 hours, then cool, dilute with water
to 500 mL, and mix. Store in tight containers, protected
from light, and in a cold place.
1-Naphthol ReagentDissolve 1 g of 1-naphthol in 25 mL
of methanol. Prepare this solution fresh.
1-Naphthol TSUse 1-Naphthol Reagent.
2-Naphthol TS (Betanaphthol TS)Dissolve 1 g of 2-naphthol in 100 mL of sodium hydroxide solution (1 in 100).
p-Naphtholbenzein TSDissolve 250 mg of p-naphtholbenzein in 100 mL of glacial acetic acid.
N-(1-Naphthyl)ethylenediamine Dihydrochloride TSDissolve 100 mg of N-(1-naphthyl)ethylenediamine dihydrochloride in 100 mL of a mixture of 7 parts of acetone and
3 parts of water.
Nesslers ReagentSee MercuricPotassium Iodide TS, Alkaline.
Neutral Red TSDissolve 100 mg of neutral red in 100 mL
of 50% alcohol.
Nickel Standard Solution TSDissolve 4.78 g of nickel (II)
sulfate heptahydrate in water, and dilute with water to
1000 mL. Immediately prior to use, dilute 10.0 mL of the
solution so obtained with water to 1000 mL. Suitable nickel
standard solutions are also available commercially.
Ninhydrin TSUse Triketohydrindene Hydrate TS.
p-Nitroaniline TSTo 350 mg of p-nitroaniline add 1.5 mL
of hydrochloric acid, and mix. Dilute with water to 50 mL,
mix, and allow to settle. Place 5 mL of the clear supernatant
in a 100-mL volumetric flask, and immerse it in an ice bath.
While it is in the ice bath, add 1 mL of hydrochloric acid,
then add, in small portions, 2 mL of sodium nitrite solution
(1 in 100), dilute with water to volume, and mix.
Nitrophenanthroline TSDissolve 150 mg of 5-nitro-1,10phenanthroline in 15 mL of freshly prepared ferrous sulfate
solution (1 in 140).
Oracet Blue B TSA 1 in 200 solution of oracet blue B in
glacial acetic acid.
Orthophenanthroline TSDissolve 150 mg of
orthophenanthroline in 10 mL of a solution of ferrous sulfate, prepared by dissolving 700 mg of ferrous sulfate in
100 mL of water. The ferrous sulfate solution must be prepared immediately before dissolving the orthophenanthroline. Store in well-closed containers.
Oxalic Acid TSDissolve 6.3 g of oxalic acid in water to
make 100 mL.
Palladium Chloride TS, BufferedWeigh 500 mg of palladium chloride into a 250-mL beaker, add 5 mL of concentrated hydrochloric acid, and warm the mixture on a steam
bath. Add 200 mL of hot water in small increments with
continued heating until solution is complete. Transfer the
solution to a 250-mL volumetric flask, and dilute with water
to volume. Transfer 50 mL to a 100-mL volumetric flask.
Add 10 mL of 1 M sodium acetate and 9.6 mL of 1 N hydrochloric acid. Dilute with water to volume.

Accessed from 10.6.1.1 by gsk on Fri Oct 30 04:44:36 EDT 2015

Reagents

1892 Test Solutions / Solutions

alcohol to 1000 mL. Store in a well-stoppered dark glass
bottle.
Potassium Iodide TSDissolve 16.5 g of potassium iodide
in water to make 100 mL. Store in light-resistant containers.
Potassium Iodide and Starch TSDissolve 0.75 g of potassium iodide in 100 mL of water. Heat to boiling, and add,
with stirring, a solution of 0.5 g of soluble starch in 35 mL
of water. Boil for 2 minutes, and allow to cool.
SENSITIVITYMix 15 mL in 0.05 mL of glacial acetic acid
and 0.3 mL of diluted iodine TS: a blue color is produced.
Potassium Iodoplatinate TSDissolve 200 mg of platinic
chloride in 2 mL of water, mix with 25 mL of potassium
iodide solution (1 in 25), and add water to make 50 mL.
Potassium Permanganate TSUse 0.1 N Potassium Permanganate (see in the section Volumetric Solutions).
Potassium Pyroantimonate TSDissolve 2 g of potassium
pyroantimonate in 85 mL of hot water. Cool quickly, and
add 50 mL of a solution containing 50 mg/mL of potassium
hydroxide in water and 1 mL of sodium hydroxide solution
(8.5 in 100). Allow to stand for 24 h, filter, and dilute with
water to 150 mL.
Potassium Sulfate TSDissolve 1 g of potassium sulfate in
water to make 100 mL.
Potassium Thiocyanate TSDissolve 9.7 g of potassium
thiocyanate in water to make 100 mL.
PyridinePyrazolone TSTo 100 mL of a saturated solution
of 1-phenyl-3-methyl-2-pyrazoline-5-one add 20 mL of a 1
in 1000 solution of 3,3-dimethyl-1,1-diphenyl-[4,4-bi2-pyrazoline]-5,5-dione in pyridine. Store in a dark bottle,
and use within 3 days.
Pyrogallol TS, AlkalineDissolve 500 mg of pyrogallol in
2 mL of water. Dissolve 12 g of potassium hydroxide in
8 mL of water. The solutions should be freshly prepared and
mixed immediately before use.
Quinaldine Red TSDissolve 100 mg of quinaldine red in
100 mL of alcohol.
Quinone TSDissolve 500 mg of p-benzoquinone in 2.5 mL
of glacial acetic acid, and dilute with alcohol to 50 mL. Prepare this solution fresh daily.
Resorcinol TSDissolve 1 g of resorcinol in hydrochloric
acid to make 100 mL.
Ruthenium Red TSDissolve 10 g of lead acetate in water,
dilute with water to 100 mL, and add 80 mg of ruthenium
red. The solution is wine-red in color. [NOTEIf necessary,
add additional ruthenium red to obtain a wine-red color.]
Saline TSDissolve 9.0 g of sodium chloride in water to
make 1000 mL.
[NOTEWhere pyrogen-free saline TS is specified in this
Pharmacopeia, saline TS that has met the requirements of
the Pyrogen Test 151 is to be used.]
Saline TS, Pyrogen-FreeSee Saline TS.
Schweitzers ReagentSee Cupric Oxide, Ammoniated, TS.
SilverAmmoniaNitrate TSDissolve 1 g of silver nitrate
in 20 mL of water. Add ammonia TS, dropwise, with constant stirring, until the precipitate is almost but not entirely
dissolved. Filter, and store in tight, light-resistant containers.
SilverAmmonium Nitrate TSSee SilverAmmoniaNitrate
TS.
Silver Diethyldithiocarbamate TSDissolve 1 g of silver
diethyldithiocarbamate in 200 mL of pyridine from a freshly
opened bottle or that which has been recently distilled.
Store in light-resistant containers, and use within 30 days.
Silver Nitrate TSUse 0.1 N Silver Nitrate (see in the section Volumetric Solutions).
Simulated Gastric Fluid TSSee Gastric Fluid, Simulated,
TS.
Simulated Intestinal Fluid TSSee Intestinal Fluid, Simulated, TS.
Sodium Acetate TSDissolve 13.6 g of sodium acetate in
water to make 100 mL.
Sodium Alizarinsulfonate TSDissolve 100 mg of sodium
alizarinsulfonate in 100 mL of water, and filter.
Sodium Aminoacetate TS (Sodium Glycinate TS)Dissolve
3.75 g of aminoacetic acid in about 500 mL of water, add

USP 38
2.1 g of sodium hydroxide, and dilute with water to
1000 mL. Mix 9 mL of the resulting solution with 1 mL of
dilute glacial acetic acid (1 in 300). This test solution has a
pH between 10.4 and 10.5.
Sodium Bisulfite TSDissolve 10 g of sodium bisulfite in
water to make 30 mL. Prepare this solution fresh.
Sodium Bitartrate TSDissolve 1 g of sodium bitartrate in
water to make 10 mL. Prepare this solution fresh.
Sodium Carbonate TSDissolve 10.6 g of anhydrous sodium carbonate in water to make 100 mL.
Sodium Chloride TS, AlkalineDissolve 2 g of sodium hydroxide in 100 mL of water, saturate the solution with sodium chloride, and filter.
Sodium Citrate TSDissolve 73.5 g of sodium citrate dihydrate in water to make 250 mL.
Sodium Citrate TS, AlkalineDissolve 50 g of sodium citrate dihydrate and 2.5 g of sodium hydroxide in water to
make 250 mL.
Sodium Cobaltinitrite TSDissolve 10 g of sodium cobaltinitrite in water to make 50 mL, and filter if necessary.
Sodium Fluoride TSDry about 500 mg of sodium fluoride
at 200 for 4 hours. Accurately weigh 222 mg of the dried
material, and dissolve in water to make 100.0 mL. Pipet
10 mL of this solution into a 1-L volumetric flask, and dilute
with water to volume. Each mL of this solution corresponds
to 0.01 mg of fluorine (F).
Sodium Hydrosulfite TS, AlkalineDissolve 25 g of potassium hydroxide in 35 mL of water, and 50 g of sodium hydrosulfite in 250 mL of water. When the test solution is required, mix 40 mL of the hydroxide solution with the
250 mL of the hydrosulfite solution. Prepare this solution
fresh.
Sodium Hydroxide TSDissolve 4.0 g of sodium hydroxide
in water to make 100 mL.
Sodium Hydroxide TS 2Transfer 8.5 g of sodium hydroxide to a 100-mL volumetric flask, and dissolve in and dilute
with water to volume.
Sodium Hydroxide TS 3Prepare a 420-mg/mL solution of
sodium hydroxide in water.
Sodium Hypobromite TSTo a solution of 20 g of sodium
hydroxide in 75 mL of water add 5 mL of bromine. After
solution has taken place, dilute with water to 100 mL. Prepare this solution fresh.
Sodium Hypochlorite TSUse Sodium Hypochlorite Solution
(see in the section Reagent Specifications).
Sodium Iodohydroxyquinolinesulfonate TSDissolve 8.8 g
of iodohydroxyquinoline sulfonic acid in 200 mL of water,
and add 6.5 mL of 4 N sodium hydroxide. Dilute with water
to 250 mL, mix, and filter.
Sodium Nitroferricyanide TSDissolve 1 g of sodium nitroferricyanide in water to make 20 mL. Prepare this solution
fresh.
Dibasic Sodium Phosphate TSDissolve 12 g of dibasic sodium phosphate in water to make 100 mL.
Sodium Phosphotungstate TSTo a solution of 20 g of sodium tungstate in 100 mL of water add sufficient phosphoric acid to impart a strongly acid reaction to litmus, and
filter. When required for use, decant the clear solution from
any sediment that may be present. Store in tight, light-resistant containers.
Sodium Sulfide TSDissolve 1 g of sodium sulfide in water
to make 10 mL. Prepare this solution fresh.
Sodium Tartrate TSDissolve 11.5 g of sodium tartrate in
water to make 100 mL.
Sodium Tetraphenylboron TSDissolve 1.2 g of sodium
tetraphenylboron in water to make 200 mL. If necessary, stir
for 5 minutes with 1 g of aluminum oxide, and filter to clarify.
Sodium Thioglycolate TSDissolve 1.5 g of sodium thioglycolate in 450 mL of water, and add 50 mL of alcohol. Use
within 3 days.
Sodium Thiosulfate TSUse 0.1 N Sodium Thiosulfate (see
in the section Volumetric Solutions).

Official from August 1, 2015

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Accessed from 10.6.1.1 by gsk on Fri Oct 30 04:44:36 EDT 2015

USP 38

Solutions / Test Solutions 1893

Change to read:

Official from August 1, 2015

Copyright (c) 2015 The United States Pharmacopeial Convention. All rights reserved.

Reagents

Standard Lead Solution TSOn the day of use, dilute

10.0 mL of lead nitrate stock solution TS with water to
100.0 mL. Each mL of standard lead solution TS contains
the equivalent to 10 g of lead.USP38
Stannous Chloride, Acid, TSDissolve 8 g of stannous
chloride in 500 mL of hydrochloric acid. Store in glass containers, and use within 3 months.
Stannous Chloride, Acid, Stronger, TSDissolve 40 g of
stannous chloride in 100 mL of hydrochloric acid. Store in
glass containers, and use within 3 months.
Starch TSMix 1 g of soluble starch with 10 mg of red
mercuric iodide and sufficient cold water to make a thin
paste. Add 200 mL of boiling water, and boil for 1 minute
with continuous stirring. Cool, and use only the clear solution. [NOTECommercially available, stabilized starch indicator solutions may be used, including mercury-free solutions
preserved with other compounds such as salicylic acid.]
Starch, Iodide-Free, TSMix 1 g of soluble starch with sufficient cold water to make a thin paste. While stirring, add
100 mL of boiling water, and allow to cool. Prepare this
solution immediately before use. Iodide-free starch TS shows
a blue color when 20 mL of potassium iodide solution (1 in
400) and 0.05 mL of an iodinepotassium iodide solution
(prepared by dissolving 127 mg of iodine and 800 mg of
potassium iodide in water and diluting with water to
100 mL) are added to 1 mL of the iodide-free starch TS.
Starch Iodide Paste TSHeat 100 mL of water in a 250-mL
beaker to boiling, add a solution of 0.75 g of potassium
iodide in 5 mL of water, then add 2 g of zinc chloride dissolved in 10 mL of water, and, while the solution is boiling,
add, with stirring, a smooth suspension of 5 g of soluble
starch in 30 mL of cold water. Continue to boil for 2 minutes, then cool. Store in well-closed containers in a cold
place.
Starch iodide paste TS must show a definite blue streak
when a glass rod, dipped in a mixture of 1 mL of 0.1 M
sodium nitrite, 500 mL of water, and 10 mL of hydrochloric
acid, is streaked on a smear of the paste.
StarchPotassium Iodide TSDissolve 500 mg of potassium iodide in 100 mL of freshly prepared starch TS. Prepare
this solution fresh.
Stronger Cupric Acetate TSSee Cupric Acetate TS,
Stronger.
Sudan III TSDissolve 0.05 g of Sudan III in 25 mL of alcohol, with warming if necessary. Cool, add 25 mL of glycerin,
and mix. Filter if undissolved material persists.
Sudan IV TSDissolve 0.5 g of Sudan IV in chloroform to
make 100 mL.
Sulfanilic Acid TSDissolve 800 mg of sulfanilic acid in
100 mL of acetic acid. Store in tight containers.
Diazotized Sulfanilic Acid TSDissolve 0.9 g of sulfanilic
acid in 9 mL of hydrochloric acid with warming, and dilute
with water to 100 mL. Cool 10 mL of this solution in iced
water, and add 10 mL of a sodium nitrite solution (4.5 in
100) previously cooled in iced water. Allow to stand at 0
for at least 15 minutes (the solution may be kept for 3 days
at this temperature). Immediately before use, add 20 mL of
sodium carbonate solution (1 in 10).
Sulfanilic-1-Naphthylamine TSDissolve 500 mg of sulfanilic acid in 150 mL of acetic acid. Dissolve 100 mg of
1-naphthylamine hydrochloride in 150 mL of acetic acid,
and mix the two solutions. The pink color that may develop
on standing can be removed by treatment with zinc.
Sulfanilic--Naphthylamine TSSee Sulfanilic-1-Naphthylamine TS.

Sulfomolybdic Acid TSDissolve, with the aid of heat,

2.5 g of ammonium molybdate in 20 mL of water, add
50 mL of 12 N sulfuric acid, and dilute with water to
100 mL. Store this solution in a polyethylene container.
Sulfuric Acid TSAdd a quantity of sulfuric acid of known
concentration to sufficient water to adjust the final concentration to between 94.5% and 95.5% (w/w) of H2SO4.
[NOTESince the acid concentration may change upon
standing or upon intermittent use, the concentration should
be checked frequently and solutions assaying more than
95.5% or less than 94.5% discarded.]
Sulfuric AcidFormaldehyde TSAdd 1 drop of formaldehyde TS to each mL of sulfuric acid, and mix. Prepare this
solution fresh.
Tannic Acid TSDissolve 1 g of tannic acid in 1 mL of alcohol, and dilute with water to 10 mL. Prepare this solution
fresh.
Tartaric Acid TSDissolve 3 g of tartaric acid in water to
make 10 mL. Prepare this solution fresh.
Tetrabromophenolphthalein Ethyl Ester TSDissolve
100 mg of tetrabromophenolphthalein ethyl ester in 90 mL
of glacial acetic acid, and dilute with glacial acetic acid to
100 mL. Prepare this solution fresh.
Tetramethylammonium Hydroxide TSUse an aqueous
solution containing, in each 100 mL, the equivalent of 10 g
of anhydrous tetramethylammonium hydroxide.
Thioacetamide TSDissolve 4 g of thioacetamide in
100 mL of water.
ThioacetamideGlycerin Base TSMix 0.2 mL of thioacetamide TS and 1 mL of glycerin base TS, and heat in a boiling water bath for 20 seconds. Use the mixture immediately.
Thorium Nitrate TSDissolve 1 g of thorium nitrate in
water to make 100 mL. Filter, if necessary.
Thymol Blue TSDissolve 100 mg of thymol blue in
100 mL of alcohol, and filter if necessary.
Thymolphthalein TSDissolve 100 mg of thymolphthalein
in 100 mL of alcohol, and filter if necessary.
Titanium Trichloride TSDissolve 15 g of titanium trichloride in 100 mL of 10% hydrochloric acid solution.
Titanium TrichlorideSulfuric Acid TSMix carefully 20 mL
of titanium trichloride TS in 13 mL of sulfuric acid. Add sufficient 30% hydrogen peroxide to produce a yellow color.
Heat until white fumes are evolved, allow to cool, and dilute
with water. Repeat the evaporation and addition of water
until a colorless solution is obtained. Dilute with water to
100 mL.
p-Toluenesulfonic Acid TSDissolve 2 g of ptoluenesulfonic acid in 10 mL of a mixture of 7 parts of acetone and 3 parts of water.
Triketohydrindene Hydrate TS (Ninhydrin TS)Dissolve
200 mg of triketohydrindene hydrate in water to make
10 mL. Prepare this solution fresh.
Trinitrophenol TS (Picric Acid TS)Dissolve the equivalent
of 1 g of anhydrous trinitrophenol in 100 mL of hot water.
Cool the solution, and filter if necessary.
Triphenyltetrazolium Chloride TSDissolve 500 mg of
triphenyltetrazolium chloride in dehydrated alcohol to make
100 mL.
Xylenol Orange TSDissolve 100 mg of xylenol orange in
100 mL of alcohol.
Zinc Uranyl Acetate TSDissolve 50 g of uranyl acetate in
a mixture of 15 mL of glacial acetic acid and water to make
500 mL. Then dissolve 150 g of zinc acetate in a mixture of
15 mL of glacial acetic acid and water to make 500 mL. Mix
the two solutions, allow to stand overnight, and pass
through a dry filter, if necessary.

Accessed from 10.6.1.1 by gsk on Fri Oct 30 04:44:36 EDT 2015

1894 Volumetric Solutions / Solutions

VOLUMETRIC SOLUTIONS
1. DEFINITIONS
1.1 Normal solutions: Normal solutions are solutions that
contain 1 gram equivalent weight of the compound in 1 L
of solution.
N = equivalent/L
N = weight (g)/equivalent weight (g) L
Normal solutions and solutions bearing a specific relationship to normal solutions, and used in volumetric determinations, are designated as follows:
1 N = normal
2 N = double normal
0.5 N = half-normal
0.1 N = tenth-normal
0.02 N = fiftieth normal
0.01 N = hundredth-normal
0.001 N = thousandth-normal
1.2 Molar solutions: Molar solutions are solutions that
contain 1 gram-molecule of the compound in 1 L of
solution.
M = mol/L

Reagents

M = weight (g)/mol weight L

Solutions containing one-tenth of a gram-molecule of the
compound in 1 L are designated tenth-molar, 0.1 M; and
other molarities are similarly indicated.
1.3 Correction factor: It is frequently difficult to prepare
standard solutions of a desired theoretical normality or molarity, and this is not essential. A solution of approximately
the desired normality or molarity is prepared and standardized by titration against a standard. The correction factor so
obtained is used in all calculations where such solutions are
used. If necessary, the concentration of the solution may be
adjusted to a given normality or molarity, by dilution or by
addition of the appropriate reagent.
The concentration of the volumetric solution does not differ from the prescribed one by more than 10%. The correction factor is determined by an appropriate number of replicates. The repeatability does not exceed 0.2% (relative
standard deviation). The correction factor should be redetermined frequently.
1.4 Blank determinations: Where it is directed that any
necessary correction be made by a blank determination,
the determination is to be conducted with the use of the
same quantities of the same reagents treated in the same
manner as the solution or mixture containing the portion of
the substance under assay or test, but with the substance
itself omitted.

2. PREPARATION AND STANDARDIZATION

2.1 Scope
When solutions of a reagent are used in different concentrations, the details of the preparation and standardization
are usually given for the concentration most frequently required. Stronger or weaker solutions are prepared and standardized in the same general manner as described, using
proportionate amounts of the reagent.
2.2 Preparation by dilution
It is possible in many instances to prepare lower concentrations accurately by making an exact dilution of a stronger
solution.

USP 38
Volumetric solutions prepared by dilution should be
restandardized as directed for the stronger solution, using
proportionate amounts of reagents.
Dilute solutions that are not stable, as, for instance, potassium permanganate 0.01 N, are preferably prepared by exactly diluting the higher normality with thoroughly boiled
and cooled water on the same day they are required for
use.
2.3 Standardization
The following directions give only one method for standardization, but other methods of standardization, capable
of yielding at least the same degree of accuracy, may be
used.
The values obtained in the standardization of volumetric
solutions are valid for all Pharmacopeial uses of these solutions, regardless of the instrumental or chemical indicators
used in the individual monographs.
Where the apparent normality or molarity of a titrant depends upon the special conditions of its use, the individual
monograph sets forth the directions for standardizing the
reagent in the specified context.
For those salts that usually are available as certified primary standards or that are available as highly purified salts
of primary standard quality, it is permissible to prepare solutions by accurately weighing a suitable quantity of the salt
and dissolving it to produce a specific volume of solution of
known concentration. Acetic, hydrochloric, and sulfuric acids
may be standardized against a sodium hydroxide solution
that recently has been standardized against a certified primary standard.
2.4 Temperature
All volumetric solutions, if practicable, are to be prepared,
standardized, and used at the standard temperature of 25.
If a titration is carried out with the volumetric solution at
a markedly different temperature, standardize the volumetric
solution used as the titrant at that different temperature, or
make a suitable temperature correction.
Acetic Acid, Double-Normal (2 N) C2H4O2, 60.05 120.10 g
in 1000 mL
Add 116 mL of glacial acetic acid to sufficient water to
make 1000 mL after cooling to room temperature.
Ammonium Thiocyanate, Tenth-Normal (0.1 N) NH4SCN,
76.12 7.612 g in 1000 mL
Dissolve about 8 g of ammonium thiocyanate in 1000 mL
of water, and standardize the solution as follows.
Accurately measure about 30 mL of 0.1 N silver nitrate VS
into a glass-stoppered flask. Dilute with 50 mL of water,
then add 2 mL of nitric acid and 2 mL of ferric ammonium
sulfate TS, and titrate with the ammonium thiocyanate solution to the first appearance of a red-brown color.

If desirable, 0.1 N ammonium thiocyanate may be replaced by 0.1 N potassium thiocyanate where the former is
directed in various tests and assays.
Bismuth Nitrate, 0.01 M Bi(NO3)3 5H2O, 485.07 1000 mL
of this solution contains 4.851 g of bismuth nitrate
pentahydrate
Dissolve 4.86 g of bismuth nitrate pentahydrate in 60 mL
of dilute nitric acid, add 0.01 N nitric acid to make
1000 mL, and standardize the solution as follows.
Accurately measure 25 mL of the prepared bismuth nitrate
solution, add 50 mL of water and 1 drop of xylenol orange
TS, and titrate the solution with 0.01 M edetate disodium
VS until the red color changes to yellow. Calculate the molarity factor.
Bromine, Tenth-Normal (0.1 N) Br, 79.90 7.990 g in
1000 mL
Dissolve 3 g of potassium bromate and 15 g of potassium
bromide in water to make 1000 mL, and standardize the
solution as follows.

Official from August 1, 2015

Copyright (c) 2015 The United States Pharmacopeial Convention. All rights reserved.