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16 by universetx608 on Sat Feb 10 00:55:15 EST 2018
USP 40
Standard solution 2—Transfer 1.0 mL of the Standard
stock solution to another 100-mL volumetric flask, dilute
with methanol to volume, and mix.
Application volume: 10 µL.
Developing solvent system—Use the upper layer of a mix-
ture of methyl isobutyl ketone, water, and glacial acetic acid
(50:25:25).
Procedure—Proceed as directed for Thin-Layer Chromatog-
raphy under Chromatography 〈621〉. Dry the plate for
15 minutes at 80°. Examine the plate under UV light at 254
nm and 365 nm. The chromatogram from the System suita-
bility solution shows two clearly separated minor spots with
RF values of about 0.18 and 0.22. Compare the intensities of
any secondary spots observed in the chromatogram of the
Test solution with those of the principal spots in the chro-
matograms of the Standard solutions. Any spot obtained
from the Test solution, except for the principal spot, is not
greater in size than the spot obtained from Standard solution
1 (0.3%), and at most two spots are more intense than the
spot obtained from Standard solution 2 (0.1%).
TEST 2—
pH 3.0 Buffer and Mobile phase—Prepare as directed in
the Assay.
System suitability solution—Dissolve about 5 mg of
Amlodipine Besylate in 5 mL of hydrogen peroxide, and
heat at 70° for 45 minutes.
Standard solution—Dissolve an accurately weighed quan-
tity of USP Amlodipine Besylate RS in Mobile phase to obtain
a solution having a known concentration of about 0.003 mg
per mL.
Test solution—Transfer about 50 mg of Amlodipine Besy-
late, accurately weighed, to a 50-mL volumetric flask, dis-
solve in and dilute with Mobile phase to volume, and mix.
Chromatographic system (see Chromatography 〈621〉)—
Prepare as directed in the Assay. Chromatograph the System
suitability solution, and record the peak responses as directed
for Procedure: the resolution, R, between amlodipine impu-
rity A and amlodipine is not less than 4.5. [NOTE—For the
purpose of identification, the relative retention times are
about 0.2 for benzene sulfonate, 0.5 for amlodipine impu-
rity A, and 1.0 for amlodipine. Amlodipine impurity A is
3-ethyl 5-methyl 2-[(2-aminoethoxy)methyl]-
4-(2-chlorophenyl)-6-methylpyridine-3,5-dicarboxylate.]
Chromatograph the Standard solution, and record the peak
responses as directed for Procedure: the standard deviation
for replicate injections is not more than 10.0%.
Procedure—Separately inject equal volumes (about 10 µL)
of the Standard solution and the Test solution into the chro-
matograph, record the chromatograms for a period of time
that is about 3 times the retention time of amlodipine, and
measure the peak responses. Calculate the percentage of
each impurity in the portion of Amlodipine Besylate taken
by the formula:
100(1/F)(CS /CT)(ri / rS)
in which F is the relative response factor, which is equal to
0.5 for amlodipine impurity A and to 1.0 for other impuri-
ties; CS and CT are the concentrations, in mg per mL, of
amlodipine besylate in the Standard solution and the Test
solution, respectively; ri is the peak response for each impu-
rity obtained from the Test solution; and rS is the peak re-
sponse for amlodipine besylate obtained from the Standard
solution: not more than 0.3% of amlodipine impurity A is
found, and not more than 0.3% of total other impurities is
found. Disregard any peak less than 0.03%, and disregard
any peak due to benzene sulfonate.
Assay—
pH 3.0 Buffer—Dissolve 7.0 mL of triethylamine in 800 mL
of water. Adjust with phosphoric acid to a pH of 3.0 ± 0.1,
and dilute with water to 1 L.
Official from December 1, 2017
Copyright (c) 2018 The United States Pharmacopeial Convention. All rights reserved.
Official Monographs / Amlodipine 2773
Mobile phase—Prepare a filtered and degassed mixture of
pH 3.0 Buffer, methanol, and acetonitrile (50:35:15). Make
adjustments if necessary (see System Suitability under Chro-
matography 〈621〉).
Standard preparation—Dissolve an accurately weighed
quantity of USP Amlodipine Besylate RS in Mobile phase to
obtain a solution having a known concentration of about
0.05 mg per mL.
Assay preparation—Transfer about 50 mg of Amlodipine
Besylate, accurately weighed, to a 50-mL volumetric flask,
dissolve in and dilute with Mobile phase to volume, and mix.
Transfer 5.0 mL of this solution to a 100-mL volumetric flask,
dilute with Mobile phase to volume, and mix.
Chromatographic system (see Chromatography 〈621〉)—The
liquid chromatograph is equipped with a 237-nm detector
and a 3.9-mm × 15-cm column that contains packing L1.
The flow rate is about 1.0 mL per minute. Chromatograph
the Standard preparation, and record the peak responses as
directed for Procedure: the standard deviation for replicate
injections is not more than 2.0%.
Procedure—Separately inject equal volumes (about 10 µL)
of the Standard preparation and the Assay preparation into
the chromatograph, record the chromatograms, and meas-
ure the responses for the major peaks. Calculate the per-
centage of C20H25ClN2O5 · C6H6O3S in the portion of
Amlodipine Besylate taken by the formula:
100(CS /CU)(rU / rS)
in which CS and CU are the concentrations, in mg per mL, of
amlodipine besylate in the Standard preparation and the As-
say preparation, respectively; and rU and rS are the peak re-
sponses obtained from the Assay preparation and the Stan-
dard preparation, respectively.
USP Monographs
.
Amlodipine Besylate Tablets
DEFINITION
Amlodipine Besylate Tablets contain NLT 90% and NMT
110% of the labeled amount of amlodipine
(C20H25ClN2O5).
IDENTIFICATION
• A. ULTRAVIOLET ABSORPTION 〈197U〉
Standard solution and Sample solution: Prepare as di-
rected in the test for Dissolution.
Acceptance criteria: Meet the requirements
• B. The retention time of the major peak of the Sample
solution corresponds to that of the Standard solution, as
obtained in the Assay.
ASSAY
• PROCEDURE
Buffer: Add 7.0 mL of triethylamine into a 1000-mL
flask containing 900 mL of water. Adjust the solution
with phosphoric acid to a pH of 3.0 ± 0.1. Dilute with
water to volume, and mix well.
Mobile phase: Methanol, acetonitrile, and Buffer
(35:15:50)
Standard solution: 0.0275 mg/mL of USP Amlodipine
Besylate RS and 0.0025 mg/mL of USP Amlodipine Re-
lated Compound A RS in Mobile phase
Sample solution: Nominally 0.02 mg/mL of amlodipine
in Mobile phase prepared as follows. Place NLT 5 Tablets
in a suitable volumetric flask, and add sufficient quan-
tity of Mobile phase to disintegrate the Tablets. Shake
for 30 min, and dilute with Mobile phase to volume.
Pass the sample through a syringe tip filter of 0.45-µm
pore size. Discard the first few mL of the filtrate.
Chromatographic system
(See Chromatography 〈621〉, System Suitability.)
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2774 Amlodipine / Official Monographs USP 40

Mode: LC CS = concentration of the Standard solution

Detector: UV 237 nm (mg/mL)
Column: 3.9-mm × 15-cm; 5-µm packing L1 D = dilution factor of the Sample solution
Flow rate: 1 mL/min Mr1 = molecular weight of amlodipine, 408.88
Injection volume: 50 µL Mr2 = molecular weight of amlodipine besylate,
Run time: NLT 3 times the retention of the 567.05
amlodipine peak V = volume of Medium, 500 mL
System suitability L = label claim (mg/Tablet)
Sample: Standard solution Tolerances: NLT 75% (Q) of the labeled amount of
[NOTE—The relative retention times for amlodipine and amlodipine (C20H25ClN2O5) is dissolved.
amlodipine related compound A are about 1.0 and 0.5 • UNIFORMITY OF DOSAGE UNITS 〈905〉: Meet the
respectively.] requirements
Suitability requirements
Resolution: NLT 8.5 between amlodipine and IMPURITIES
amlodipine related compound A • ORGANIC IMPURITIES
Tailing factor: NMT 2.0 for both amlodipine and Buffer, Mobile phase, Standard solution, Chromato-
amlodipine related compound A graphic system, and System suitability: Proceed as
Relative standard deviation: NMT 5.0% for directed in the Assay.
amlodipine related compound A Sample solution: Place a suitable number of Tablets
Analysis into a 25-mL volumetric flask to obtain a solution with
Samples: Standard solution and Sample solution a final nominal concentration of 0.4 mg/mL of
Calculate the percentage of the labeled amount of amlodipine. Add about 10 mL of Mobile phase to the
amlodipine (C20H25ClN2O5) in the portion of Tablets flask. Swirl to disintegrate the Tablets, then sonicate for
taken: 5 min to completely dissolve, and cool the sample to
room temperature. Dilute with Mobile phase to volume.
Result = (rU/rS) × (CS/CU) × (Mr1/Mr2) × 100 Stir for an additional 15 min using a magnetic stir bar,
and pass the sample through a syringe tip filter of 0.45-
rU = peak response from the Sample solution µm pore size, discarding the first 5 mL.
rS = peak response from the Standard solution Analysis
CS = concentration of USP Amlodipine Besylate RS Samples: Standard solution and Sample solution
in the Standard solution (mg/mL) Calculate the percentage of amlodipine related com-
CU = nominal concentration of amlodipine in the pound A in the portion of Tablets taken:
Sample solution (mg/mL)
Mr1 = molecular weight of amlodipine, 408.88 Result = (rU/rS) × (CS/CU) × (Mr1/Mr2) × 100
Mr2 = molecular weight of amlodipine besylate,
567.05 rU = peak response of amlodipine related
Acceptance criteria: 90%–110% of the labeled amount compound A from the Sample solution
of amlodipine (C20H25ClN2O5) rS = peak response of amlodipine related
USP Monographs

compound A from the Standard solution

PERFORMANCE TESTS CS = concentration of USP Amlodipine Related
• DISSOLUTION 〈711〉 Compound A RS in the Standard solution
[NOTE—Do not expose any of the solutions to stainless (mg/mL)
steel because of the degradation of amlodipine.] CU = nominal concentration of amlodipine in the
Medium: 0.01 N hydrochloric acid; 500 mL Sample solution (mg/mL)
Apparatus 2: 75 rpm. [NOTE—Use paddles covered Mr1 = molecular weight of amlodipine related
with Teflon or made of any inert material except stain- compound A, 406.86
less steel.] Mr2 = molecular weight of amlodipine related
Time: 30 min compound A fumarate, 522.93
Standard solution: Make appropriate dilutions of USP Calculate the percentage of amlodipine glucose/
Amlodipine Besylate RS in Medium to obtain the follow- galactose adduct or amlodipine lactose adduct, if
ing concentrations: 0.00695 mg/mL for Tablets labeled present, and any unspecified degradation product in
to contain 2.5 mg, 0.0139 mg/mL for Tablets labeled to the portion of Tablets taken:
contain 5 mg, and 0.0278 mg/mL for Tablets labeled to
contain 10 mg. These solutions are stable for 1 day. Result = (rU/rS) × (CS/CU) × (Mr1/Mr2) × 100
Sample solution: Pass a portion of the solution under
test through a suitable filter of 0.45-µm pore size. rU = peak response of the amlodipine glucose/
Analysis galactose adduct, amlodipine lactose adduct,
Samples: Standard solution and Sample solution or any unspecified degradation product from
Determine the amount of amlodipine (C20H25ClN2O5) the Sample solution
dissolved by using UV absorption at the wavelength of rS = peak response of amlodipine from the
maximum absorbance at about 239 nm on portions of Standard solution
the Sample solution in comparison with the Standard CS = concentration of USP Amlodipine Besylate RS
solution, using a 1-cm quartz cell and the Medium as from the Standard solution (mg/mL)
the blank. CU = nominal concentration of amlodipine in the
Calculate the percentage of the labeled amount of Sample solution (mg/mL)
amlodipine (C20H25ClN2O5) dissolved: Mr1 = molecular weight of amlodipine, 408.9
Mr2 = molecular weight of amlodipine besylate,
Result = (AU/AS) × CS × D × (Mr1/Mr2) × V × (1/L) × 100 567.05
Acceptance criteria: See Table 1.
AU = absorbance of the Sample solution
AS = absorbance of the Standard solution

Official from December 1, 2017

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Accessed from 59.99.44.16 by universetx608 on Sat Feb 10 00:55:15 EST 2018

USP 40 Official Monographs / Ammonium 2775

Table 1 is equivalent to 48.04 mg of ammonium carbonate

Relative Acceptance
[(NH4)2CO3].
Retention Criteria,
Acceptance criteria: In each 100 mL, NLT 1.7 g and
Name Time NMT (%)
NMT 2.1 g of total ammonia (NH3) and Ammonium
Carbonate corresponding to NLT 3.5 g and NMT 4.5 g
Amlodipine related com- of ammonium carbonate [(NH4)2CO3]
pound Aa . 0.50 1.0
Amlodipine lactose ad- OTHER COMPONENTS
ductb . 0.80 0.5 • ALCOHOL DETERMINATION, Method I 〈611〉: 62.0%–68.0%
Amlodipine glucose/ga-
lactose ADDITIONAL REQUIREMENTS
adductb 0.90 0.5 • PACKAGING AND STORAGE: Preserve in tight, light-resistant
containers at a temperature not exceeding 30°.
.

Amlodipine besylate 1.0 —

Any unspecified degra-
dation —
product 0.2
a 3-Ethyl, 5-methyl [2-(2-aminoethoxymethyl)-4-(2-chlorophenyl)-6-meth-
Ammonium Chloride
.

yl-3,5-pyridinedicarboxylate] fumarate.
b Formulation-specific impurities.
.

NH4Cl 53.49
ADDITIONAL REQUIREMENTS Ammonium chloride.
• PACKAGING AND STORAGE: Preserve in tight, light-resistant Ammonium chloride [12125-02-9].
containers. Store at controlled room temperature.
• USP REFERENCE STANDARDS 〈11〉 » Ammonium Chloride contains not less than
USP Amlodipine Besylate RS 99.5 percent and not more than 100.5 percent of
USP Amlodipine Related Compound A RS NH4Cl, calculated on the dried basis.
3-Ethyl, 5-methyl [2-(2-aminoethoxymethyl)-
4-(2-chlorophenyl)-6-methyl-3,5-pyridinedicarboxylate] Packaging and storage—Preserve in tight containers.
fumarate. Identification—A solution (1 in 10) responds to the tests
C20H23ClN2O5 · C4H4O4 522.93 for Ammonium 〈191〉 and for Chloride 〈191〉.
pH 〈791〉: between 4.6 and 6.0, in a solution (1 in 20).
Loss on drying 〈731〉—Dry it over silica gel for 4 hours: it
loses not more than 0.5% of its weight.
Residue on ignition 〈281〉—Add 1 mL of sulfuric acid to
.

Aromatic Ammonia Spirit about 2 g, accurately weighed, and heat the mixture gently
until volatilization is complete: the residue is white, and
DEFINITION when ignited, not more than 0.1% of nonvolatile substance

Aromatic Ammonia Spirit is a hydroalcoholic solution that
contains, in each 100 mL, NLT 1.7 g and NMT 2.1 g of
total ammonia (NH3) and Ammonium Carbonate corre-
sponding to NLT 3.5 g and NMT 4.5 g of ammonium car-
bonate [(NH4)2CO3].
USP Monographs
remains.
Limit of thiocyanate—Acidify 10 mL of a solution (1 in
10) with hydrochloric acid, and add a few drops of ferric
chloride TS: no orange-red color is produced.

ASSAY
• TOTAL AMMONIA (NH3) Delete the following:
Sample: 10.0 mL •Heavy metals, Method I 〈231〉:
Titrimetric system . 0.001%.• (Official 1-Jan-2018)
(See Titrimetry 〈541〉, Residual Titrations.) Assay—Transfer about 100 mg of Ammonium Chloride, ac-
Mode: Residual titration curately weighed, to a conical flask, add 10 mL of water,
Titrant: 0.5 N sodium hydroxide VS and swirl to dissolve. Add 10 mL of glacial acetic acid,
Analysis: Transfer the Sample to a 250-mL conical flask 75 mL of methanol, and 0.5 mL of eosin Y TS. Titrate, with
containing 50 mL of water. Add 30.0 mL of 0.5 N sulfu- shaking, with 0.1 N silver nitrate VS to a pink endpoint.
ric acid VS, and boil until the solution becomes clear. Each mL of 0.1 N silver nitrate is equivalent to 5.349 mg of
Cool, add methyl red TS, and titrate the excess acid NH4Cl.
with Titrant. Perform a blank determination. Each mL of
0.5 N sulfuric acid is equivalent to 8.515 mg of ammo-
nia (NH3).
• AMMONIUM CARBONATE
Sample: 10.0 mL Ammonium Chloride Injection
.

Titrimetric system
(See Titrimetry 〈541〉, Residual Titrations.)
Mode: Residual titration
Titrant: 0.5 N sulfuric acid VS
Analysis: Transfer the Sample to a 300-mL flask. Add
30 mL of 0.5 N sodium hydroxide, and boil the mix-
ture, replacing the water lost by evaporation, until the
vapors no longer turn moistened red litmus paper blue.
Cool, dilute with 100 mL of cold, carbon dioxide-free
water, add 6 drops of phenolphthalein TS, then add just
enough 0.5 N sulfuric acid VS to discharge the color of
the phenolphthalein. Add methyl orange TS, and titrate
with Titrant. Perform a blank determination. Each mL of
Titrant consumed in the titration with methyl orange TS
» Ammonium Chloride Injection is a sterile solu-
tion of Ammonium Chloride in Water for Injec-
tion. It contains not less than 95.0 percent and
not more than 105.0 percent of the labeled
amount of NH4Cl. Hydrochloric acid may be
added to adjust the pH.
Packaging and storage—Preserve in single-dose or multi-
ple-dose containers, preferably of Type I or Type II glass.
Labeling—The label states the content of ammonium chlo-
ride in terms of weight and of milliequivalents in a given
volume. The label states also the total osmolar concentra-
tion in mOsmol per L or per mL. The label states that the

Official from December 1, 2017

Copyright (c) 2018 The United States Pharmacopeial Convention. All rights reserved.