USP 36 Official Monographs / Amlodipine 2467

Assay preparation—Transfer about 50 mg of Amlodipine Flow rate: 1 mL/min

Besylate, accurately weighed, to a 50-mL volumetric flask, Injection size: 50 µL
dissolve in and dilute with Mobile phase to volume, and mix. System suitability
Transfer 5.0 mL of this solution to a 100-mL volumetric flask, Sample: System suitability solution
dilute with Mobile phase to volume, and mix. [NOTE—The run time is about three times the retention
Chromatographic system (see Chromatography 〈621〉)—The of the amlodipine peak.]
liquid chromatograph is equipped with a 237-nm detector Suitability requirements
and a 3.9-mm × 15-cm column that contains packing L1. Resolution: NLT 8.5 between amlodipine and
The flow rate is about 1.0 mL per minute. Chromatograph amlodipine related compound A
the Standard preparation, and record the peak responses as Tailing factor: NMT 2.0 for both amlodipine and
directed for Procedure: the standard deviation for replicate amlodipine related compound A
injections is not more than 2.0%. Relative standard deviation: NMT 1.0% for
Procedure—Separately inject equal volumes (about 10 µL) amlodipine and NMT 5.0% for amlodipine related
of the Standard preparation and the Assay preparation into compound A
the chromatograph, record the chromatograms, and meas- Analysis
ure the responses for the major peaks. Calculate the per- Samples: Standard solution and Sample solution
centage of C20H25ClN2O5 · C6H6O3S in the portion of Calculate the percentage of the labeled amount of
Amlodipine Besylate taken by the formula: amlodipine (C20H25N2O5Cl) in the portion of Tablets
taken:
100(CS /CU)(rU / rS)
Result = (rU/rS) × (CS/CU) × 100
in which CS and CU are the concentrations, in mg per mL, of rU = peak response from the Sample solution
amlodipine besylate in the Standard preparation and the As- rS = peak response from the Standard solution
say preparation, respectively; and rU and rS are the peak re- CS = concentration of USP Amlodipine Besylate RS
sponses obtained from the Assay preparation and the Stan- in the Standard solution (mg/mL)
dard preparation, respectively. CU = nominal concentration of amlodipine in the
Sample solution (mg/mL)
Acceptance criteria: 90%–110% of the labeled amount
of amlodipine (C20H25N2O5Cl)
Amlodipine Besylate Tablets
.

PERFORMANCE TESTS
• DISSOLUTION 〈711〉
DEFINITION [NOTE—Do not expose any of the solutions to stainless
Amlodipine Besylate Tablets contain NLT 90% and NMT steel because of the degradation of amlodipine.]
110% of the labeled amount of amlodipine Medium: 0.01 N hydrochloric acid; 500 mL
(C20H25N2O5Cl). Apparatus 2: 75 rpm. [NOTE—Use paddles covered with
Teflon or made of any inert material except stainless
IDENTIFICATION steel.]
• A. ULTRAVIOLET ABSORPTION 〈197U〉 Time: 30 min
Standard solution and Sample solution: Prepare as di- Standard solution: Make appropriate dilutions of USP
rected in the test for Dissolution. Amlodipine Besylate RS in Medium to obtain the follow-
Acceptance criteria: Meet the requirements ing concentrations: 0.00695 mg/mL for Tablets labeled
• B. The retention time of the major peak of the Sample to contain 2.5 mg; 0.0139 mg/mL for Tablets labeled to
solution corresponds to that of the Standard solution, as contain 5 mg; 0.0278 mg/mL for Tablets labeled to
obtained in the Assay. contain 10 mg. These solutions are stable for one day.
ASSAY Sample solution: Pass a portion of the solution under
• PROCEDURE test through a suitable filter of 0.45-µm pore size.
Buffer: Add 7.0 mL of triethylamine into a 1000-mL flask Analysis: Determine the amount of amlodipine
containing 900 mL of water. Adjust the solution with (C20H25N2O5Cl) dissolved by using UV absorption at the
phosphoric acid to a pH of 3.0 ± 0.1. Dilute with water wavelength of maximum absorbance at about 239 nm
to volume, and mix well. on portions of the Sample solution in comparison with
Mobile phase: Methanol, acetonitrile, and Buffer the Standard solution, using a 1-cm quartz cell and the
(35:15:50) Medium as blank.
System suitability solution: 0.02 mg/mL of USP Calculate the percentage of the labeled amount of
Amlodipine Besylate RS and 0.002 mg/mL of USP amlodipine (C20H25ClN2O5) dissolved:
Amlodipine Related Compound A RS in Mobile phase
Standard solution: 0.02 mg/mL of amlodipine prepared Result = (AU/AS) × (CS/L) × D × (Mr1/Mr2) × V × 100
from USP Amlodipine Besylate RS in Mobile phase AU = absorbance of the Sample solution
Sample stock solution: Place 5 Tablets into a 500-mL AS = absorbance of the Standard solution
volumetric flask. Add 50 mL of Mobile phase to the CS = concentration of the Standard solution
flask, and swirl to disintegrate the Tablets. Add 300 mL (mg/mL)
of Mobile phase, insert the stopper into the flask, and L = label claim (mg/Tablet)
shake on a reciprocating shaker for 30 min. Dilute with D = dilution factor of the Sample solution
Mobile phase to volume, and mix well. Mr1 = molecular weight of amlodipine, 408.88
Sample solution: 0.02 mg/mL of amlodipine from the Mr2 = molecular weight of amlodipine besylate,
Sample stock solution in Mobile phase. Pass the sample 567.06
through a syringe tip filter of 0.45-µm pore size. V = volume of Medium, 500 mL
Chromatographic system Tolerances: NLT 75% (Q) of the labeled amount of
(See Chromatography 〈621〉, System Suitability.) amlodipine (C20H25N2O5Cl) is dissolved.
Mode: LC • UNIFORMITY OF DOSAGE UNITS 〈905〉: Meet the
Detector: UV 237 nm requirements
Column: 3.9-mm × 15-cm; 5-µm packing L1
2468 Amlodipine / Official Monographs USP 36

IMPURITIES Acceptance criteria: See Table 1.

• ORGANIC IMPURITIES
Buffer, Mobile phase, System suitability solution, Table 1
Chromatographic system, and System suitabil-
ity: Proceed as directed in the Assay. Relative Acceptance
Standard solution: Use the System suitability solution. Retention Criteria,
Sample solution: Place a suitable number of Tablets into Name Time NMT (%)
a 25-mL volumetric flask to obtain a solution having a Amlodipine related compound
0.50 1.0
final nominal concentration of 0.4 mg/mL of Aa
amlodipine. Add about 10 mL of Mobile phase to the Amlodipine lactose adductb 0.80 0.5
flask. Swirl to disintegrate the Tablet(s), followed by Amlodipine glucose/galactose
sonication for 5 min to completely dissolve, and then 0.90 0.5
adductb
cool the sample to room temperature. Dilute with Mo- Amlodipine besylate 1.0 —
bile phase to volume. Stir for an additional 15 min using
a magnetic stir bar, and pass the sample through a sy- Any other individual unspeci-
— 0.20
ringe tip filter of 0.45-µm pore size, discarding the first fied degradation product
a 3-Ethyl, 5-methyl [2-(2-aminoethoxymethyl)-4-(2-chlorophenyl)-6-meth-
5 mL.
yl-3,5-pyridinedicarboxylate].
Analysis b Formulation-specific impurities.
Samples: Standard solution and Sample solution
Calculate the percentage of amlodipine related com- ADDITIONAL REQUIREMENTS
pound A in the portion of Tablets taken: • PACKAGING AND STORAGE: Preserve in tight, light-resistant
containers. Store at controlled room temperature.
Result = (rU/rS) × (CS/CU) × (Mr1/Mr2) × 100 • USP REFERENCE STANDARDS 〈11〉
USP Amlodipine Besylate RS
rU = peak response of amlodipine related USP Amlodipine Related Compound A RS
compound A from the Sample solution 3-Ethyl, 5-methyl [2-(2-aminoethoxymethyl)-
rS = peak response of amlodipine related 4-(2-chlorophenyl)-6-methyl-3,5-pyridinedicarboxylate
compound A from the Standard solution fumarate.
CS = concentration of USP Amlodipine Related C20H23ClN2O5 · C4H4O4 522.93
Compound A RS in the Standard solution
(mg/mL)
CU = nominal concentration of amlodipine in the
Sample solution (mg/mL)
Mr1 = molecular weight of amlodipine related
Aromatic Ammonia Spirit
.

compound A, 406.86
Mr2 = molecular weight of amlodipine related
compound A fumarate, 522.93 » Aromatic Ammonia Spirit is a hydroalcoholic
Calculate the percentage of amlodipine glucose/ solution that contains, in each 100 mL, not less
galactose adduct or amlodipine lactose adduct, if
present, in the portion of Tablets taken: than 1.7 g and not more than 2.1 g of total NH3,
and Ammonium Carbonate corresponding to not
Result = (rU/rS) × (CS/CU) × (Mr1/Mr2) × 100 less than 3.5 g and not more than 4.5 g of
rU = peak response of the amlodipine glucose/ (NH4)2CO3.
galactose adduct or amlodipine lactose Packaging and storage—Preserve in tight, light-resistant
adduct in the Sample solution containers, at a temperature not exceeding 30°.
rS = peak response of amlodipine in the Standard
solution Alcohol content, Method I 〈611〉: between 62.0% and
CS = concentration of USP Amlodipine Besylate RS 68.0% of C2H5OH.
in the Standard solution (mg/mL) Assay for total NH3—Transfer 10.0 mL to a 250-mL coni-
CU = nominal concentration of amlodipine in the cal flask containing about 50 mL of water. Add 30.0 mL of
Sample solution (mg/mL) 0.5 N sulfuric acid VS, and boil until the solution becomes
Mr1 = molecular weight of amlodipine, 408.9 clear. Cool, add methyl red TS, and titrate the excess acid
Mr2 = molecular weight of amlodipine besylate, with 0.5 N sodium hydroxide VS. Perform a blank determi-
567.05 nation (see Residual Titrations under Titrimetry 〈541〉). Each
Calculate the percentage of any other individual mL of 0.5 N sulfuric acid is equivalent to 8.515 mg of NH3.
unspecified degradation product in the portion of Assay for ammonium carbonate—Transfer 10.0 mL to a
Tablets taken: flask of about 300-mL capacity. Add 30 mL of 0.5 N sodium
hydroxide, and boil the mixture, replacing the water lost by
Result = (rU/rS) × (CS/CU) × 100 evaporation, until the vapors no longer turn moistened red
litmus paper blue. Cool, dilute with 100 mL of cold, carbon
rU = peak response of each impurity from the dioxide-free water, add about 6 drops of phenolphthalein
Sample solution TS, then add just enough 0.5 N sulfuric acid VS to discharge
rS = peak response of amlodipine from the the color of the phenolphthalein. Add methyl orange TS,
Standard solution and titrate with 0.5 N sulfuric acid VS. Perform a blank de-
CS = concentration of amlodipine in the Standard termination (see Residual Titrations under Titrimetry 〈541〉).
solution (mg/mL) Each mL of 0.5 N sulfuric acid consumed in the titration
CU = nominal concentration of amlodipine in the with methyl orange TS is equivalent to 48.04 mg of
Sample solution (mg/mL) (NH4)2CO3.