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Levothyroxine / Official Monographs

Relative standard deviation: NMT 2.0% of levothyroxine

Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of C15H10I4NNaO4 in the
Levothyroxine Sodium:
Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
= peak response of the Sample solution
= peak response of the Standard solution
= concentration of USP Levothyroxine RS in the
Standard solution (g/mL)
= nominal concentration of levothyroxine in the
CU
Sample solution (g/mL)
= molecular weight of levothyroxine sodium,
Mr1
798.85
= molecular weight of levothyroxine, 776.87
Mr2
Acceptance criteria: 97.0%103.0%
rU
rS
CS

IMPURITIES
Inorganic Impurities
LIMIT OF INORGANIC IODIDES
Extracting solution: Sulfuric acid in water, 1 in 100
Reference solution: 0.131 mg/mL of potassium iodide in
water (equivalent to 0.100 mg/mL of iodide). Dilute with
Extracting solution to 0.06 g/mL of iodide.
[NOTEPrepare this solution on the day of use.]
Sample solution: 0.075 mg/mL of Levothyroxine Sodium
in the Extracting solution
[NOTESonicate for 5 min.]
Electrode system: Use an iodide-specific, ion-indicating
electrode and a silversilver chloride reference electrode
connected to a pH meter capable of measuring potentials
with a minimum reproducibility of 1 mV (see pH 791).
Analysis: Transfer the Reference solution to a beaker
containing a magnetic stirring bar. Rinse and dry the
electrodes, insert in the solution, stir for 5 min or until the
reading stabilizes, and read the potential, in mV. Repeat
this process using the Sample solution.
Acceptance criteria: The Sample solution has a higher
potential, in mV, than the Reference solution: the limit is
0.08%.
Organic Impurities
PROCEDURE 1: LIMIT OF LIOTHYRONINE SODIUM
Mobile phase, Standard solution, Sample solution,
Chromatographic system, and System suitability:
Proceed as directed in the Assay.
Analysis
Calculate the percentage of C15H11I3NNaO4 in the
Levothyroxine Sodium:
Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
rU
rS
CS

= peak response from the Sample solution

= peak response from the Standard solution
= concentration of USP Liothyronine RS in the
Standard solution (mg/mL)
CU
= nominal concentration of liothyronine in the
Sample solution (mg/mL)
Mr1
= molecular weight of liothyronine sodium, 672.96
= molecular weight of liothyronine, 650.98
Mr2
Acceptance criteria: NMT 2.0%
PROCEDURE 2
Diluent: Acetonitrile and water (1:1)
Solution A: Dilute 5 mL of phosphoric acid with Diluent to
100.0 mL.
Mobile phase: Dissolve 1.0 g of sodium 1heptanesulfonate in 200 mL of water. Add 200 mL of
acetonitrile, 400 mL of methanol, and 1.0 mL of
phosphoric acid. Dilute with water to 1 L.
Standard stock solution 1: Transfer 25 mg of USP
Levothyroxine RS to a 100-mL volumetric flask. Add 50 mL

USP 32
of Diluent and 1 drop of 10 N sodium hydroxide, and
sonicate until dissolved. Add 7 mL of Solution A, and dilute
with Diluent to volume.
Standard stock solution 2: Transfer 25 mg of USP
Liothyronine RS to a 100-mL volumetric flask. Add 50 mL
of Diluent and 1 drop of 10 N sodium hydroxide, and
sonicate until dissolved. Add 7 mL of Solution A, and dilute
with Diluent to volume.
System suitability solution: 5.0 mL of the Standard stock
solution 1 and 5.0 mL of the Standard stock solution 2 to a
100-mL volumetric flask. Add 7 mL of Solution A, and dilute
with Diluent to volume.
Standard solution: Pipet 4.0 mL of the System suitability
solution to a 100-mL volumetric flask. Add 7 mL of Solution
A, and dilute with Diluent to volume.
Blank: Add 7 mL of Solution A to a 100-mL volumetric
flask, and dilute with Diluent to volume.
Sample solution: Transfer 25 mg of Levothyroxine Sodium
to a 100-mL volumetric flask. Add 50 mL of Diluent, and
sonicate until dissolved. Add 7 mL of Solution A, and dilute
with Diluent to volume.
Chromatographic system
(See Chromatography 621, System Suitability.)
Mode: LC
Detector: UV 225 nm
Column: 4.6-mm 15-cm; 5-m packing L1
Flow rate: 1.5 mL/min
Injection size: 15 L
Temperature: 35
System suitability
Samples: System suitability solution and Standard solution
Suitability requirements
Resolution: NLT 5.0 between levothyroxine and
liothyronine, System suitability solution
Relative standard deviation: NMT 2.0% for the
levothroxine peak in the Standard solution
Analysis
Samples: Standard solution, Blank, and Sample solution
[NOTERecord the chromatograms for at least six times
the retention time of the levothyroxine peak. Verify that
no peaks elute in the Blank solution at the expected
retention times for levothyroxine and related
compounds.]
Calculate the area percentage of each relative compound
in the portion of Levothyroxine taken:
Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100/(1 0.01L)
rU

= peak area for each impurity from the Sample

solution
= peak area for levothyroxine from the Standard
rS
solution
= concentration of levothyroxine in the Standard
CS
solution (mg/ml)
= nominal concentration of levothyroxine sodium
CU
in the Sample solution (mg/ml)
= molecular weight of levothyroxine sodium,
Mr1
798.85
= molecular weight of levothyroxine, 776.87
Mr2
L
= percentage of water in Levothyroxine Sodium,
as determined separately in the test for Water
Determination 921
[NOTEThe relative response factor for the impurities
listed in Impurity Table 1 is 1.00. Any unspecified
impurity peaks should be assigned a relative response
factor of 1.00.]
Disregard peaks corresponding to those of the Blank
solution, and disregard peaks corresponding to less than
0.03%.

Copyright 2008 The United States Pharmacopeial Convention. All Rights Reserved.
For Discussion Purposes Only Not for Dissemination