Printed on: Fri Jan 05 2024, 10:07:27 PM(EST) Status: Currently Official on 06-Jan-2024 DocId: GUID-9B183E8E-D12D-4AB0-9AC7-403F0B377E11_4_en-US
Printed by: USP NF Official Date: Official as of 01-Oct-2023 Document Type: USP @2024 USPC
Do Not Distribute DOI Ref: qm7iz DOI: [Link]
1
Injection volume: 50 µL
Digoxin Tablets System suitability
DEFINITION Sample: System suitability solution
Digoxin Tablets contain NLT 90.0% and NMT 105.0% of the Suitability requirements
labeled amount of digoxin (C41H64O14). Resolution: NLT 4.0 between digoxin and digoxigenin
Column efficiency: NLT 1200 theoretical plates
IDENTIFICATION Tailing factor: NMT 2.0
• A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Relative standard deviation: NMT 2.0%
á201ñ Analysis
Diluent: Dehydrated alcohol Samples: Standard solution and Sample solution
Standard solution: 0.25 mg/mL of USP Digoxin RS in Calculate the percentage of digoxin (C41H64O14) in the
Diluent portion of Tablets taken:
Sample solution: Transfer a quantity of finely powdered
Tablets, equivalent to 0.5 mg of digoxin, into a 10-mL Result = (rU/rS) × (CS/CU) × 100
centrifuge tube. Add 2 mL of Diluent, sonicate for 10–
15 min, and centrifuge. Decant and use the supernatant. rU = peak response of digoxin from the Sample solution
Chromatographic system rS = peak response of digoxin from the Standard
(See Chromatography á621ñ, Thin-Layer Chromatography.) solution
Mode: TLC CS = concentration of USP Digoxin RS in the Standard
Adsorbent: 0.25-mm layer of chromatographic silica gel solution (µg/mL)
mixture to which octadecylsilane (C18) is permanently CU = nominal concentration of ▲digoxin▲ (ERR 1-Oct-2023)
bonded in the Sample solution (µg/mL)
al
Application volume: 10 µL
Developing solvent system: Methanol and water (7:3) Acceptance criteria: 90.0%–105.0%
Spray reagent: Mix 10 mL of a freshly prepared solution
PERFORMANCE TESTS
of chloramine T (3 in 100) and 40 mL of a 1 in 4 solution
• DISSOLUTION á711ñ
of trichloroacetic acid in dehydrated alcohol. [NOTE—Mix
Test 1
before use.]
Analysis
Samples: Standard solution and Sample solution
Apply the Samples on a line parallel to and about 2.5 cm
ci [NOTE—Throughout this procedure, use scrupulously
clean glassware, rinsed successively with hydrochloric
acid, water, and alcohol, and carefully dried. Take
precautions to prevent contamination from fluorescent
from the bottom edge of a reversed-phase thin-layer
particles and from metal and rubber surfaces.]
chromatographic plate. Allow the spots to dry, and place
ffi
Medium: 0.1 N hydrochloric acid; 500 mL
the plates in a developing chamber. Develop the
[NOTE—Use the same batch of Medium throughout
chromatogram until the solvent front has moved about
the test.]
15 cm above the line of application. Remove the plate,
Apparatus 1: 120 rpm
and allow the solvent to evaporate. Spray the plate with
Time: 60 min
Spray reagent, and heat in an oven at 110° for 10 min.
Solution A: 2 mg/mL of ascorbic acid in methanol
Examine the plate under long-wavelength UV light.
O
Solution B: On the day of use, dilute 2.0 mL of recently
Acceptance criteria: The RF value of the principal spot of
assayed 30% hydrogen peroxide with methanol to 100 mL.
the Sample solution corresponds to that of the Standard Store in a refrigerator. Just before use, dilute 2.0 mL of this
solution. solution with methanol to 100 mL.
• B. The retention time of the major peak of the Sample Standard solutions: Transfer 25 mg of USP Digoxin RS
solution corresponds to that of the Standard solution, as into a 500-mL volumetric flask. Dissolve in a minimum
obtained in the Assay. amount of alcohol, then add diluted alcohol (4 in 5) to
ASSAY volume. Dilute 10.0 mL of this solution with diluted alcohol
(4 in 5) to 100.0 mL. Just before use, dilute suitable aliquots
Change to read: of the resulting solution with Medium to 50.0 mL to prepare
Standard solutions equivalent to 20%, 40%, 60%, 80%, and
• PROCEDURE 100%, respectively, of the labeled amount of digoxin in
Mobile phase: Acetonitrile and water (13:37) 500 mL.
System suitability solution: 40 µg/mL each of USP Digoxin Sample solution: Promptly after withdrawal, pass a portion
RS and digoxigenin in diluted alcohol of the solution under test through a filter having a 0.8-µm
Standard solution: 40 µg/mL of USP Digoxin RS in diluted or finer pore size, discarding the first 10 mL of the filtrate.
alcohol. [NOTE—Use a sonic bath to aid dissolution.] Analysis: Transfer to individual glass-stoppered flasks
Sample solution: Weigh and finely powder NLT 20 Tablets. duplicate 1.0-mL portions of the Sample solution, 1.0-mL
Transfer a portion of the powder, equivalent to 1 mg of portions of each of the Standard solutions, and 1.0 mL of
digoxin, to a glass-stoppered, 50-mL conical flask. Add the Medium to provide a blank. Begin with the Standard
25.0 mL of diluted alcohol with swirling, sonicate for solutions, and keep all flasks in the same sequence
30 min, and cool. Pass a portion of this solution through a throughout, so that the elapsed time from addition of
0.8-µm pore size membrane filter, discarding the first 10 mL reagents to reading of fluorescence is the same for each
of the filtrate. flask in the set. Treating one flask at a time, add the
Chromatographic system following three reagents, in the order named, in as rapid a
(See Chromatography á621ñ, System Suitability.) sequence as possible, swirling after each addition: 1.0 mL
Mode: LC of Solution A, 5.0 mL of hydrochloric acid, and 1.0 mL of
Detector: UV 218 nm Solution B. Insert the stoppers in the flasks. After 2 h,
Column: 4.6-mm × 25-cm; 5-μm packing L1 measure the fluorescence at about 485 nm, the excitation
Temperature: 25° wavelength being about 372 nm. To check the stability of
Flow rate: 2 mL/min the fluorometer, repeat the measurement of fluorescence
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Printed on: Fri Jan 05 2024, 10:07:27 PM(EST) Status: Currently Official on 06-Jan-2024 DocId: GUID-9B183E8E-D12D-4AB0-9AC7-403F0B377E11_4_en-US
Printed by: USP NF Official Date: Official as of 01-Oct-2023 Document Type: USP @2024 USPC
Do Not Distribute DOI Ref: qm7iz DOI: [Link]
2
on one or more treated Standard solutions. Correct each Table 2 (continued)
reading for the blank, and plot a standard curve of Time Flow Rate
fluorescence versus percentage dissolution. (min) (mL/min)
Determine the percentage dissolution of digoxin in the 2.5 0.6
Sample solution by reading from the standard graph.
Tolerances: NLT 80% (Q) of the labeled amount 3.0 2.0
of C41H64O14 is dissolved. The requirement is met if the 6.0 2.0
quantities dissolved from the Tablets tested conform to
Table 1 instead of the table shown under Dissolution á711ñ.
System suitability
Table 1 Sample: Standard solution
Suitability requirements
Stage Number Tested Acceptance Criteria
Tailing factor: NMT 2.0
S1 6 Each unit is NLT Q + 5%. Relative standard deviation: NMT 5.0%
Analysis
S2 6 Average of 12 units (S1 +
S2) is equal to or greater
Samples: Standard solution and Sample solution
than Q, and no unit is Calculate the percentages of the labeled amount of digoxin
less than (C41H64O14) dissolved:
Q − 5%.
Result = (rU/rS) × CS × D × V × (1/L) × 100
Test 2
[NOTE—If the product complies with this test, the rU = peak response of digoxin from the Sample solution
rS = peak response of digoxin from the Standard
al
labeling indicates that it meets USP Dissolution Test 2.]
Medium: Water, deaerated, 500 mL solution
Apparatus 1: 120 rpm CS = concentration of USP Digoxin RS in the Standard
Time: 20 min solution (mg/mL)
Solution A: Mix 890 mL of water with 110 mL of D = dilution factor for the Sample solution
acetonitrile. V = volume of Medium, 500 mL
Solution B: Mix 100 mL of water with 900 mL of
acetonitrile.
Mobile phase: Mix 650 mL of Solution A and 350 mL of
ci L = label claim (mg)
Tolerances: NLT 80% (Q) of the labeled amount of
Solution B. digoxin (C41H64O14) is dissolved. The requirement is met if
Standard stock solution: Transfer 25 mg of USP Digoxin RS the quantities dissolved from the Tablets tested conform to
ffi
to a 200-mL volumetric flask and dissolve with 10 mL of Table 3 instead of the table shown under Dissolution á711ñ.
methanol. Fill up to the volume with Medium.
Standard solution: Dilute the Standard stock solution with Table 3
Medium in order to obtain a final concentration of L/ Stage Number Tested Acceptance Criteria
500 mg/mL, where L is the product label claim in
S1 6 NLT 80% (Q) of the la-
milligrams.
O
beled amount of digox-
Sample solution: Pass 10 mL of the solution under test in
through a suitable polyvinylidene fluoride (PVDF) filter of
S2 12 Average of 12 units (S1 +
0.45-µm pore size, discarding the first 4 mL. Dilute with S2) is equal to or greater
Medium. than Q, and no unit is
Chromatographic system less than
(See Chromatography á621ñ, System Suitability.) Q − 5%.
Mode: LC
Detector: UV 220 nm • UNIFORMITY OF DOSAGE UNITS á905ñ: Meet the
Column: 3.0-mm × 5-cm; 3.5-µm packing L11 requirements
Temperatures
Column: 40° ADDITIONAL REQUIREMENTS
Autosampler: 5° • PACKAGING AND STORAGE: Preserve in tight containers.
Injection volume: 500 µL • LABELING: The labeling states the Dissolution test used only
Flow rate: See Table 2. if Test 1 is not used.
• USP REFERENCE STANDARDS á11ñ
Table 2 USP Digoxin RS
Time Flow Rate
(min) (mL/min)
0.0 0.6
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