Printed on: Sat Jun 26 2021, 05:42:48 PM Official Status: Currently Official on 26-Jun-2021 DocId: 1_GUID-36929891-2847-468A-A910-66454A3CA97D_4_en-US

(EST)
Printed by: Nguyen Nhan Official Date: Official as of 01-Aug-2018 Document Type: USP & DS @2021 USPC
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CS = concentration of cholecalciferol in the Standard

Cholecalciferol Tablets solution (µg/mL)
DEFINITION CU = nominal concentration of cholecalciferol in the
Cholecalciferol Tablets contain NLT 90.0% and NMT 115.0% Sample solution (µg/mL)
of the labeled amount of cholecalciferol (C27H44O).
Acceptance criteria: 90.0%–115.0%
IDENTIFICATION PERFORMANCE TESTS
• A. The retention time of the major peak of the Sample • DISSOLUTION á711ñ
solution corresponds to that of the Standard solution, as [NOTE—Perform this test under light conditions that
obtained in the Assay. minimize photodegradation.]
• B. The UV spectrum of the major peak of the Sample Medium: 0.1% (w/v) octoxynol-9 in water; 500 mL
solution corresponds to that of the Standard solution, as Apparatus 2: 75 rpm
obtained in the Assay. Time: 30 min
ASSAY Mobile phase: Acetonitrile and water (70:30)
• PROCEDURE Standard stock solution: 0.5 mg/mL of USP
[NOTE—Use amber, low-actinic glassware.] Cholecalciferol RS in isopropyl alcohol
Diluent: Water, dimethylformamide, and isopropyl alcohol Standard solution: Dilute Standard stock solution with
(5:10:35, v/v) Medium to obtain the concentration equivalent to that
Mobile phase: Acetonitrile and water (70:30) expected in the Sample solution.
Standard stock solution: 0.5 mg/mL of USP Sample solution: Withdraw a portion of the solution under
Cholecalciferol RS in isopropyl alcohol test, pass through a 0.45-µm PVDF filter, and use the
Ergocalciferol stock solution: 0.5 mg/mL of USP pooled sample as the test specimen.

al
Ergocalciferol RS in isopropyl alcohol Chromatographic system
System suitability solution: Combine equal volumes of (See Chromatography á621ñ, System Suitability.)
Standard stock solution and Ergocalciferol stock solution and Mode: LC
dilute with Diluent to obtain a concentration of about 1 µg/ Detector: UV 265 nm
mL each of cholecalciferol and ergocalciferol.
ci Column: 4.6-mm × 15-cm; 2.7-µm packing L96
Standard solution: 1 µg/mL of USP Cholecalciferol RS in Column temperature: 40°
Diluent from the Standard stock solution, prepared Flow rate: 1 mL/min
accurately Injection volume: 100 µL
Sample solution: Finely powder NLT 20 Tablets. Transfer a System suitability
portion of the powder, nominally equivalent to 50 µg of Sample: Standard solution
ffi
cholecalciferol, into a 50-mL volumetric flask, add 5 mL of Suitability requirements
water and sonicate for 10 min with intermittent shaking. Relative standard deviation: NMT 2.0%
Add 10 mL of dimethylformamide, sonicate for 15 min with Analysis
intermittent shaking, cool to room temperature, and dilute Samples: Standard solution and Sample solution
with isopropyl alcohol to volume. Mix well and pass Calculate the percentage of the labeled amount of
through a 0.45-µm nylon filter. cholecalciferol dissolved:
O

Chromatographic system
(See Chromatography á621ñ, System Suitability.) Result = (r U/r S) × (C S × V/L) × 100
Mode: LC
Detector: UV 265 nm rU = peak area of cholecalciferol from the Sample
Column: 4.6-mm × 15-cm; 2.7-µm packing L96 solution
Column temperature: 40° rS = peak area of cholecalciferol from the Standard
Flow rate: 1 mL/min solution
Injection volume: 20 µL CS = concentration of cholecalciferol in the Standard
System suitability solution (µg/mL)
Samples: System suitability solution and Standard solution V = volume of Medium, 500 mL
[NOTE—The relative retention times for cholecalciferol L = label claim of cholecalciferol (µg/Tablet)
and ergocalciferol are 1.0 and 1.1, respectively.]
Suitability requirements Tolerances: NLT 75% (Q) of the labeled amount of
Resolution: NLT 1.5 between the cholecalciferol and cholecalciferol is dissolved.
ergocalciferol peaks, System suitability solution • UNIFORMITY OF DOSAGE UNITS á905ñ: Meet the
Relative standard deviation: NMT 2.0%, Standard requirements
solution ADDITIONAL REQUIREMENTS
Analysis • PACKAGING AND STORAGE: Preserve in tight, light-resistant
Samples: Standard solution and Sample solution containers.
Calculate the percentage of the labeled amount of • LABELING: Label the Tablets to indicate the content of
cholecalciferol (C27H44O) in the portion of Tablets taken: cholecalciferol in µg.
• USP REFERENCE STANDARDS á11ñ
Result = (r U/r S) × (C S/C U) × 100 USP Cholecalciferol RS
USP Ergocalciferol RS
rU = peak area of cholecalciferol from the Sample
solution
rS = peak area of cholecalciferol from the Standard
solution

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