Printed on: Fri May 19 2023, 05:02:00 AM(EST) Status: Currently Official on 19-May-2023 DocId: GUID-A1199B07-4EA5-476C-ADC6-EF9D4E9231B1_4_en-US
Printed by: Anandkumar Joshi Official Date: Official as of 01-May-2020 Document Type: USP @2023 USPC
Do Not Distribute DOI Ref: dn2lv DOI: https://doi.org/10.31003/USPNF_M11170_04_01
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Relative standard deviation: NMT 2.0%
Caffeine Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of caffeine (C8H10N4O2) in the
portion of Caffeine taken:
Result = (r U/r S) × (C S/C U) × 100
C8H10N4O2 · H2O 212.21
rU = peak response of caffeine from the Sample solution
C8H10N4O2 194.19 rS = peak response of caffeine from the Standard
1H-Purine-2,6-dione, 3,7-dihydro-1,3,7-trimethyl-; solution
1,3,7-Trimethylxanthine CAS RN®: 58-08-2; UNII: CS = concentration of USP Caffeine RS in the Standard
3G6A5W338E. solution (mg/mL)
Monohydrate CAS RN®: 5743-12-4. CU = concentration of Caffeine in the Sample solution
DEFINITION (mg/mL)
Caffeine is anhydrous or contains one molecule of water of
hydration. It contains NLT 98.5% and NMT 101.0% Acceptance criteria: 98.5%–101.0% on the anhydrous
of C8H10N4O2, calculated on the anhydrous basis. basis
IMPURITIES
IDENTIFICATION • RESIDUE ON IGNITION á281ñ: NMT 0.1%
Change to read: • ORGANIC IMPURITIES
Mobile phase, Standard solution, Sample solution,
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• A. ▲SPECTROSCOPIC IDENTIFICATION TESTS á197ñ, Infrared Chromatographic system, and System suitability:
Spectroscopy: 197M▲ (CN 1-May-2020) Proceed as directed in the Assay.
• B. The retention time of the caffeine peak of the Sample Analysis
solution corresponds to that of the Standard solution, as Sample: Sample solution
obtained in the Assay. ci Calculate the percentage of each impurity in the portion of
Caffeine taken:
ASSAY
• PROCEDURE Result = (r U/r T) × 100
Buffer: 0.82 g/L of anhydrous sodium acetate
Mobile phase: Acetonitrile, tetrahydrofuran, and Buffer rU = peak response for each impurity from the
(25:20:955). Adjust with glacial acetic acid to a pH of 4.5. Sample solution
ffi
System suitability solution: 0.02 mg/mL of theophylline in rT = sum of the responses of all the peaks from the
Mobile phase. Shake, and sonicate if necessary, to dissolve. Sample solution
Standard solution: Transfer 5.0 mg of USP Caffeine RS to a
25-mL volumetric flask. Add 5.0 mL of the System suitability Acceptance criteria
solution and 10 mL of Mobile phase. Shake, and sonicate if Individual impurities: NMT 0.1%
necessary. Dilute with Mobile phase to volume, and filter. Total impurities: NMT 0.1%
O
Sample solution: 0.2 mg/mL of Caffeine in Mobile phase.
[NOTE—Shake, and sonicate if necessary, to dissolve.] SPECIFIC TESTS
Chromatographic system • WATER DETERMINATION, Method III á921ñ: Dry a sample at
(See Chromatography á621ñ, System Suitability.) 80° for 4 h: the anhydrous form loses NMT 0.5% of its
Mode: LC weight, and the hydrous form loses NMT 8.5% of its
Detector: UV 275 nm weight.
Column: 4.6-mm × 15-cm; packing L1 ADDITIONAL REQUIREMENTS
Flow rate: 1 mL/min • PACKAGING AND STORAGE: Preserve hydrous Caffeine in
Injection size: 10 µL tight containers. Preserve anhydrous Caffeine in well-closed
System suitability containers.
Sample: Standard solution • LABELING: Label it to indicate whether it is anhydrous or
[NOTE—The relative retention times for theophylline hydrous.
and caffeine are 0.69 and 1.0, respectively.] • USP REFERENCE STANDARDS á11ñ
Suitability requirements USP Caffeine RS
Resolution: NLT 6.0 between theophylline and caffeine
Tailing factor: NMT 2.0 for theophylline and caffeine
peaks
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