22.07.
2025, 09:06 USP-NF Biotin
Printed on: Tue Jul 22 2025, 09:06:41 am
Printed by: Artem Kulikov
Status: Currently Official on 22-Jul-2025
Official Date: Official as of 01-May-2020
Document Type: USP Monographs
DocId: GUID-904A2528-89D8-4AE7-A042-3BABDE60AA16_2_en-US
DOI: https://doi.org/10.31003/USPNF_M9515_02_01
DOI Ref: wd53x
Printed from: https://online.uspnf.com/uspnf/document/1_GUID-904A2528-89D8-4AE7-A042-3BABDE60AA16_2_en-US
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Biotin
C10H16N2O3S 244.31
1H-Thieno[3,4-d]imidazole-4-pentanoic acid, hexahydro-2-oxo-, [3aS-(3aα,4β,6aα)]-;
(3aS,4S,6aR)-Hexahydro-2-oxo-1H-thieno[3,4-d]imidazole-4-valeric acid CAS RN®: 58-85-5; UNII: 6SO6U10H04.
DEFINITION
Biotin contains NLT 97.5% and NMT 102.0% of biotin (C10H16N2O3S).
IDENTIFICATION
Change to read:
• A. ▲Spectroscopic Identification Tests 〈197〉, Infrared Spectroscopy: 197K▲ (CN 1-May-2020)
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• B. It meets the requirements in Specific Tests for Optical Rotation〈781S〉, Specific Rotation.
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• C. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.
ASSAY
• Procedure
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Buffer solution: Dissolve 1 g of sodium perchlorate monohydrate in 500 mL of water, add 1 mL of phosphoric acid, and dilute with water to
1000 mL.
Mobile phase: Acetonitrile and Buffer solution (8.5:91.5)
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Diluent: Acetonitrile and water (1:4)
Standard solution: 0.1 mg/mL of USP Biotin RS in Diluent. Sonicate if necessary to dissolve.
Sample solution: 0.1 mg/mL of Biotin in Diluent. Sonicate if necessary to dissolve.
Chromatographic system
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(See Chromatography 〈621〉, System Suitability.)
Mode: LC
Detector: UV 200 nm
Column: 4.6-mm × 15-cm; 3-µm packing L7
Flow rate: 1.2 mL/min
Injection volume: 50 µL
System suitability
Sample: Standard solution
Suitability requirements
Tailing factor: NMT 1.5
Relative standard deviation: NMT 2.0% for replicate injections
Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of biotin (C10H16N2O3S) in the portion of Biotin taken:
Result = (rU/rS) × (CS/CU) × 100
rU = peak response from the Sample solution
rS = peak response from the Standard solution
CS = concentration of USP Biotin RS in the Standard solution (mg/mL)
CU = concentration of Biotin in the Sample solution (mg/mL)
https://online.uspnf.com/uspnf/document/1_GUID-904A2528-89D8-4AE7-A042-3BABDE60AA16_2_en-US?source=Quick Search&highlight=Biotin 1/2
22.07.2025, 09:06 USP-NF Biotin
Acceptance criteria: 97.5%–102.0%
IMPURITIES
• Related Compounds
Buffer solution, Mobile phase, Diluent, Standard solution, Sample solution, Chromatographic system, and System suitability: Proceed as
directed in the Assay.
Analysis
Sample: Sample solution
Measure the peak responses of the Sample solution.
Calculate the percentage of each impurity in the portion of Biotin taken:
Result = (rU/rT) × 100
rU = peak response of each impurity from the Sample solution
rT = sum of the peak responses of all the peaks from the Sample solution
Acceptance criteria
Individual impurity: NMT 1.0%
Total impurities: NMT 2.0%
SPECIFIC TESTS
• Optical Rotation, Specific Rotation〈781S〉
Sample solution: 20 mg/mL in 0.1 N sodium hydroxide
Acceptance criteria: +89° to +93°
ADDITIONAL REQUIREMENTS
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• Packaging and Storage: Store in tight containers.
• USP Reference Standards 〈11〉
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USP Biotin RS
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Auxiliary Information - Please check for your question in the FAQs before contacting USP.
Topic/Question Contact Expert Committee
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Natalia Davydova NBDS2020 Non-botanical Dietary
BIOTIN
Scientific Liaison Supplements
RS Technical Services NBDS2020 Non-botanical Dietary
REFERENCE STANDARDS SUPPORT
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RSTECH@usp.org Supplements
Chromatographic Database Information: Chromatographic Database
Most Recently Appeared In:
Pharmacopeial Forum: Volume No. PF 39(6)
Current DocID: GUID-904A2528-89D8-4AE7-A042-3BABDE60AA16_2_en-US
DOI: https://doi.org/10.31003/USPNF_M9515_02_01
DOI ref: wd53x
https://online.uspnf.com/uspnf/document/1_GUID-904A2528-89D8-4AE7-A042-3BABDE60AA16_2_en-US?source=Quick Search&highlight=Biotin 2/2
