Biotin

This article is compiled based on the United States Pharmacopeia (USP) – 2025 Edition

Issued and maintained by the United States Pharmacopeial Convention (USP)

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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.

1 DEFINITION

Biotin contains NLT 97.5% and NMT 102.0% of biotin (C₁₀H₁₆N₂O₃S).

2 IDENTIFICATION

A. Spectroscopic Identification Tests 〈197〉, Infrared Spectroscopy: 197K (CN 1-May-2020)

B. It meets the requirements in Specific Tests for Optical Rotation 〈781S〉, Specific Rotation.

C. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.

3 ASSAY

Procedure

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)

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

(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)

Acceptance criteria: 97.5%–102.0%

4 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 = (rᵢ/rₜ) × 100

rᵢ = 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%

5 SPECIFIC TESTS

Optical Rotation, Specific Rotation 〈781S〉

Sample solution: 20 mg/mL in 0.1 N sodium hydroxide

Acceptance criteria: +89° to +93°

6 ADDITIONAL REQUIREMENTS

Packaging and Storage: Store in tight containers.

USP Reference Standards 〈11〉

USP Biotin RS