Biotin Capsules

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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1 DEFINITION

Biotin Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of biotin (C₁₀H₁₆N₂O₃S).

2 IDENTIFICATION

A. 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: 0.05 mg/mL of USP Biotin RS in Diluent. Sonicate, if necessary, to dissolve.

Sample solution: Weigh NLT 20 Capsules in a tared weighing bottle. Open the Capsules, without the loss of shell material, and transfer the contents to a 100-mL beaker. Remove any contents adhering to the empty shells by washing, if necessary, with several portions of ether. Discard the washings, and dry the Capsule shells with the aid of a current of dry air until the odor of ether is no longer perceptible. Weigh the empty Capsule shells in the tared weighing bottle, and calculate the average net weight per Capsule. Transfer a portion of the Capsule contents, equivalent to a nominal amount of 5 mg of biotin to a 100-mL volumetric flask, add 60 mL of water, and shake in a water bath at 65° for 20 min. Sonicate for 5 min, shake by mechanical means for 15 min, and cool to room temperature. Add 20 mL of acetonitrile, dilute with water to volume, and filter.

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 Capsules 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 = nominal concentration of biotin in the Sample solution (mg/mL)

Acceptance criteria: 90.0%–110.0%

4 PERFORMANCE TESTS

Dissolution 〈711〉

Buffer solution: 0.02 N anhydrous disodium hydrogen phosphate adjusted with phosphoric acid to a pH of 7.4

Medium: Buffer solution; 500 mL

Apparatus 1: 100 rpm

Time: 1 h

Standard solution: Dissolve a suitable amount of USP Biotin RS in Buffer solution to obtain a concentration similar to that expected in the Sample solution.

Sample solution: Withdraw a portion of the solution under test, pass through a suitable filter, and use the pooled sample as the test specimen.

Analysis

Proceed as directed in the Assay, making any necessary adjustments.

Calculate the percentage of the labeled amount of biotin (C₁₀H₁₆N₂O₃S) dissolved:

Result = (rᵤ/rₛ) × (Cₛ × V/L) × 100

rᵤ = peak area from the Sample solution

rₛ = peak area from the Standard solution

Cₛ = concentration of USP Biotin RS in the Standard solution (µg/mL)

V = volume of Medium, 500 mL

L = labeled amount of biotin (µg/Capsule)

Tolerances: NLT 75% (Q) of the labeled amount of biotin (C₁₀H₁₆N₂O₃S) is dissolved.

Uniformity of Dosage Units 〈905〉: Meet the requirements

5 ADDITIONAL REQUIREMENTS

Packaging and Storage: Preserve in tight, light-resistant containers.

USP Reference Standards 〈11〉

USP Biotin RS