Alprazolam Tablets

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

Alprazolam Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of alprazolam (C₁₇H₁₃CIN₄).

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.

B. The UV spectrum of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.

3 ASSAY

3.1 PROCEDURE

Solution A: 0.77 g/L of ammonium acetate prepared as follows. Dissolve 0.77 g of ammonium acetate in each liter of water and adjust with acetic acid to a pH of 4.7.

Solution B: Acetonitrile

Mobile phase: See Table 1.

Table 1

Diluent: Acetonitrile and water (40:60)

System suitability solution: 50 µg/mL of USP Alprazolam RS, 1 µg/mL of USP Alprazolam Related Compound A RS, and 1 µg/mL of USP Chlordiazepoxide Related Compound A RS in Diluent

Standard solution: 50 µg/mL of USP Alprazolam RS in Diluent

Sample solution: Nominally 50 µg/mL of alprazolam from Tablets prepared as follows. Transfer a suitable portion of powder from NLT 10 Tablets to an appropriate volumetric flask. Add 80% of the total flask volume of Diluent. Sonicate for NLT 10 min. Dilute with Diluent to volume. Centrifuge a portion and use the clear supernatant. [NOTE-The use of a centrifuge speed of 3500 rpm for 10 min may be suitable.]

3.2 Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 260 nm. For Identification B, use a diode array detector in the range of 200-400 nm.

Column: 4.6-mm x 15-cm; 3-µm packing L7

Flow rate: 1 mL/min

Injection volume: 25 µL

3.3 System suitability

Samples: System suitability solution and Standard solution

[NOTE-The relative retention times for chlordiazepoxide related compound A, alprazolam related compound A, and alprazolam are 0.7, 0.8, and 1.0, respectively.]

Resolution: NLT 1.5 between chlordiazepoxide related compound A and alprazolam related compound A, System suitability solution

Suitability requirements

Tailing factor: NMT 1.5, Standard solution

Relative standard deviation: NMT 2.0%, Standard solution

3.4 Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of the labeled amount of alprazolam (C₁₇H₁₃CIN₄) in the portion of Tablets taken:

Result = (r_U/r_S) x (C_S/C_U) x 100

r_U = peak response from the Sample solution

r_S = peak response from the Standard solution

C_S = concentration of USP Alprazolam RS in the Standard solution (µg/mL)

C_U = nominal concentration of alprazolam in the Sample solution (µg/mL)

Acceptance criteria: 90.0%-110.0%

4 PERFORMANCE TESTS

4.1 DISSOLUTION (711), Procedure, Apparatus 1 and Apparatus 2. Immediate-release dosage forms. Procedure for a pooled sample for immediate-release dosage forms

Buffer stock solution: Dissolve 80 g of monobasic potassium phosphate and 20 g of dibasic potassium phosphate in 1 L of water. Add, with mixing, phosphoric acid or potassium hydroxide solution (45 in 100), as necessary to adjust the solution, such that the resulting solution has a pH of 6.0 ± 0.1.

Buffer: Prepare a 1-in-10 dilution of the Buffer stock solution to obtain a solution that has a pH of 6.0 ± 0.1.

Medium: Buffer, 500 mL

Apparatus 1: 100 rpm

Time: 30 min

Mobile phase: Acetonitrile, tetrahydrofuran, and Buffer (35:5:60)

Standard stock solution: 0.05 mg/mL of USP Alprazolam RS in methanol

Standard solution: Add 50 mL of Buffer stock solution and 250 mL of water to a 500-mL flask. Add to the flask 5.0 mL of Standard stock

solution for every 0.25 mg of alprazolam contained in the Tablet being assayed. Dilute with water to volume.

Sample solution: Pass a portion of the solution under test through a suitable filter.

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 254 nm

Column: 4.6-mm x 10-cm; packing LZ

Flow rate: 1 mL/min

System suitability

Sample: Standard solution

Suitability requirements

Column efficiency: NLT 500 theoretical plates

Relative standard deviation: NMT 3.0% for replicate injections

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of the labeled amount of alprazolam (C₁₇H₁₃CIN₄) dissolved.

Tolerances: NLT 80% (Q) of the labeled amount of alprazolam (C₁₇H₁₃CIN₄) is dissolved.

Change to read:

4.2 UNIFORMITY OF DOSAGE UNITS (905): Meet the requirements (CN 1-Aug-2023)

Mobile phase: Acetonitrile, chloroform, butyl alcohol, glacial acetic acid, and water (850: 80: 50:0.5:20)

Internal standard solution: 0.032 mg/mL of triazolam in acetonitrile

Standard solution: 0.025 mg/mL of USP Alprazolam RS in Internal standard solution

Sample solution: Transfer 1 Tablet to a container. Add 0.4 mL of water directly onto the Tablet, allow the Tablet to stand for 2 min, and then swirl the container to disperse the Tablet. For every 0.25 mg of alprazolam contained in the Tablet, add 10.0 mL of Internal standard solution to the container. Shake, and centrifuge if necessary.

(See Chromatography (621), System Suitability.)

Chromatographic system

Mode: LC

Detector: UV 254 nm

Column: 4.6-mm × 30-cm; packing 13

Flow rate: 2 mL/min

Injection volume: 20 µL

System suitability

Sample: Standard solution

Suitability requirements

Resolution: NLT 2.0 between triazolam and alprazolam

Relative standard deviation: NMT 2.0% for replicate injections

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of the labeled amount of alprazolam (C₁₇H₁₃CIN₄) in the Tablet taken:

Result = (R_U/R_S) x C x V x (100/L)

R_U = peak area response ratio of alprazolam relative to the internal standard from the Sample solution

R_S = peak area response ratio of alprazolam relative to the internal standard from the Standard solution

C = concentration of USP Alprazolam RS in the Standard solution (mg/mL)

V = volume of the Internal standard solution used to prepare the Sample solution (mL)

L = label claim (mg/Tablet)

5 ADDITIONAL REQUIREMENTS

PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at controlled room temperature.

USP REFERENCE STANDARDS (11)

USP Alprazolam RS

USP Alprazolam Related Compound A RS

2-(2-Acetylhydrazino)-7-chloro-5-phenyl-3H-1,4-benzodiazepine.

C₁₇H₁₅CIN₄O 326.78

USP Chlordiazepoxide Related Compound A RS

7-Chloro-2-oxo-5-phenyl-2,3-dihydro-1H-benzo[e][1,4]diazepine 4-oxide;

Also known as 7-Chloro-1,3-dihydro-5-phenyl-2H-1,4-benzodiazepin-2-one 4-oxide.

C₁₅H₁₁CIN₂O₂ 286.71