Anagrelide 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

Anagrelide Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of anagrelide (C₁₀H₇CI₂N₃O).

2 IDENTIFICATION

A. The UV absorption spectrum of the major peak of the Sample solution exhibits maxima and minima at the same wavelengths as those of the corresponding peak of the Standard solution, as obtained in the Assay.

B. 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

3.1 PROCEDURE

Solution A: 1.0 g/L of sodium hexanesulfonate. Add 1.0 mL of phosphoric acid and filter.

Mobile phase: Acetonitrile and Solution A (7:13)

Diluent: Acetonitrile and water (1:1)

Standard stock solution: 0.25 mg/mL of USP Anagrelide Hydrochloride RS in acetonitrile. Initially add acetonitrile (about 80% of the volume of the flask) and a small quantity of 2 N hydrochloric acid (about 0.2 mL for every 100 mL of the final volume), Sonicate to dissolve, and dilute with acetonitrile to volume.

Standard solution: 0.01 mg/mL of anagrelide free base in Diluent from Standard stock solution

Sample solution: Nominally 0.01 mg/mL of anagrelide free base prepared from the contents of NLT 20 Capsules. Add Diluent (80% of the volume of the flask), sonicate for 10 min, and stir for 15 min. Further dilute with Diluent to volume, centrifuge for 15 min at 4000 rpm, and use the supernatant for analysis.

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 254 nm. For Identification A, use a diode array detector in the range of 200-400 nm.

Column: 4.6-mm x 15-cm; 4-µm packing L11

Column temperature: 60°

Flow rate: 1.0 mL/min

Injection volume: 20 µL

System suitability

Sample: Standard solution

Suitability requirements

Tailing factor: NMT 2.0

Relative standard deviation: NMT 2.0%

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of the labeled amount of anagrelide (C₁₀H₇CI₂N₃O) in the portion of Capsules taken:

Result = (r_u /r_s ) × (C_s /C_u ) × 100

r_u = peak response of anagrelide from the Sample solution

r_s = peak response of anagrelide from the Standard solution

C_s = concentration of anagrelide in the Standard solution (mg/mL)

C_u = nominal concentration of anagrelide in the Sample solution (mg/mL)

Acceptance criteria: 90.0%-110.0%

4 PERFORMANCE TESTS

Change to read:

DISSOLUTION (711)

Medium: 0.1 N hydrochloric acid: 900 mL

Apparatus 1: 100 rpm

Time: 15 min

Mobile phase: Prepare as directed in the Assay

Standard solution: Transfer about 30.32 mg of USP Anagrelide Hydrochloride RS, equivalent to 25.00 mg of anagrelide, to a 100-mL volumetric flask. Add about 80 mL of acetonitrile and 3 drops of 2 N hydrochloric acid. Sonicate until dissolved. Dilute with acetonitrile to volume. Dilute this solution with Medium to obtain a final concentration of about (L/1000) mg/ml, where L is the label claim in mg/Capsule.

Sample solution: Pass a portion of the solution under test through a suitable filter of 0.45-um pore size.

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 274 nm

Column: 4.6-mm x 15-cm; 5-µm packing L7

Sample cooler temperature: 5"

Flow rate: 1.0 mL/min

Injection volume: 20 µL

System suitability

Sample: Standard solution

Suitability requirements

Tailing factor: NMT 2.0

Relative standard deviation: NMT 2.0%

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of the labeled amount of anagrelide dissolved:

Result = (r_u /r_s ) × C_{s (ERR 1-Oct-2019)} × (M_r1 /M_r2 ) × (V/L) × 100

r_u = peak response of anagrelide from the Sample solution

r_s = peak response of anagrelide from the Standard solution

C_s = concentration of USP Anagrelide Hydrochloride RS in the Standard solution (mg/mL)

M_r1 = molecular weight of anagrelide, 256.09

M_r2 = molecular weight of anagrelide hydrochloride, 292.55

V = volume of Medium, 900 mL

L = label claim (mg/Capsule)

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

UNIFORMITY OF DOSAGE UNITS (905): Meet the requirements

5 IMPURITIES

5.1 ORGANIC IMPURITIES

Buffer: 6.8 g/L of monobasic potassium phosphate. Adjust with diluted phosphoric acid to a pH of 3.50 ± 0.05. Mix well and filter.

Mobile phase: Acetonitrile and Buffer (27:73)

Diluent: Acetonitrile and water (7:13)

Related compound A stock solution: 10 µg/mL of USP Anagrelide Related Compound A RS in Diluent

Related compound C stock solution: 10 µg/mL of USP Anagrelide Related Compound C. RS in Diluent

System suitability solution: 0.2 µg/mL each of USP Anagrelide Related Compound A RS and USP Anagrelide Related Compound C. RS and 0.02 mg/mL of USP Anagrelide Hydrochloride RS prepared as follows. Initially dissolve USP Anagrelide Hydrochloride RS in Diluent (about 80% of the volume of the flask), sonicate for 10 min, and stir for 15 min. Add appropriate quantities of Related compound A stock solution and Related compound C stock solution, and dilute with Diluent to volume.

Standard stock solution: Prepare as directed in the Assay.

Standard solution: 0.10 µg/mL of anagrelide free base in Diluent from Standard stock solution

Sample solution: Nominally 0.02 mg/mL of anagrelide free base from NLT 20 Capsules. Initially add Diluent to about 80% of the volume of the flask, sonicate for 10 min, stir for about 15 min, and dilute with Diluent to volume. Centrifuge the solution at about 4000 rpm for 15 min, and use the supernatant for analysis.

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 254 nm

Column: 4.6-mm × 15-cm; 4-µm packing L11

Column temperature: 45°

Flow rate: 1 mL/min

Injection volume: 30 µL

5.2 System suitability

Samples: System suitability solution and Standard solution

Suitability requirements

Resolution: NLT 2.0 between anagrelide hydrochloride and anagrelide related compound C, and between anagrelide hydrochloride and

anagrelide related compound A, System suitability solution

Tailing factor: NMT 2.0, Standard solution

Relative standard deviation: NMT 2.0%, Standard solution

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of each impurity in the portion of Capsules taken:

Result = (r_u/r_s) x (C_s/C_u) x (1/F) x 100

r_u = peak response of each individual impurity from the Sample solution

r_s = peak response of anagrelide from the Standard solution

C_s = concentration of anagrelide in the Standard solution (mg/mL)

C_u = nominal concentration of anagrelide in the Sample solution (mg/mL)

F = relative response factor (see Table 1)

Acceptance criteria: See Table 1.

Table 1

^a This is a process-related impurity and controlled in the drug substance.

^b [2-Amino-5,6-dichloroquinazoline-3(4H)-yl]acetic acid.

^c 6,7,8-Trichloro-3,5-dihydroimidazo(2,1-b]quinazolin-2(1H)-one.

6 ADDITIONAL REQUIREMENTS

PACKAGING AND STORAGE: Preserve in tight containers, protected from light. Store at controlled room temperature.

USP REFERENCE STANDARDS (11)

USP Anaorelide Hydrochloride RS

USP Anagrelide Related Compound A RS

Ethyl 2-(6-amino-2,3-dichlorobenzylamino)acetate.

C₁₁H₁₄Cl₂N₂O₂                    277.15

Ethyl 2-(5,6-dichloro-2-imino-1,2-dihydroquinazolin-3(4H)-yl)acetate hydrobromide.

C₁₂H₁₃Cl₂N₃O₂  · HBr         383.07