Anagrelide Hydrochloride

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

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

DOWNLOAD PDF HERE

C₁₀H₇Cl₂N₃O · HCl · H₂O                 310.56

Anhydrous 292.55 CAS RN: 58579-51-4; UNII: VNS4435G39.

Imidazo[2,1-b]quinazolin-2(3H)-one, 6,7-dichloro-1,5-dihydro-, monohydrochloride, monohydrate;

6,7-Dichloro-1,5-dihydroimidazo[2,1-b]-quinazolin-2(3H)-one monohydrochloride, monohydrate CAS RN: 823178-43-4.

1 DEFINITION

Anagrelide Hydrochloride contains NLT 98.0% and NMT 102.0% of anagrelide hydrochloride (C₁₀H₇Cl₂N₃O · HCl), calculated on the anhydrous basis.

2 IDENTIFICATION

Change to read:

A. SPECTROSCOPIC IDENTIFICATION TESTS (197), Infrared Spectroscopy: 197K _{(CN 1-MAY-2020)}

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

C. IDENTIFICATION TESTS GENERAL, Chloride (191); Meets the requirements

3 ASSAY

3.1 PROCEDURE

Use freshly prepared standard and sample solutions and inject within 2 h..

Solution A: 6.8 g/L of monobasic potassium phosphate in water. Adjust with phosphoric acid to a a pH of 2.5.

Mobile phase: Acetonitrile and Solution A (1:3)

Diluent: Acetonitrile and water (1:1)

Standard stock solution: 0.5 mg/mL of anagrelide hydrochloride in acetonitrile prepared as follows. Transfer USP Anagrelide Hydrochloride RS into a suitable volumetric flask, add a small amount of 2 N hydrochloric acid (3 drops per every 50 mL of the final volume) and acetonitrile equivalent to fill about 80% of the final volume. Sonicate to dissolve, and dilute with acetonitrile to volume.

Standard solution: 0.05 mg/ml. of anagrelide hydrochloride in Diluent from Standard stock solution

Sample stock solution: Weigh Anagrelide Hydrochloride, equivalent to 25 mg of anhydrous salt, into a 50-mL volumetric flask, add 3 drops of 2 N hydrochloric acid and 40 mL of acetonitrile. Sonicate to dissolve, and dilute with acetonitrile to volume.

Sample solution: Transfer 5 mL of Sample stock solution to a 50-ml volumetric flask, and dilute with Diluent to volume.

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 254 nm

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

Flow rate: 1.2 mL/min

Injection volume: 20 µL

System suitability

Sample: Standard solution

Suitability requirements

Column efficiency: NLT 3000 theoretical plates

Tailing factor: NMT 2.0

Relative standard deviation: NMT 2.0%

3.2 Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of anagrelide hydrochloride (C₁₀H₇Cl₂N₃O · HCl) in the portion of Anagrelide Hydrochloride 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 USP Anagrelide Hydrochloride RS in the Standard solution (mg/mL)

C_u = concentration of Anagrelide Hydrochloride in the Sample solution (mg/mL)

Acceptance criteria: 98.0%–102.0% on the anhydrous basis

4 IMPURITIES

RESIDUE ON IGNITION (281): NMT 0.1%

ORGANIC IMPURITIES

Use freshly prepared standard and sample solutions and inject within 2 h.

Mobile phase: Proceed as directed in the Assay.

Diluent A: Use the Diluent as described in the Assay.

Diluent B: Acetonitrile and water (1:3)

Standard stock solution A: 0.05 mg/mL of USP Anagrelide Related Compound A RS in Diluent A

Standard stock solution B: 0.05 mg/mL of anagrelide related compound B in acetonitrile. Transfer USP Anagrelide Related Compound B RS into a suitable volumetric flask, add acetonitrile equivalent to fill about 50% of the final volume and a small amount of 2 N hydrochloric acid (3 drops per 200 mL of the final volume). Sonicate to dissolve, heat in the hot water bath if necessary, and dilute with acetonitrile to volume.

Standard stock solution C: 0.1 mg/ml. of anagrelide hydrochloride in acetonitrile. Transfer USP Anagrelide Hydrochloride RS into a suitable volumetric flask, add acetonitrile equivalent to fill about 80% of the final volume and a small amount of 0.12 N hydrochloric acid (1 mL per 100 ml. of the final volume). Sonicate to dissolve, and dilute with acetonitrile to volume.

System suitability solution: 0.25 µg/mL of each of anagrelide related compound A and anagrelide related compound B in Mobile phase from

Standard stock solution A and Standard stock solution B

Standard solution: 0.05 µg/mL of anagrelide hydrochloride in Mobile phase from Standard stock solution C

Sample stock solution: Weigh Anagrelide Hydrochloride, equivalent to 25 mg of anhydrous salt, into a 50-mL volumetric flask. Add 45 mL of acetonitrile, sonicate, and swirl the flask until the preparation turns into a cloudy liquid. Add 1 drop of 0.12 N hydrochloric acid, swirl the flask until the liquid turns to clear, and dilute with acetonitrile to volume.

Sample solution: Transfer 5 mL of Sample stock solution into a 50-mL volumetric flask, and dilute with Diluent B to volume. FICIA

Chromatographic system.

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 254 nm

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

Autosampler temperature: 5"

Flow rate: 1.2 mL/min

Injection volume: 50 µL

System suitability

Samples: System suitability solution and Standard solution

Suitability requirements

Resolution: NLT 2.0 between anagrelide related compound B and anagrelide related compound A, System suitability solution

Column efficiency: NLT 3000 theoretical plates, Standard solution

Tailing factor: NMT 2.0, Standard solution

Relative standard deviation: NMT 10.0%, Standard solution

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of each impurity in the portion of Anagrelide Hydrochloride, on the anhydrous basis, taken:

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

r_u = peak response of each impurity 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)

C_u = concentration of Anagrelide Hydrochloride (anhydrous) in the Sample solution (mg/mL)

F = relative response factor for each individual impurity (see Table 1)

Acceptance criteria See Table 1. Disregard any impurity peak less than 0.05%.

Table 1

^a (2-Amino-5,6-dichloroquinazolin-3(4H)-yl)acetic acid.

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

^c Methyl 2-(5,6 dichloro-2-imino-1,2-dihydroquinazolin-3(4H)-yl)acetate.

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

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

5 SPECIFIC TESTS

Water Determination, Method I〈921〉: 4.5%–7.5%

6 ADDITIONAL REQUIREMENTS

Packaging and Storage: Preserve in tight, light-resistant containers. Store in a cold place.

USP Reference Standards 〈11〉

USP Anagrelide Hydrochloride RS

USP Anagrelide Related Compound A RS

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

C₁₁H₁₄Cl₂N₂O₂            277.15

USP Anagrelide Related Compound B RS

(2-Amino-5,6-dichloroquinazolin-3(4H)-yl)acetic acid.

C₁₀H₉Cl₂N₃O₂             274.10