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2552 Anagrelide / Official Monographs USP 39

compound B in Mobile phase from Standard stock solu- Table 1 (Continued)

tion A and Standard stock solution B Relative Relative Acceptance
Standard solution: 0.05 µg/mL of anagrelide hydro- Retention Response Criteria,
chloride in Mobile phase from Standard stock solution C Name Time Factor NMT (%)
Sample stock solution: Weigh Anagrelide Hydrochlo-
ride, equivalent to 25 mg of anhydrous salt, into a Anagrelide 1.00 1.0 —
50-mL volumetric flask. Add 45 mL of acetonitrile, soni- Anagrelide related
cate, and swirl the flask until the preparation turns into compound Cd . 1.41 0.32 0.15
a cloudy liquid. Add 1 drop of 0.12 N hydrochloric Anagrelide trichloro
acid, swirl the flask until the liquid turns to clear, and derivativee . 2.44 1.0 0.15
dilute with acetonitrile to volume. Any unspecified
Sample solution: Transfer 5 mL of Sample stock solution —
impurity 1.0 0.1
into a 50-mL volumetric flask, and dilute with Diluent B Total impurities — — 1.0
to volume. a (2-Amino-5,6-dichloroquinazolin-3(4H)-yl)aceticacid.
Chromatographic system
.

b Ethyl 2-(6-amino-2,3-dichlorobenzylamino)acetate.
(See Chromatography 〈621〉, System Suitability.)
.

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

Mode: LC
.

d Ethyl 2-(5,6-dichloro-2-imino-1,2-dihydroquinazolin-3(4H)-yl)acetate
Detector: UV 254 nm
.

hydrobromide.
Column: 4.6-mm × 15-cm; 4-µm packing L11 e 6,7,8-Trichloro-3,5-dihydroimidazo[2,1-b]quinazolin-2(1H)-one.
Autosampler temperature: 5°
.

Flow rate: 1.2 mL/min SPECIFIC TESTS

Injection volume: 50 µL
System suitability
Samples: System suitability solution and Standard
solution
Suitability requirements
Resolution: NLT 2.0 between anagrelide related com-
pound 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 por-
tion of Anagrelide Hydrochloride, on the anhydrous
USP Monographs
• WATER DETERMINATION, Method I 〈921〉: 4.5%–7.5%
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

basis, taken: Anagrelide Capsules

Result = (rU/rS) × (CS/CU) × (1/F) × 100 DEFINITION
Anagrelide Capsules contain NLT 90.0% and NMT 110.0%
rU = peak response of each impurity from the of the labeled amount of C10H7Cl2N3O.
Sample solution
rS = peak response of anagrelide from the Standard IDENTIFICATION
solution • The retention time of the major peak of the Sample solu-
CS = concentration of USP Anagrelide tion corresponds to that of the Standard solution, as ob-
Hydrochloride RS in the Standard solution tained in the Assay.
(mg/mL)
CU = concentration of Anagrelide Hydrochloride ASSAY
(anhydrous) in the Sample solution (mg/mL) • PROCEDURE
F = relative response factor for each individual Solution A: 1.0 g/L of sodium hexanesulfonate. Add
impurity (see Table 1) 1.0 mL of phosphoric acid and filter.
Acceptance criteria See Table 1. Disregard any impurity Mobile phase: Acetonitrile and Solution A (7:13)
peak less than 0.05%. Diluent: Acetonitrile and water (1:1)
Standard stock solution: 0.25 mg/mL of USP Anagre-
lide Hydrochloride RS in acetonitrile. Initially add aceto-
Table 1 nitrile (about 80% of the volume of the flask) and a
Relative Relative Acceptance small quantity of 2 N hydrochloric acid (about 0.2 mL
Retention Response Criteria, for every 100 mL of the final volume). Sonicate to dis-
Name Time Factor NMT (%) solve, and dilute with acetonitrile to volume.
Anagrelide related Standard solution: 0.01 mg/mL of anagrelide free base
compound Ba 0.40 0.43 0.3 in Diluent from Standard stock solution
Sample solution: 0.01 mg/mL of anagrelide free base
.

Anagrelide related
compound Ab 0.55 0.37 0.15 prepared from the contents of NLT 20 Capsules. Add
Diluent (80% of the volume of the flask), sonicate for
.

Anagrelide open ring

10 min, and stir for 15 min. Further dilute with Diluent
methyl ester (if
to volume, centrifuge for 15 min at 4000 rpm, and use
present)c 0.80 0.51 0.25
.

the supernatant for analysis.

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.
.

Official from May 1, 2016

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Accessed from 10.6.1.1 by pdxcamed on Fri Jun 24 03:50:39 EDT 2016

USP 39 Official Monographs / Anagrelide 2553

Chromatographic system V = volume of Medium, 900 mL

(See Chromatography 〈621〉, System Suitability.) L = label claim (mg/Capsule)
Mode: LC Tolerances: NLT 80% (Q) of the labeled amount of
Detector: UV 254 nm anagrelide is dissolved.
Column: 4.6-mm × 15-cm; 4-µm packing L11 • UNIFORMITY OF DOSAGE UNITS 〈905〉: Meet the
Column temperature: 60° requirements
Flow rate: 1.0 mL/min
Injection size: 20 µL IMPURITIES
System suitability Organic Impurities
Sample: Standard solution • PROCEDURE
Suitability requirements Buffer solution: 6.8 g/L of monobasic potassium phos-
Column efficiency: NLT 3000 theoretical plates phate. Adjust with diluted phosphoric acid to a pH of
Tailing factor: NMT 2.0 3.50 ± 0.05. Mix well and filter.
Relative standard deviation: NMT 2.0% Mobile phase: Acetonitrile and Buffer solution (27:73)
Analysis Diluent: Acetonitrile:water (7:13)
Samples: Standard solution and Sample solution Related compound A stock solution: 10 µg/mL of
Calculate the percentage of C14H13N5O5S2, based on the USP Anagrelide Related Compound A RS in Diluent
label claim, in the portion of Capsules taken: Related compound C stock solution: 10 µg/mL of
USP Anagrelide Related Compound C RS in Diluent
Result = (rU/rS) × (CS/CU) × 100 System suitability solution: 0.2 µg/mL of each USP
Anagrelide Related Compound A RS and USP Anagre-
rU = peak response of anagrelide from the Sample lide Related Compound C RS and 0.02 mg/mL of USP
solution Anagrelide Hydrochloride RS. Initially dissolve USP
rS = peak response of anagrelide from the Standard Anagrelide Hydrochloride RS in Diluent (about 80% of
solution the volume of the flask), sonicate for 10 min, and stir
Cs = concentration of anagrelide in the Standard for 15 min. Add appropriate amounts of Related com-
solution (mg/mL) pound A stock solution and Related compound C stock
CU = nominal concentration of anagrelide in the solution, and dilute with Diluent to volume.
Sample solution (mg/mL) Standard stock solution: 0.1 mg/mL of anagrelide free
Acceptance criteria: 90.0%–110.0% base by dissolving USP Anagrelide Hydrochloride RS in
acetonitrile (about 80% of the volume of the flask).
PERFORMANCE TESTS Add a small quantity of 2 N hydrochloric acid (about
• DISSOLUTION 〈711〉 0.2 mL for every 100 mL of the final volume), sonicate
Medium: 0.1 N hydrochloric acid; 900 mL to dissolve, and dilute with acetonitrile to volume.
Apparatus 1: 100 rpm Standard solution: 0.10 µg/mL of anagrelide free base
Time: 15 min in Diluent from Standard stock solution
Mobile phase: Proceed as directed in the Assay. Sample solution: 0.02 mg/mL of anagrelide free base
Standard solution: Transfer about 30.32 mg of USP from NLT 20 Capsules. Initially add Diluent to about

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 Capsule label claim in mg.
Sample solution: Pass a portion of the solution under
test through a suitable filter of 0.45-µm pore size.
Chromatographic system
(See Chromatography 〈621〉, System Suitability.)
Mode: LC
Detector: UV 274 nm
Column: 4.6-mm × 15-cm; 5-µm packing L7
Sample cooler temperature: 5°
Flow rate: 1.0 mL/min
Injection size: 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%
Calculate the percentage of anagrelide dissolved:
Result = (rU/rS) × WS(100 – WC)/(25 × 100) × Mr1/Mr2 ×
V/L x 100
rU = peak response from the Sample solution
rS = peak response from the Standard solution
WS = weight of the USP Anagrelide Hydrochloride
RS taken (mg)
WC = water content of the USP Anagrelide
Hydrochloride RS (%)
Mr1 = molecular weight of anagrelide, 256.10
Mr2 = molecular weight of anagrelide hydrochloride,
292.56
USP Monographs
80% of the volume of the flask, sonicate for 10 min,
stir for about 15 min, and dilute with Diluent to vol-
ume. Centrifuge (about 4000 rpm) the solution 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: 1.0 mL/min
Injection size: 30 µL
System suitability
Samples: System suitability solution and Standard
solution
Suitability requirements
Resolution: NLT 2.0 between anagrelide hydrochlo-
ride and anagrelide related compound C, and be-
tween anagrelide hydrochloride and anagrelide re-
lated compound A, System suitability solution
Column efficiency: NLT 3000 theoretical plates,
Standard solution
Tailing factor: NMT 2.0, Standard solution
Relative standard deviation: NMT 2.0% for the
anagrelide peak, Standard solution
Calculate the percentage of each impurity in the por-
tion of Capsules taken:
Result = (rU/rS) × (CS/CU) × (1/F) × 100
rU = peak response of each individual impurity
from the Sample solution
rS = peak response of anagrelide from the
Standard solution
CU = nominal concentration of anagrelide in the
Sample solution (mg/mL)

Official from May 1, 2016

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 Accessed from 10.6.1.1 by pdxcamed on Fri Jun 24 03:50:39 EDT 2016

2554 Anagrelide / Official Monographs USP 39

CS = concentration of anagrelide in the Standard Solution B: Acetonitrile, methanol, trifluoroacetic acid,

solution (mg/mL) and water (150: 450: 0.5: 400)
F = relative response factor (see Impurity Table 1) Mobile phase: See Table 1.
Acceptance criteria: The individual and total impurities
meet the limits in Impurity Table 1. [NOTE—Anagrelide Table 1
related compound A (RRT = 0.86) and anagrelide re-
lated compound C (RRT = 1.15) are process related Time Solution A Solution B
and controlled in the drug substance.] (min) (%) (%)
0 100 0
Impurity Table 1 10 100 0
40 0 100
Relative Relative Acceptance
Retention Response Criteria, 41 100 0
Name Time Factor NMT (%) 56 100 0
Anagrelide
hydrochloride 1.0
— — [NOTE—These gradient elution times are established on
an HPLC system with a dwell time of approximately 0
Anagrelide related min. The gradient elution times in the table can be
compound Ba . 0.3 0.34 1.0 adjusted by subtracting the dwell time to achieve the
Anagrelide trichloro separation described.]
derivativeb . 1.8–2.3 1.0 0.15 Standard solution: 0.5 mg/mL of USP Anastrozole RS
Any other
— —
prepared as follows. Transfer USP Anastrozole RS into a
individual impurity 0.2 suitable volumetric flask. Dissolve in acetonitrile, using
Total Impurities — — 1.5 40% of the final volume, and then dilute with Solution
a [2-Amino-5,6-dichloroquinazoline-3(4H)-yl]acetic acid. A to volume.
Sample solution: 0.5 mg/mL of Anastrozole prepared
.

b 6,7,8-Trichloro-3,5-dihydroimidazo[2,1-b]quinazolin-2(1H)-one.
as follows. Transfer 25 mg of Anastrozole to a 50-mL
.

ADDITIONAL REQUIREMENTS volumetric flask, add 20 mL of acetonitrile to dissolve.

• PACKAGING AND STORAGE: Preserve in tight containers, Dilute with Solution A to volume.
protected from light. Store at controlled room Chromatographic system
temperature. (See Chromatography 〈621〉, System Suitability.)
• USP REFERENCE STANDARDS 〈11〉 Mode: LC
USP Anagrelide Hydrochloride RS Detector: UV 215 nm
USP Anagrelide Related Compound A RS Column: 3.2-mm × 10-cm; 5-µm packing L42
Ethyl 2-(6-amino-2,3-dichlorobenzylamino)acetate. Flow rate: 0.75 mL/min
C11H14Cl2N2O2 277.15 Injection volume: 10 µL
USP Anagrelide Related Compound C RS System suitability
Ethyl 2-(5,6-dichloro-2-imino-1,2-dihydroquinazolin- Sample: Standard solution
USP Monographs

3(4H)-yl)acetate hydrobromide. Suitability requirements

Tailing factor: Between 0.9 and 1.4
Relative standard deviation: NMT 0.73%
Analysis
.

Samples: Standard solution and Sample solution

Anastrozole Calculate the percentage of anastrozole (C17H19N5) in
the portion of Anastrozole taken:
Result = (rU/rS) × (CS/CU) × 100
rU = peak area of anastrozole from the Sample
solution
rS = peak area of anastrozole from the Standard
solution
CS = concentration of USP Anastrozole RS in the
C17H19N5 293.37 Standard solution (mg/mL)
1,3-Benzenediacetonitrile, α,α,α′,α′-tetramethyl-5-(1H-1,2, CU = concentration of Anastrozole in the Sample
4-triazol-1-ylmethyl)-; solution (mg/mL)
α,α,α′,α′-Tetramethyl-5-(1H-1,2,4-triazol-1-ylmethyl)-m- Acceptance criteria: 98.0%–102.0% on the anhydrous
benzenediacetonitrile [120511-73-1]. and solvent-free basis

DEFINITION IMPURITIES
Anastrozole contains NLT 98.0% and NMT 102.0% of anas- • RESIDUE ON IGNITION 〈281〉: NMT 0.1%
trozole (C17H19N5), calculated on the anhydrous and sol-
vent-free basis. Delete the following:
IDENTIFICATION •• HEAVY METALS, Method II 〈231〉: NMT 10 ppm• (Official 1-
• A. INFRARED ABSORPTION 〈197K〉 .

• B. The retention time of the major peak of the Sample Jan-2018)

solution corresponds to that of the Standard solution, as • ORGANIC IMPURITIES

obtained in the Assay. Solution A, Solution B, and Chromatographic system:
Proceed as directed in the Assay.
ASSAY Standard stock solution: 0.2 mg/mL of USP Anas-
• PROCEDURE trozole RS prepared as follows. Dissolve in acetonitrile,
Solution A: Acetonitrile, methanol, trifluoroacetic acid, using 40% of the final volume, and then dilute with
and water (100: 300: 0.5: 600) Solution A to volume.
Standard solution: 0.02 mg/mL of USP Anastrozole RS
in Solution A from the Standard stock solution

Official from May 1, 2016

Copyright (c) 2016 The United States Pharmacopeial Convention. All rights reserved.