Plerixafor

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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C₂₈H₅₄N₈ 502.80

1,4,8,11-Tetraazacyclotetradecane, 1,1'-[1,4-phenylenebis(methylene)]bis-;

1,4-Bis[(1,4,8,11-tetraazacyclotetradecan-1-yl)methyl]benzene CAS RN®: 110078-46-1; UNII: S915P5499N.

1 DEFINITION

Plerixafor contains NLT 97.0% and NMT 103.0% of plerixafor (C₂₈H₅₄N₈), calculated on the anhydrous basis.

2 IDENTIFICATION

A. Spectroscopic Identification Tests 〈197〉, Infrared Spectroscopy: 197A or 197K

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

Procedure

Solution A: 0.1% (v/v) trifluoroacetic acid in water

Solution B: Methanol and water (50:50). To each liter of the solution, add 1 mL of trifluoroacetic acid.

Mobile phase: See Table 1.

Table 1

Diluent: 0.05 N hydrochloric acid

Standard solution: 1 mg/mL of USP Plerixafor RS in Diluent. Sonicate to dissolve.

Sample solution: 1 mg/mL of Plerixafor in Diluent. Sonicate to dissolve.

Chromatographic system

(See Chromatography 〈621〉, System Suitability.)

Mode: LC

Detector: UV 230 nm

Columns

Guard: 4.6-mm × 12.5-mm; 5-μm packing L7

Analytical: 4.6-mm × 15-cm; 3.5-μm packing L7

Column temperature: 40°

Flow rate: See Table 1.

Injection volume: 20 μL

System suitability

Sample: Standard solution

Suitability requirements

Tailing factor: NMT 5.0

Relative standard deviation: NMT 1.10%

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of plerixafor (C₂₈H₅₄N₈) in the portion of Plerixafor taken:

Result = (r_U/r_S) × (C_S/C_U) × 100

r_U = peak response of plerixafor from the Sample solution

r_S = peak response of plerixafor from the Standard solution

C_S = concentration of USP Plerixafor RS in the Standard solution (mg/mL)

C_U = concentration of Plerixafor in the Sample solution (mg/mL)

Acceptance criteria: 97.0%–103.0% on the anhydrous basis

4 IMPURITIES

Residue on Ignition 〈281〉: NMT 0.1%

Organic Impurities

Solution A, Solution B, Mobile phase, Diluent, Standard solution, Sample solution, and Chromatographic system: Proceed as directed in the Assay.

System suitability solution: 1 mg/mL of USP Plerixafor System Suitability Mixture RS in Diluent. Sonicate to dissolve.

Sensitivity solution: 0.0005 mg/mL of USP Plerixafor RS in Diluent from the Standard solution

System suitability

[Note - Column conditioning may be performed as needed to meet the system suitability requirements using 1.6 mg/mL of USP Plerixafor System Suitability Mixture RS in water as a column conditioning solution. Flush the column with 0.1% v/v trifluoroacetic acid in methanol for 1 h. Equilibrate the column with the starting Mobile phase gradient for 30 min. Carry out multiple injections of the column conditioning solution until the resolution criterion of NLT 1.0 between plerixafor 4-benzyl analog and plerixafor 8-benzyl analog is met.]

Samples: Standard solution, System suitability solution, and Sensitivity solution

[Note - The relative retention times in Table 2 are provided as information that could aid in peak assignment.]

Table 2

^a{4-[(1,4,8,11-Tetraazacyclotetradecan-1-yl)methyl]phenyl}methanol.

^b 1,4-Bis{4-[(1,4,8,11-tetraazacyclotetradecan-1-yl)methyl]benzyl}-1,4,8,11-tetraazacyclotetradecane.

^c 1,8-Bis{4-[(1,4,8,11-tetraazacyclotetradecan-1-yl)methyl]benzyl}-1,4,8,11-tetraazacyclotetradecane.

^d 1,11-Bis{4-[(1,4,8,11-tetraazacyclotetradecan-1-yl)methyl]benzyl}-1,4,8,11-tetraazacyclotetradecane.

Suitability requirements

Resolution: NLT 1.0 between plerixafor 4-benzyl analog and plerixafor 8-benzyl analog; NLT 1.0 between plerixafor and plerixafor benzyl alcohol, System suitability solution

Relative standard deviation: NMT 1.0%, Standard solution

Signal-to-noise ratio: NLT 10, Sensitivity solution

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of each specified and unspecified impurity in the portion of Plerixafor taken:

Result = (r_U/r_S) × (C_S/C_U) × (1/F) × 100

r_U = peak response of each specified or unspecified impurity from the Sample solution

r_S = peak response of plerixafor from the Standard solution

C_S = concentration of USP Plerixafor RS in the Standard solution (mg/mL)

C_U = concentration of Plerixafor in the Sample solution (mg/mL)

F = relative response factor (see Table 3)

Acceptance criteria: See Table 3. The reporting threshold is 0.05%.

Table 3

^a Sum of the results obtained in the Organic Impurities and Limit of Cyclam tests.

Limit of Cyclam

Solution A: 0.1% (v/v) trifluoroacetic acid in water

Solution B: 0.1% (v/v) trifluoroacetic acid in acetonitrile

Mobile phase: See Table 4.

Table 4

Derivatization solution: 20 mg/mL of cupric acetate monohydrate in water

Standard solution: 0.005 mg/mL of USP Cyclam RS in water

Sample solution: 1 mg/mL of Plerixafor in water

Derivatized standard solution: 0.0025 mg/mL of cyclam prepared as follows. Mix 2 mL of the Standard solution with 2 mL of the Derivatization solution in a suitable head space vial. Crimp to seal the vial and place the vial in a 55° water bath for 15 min. Cool the vial and use for analysis.

Derivatized sample solution: 0.5 mg/mL of plerixafor prepared as follows. Mix 2 mL of the Sample solution with 2 mL of the Derivatization solution in a suitable head space vial. Crimp to seal the vial and place the vial in a 55° water bath for 15 min. Cool the vial and use for analysis.

Chromatographic system

(See Chromatography 〈621〉, System Suitability.)

Mode: LC

Detector: UV 266 nm

Column: 4.6-mm × 15-cm; 5-μm packing L1

Flow rate: 1 mL/min

Injection volume: 20 μL

System suitability

Sample: Derivatized standard solution

Suitability requirements

Relative standard deviation: NMT 10.0%

Analysis

Samples: Derivatized standard solution and Derivatized sample solution

Calculate the percentage of cyclam in the portion of Plerixafor taken:

Result = (r_U/r_S) × (C_S/C_U) × 100

r_U = peak response of cyclam from the Derivatized sample solution

r_S = peak response of cyclam from the Derivatized standard solution

C_S = concentration of USP Cyclam RS in the Derivatized standard solution (mg/mL)

C_U = concentration of Plerixafor in the Derivatized sample solution (mg/mL)

Acceptance criteria: NMT 0.10%

5 SPECIFIC TESTS

Water Determination 〈921〉, Method I, Method Ic: NMT 1.5%

Bacterial Endotoxins Test 〈85〉: Where the label states that Plerixafor is sterile or must be subjected to further processing during the preparation of injectable dosage forms, the level of bacterial endotoxins is such that the requirement under the relevant dosage form monograph(s) in which Plerixafor is used can be met.

Microbial Enumeration Tests 〈61〉 and Tests for Specified Microorganisms 〈62〉: The total aerobic microbial count does not exceed 102 cfu/g. The total yeasts and molds count does not exceed 102 cfu/g.

6 ADDITIONAL REQUIREMENTS

Packaging and Storage: Preserve in tight containers at controlled room temperature.

Labeling: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms.

USP Reference Standards 〈11〉

USP Cyclam RS

1,4,8,11-Tetraazacyclotetradecane.

C₁₀H₂₄N₄ 200.33

USP Plerixafor RS

USP Plerixafor System Suitability Mixture RS

Contains a mixture of the following 5 compounds:

Plerixafor.

Plerixafor 4-benzyl analog: 1,4-Bis{4-[(1,4,8,11-tetraazacyclotetradecan-1-yl)methyl]benzyl}-1,4,8,11-tetraazacyclotetradecane.

Plerixafor 8-benzyl analog: 1,8-Bis{4-[(1,4,8,11-tetraazacyclotetradecan-1-yl)methyl]benzyl}-1,4,8,11-tetraazacyclotetradecane.

Plerixafor 11-benzyl analog: 1,11-Bis{4-[(1,4,8,11-tetraazacyclotetradecan-1-yl)methyl]benzyl}-1,4,8,11-tetraazacyclotetradecane.

Plerixafor benzyl alcohol: {4-[(1,4,8,11-Tetraazacyclotetradecan-1-yl)methyl]phenyl}methanol.