Cyclosporine

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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C₆₂H₁₁₁N₁₁O₁₂               1202.61

Cyclo[[(E)-(2S,3R,4R)-3-hydroxy-4-methyl-2-(methylamino) oct-6-enoyl]-L-2-aminobutyryl-N-methylglycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl];

[R-[R*R*-(E)]]-Cyclic (L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl-3-hydroxy-N,4-dimethyl-L-2-amino-6-octenoyl-L-a-aminobutyryl-N-methylglycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl) CAS RN: 59865-13-3; UNII: 83HN0GTJ6D.

1 DEFINITION

Cyclosporine contains NLT 97.0% and NMT 101.5% of cyclosporine A (C₆₂H₁₁₁N₁₁O₁₂), calculated on the dried basis. 

2 IDENTIFICATION

Add the following:

A. SPECTROSCOPIC IDENTIFICATION TESTS (197), Infrared Spectroscopy: 197A or 197K (USP 1-Dec-2021)

Change to read:

B._{▲ USP 1-DEC-2021)} 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

Change to read:

3.1 PROCEDURE

Mobile phase: Acetonitrile, tert-butyl methyl ether, water, and phosphoric acid (430:50:520:1)

Diluent: Acetonitrile and water (1:1)

System suitability solution: 1.25 mg/mL of USP Cyclosporine Resolution Mixture RS in Diluent

Standard solution: 1.25 mg/mL of USP Cyclosporine RS in Diluent

Sample solution: 1.25 mg/mL of Cyclosporine in Diluent

3.1.1 Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 210 nm

Column: 4-mm × 25-cm; 3- to 5-μm packing L1. ^▲If needed, tubing may be_{▲ (USP 1-Dec-2021)} connected to the column inlet ^▲to support the Column temperature.^{▲ (USP 1-Dec-2021)}

Column temperature: 80°. ^▲If used,_{▲ (USP 1-Dec-2021)} the tubing ^▲may be_{▲ (USP 1-Dec-2021)} maintained at 80°, to ensure that the Mobile phase entering the column is heated to 80°.

Flow rate: 1.2 mL/min

Injection volume: 20 µL

3.1.2 System suitability

Samples: System suitability solution and Standard solution

[NOTE-The relative retention times for cyclosporine U and cyclosporine are 0.95 and 1.0, respectively.]

3.1.3 Suitability requirements

Resolution: NLT 1.0 between cyclosporine U and cyclosporine, System suitability solution

Relative standard deviation: NMT 1.0%, Standard solution

3.1.4 Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of cyclosporine (C₆₂H₁₁₁N₁₁O₁₂  ) in the portion of Cyclosporine taken:

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

r_U = peak area of cyclosporine from the Sample solution

r_S = peak area of cyclosporine from the Standard solution

C_S = concentration of ^▲USP Cyclosporine RS in _{▲(USP 1-Dec-2021)} the Standard solution (mg/mL)

C_U = concentration of ^▲Cyclosporine in _{▲(USP 1-Dec-2021)} the Sample solution (mg/mL)

P = potency of cyclosporine in USP Cyclosporine RS (mg/mg)

Acceptance criteria: 97.0%-101.5% on the dried basis

4 IMPURITIES

Change to read:

4.1 ORGANIC IMPURITIES

Mobile phase, Diluent, System suitability solution, Sample solution, and Chromatographic system: Proceed as directed in the Assay.

Standard solution: 0.01 mg/mL of USP Cyclosporine RS in Diluent

4.1.1 System suitability

Samples: System suitability solution and Standard solution

[NOTE-The relative retention times for cyclosporine U and cyclosporine are 0.95 and 1.0, respectively.]

4.1.2 Suitability requirements

Resolution: NLT 1.0 between cyclosporine U and cyclosporine, System suitability solution

Relative standard deviation: NMT 10.0%, Standard solution

4.1.3 Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of ^▲any individual _{▲(USP 1-Dec-2021)} impurity in the portion of Cyclosporine taken:

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

r_U = peak area of ^▲each _▲(USP 1-Dec-2021) individual impurity from the Sample solution

r_S = peak area of cyclosporine from the Standard solution r_{s} = 1

C_S = concentration of ^▲USP Cyclosporine RS in _{▲(USP 1-Dec-2021)} the Standard solution (mg/mL)

C_U = concentration of ^▲Cyclosporine in _{▲(USP 1-Dec-2021)} the Sample solution (mg/mL)

P = potency of cyclosporine in USP Cyclosporine RS (mg/mg)

Acceptance criteria: The reporting threshold is 0.05%.

Any individual impurity: NMT 0.7%

Total impurities: NMT 1.5%

5 SPECIFIC TESTS

Change to read:

LOSS ON DRYING (731)

(USP 1-Dec-2021)

Analysis: Dry in a capillary-stoppered bottle under vacuum at a pressure not exceeding 5 mm of mercury at 60° for 3 h.

Acceptance criteria: NMT 2.0%

6 ADDITIONAL REQUIREMENTS

PACKAGING AND STORAGE: Preserve in tight, light-resistant containers.

Change to read:

USP REFERENCE STANDARDS (11)

USP Cyclosporine RS

USP Cyclosporine Resolution Mixture RS

Contains a mixture of the following two compounds [in a 100:1 (w/w) mixture of cyclosporine and cyclosporine U]:

Cyclosporine.

Cyclosporine U: Cyclo[[(E)-(2S,3R,4R)-3-hydroxy-4-methyl-2-(methylamino) oct-6-enoyl]-1-2-aminobutyryl-N-methylglycyl-N-methyl-L-leucyl-L-valyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl]._{▲ (USP 1-Dec-2021)}

C₆₂H₁₁₁N₁₁O₁₂          ^▲1188.61 _{▲(USP 1-Dec-2021)}