Argatroban Injection

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

Argatroban Injection is a sterile solution of Argatroban. It contains NLT 90.0% and NMT 110.0% of the labeled amount of argatroban (C₂₃H₃₆N₆O₅S.H₂O).

2 IDENTIFICATION

A. The retention times of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay.

B. The UV spectra of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay.

3 ASSAY

3.1 PROCEDURE

Protect solutions containing argatroban from light.

Buffer: Dissolve 1 g of ammonium acetate in 750 mL of water. Adjust with glacial acetic acid to a pH of 5.5. Dilute with water to 1000 mL.

Solution A: Methanol and Buffer (50:50)

Solution B: Methanol

Mobile phase: See Table 1.

Table 1

Standard solution: 2 mg/mL of USP Argatroban RS prepared as follows. Transfer a quantity of USP Argatroban RS to a suitable volumetric flask, and add 5% of the flask volume of methanol to dissolve. Dilute with Solution A to volume.

Sample solution: Nominally 2 mg/mL of argatroban (monohydrate) prepared as follows. Transfer a volume of the Injection to a suitable volumetric flask, and dilute with Solution A to volume.

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 259 nm. For Identification B, use a diode array detector in the range of 200-400 nm.

Column: 4.6-mm × 25-cm; 5-µm packing L1

Column temperature: 50°

Flow rate: 1.0 mL/min

Injection volume: 10 µL

System suitability

Sample: Standard solution

[NOTE-The relative retention times for argatroban (R-isomer) and argatroban (S-isomer) are 1.00 and 1.06, respectively.]

Suitability requirements

Resolution: NLT 1.3 between argatroban (R-isomer) and argatroban (S-isomer)

Tailing factor: NMT 1.5 for argatroban (R-isomer) and argatroban (S-isomer)

Relative standard deviation: NMT 1.0% for the sum of the peak responses of argatroban (R-isomer) and argatroban (S-isomer) Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of the labeled amount of argatroban (C₂₃H₃₆N₆O₅S.H₂O) in the portion of Injection taken:

Result = (r_U/r_S) × (C_S/C_U) × (M_r1/M_r2) × 100

r_U = sum of the peak responses of argatroban (R-isomer) and argatroban (S-isomer) from the Sample solution

r_S = sum of the peak responses of argatroban (R-isomer) and argatroban (S-isomer) from the Standard solution

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

C_U = nominal concentration of argatroban (monohydrate) in the Sample solution (mg/mL)

M_r1 = molecular weight of argatroban (monohydrate), 526.65

M_r2 = molecular weight of argatroban (anhydrous), 508.63 12

Acceptance criteria: 90.0%-110.0%

4 IMPURITIES

4.1 ORGANIC IMPURITIES

Protect solutions containing argatroban from light.

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

Sensitivity solution: 2.0 µg/mL of USP Argatroban RS from the Standard solution diluted with Solution A

System suitability

Samples: Standard solution and Sensitivity solution

Suitability requirements

Resolution: NLT 1.3 between argatroban (R-isomer) and argatroban (S-isomer), Standard solution

Relative standard deviation: NMT 1.0% for the sum of the peak responses of argatroban (R-isomer) and argatroban (S-isomer), Standard solution

Signal-to-noise ratio: NLT 10 for argatroban (R-isomer) and argatroban (S-isomer), Sensitivity solution

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of dehydroargatroban and any unspecified degradation product in the portion of the Injection taken:

Result = (r_U/r_S) × (C_S/C_U) × (M_r1/M_r2) × (1/F) x 100

r_U = peak response of dehydroargatroban or any unspecified degradation product from the Sample solution

r_S = sum of the peak responses of argatroban (R-isomer) and argatroban (S-isomer) from the Standard solution s

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

C_U = nominal concentration of argatroban (monohydrate) in the Sample solution (mg/mL)

M_r1 = molecular weight of argatroban (monohydrate), 526.65

M_r2 = molecular weight of argatroban (anhydrous), 508.63

F = relative response factor (see Table 2)

Acceptance criteria: See Table 2. The reporting threshold is 0.05%.

Table 2

^a The relative response factor is calculated relative to argatroban monohydrate.

^b (2R,4R)-4-Methyl-1-{[(3-methylquinolin-8-yl) sulfonyl]-L-arginyl)piperidine-2-carboxylic acid.

^c (2R,4R)-4-Methyl-1-{[((R)-3-methyl-1,2,3,4-tetrahydroquinolin-8-yl) sulfonyl]-L-arginyl)piperidine-2-carboxylic acid.

^d (2R,4R)-4-Methyl-1-{[((S)-3-methyl-1,2,3,4-tetrahydroquinolin-8-yl) sulfonyl]-L-arginyl)piperidine-2-carboxylic acid.

^e Excluding dehydroargatroban.

5 SPECIFIC TESTS

PH (791): 6-8.5. [NOTE-This pH test is applicable to formulations which contain Sorbitol.]

PARTICULATE MATTER IN INJECTIONS (788): Meets the requirements

STERILITY TESTS (71): Meets the requirements

BACTERIAL ENDOTOXINS TEST (85): Meets the requirements

OTHER REQUIREMENTS: Meets the requirements in Injections and Implanted Drug Products (1).

6 ADDITIONAL REQUIREMENTS

PACKAGING AND STORAGE: Preserve in Type 1 glass vials. Store at controlled room temperature. Protect from light. Do not freeze.

USP REFERENCE STANDARDS (11).

USP Argatroban RSA (USP 1-Dec-2022)