Argatroban

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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(ERR 1-Mar-2019)

C₂₃H₃₆N₆O₅S.H₂O 526.65

2-Piperidinecarboxylic acid, 1-[(S)-5-[(aminoiminomethyl)amino]-1-oxo-2-{[(1,2,3,4-tetrahydro-3-methyl-8-quinolinyl) sulfonyl)amino}pentyl]-4-methyl-, (2R,4R)-monohydrate;

(2R,4R)-4-Methyl-1-(N2-[(1,2,3,4-tetrahydro-3-methyl-8-quinolyl) sulfonyl]-L-arginyl)pipecolic acid, monohydrate CAS RN^®: 141396-28-3; UNII: IY90U61Z3S.

1 DEFINITION

Argatroban contains NLT 98.0% and NMT 102.0% of argatroban (C23H36NOS), calculated on the anhydrous and solvent-free basis.

2 IDENTIFICATION

Change to read:

A. ASPECTROSCOPIC IDENTIFICATION TESTS (197), Infrared Spectroscopy: 197K (CN 1-MAY-2020)

B. The retention times 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

[NOTE-It is recommended to keep all solutions containing argatroban at about 4°.]

Solution A: 10 mM ammonium acetate and 5 mM sodium 1-heptanesulfonate

Solution B: Acetonitrile and methanol (500:300)

Mobile phase: See Table 1.

Table 1

Standard solution: 4 mg/mL of USP Argatroban RS in methanol

Sample solution: 4 mg/mL of Argatroban in methanol

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 259 nm

Column: 4.6-mm × 25-cm; 3-µm packing L1

Temperatures

Column: 50°

Autosampler: 4°

Flow rate: 0.6 mL/min

Injection volume: 10 µL

System suitability

Sample: Standard solution

Suitability requirements

Tailing factor: NMT 1.5 for both peaks

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

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of argatroban (C₂₃H₃₆N₆O₅S) in the portion of Argatroban taken:

Result = (r_U/r_S) × (C_S/C_U) × 100

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

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

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

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

Acceptance criteria: 98.0%-102.0% on the anhydrous and solvent-free basis

4 IMPURITIES

4.1 RESIDUE ON IGNITION (281)

NMT 0.1%

4.2 ORGANIC IMPURITIES

[NOTE-It is recommended to keep all solutions containing argatroban at about 4°.]

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

Sensitivity solution: 2 µg/mL of USP Argatroban RS in methanol

Standard solution: 4 µg/mL each of USP Argatroban RS, USP Argatroban Related Compound A RS, and USP Argatroban Related Compound B

RS in methanol

System suitability

Samples: Sensitivity solution and Standard solution

Suitability requirements

Resolution: NLT 1.2 between (R)-argatroban and (S)-argatroban, Standard solution

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

Relative standard deviation: NMT 5% for all peaks. For argatroban, use the sum of the peak responses of (R)-argatroban and (S)-argatroban, Standard solution.

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of argatroban related compound A and argatroban related compound B in the portion of Argatroban taken:

Result = (r_U/r_S) × (C_S/C_U) × 100

r_U = peak response of argatroban related compound A or argatroban related compound B from the Sample solution

r_S = peak response of argatroban related compound A or argatroban related compound B from the Standard solution

C_S = concentration of USP Argatroban Related Compound A RS or USP Argatroban Related Compound B RS in the Standard solution (mg/mL)

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

Calculate the percentage of any unspecified impurity in the portion of Argatroban taken:

Result = (r_U/r_S) × (C_S/C_U) × 100

r_U = peak response of any unspecified impurity from the Sample solution

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

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

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

Acceptance criteria: See Table 2. Disregard any peak below 0.05%.

Table 2

^a (2R,4R)-1-[N8-Nitro-N2-(3-methylquinoline-8-sulfonyl)-L-arginyl]-4-methylpiperidine-2-carboxylic acid.

^b Ethyl (4R)-1-[N-nitro-L-arginyl]-4-methylpiperidine-2-carboxylate.

^c (2R,4R)-4-Methyl-1-(N2-[((R)-1,2,3,4-tetrahydro-3-methyl-8-quinolyl) sulfonyl]-L-arginyl)pipecolic acid.

^d (2R,4R)-4-Methyl-1-{N²-[((S)-1,2,3,4-tetrahydro-3-methyl-8-quinolyl) sulfonyl]-L-arginyl)pipecolic acid.

^e Total impurities include specified and unspecified impurities and argatroban related compound C from the test for Content of Argatroban Related Compound C.

4.3 CONTENT OF ARGATROBAN RELATED COMPOUND C

[NOTE-It is recommended to keep all solutions containing argatroban at about 4°.]

Buffer: 10 mM ammonium acetate and 5 mM sodium 1-heptanesulfonate

Solution A: Acetonitrile, dehydrated alcohol, and Buffer (80:240:680)

Mobile phase: See Table 3.

Table 3

Sensitivity solution: 4 µg/mL of USP Argatroban Related Compound C RS in methanol

System suitability solution: 10 mg/mL of USP Argatroban RS and 0.1 mg/mL of USP Argatroban Related Compound C RS in methanol

Sample solution: 10 mg/mL of Argatroban in methanol

Chromatographic system: Proceed as directed in the Assay.

System suitability

Samples: Sensitivity solution and System suitability solution

Suitability requirements

Resolution: NLT 1.4 between argatroban related compound C and (R)-argatroban, System suitability solution

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

Relative standard deviation: NMT 2.5% for argatroban related compound C, System suitability solution

Analysis

Sample: Sample solution

Calculate the percentage of argatroban related compound C in the portion of Argatroban taken:

Result = (r_U/r_T) × 100

r_U = peak response of argatroban related compound C from the Sample solution

r_T = total of all peak responses from the Sample solution

Acceptance criteria: See Table 4.

Table 4

^a (2R,4R)-1-[N⁸-Amino-N²-(3-methyl-1,2,3,4-tetrahydroquinoline-8-sulfonyl)-L-arginyl]-4-methylpiperidine-2-carboxylic acid.

4.4 CONTENT OF STEREOISOMERS

[NOTE-It is recommended to keep all solutions containing argatroban at about 4°.]

Mobile phase: Methanol and water (520:480)

Standard solution: 0.16 mg/mL of USP Argatroban RS in methanol

Sample solution: 0.16 mg/mL of Argatroban in methanol

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 259 nm

Column: 4.6-mm × 25-cm; 3-µm packing L1

Column temperature: 50°

Flow rate: 0.6 mL/min

Injection volume: 10 µL

Run time: NLT 1.4 times the retention time of (R)-argatroban

System suitability

Sample: Standard solution

Suitability requirements

Resolution: NLT 1.2 between (R)-argatroban and (S)-argatroban

Relative standard deviation: NMT 2.0% for (R)-argatroban and (S)-argatroban

Analysis

Sample: Sample solution

Calculate the percentage of (R)-argatroban and (S)-argatroban in the portion of Argatroban taken:

Result = [(r_U or r_S)/(r_U + r_S)] × 100

r_U = peak response of (R)-argatroban from the Sample solution

r_S = peak response of (S)-argatroban from the Sample solution

Acceptance criteria: See Table 5.

Table 5

^a (2R,4R)-4-Methyl-1-{N²-[((R)-1,2,3,4-tetrahydro-3-methyl-8-quin olyl) sulfonyl]-L-arginyl)pipecolic acid.

^b (2R,4R)-4-Methyl-1-(N²-[((S)-1,2,3,4-tetrahydro-3-methyl-8-quin olyl) sulfonyl]-L-arginyl)pipecolic acid.

5 SPECIFIC TESTS

WATER DETERMINATION (921), Method la: 3.0%-6.0%

BACTERIAL ENDOTOXINS TEST (85): NMT 2.0 USP Endotoxin Units/mg of argatroban

MICROBIAL ENUMERATION TESTS (61) and TESTS FOR SPECIFIED MICROORGANISMS (62): The total aerobic microbial count is NMT 102 cfu/g, and the total

combined molds and yeasts count is NMT 102 cfu/g.

6 ADDITIONAL REQUIREMENTS

6.1 PACKAGING AND STORAGE

Preserve in tight, light-resistant containers, and store at controlled room temperature.

6.2 USP REFERENCE STANDARDS (11)

USP Argatroban RS

USP Argatroban Related Compound A RS

(2R,4R)-1-[N⁸-Nitro-N2-(3-methylquinoline-8-sulfonyl)-L-arginyl]-4-methylpiperidine-2-carboxylic acid.

C₂₃H₃₁N₇O₇S  549.60

USP Argatroban Related Compound B RS

Ethyl (4R)-1-[N⁸-nitro-L-arginyl]-4-methylpiperidine-2-carboxylate dihydrochloride.

C₁₅H₂₈N₆O₅S.2HCI  445.34

USP Argatroban Related Compound C RS

(2R,4R)-1-[N⁸-Amino-N²-(3-methyl-1,2,3,4-tetrahydroquinoline-8-sulfonyl)-L-arginyl]-4-methylpiperidine-2-carboxylic acid.

C₂₃H₃₇N₇O₅S  523.65