Doxorubicin Hydrochloride

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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5,12-Naphthacenedione, 10-[(3-amino-2,3,6-trideoxy-a--lyxo-hexopyranosyl)oxy]-7,8,9,10-tetrahydro-6,8,11-trihydroxy-8-(hydroxylacetyl) methoxy-, hydrochloride (8S-cis)-;

(8S,10S)-10-[(3-Amino-2,3,6-trideoxy-a--lyxo-hexopyranosyl)oxy]-8-glycoloyl-7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12-

naphthacenedione hydrochloride CAS RN^®: 25316-40-9; UNII: 82F2G7BL4E.

1 DEFINITION

Doxorubicin Hydrochloride contains NLT 98.0% and NMT 102.0% of doxorubicin hydrochloride (C₂₇H₂₉NO₁₁· HCl), calculated on the anhydrous, solvent-free basis.

[CAUTION-Great care should be taken to prevent inhaling particles of doxorubicin hydrochloride and exposing the skin to it.]

2 IDENTIFICATION

A. The retention time of the doxorubicin peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.

B. SPECTROSCOPIC IDENTIFICATION TESTS (197), Infrared Spectroscopy: 197K

C. IDENTIFICATION TESTS GENERAL, Chloride (191)

3 ASSAY

Change to read:

PROCEDURE

Solution A: 0.1% Trifluoroacetic acid prepared by transferring 1.0 mL of trifluoroacetic acid to 1 L of water

Solution B: Acetonitrile, methanol, and trifluoroacetic acid (800:200:1)

Mobile phase: See Table 1.

Diluent: Solution A and Solution B (50:50)

[NOTE-Protect solutions containing doxorubicin from light.]

System suitability solution: 0.1 mg/ml, each of USP Doxorubicin Hydrochloride RS and USP Epirubicin Hydrochloride RSA (ERR 1-May-2024) In

Diluent

Standard solution: 0.1 mg/ml, of SP Domubicin Hydrochloride RS in Dient

Sample solution: 0.1 mg/ml, of Doxorubicin Hydrachlonde in Cluent

Chromatographic system

(See Chromatopanky (1211. Smm Sumalday)

Mode: LC

Detector: UV 254 m

Column: 2.1mm x 10cm, 1.7-um packing Li

Temperatures

Column: 35

Autosampler: 4

Flow rate: 0.5 ml/min

Injection volume: 2 με

System suitability

Samples: System sutatility solution and Standard solution

[Nom-The relative retention times for doxorubicin and epirubicin are 1.0 and 1.05, respectively]

Suitability requirements

Resolution: NLT 1.5 between doxorubicin and eprubicin, System surhabdity solution

Tailing factor: 0.8-1.5. Standard solution

Relative standard deviation: NMT 73%, Standard solution

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of doxorubicin hydrochloride (CHNO, HCI) in the portion of Doxorubicin Hydrochloride taken:

Result = (r_u/r_s) × (C_s/C_u) × P × F × 100

r_u = peak response of doxorubicin from the Sample solution

rs = peak response of doxorubicin from the Standard solution

C_s = concentration of USP Dosunibicin Hydrochloride RS in the Standard solution (mg/mL)

C_u = concentration of Doxorubicin Hydrochloride in the Sample solution (mg/ml)

P = potency of doxorubicin hydrochloride in USP Doxorubicin Hydrochlonde RS (µg/mg)

F = conversion factor, 0.001 mg/ول

Acceptance criteria: 98.0%-102.0% on the anhydrous, sofvent-free basis

4 IMPURITIES

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

Nore-Protect solutions containing doxorubicin from light.

Standard solution: 0.002 mg/ml, each of bicin Hydrochloride RS USP Respubicinome RS, JUSP Daunorubicon Hydrochinnde RS and USP Daunorubicinone RS in

Sample solution: 0.4 mg/mL of Doxorubicin Hydrochloride in Diluent

System suitability

Samples: System suitability solution and Standard solution

[Non-See Table 2 for the relative retention times.]

Suitability requirements

Resolution: NLT 1.5 between doxorubicin and spirubicin, System suhability solution

Relative standard deviation: NMT 5.0 Standard solution

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of doxorubicinone in the portion of Doxorubicin Hydrochloride taken

Result = (r_u/r_s) × (C_s/C_u) × P × F × 100

r_u = peak response of doxorubicinone from the Sample solution

rs = peak response of doxorubicinome from the Standard solution

C_s = Cconcentration of USP Doxorubicinone RS in the Standard solution (mg/m)

C_u = concentration of Doxorubicin Hydrochloride in the Sample solution (mg/ml)

P = potency of doxorubicinone in the USP Doxorubicinone RS (mg/mg)

Calculate the permentage of daunorubicinione in the portion of Doxorubicin Hydrochloride taken

Result = (r_u/r_s) × (C_s/C_u) × P × F × 100

r_u = peak response of daunorubicinone from the Sample solution

rs​​​​​​​ = peak response of dounorubicinone from the Standard solution

C_s = concentration of USP Daunorubicinone RS in the Standard solution (mg/ml)

C_u = concentration of Doxorubicin Hydrochloride in the Sample solution (mg/mL)

P = potency of daunorubicinone in USP Daunorubicinone RS (mg/mg)

Calculate the percentage of daunonibicin in the portion of Doxorubicin Hydrochloride taken

Result = (r_u/r_s) × (C_s/C_u) × P × F × 100

r_u = peak response of daunorubicin from the Sample solution

rs​​​​​​​ = peak response of daunarubicin from the Standard solution

C_s = concentration of USP Daunenidicin Hydrochloride RS in the Standard solution (mg/ml)

C_u = concentration of Doxorubicin Hydrochloride in the Sample solution (mg/ml)

P = potency of dautorubicin in USP Daunorubicin Hydrochloride RS (pg/mg)

F = convension factor, 0.001 mg/ug

Calculate the percentage of any individual unspecified impurity in the portion of Doxorubicin Hydrochloride taken

Result = (r_u/r_s) × (C_s/C_u) × P × F × 100

r_u = peak response of any individual unspecified impurity from the Sample solution

rs​​​​​​​ = peak response of doxorubicin from the Standard solution

C_s = concentration of USP Doxorubicin Hydrochloride Rain the Standard solution (mg/mL)

C_u = concentration of Doxorubicin Hydrochloride in the Sample solution (mg/mL)

P = potency of doxorubicin hydrochloride in USP Duxordion Hutochloride RS5 (g/mg)

F = conversion factor 0.001 mg/g

Acceptance criteria: See Table 2

^aFor resolution measurement only. Not to be reported not to be included in total impurities

^b(88,105)-6,8,10,11-Tetrahydoxy-8-hydroxyacetyl-1-methoxy-7,8,9,10-tetrahydrotetracene-5,12-dione

^c(82.105)-8-Acetyl-6,8,10,11-tetrahydroxy-1-methoxy-7,8,9,10-tetrahydrotetracene-5, 12-dione

LANTIFACTURE AND ALCOHOL

Internal standard solution: 1 mg/ml of dioxane in water

Standard solution: 0.2 mg/ml. of 15P Acetone RS 0.3 mg/ml, of dehydrated alcohol in internal standard solution

Sample solution: 200 mg of Doxorubicin Hydrochloride in 30 ml (3.0 g) of internal standard solution

Chromatographic system

(See Chromatoglashy/621) Sutem Suitability)

Mode: GC

Column: 4mm 2m packed with 8-10% liquid phase 16 and 25% potassium hydroxide on 100-to 120 mesh support STA

Detector: Flame ionization

Column temperature: 60

Carrier gas: Helum

Nore-Adjust the column temperature and comer gas flow rate so that dioxane elutes in about 6 min

Injection volume: 1 L

Flow rabe: Adjust the column temperature and carrier gas flow rate so that dioxane elutes in about 6 min.

System suitability

Sample: Standard solution

Nore-The relative retention tames for acetone, alcohol, and dioxane are about 02,05, and 10, respectively

Suitability requirements

Resolution: NLT 2.0 between adjacent peaks

Tailing factor: NMT 1.5 for the alcohol pesk

Relative standard deviation: NMT 4.0% for the peak response ratios of acetone and alcohol to the internal standard

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage, by weight, of acetone and alcohol respectively, in the portion of Doxorubicar Hydrochloride taken

Result = (R_U/R_S) × (C_A/C_D) × (D_U/W_U) × 100

R_U = peak response ratio of the analyte (acetone or alcohol) to dioxane from the Sample solution

R_S​​​​​​​ = peak response ratio of the analyte (acartone or alcohol) to dioxane from the Standard solution

C_A = concentration of the analyte (acetone or alcohol in the Standard solution (mg/ml)

C_D = concentration of dioxane in the Standard solution (mg/mL)

D_U = Weight of dioxane in the Sample solution (mg)

W_U = Weight of Doxorubicin Hydrochloride in the Sample solution (mg)

Acceptance criteria

Acetone: NMT 0.5%

Total of soetone and alcohol: NMT 2.5%

5 SPECIFIC TESTS

COSTALNTY (095) Meets the requirements, except where it is labeled as amorphous, most particles do not exhibit birefringence and

extinction positions

Sample solution: 5 mg/mL

Acceptance criteria: 4.0-5.5

Ware Datos Method 11221 NMT 4.0%

6 ADDITIONAL REQUIREMENTS

PACKAGING AND STowce Preserve in tight containers, and store at controlled room temperature except where it is labeled as amorphous, in which case it should be stored in the freezer

LABLELING: The amorphous form is so labeled.