Printed on: Sat Jul 10 2021, 07:06:58 PM Official Status: Currently Official on 11-Jul-2021 DocId: 1_GUID-41785247-DA22-407F-B060-7182FDA4FD3A_3_en-US

(EST)
Printed by: Nguyen Nhan Official Date: Official as of 01-May-2018 Document Type: USP @2021 USPC
1

[NOTE—The relative retention times for doxorubicin

Doxorubicin Hydrochloride for and epirubicin are 1.0 and 1.05, respectively.]
Injection Suitability requirements
Resolution: NLT 1.5 between doxorubicin and
DEFINITION epirubicin, System suitability solution
Doxorubicin Hydrochloride for Injection is a sterile mixture of Tailing factor: 0.8–1.5, Standard solution
Doxorubicin Hydrochloride and Lactose. It contains NLT Relative standard deviation: NMT 0.73%, Standard
90.0% and NMT 115.0% of the labeled amount of solution
doxorubicin hydrochloride (C27H29NO11 · HCl). Analysis
[CAUTION—Great care should be taken to prevent inhaling Samples: Standard solution and Sample solution
particles of Doxorubicin Hydrochloride and exposing Calculate the percentage of the labeled amount of
the skin to it.] doxorubicin hydrochloride (C27H29NO11 · HCl) in the
portion of Doxorubicin Hydrochloride for Injection taken:
IDENTIFICATION
• A. The retention time of the doxorubicin peak of the Result = (r U/r S) × (C S/C U) × P × F × 100
Sample solution corresponds to that of the Standard
solution, as obtained in the Assay. rU = peak response of doxorubicin from the Sample
• B. The UV spectrum of the doxorubicin peak of the Sample solution
solution corresponds to that of the Standard solution, as rS = peak response of doxorubicin from the Standard
obtained in the Assay. solution
ASSAY CS = concentration of USP Doxorubicin
• PROCEDURE Hydrochloride RS in the Standard solution
(mg/mL)

al
Solution A: 0.1% Trifluoroacetic acid prepared by diluting
1.0 mL of trifluoroacetic acid with water to 1 L CU = nominal concentration of doxorubicin
Solution B: Acetonitrile, methanol, and trifluoroacetic acid hydrochloride in the Sample solution (mg/mL)
(800:200:1) P = potency of doxorubicin in USP Doxorubicin
Mobile phase: See Table 1. Hydrochloride RS (µg/mg)
F = conversion factor, 0.001 mg/µg

Time
(min)
Table 1
Solution A
(%)
ci
Solution B
(%)
Acceptance criteria: 90.0%–115.0%
IMPURITIES
0 90 10 • ORGANIC IMPURITIES
Mobile phase, Diluent, System suitability solution, and
ffi
15 25 75 Chromatographic system: Proceed as directed in the
16 25 75 Assay.
[NOTE—Protect solutions containing doxorubicin from
16.1 90 10 light.]
18 90 10 Standard solution: 0.002 mg/mL each of USP Doxorubicin
Hydrochloride RS, USP Doxorubicinone RS, and USP
O

Daunorubicinone RS in Diluent
Diluent: Solution A and Solution B (50:50)
Sample solution: Nominally 0.4 mg/mL of doxorubicin
[NOTE—Protect solutions containing doxorubicin from
hydrochloride in Diluent from Doxorubicin Hydrochloride
light.]
for Injection prepared as follows. Add 5 mL of Diluent into
System suitability solution: 0.1 mg/mL each of USP
the container of Doxorubicin Hydrochloride for Injection,
Doxorubicin Hydrochloride RS and USP Epirubicin
and transfer the contents to a volumetric flask of
Hydrochloride RS in Diluent
appropriate size. Rinse the container with additional Diluent
Standard solution: 0.1 mg/mL of USP Doxorubicin
NLT 3 times. Dilute with Diluent to volume, and mix.
Hydrochloride RS in Diluent
System suitability
Sample solution: Nominally 0.1 mg/mL of doxorubicin
Samples: System suitability solution and Standard solution
hydrochloride in Diluent from Doxorubicin Hydrochloride
[NOTE—See Table 2 for the relative retention times.]
for Injection prepared as follows. Add 5 mL of Diluent into
Suitability requirements
the container of Doxorubicin Hydrochloride for Injection,
Resolution: NLT 1.5 between doxorubicin and
and transfer the contents to a volumetric flask of
epirubicin, System suitability solution
appropriate size. Rinse the container with additional Diluent
Relative standard deviation: NMT 5.0%, Standard
NLT 3 times. Dilute with Diluent to volume, and mix.
solution
Chromatographic system
Analysis
(See Chromatography á621ñ, System Suitability.)
Samples: Standard solution and Sample solution
Mode: LC
Calculate the percentage of doxorubicinone in the portion
Detector: UV 254 nm. For Identification test B, use a
of Doxorubicin Hydrochloride for Injection taken:
photo-diode array detector in the range of 190–400 nm.
Column: 2.1-mm × 10-cm; 1.7-µm packing L1 Result = (r U/r S) × (C S/C U) × P × 100
Temperatures
Autosampler: 4° rU = peak response of doxorubicinone from the
Column: 35° Sample solution
Flow rate: 0.5 mL/min rS = peak response of doxorubicinone from the
Injection volume: 2 µL Standard solution
System suitability CS = concentration of USP Doxorubicinone RS in the
Samples: System suitability solution and Standard solution Standard solution (mg/mL)

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Printed on: Sat Jul 10 2021, 07:06:58 PM Official Status: Currently Official on 11-Jul-2021 DocId: 1_GUID-41785247-DA22-407F-B060-7182FDA4FD3A_3_en-US
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Printed by: Nguyen Nhan Official Date: Official as of 01-May-2018 Document Type: USP @2021 USPC
2

CU = nominal concentration of doxorubicin
hydrochloride in the Sample solution (mg/mL)
P = potency of doxorubicinone in USP
Doxorubicinone RS (mg/mg)
Calculate the percentage of daunorubicinone in the portion
of Doxorubicin Hydrochloride for Injection taken:
Result = (r U/r S) × (C S/C U) × P × 100
rU = peak response of daunorubicinone from the
Sample solution
rS = peak response of daunorubicinone from the
Standard solution
CS = concentration of USP Daunorubicinone RS in the
Standard solution (mg/mL)
CU = nominal concentration of doxorubicin
hydrochloride in the Sample solution (mg/mL)
P = potency of daunorubicinone in USP
Daunorubicinone RS (mg/mg)
Calculate the percentage of any individual unspecified
degradation product in the portion of Doxorubicin
a For resolution measurement only. Not to be reported; not to be included in
total impurities.
b (8S,10S)-6,8,10,11-Tetrahydroxy-8-(hydroxyacetyl)-1-methoxy-7,8,9,10-
tetrahydrotetracene-5,12-dione.
c (8S,10S)-8-Acetyl-6,8,10,11-tetrahydroxy-1-methoxy-7,8,9,10-
tetrahydrotetracene-5,12-dione.
SPECIFIC TESTS
• CONSTITUTED SOLUTION: At the time of use, it meets the
requirements in Injections and Implanted Drug Products á1ñ,
Specific Tests, Completeness and clarity of solutions.
• STERILITY TESTS á71ñ: It meets the requirements when
tested as directed in Test for Sterility of the Product to Be
Examined, Membrane Filtration, the entire contents of all the
containers being collected aseptically with the aid of
200 mL of Fluid A before filtering.
• WATER DETERMINATION, Method I á921ñ
Sample solution: Prepare as directed for a hygroscopic
specimen.
Acceptance criteria: NMT 4.0%
• PH á791ñ
Sample solution: Constitute as directed in the labeling
using water as the diluent.

al
Hydrochloride for Injection taken: Acceptance criteria: 4.5–6.5
• BACTERIAL ENDOTOXINS TEST á85ñ
Result = (r U/r S) × (C S/C U) × P × F × 100 Sample solution: 1.1 mg/mL of doxorubicin hydrochloride
from Doxorubicin Hydrochloride for Injection
rU = peak response of each unspecified degradation Acceptance criteria: NMT 2.2 USP Endotoxin Units/mg of
doxorubicin hydrochloride
rS

CS
product from the Sample solution
= peak response of doxorubicin from the Standard
solution
= concentration of USP Doxorubicin
ci • UNIFORMITY OF DOSAGE UNITS á905ñ: Meets the
requirements
• LABELING á7ñ, LABELS AND LABELING FOR INJECTABLE
Hydrochloride RS in the Standard solution PRODUCTS: Meets the requirements.
(mg/mL)
ffi
ADDITIONAL REQUIREMENTS
CU = nominal concentration of doxorubicin • PACKAGING AND STORAGE: Preserve as described in
hydrochloride in the Sample solution (mg/mL) Packaging and Storage Requirements á659ñ, Injection
P = potency of doxorubicin in USP Doxorubicin Packaging, Packaging for constitution, except that
Hydrochloride RS (µg/mg) multiple-dose containers may provide for the withdrawal of
F = conversion factor, 0.001 mg/µg NMT 100 mL when constituted as directed in the labeling.
O

Store unreconstituted vials at controlled room temperature.

Acceptance criteria: See Table 2. Protect from light.
• USP REFERENCE STANDARDS á11ñ
Table 2 USP Daunorubicinone RS
Relative Acceptance (8S,10S)-8-Acetyl-6,8,10,11-tetrahydroxy-1-methoxy-
Retention Criteria, 7,8,9,10-tetrahydrotetracene-5,12-dione.
Name Time NMT (%)
C21H18O8 398.36
Doxorubicin 1.0 — USP Doxorubicin Hydrochloride RS
Epirubicin a
1.05 — USP Doxorubicinone RS
(8S,10S)-6,8,10,11-Tetrahydroxy-8-(hydroxyacetyl)-1-
Doxorubicinoneb 1.08 0.5 methoxy-7,8,9,10-tetrahydrotetracene-5,12-dione.
Daunorubicinonec 1.35 0.5 C21H18O9 414.36
USP Epirubicin Hydrochloride RS
Any individual
unspecified
—
degradation
product 0.5

Total impurities — 2.0

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