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3838 Etoposide / Official Monographs USP 39

Table 2 Standard stock solution: 2.0 mg/mL of USP Etoposide

Relative Acceptance
RS in acetonitrile
Retention Criteria,
Standard solution: 0.2 mg/mL of USP Etoposide RS in
Name Time NMT (%)
Mobile phase from the Standard stock solution
Sample solution: Equivalent to 0.2 mg/mL of etoposide
Etoposide 1.0 — in Mobile phase from the Injection
Picroetoposide 1.43 2.0 Chromatographic system
Any unspecified impurity — — (See Chromatography 〈621〉, System Suitability.)
Total impurities — 3.0 Mode: LC
Detector: UV 254 nm
Column: 3.9-mm × 30-cm; packing L11
ADDITIONAL REQUIREMENTS Flow rate: 1 mL/min
• PACKAGING AND STORAGE: Preserve in tight containers in a Injection volume: 20 µL
cold place. Do not freeze. Run time: NLT 1.5 times the retention time of
• USP REFERENCE STANDARDS 〈11〉 etoposide
USP Etoposide RS System suitability
USP Etoposide Resolution Mixture RS Samples: System suitability solution and Standard
solution
Suitability requirements
Resolution: NLT 1.35 between the etoposide and α-
.

etoposide peaks, System suitability solution

Etoposide Injection Relative standard deviation: NMT 2.0% for etopo-
side, Standard solution
DEFINITION Analysis
Etoposide Injection contains NLT 90.0% and NMT 110.0% Samples: Standard solution and Sample solution
of the labeled amount of etoposide (C29H32O13) in a sterile Calculate the percentage of etoposide (C29H32O13) in
solution in a nonaqueous medium intended for dilution the portion of Injection taken:
with a suitable parenteral vehicle before intravenous
infusion. Result = (rU/rS) × (CS/CU) × 100
IDENTIFICATION rU = peak response of etoposide from the Sample
• A. solution
Diluent: Chloroform and methanol (9:1) rS = peak response of etoposide from the Standard
Standard solution: 0.8 mg/mL of USP Etoposide RS in solution
Diluent CS = concentration of USP Etoposide RS in the
Sample solution: Equivalent to 0.8 mg/mL of etoposide Standard solution (mg/mL)
in Diluent from the Injection CU = nominal concentration of etoposide in the
Chromatographic system Sample solution (mg/mL)
USP Monographs

(See Chromatography 〈621〉, Thin-Layer Chromato- Acceptance criteria: 90.0%–110.0%

graphy.)
Mode: TLC IMPURITIES
Adsorbent: 0.25-mm layer of chromatographic silica • ORGANIC IMPURITIES
gel mixture Buffer: Prepare as directed in the Assay.
Application volume: 10 µL Solution A: Acetonitrile and Buffer (20:80)
Developing solvent system: Chloroform, acetone, al- Solution B: Acetonitrile and Buffer (60:40)
cohol, and water (80: 25: 2.5: 0.5) Mobile phase: See Table 1.
Spray reagent: Add 10 mL of sulfuric acid with cool-
ing and stirring to 70 mL of dehydrated alcohol in a Table 1
100-mL volumetric flask. Dilute with dehydrated alco-
Time Solution A Solution B
hol to volume, and mix.
(min) (%) (%)
Analysis:
Samples: Standard solution and Sample solution 0 100 0
Allow the chromatogram to develop until the solvent 15 100 0
front has moved 17 cm from the origin. Remove the 30 40 60
plate, and allow it to air-dry in a fume hood for 5 40 40 60
min. Replace the plate in the tank, and develop again 42 0 100
to a distance of 17 cm from the origin. Remove the
45 0 100
plate, and air-dry it in a fume hood for about 20 min.
Spray the plate with the Spray reagent, and heat in a 47 100 0
forced-air oven at 120° for about 15 min. 50 100 0
Acceptance criteria: The appearance and RF value of
the principal spot from the Sample solution corresponds Diluent: Acetonitrile and 0.02 M sodium acetate previ-
to that from the Standard solution. ously adjusted with acetic acid to a pH of 4.0 (30:70)
• B. The retention time of the major peak of the Sample Standard stock solution: 2.0 mg/mL of USP Etoposide
solution corresponds to that of the Standard solution, as RS in Diluent
obtained in the Assay. System suitability stock solution: 0.2 mg/mL of n-pro-
pylparaben in Diluent
ASSAY System suitability solution: Transfer 5.0 mL of the Sys-
• PROCEDURE tem suitability stock solution and 5.0 mL of the Standard
Buffer: 2.72 g/L of sodium acetate in water. Adjust with stock solution to a 50-mL volumetric flask, and dilute
glacial acetic acid to a pH of 4.0. with Diluent to volume. Transfer 5.0 mL of this solution
Mobile phase: Acetonitrile and Buffer (26:74) to a 100-mL volumetric flask, and dilute with Diluent to
System suitability solution: 0.3 mg/mL of USP Etopo- volume.
side Resolution Mixture RS in Mobile phase Standard solution: 10 µg/mL of USP Etoposide RS from
the Standard stock solution in Diluent

Official from May 1, 2016

Copyright (c) 2016 The United States Pharmacopeial Convention. All rights reserved.
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USP 39 Official Monographs / Eucalyptol 3839

Sample solution: Nominally equivalent to 2.0 mg/mL CU = concentration of the Sample solution (mg/mL)
of etoposide in Diluent Acceptance criteria: 90.0%–110.0%
Chromatographic system
(See Chromatography 〈621〉, System Suitability.)
Mode: LC Change to read:
Detector: UV 254 nm
Column: 3.9-mm × 15-cm; less than 5-µm packing • OTHER REQUIREMENTS: It meets the requirements in •In- .

L11 jections and Implanted Drug Products 〈1〉• (CN 1-May-2016).

Flow rate: 1.5 mL/min ADDITIONAL REQUIREMENTS
Injection volume: 25 µL • PACKAGING AND STORAGE: Preserve in single-dose or mul-
Run time: NLT 40 min tiple-dose containers of Type I glass.
System suitability • LABELING: Label it to indicate that it must be diluted with
[NOTE—Run time is 15 min in isocratic conditions.] suitable parenteral vehicle before intravenous infusion.
Sample: System suitability solution • USP REFERENCE STANDARDS 〈11〉
Suitability requirements USP Endotoxin RS
Resolution: NLT 1.1 between propylparaben and USP Etoposide RS
etoposide USP Etoposide Resolution Mixture RS
Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of each impurity in the por-
tion of Injection taken: .

Result = (rU/rS) × (CS/CU) × 100 Eucalyptol

rU = peak response of each impurity from the
Sample solution
rS = peak response of etoposide from the Standard
solution
CS = concentration of USP Etoposide RS in the
Standard solution (mg/mL) C10H18O 154.25
CU = nominal concentration of etoposide in the 1,3,3-Trimethyl-2-oxabicyclo[2.2.2]octane;
Sample solution (mg/mL) 1,8-Epoxy-p-menthane [470-82-6].
Acceptance criteria DEFINITION
Total impurities: NMT 3.0% Eucalyptol is obtained from oil of eucalyptus and from other
SPECIFIC TESTS sources. It contains NLT 98.0% and NMT 100.0% of
• PH 〈791〉 C10H18O.
Sample solution: 5.0 mL of Injection in 45 mL of water IDENTIFICATION

USP Monographs
Acceptance criteria: 3.0–4.0 • A. INFRARED ABSORPTION 〈197F〉
• ALCOHOL DETERMINATION, Method II 〈611〉 (if present): • B. Add 1 mL of phosphoric acid to 1 mL of Eucalyptol
90.0%–110.0% of the labeled amount of alcohol contained in a test tube maintained in an ice bath. A
(C2H5OH), using n-propyl alcohol as the internal standard solid white crystalline mass is formed, from which euca-
• BACTERIAL ENDOTOXINS TEST 〈85〉 lyptol separates upon addition of warm water.
Sample solution: Dilute the Injection with sterile water
to obtain 0.31 mg/mL of etoposide activity ASSAY
Acceptance criteria: NMT 2.0 USP Endotoxin Units/mg • PROCEDURE
of etoposide System suitability solution: 0.2 mg/mL of limonene
• BENZYL ALCOHOL CONTENT (if present) and 0.9 mg/mL of USP Eucalyptol RS in methanol
Buffer, Mobile phase, System suitability solution, Sample solution: 0.9 mg/mL of Eucalyptol in methanol
Sample solution, Chromatographic system, and Sys- Blank: Methanol
tem suitability: Proceed as directed in the Assay. Chromatographic system
Standard solution: Transfer 0.75 mL of freshly distilled (See Chromatography 〈621〉, System Suitability.)
benzyl alcohol, accurately weighed, to a 50-mL volu- Mode: GC
metric flask, dissolve in and dilute with Mobile phase to Detector: Flame ionization
volume, and mix. Transfer 1.0 mL of this solution to a Column: 0.32-mm × 60-m fused-silica capillary col-
50-mL volumetric flask, dilute with Mobile phase to vol- umn coated with phase G16
ume, and mix. Temperature
Analysis Injector port: 250°
Samples: Sample solution and Standard solution Detector: 250°
Calculate the percentage of the labeled amount of Column: See Table 1.
benzyl alcohol in the portion of Injection taken:
Result = (rU/rS) × (CS/CU) × 100 Table 1
Hold Time
rU = peak response of benzyl alcohol from the Initial Temperature Final at Final
Sample solution Temperature Ramp Temperature Temperature
rS = peak response of benzyl alcohol from the (°) (°/min) (°) (min)
Standard solution 60 6 200 —
CS = concentration of benzyl alcohol in the
Standard solution (mg/mL)

Official from May 1, 2016

Copyright (c) 2016 The United States Pharmacopeial Convention. All rights reserved.