Printed on: Wed Feb 08 2023, 11:48:48 PM(EST) Status: Currently Official on 09-Feb-2023 DocId: GUID-C0933C5C-368A-4492-BC41-930252AE330A_5_en-US

Printed by: Dang Van Vu Official Date: Official as of 01-Jan-2021 Document Type: USP @2023 USPC
Do Not Distribute DOI Ref: 37uvj DOI: https://doi.org/10.31003/USPNF_M32760_05_01
1

System suitability solution: 0.3 µg/mL of USP Fentanyl

Fentanyl Related Compound E RS and 0.1 mg/mL of USP Fentanyl
RS prepared by diluting System suitability stock solution and
Change to read: Standard stock solution with Diluent
Sample stock solution: 1 mg/mL of Fentanyl prepared as
follows. Transfer a suitable amount of Fentanyl to an
adequate volumetric flask. Add 25% of final volume of
acetonitrile, dissolve by shaking, and dilute with Diluent to
volume.
Sample solution: 0.1 mg/mL of Fentanyl from the Sample
stock solution in Diluent
Chromatographic system
C22H28N2O ▲
336.48▲ (ERR 1-Jan-2021) (See Chromatography á621ñ, System Suitability.)
Propanamide, N-phenyl-N-[1-(2-phenylethyl)-4-piperidinyl]; Mode: LC
N-(1-Phenethylpiperidin-4-yl)-N-phenylpropionamide Detector: UV 206 nm
CAS RN®: 437-38-7. Column: 4.6-mm × 15-cm; 3.5-µm packing L7
DEFINITION Column temperature: 35°
Fentanyl contains NLT 98.0% and NMT 102.0% of fentanyl Flow rate: 1 mL/min
(C22H28N2O), calculated on the dried basis. Injection volume: 30 µL
[CAUTION—Great care should be taken to prevent inhaling System suitability
particles of Fentanyl and skin exposure.] Samples: System suitability solution and Standard solution
Suitability requirements

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IDENTIFICATION Resolution: NLT 1.2 between fentanyl and fentanyl
• A. SPECTROSCOPIC IDENTIFICATION TESTS á197ñ, Infrared related compound E peaks, System suitability solution
Spectroscopy: 197K or 197A Tailing factor: 0.5–2.0, Standard solution
• B. The retention time of the major peak of the Sample Relative standard deviation: NMT 0.73%, Standard
solution corresponds to that of the Standard solution, as solution
obtained in the Assay. Analysis
ASSAY
• PROCEDURE
Solution A: Add 3 mL of triethylamine to about 950 mL of
ci Samples: Standard solution and Sample solution
Calculate the percentage of fentanyl (C22H28N2O) in the
portion of Fentanyl taken:
water in a 1000-mL volumetric flask. Adjust with perchloric Result = (rU/rS) × (CS/CU) × 100
ffi
acid to a pH of 2.62 ± 0.02, and dilute with water to
volume. rU = peak response of fentanyl from the Sample
Solution B: Acetonitrile solution
Diluent: Solution A and Solution B (9:1) rS = peak response of fentanyl from the Standard
Mobile phase: See Table 1. solution
CS = concentration of USP Fentanyl RS in the Standard
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Table 1 solution (mg/mL)

Time Solution A Solution B CU = concentration of Fentanyl in the Sample solution
(min) (%) (%) (mg/mL)
0 90 10
Acceptance criteria: 98.0%–102.0% on the dried basis
2 90 10
IMPURITIES
3.5 85 15
• RESIDUE ON IGNITION á281ñ: NMT 0.5%
5.1 82 18 • ORGANIC IMPURITIES
[NOTE—The use of high-purity (such as HPLC-grade)
7.6 72 28
ammonium formate and ultratrace ammonium hydroxide
11.5 63 37 is recommended.]
Solution A: Acetonitrile and 10 mM ammonium formate
15 40 60
(5:95). Adjust with ammonium hydroxide to a pH of 9.5.
19 40 60 Solution B: Acetonitrile and 10 mM ammonium formate
20 90 10
(95:5). Adjust with ammonium hydroxide to a pH of 9.5.
Mobile phase: See Table 2.
25 90 10
Table 2
System suitability stock solution: 0.1 mg/mL of USP Time Solution A Solution B
Fentanyl Related Compound E RS prepared as follows. (min) (%) (%)
Transfer a suitable amount of USP Fentanyl Related 0 85 15
Compound E RS to an adequate volumetric flask and
dissolve in 50% of the final volume using acetonitrile. Dilute 9 65 35
with water to volume. 33 50 50
Standard stock solution: 1 mg/mL of USP Fentanyl RS
prepared by dissolving in a suitable volumetric flask 40% 38 50 50
filled with acetonitrile and diluting with water to volume 38.1 85 15
Standard solution: 0.1 mg/mL of USP Fentanyl RS from the
45 85 15
Standard stock solution in Diluent

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Printed on: Wed Feb 08 2023, 11:48:48 PM(EST) Status: Currently Official on 09-Feb-2023 DocId: GUID-C0933C5C-368A-4492-BC41-930252AE330A_5_en-US
Printed by: Dang Van Vu Official Date: Official as of 01-Jan-2021 Document Type: USP @2023 USPC
Do Not Distribute DOI Ref: 37uvj DOI: https://doi.org/10.31003/USPNF_M32760_05_01
2

Diluent: 0.025% trifluoroacetic acid in a mixture of rS = peak response of fentanyl related compound G
acetonitrile and water (5:95) from the Standard solution
Sensitivity solution: 0.5 µg/mL of USP Fentanyl RS CS = concentration of USP Fentanyl Related
prepared as follows. Transfer a suitable amount of USP Compound G RS in the Standard solution
Fentanyl RS to an adequate volumetric flask. Dissolve in (mg/mL)
10% of final volume using acetonitrile. Sonicate to dissolve, CU = concentration of Fentanyl in the Sample solution
if necessary. Dilute with Diluent to volume. (mg/mL)
Standard solution: 2.0 µg/mL each of USP Fentanyl Related F = relative response factor (see Table 3)
Compound E RS and USP Fentanyl Related Compound G
RS prepared as follows. Transfer suitable amounts of USP Acceptance criteria: See Table 3. The reporting threshold is
Fentanyl Related Compound E RS and USP Fentanyl Related 0.05%.
Compound G RS to an adequate volumetric flask. Dissolve
in 10% of final volume using acetonitrile. Sonicate to Table 3
dissolve, if necessary. Dilute with Diluent to volume. Relative Relative Acceptance
Sample solution: 1.0 mg/mL of Fentanyl prepared as Retention Response Criteria,
follows. Transfer a suitable amount of Fentanyl to an Name Time Factora NMT (%)
adequate volumetric flask. Dissolve in 10% of final volume Fentanyl
using acetonitrile. Sonicate to dissolve, if necessary. Dilute N-oxideb 0.26 0.69 0.15
with Diluent to volume.
Fentanyl related
Chromatographic system compound G 0.78 1.0 0.15
(See Chromatography á621ñ, System Suitability.)
Mode: LC Fentanyl pyruvyl
Detector: UV 240 nm analogc 0.92 3.0 0.15

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Column: 4.6-mm × 15-cm; 3.5-µm packing L1 Fentanyl 1.00 — —
Column temperature: 45°
Fentanyl related
Flow rate: 1.0 mL/min compound E 1.14
—
0.15
Injection volume: 40 µL
System suitability Fentanyl butyryl
Samples: Sensitivity solution and Standard solution
Suitability requirements
Relative standard deviation: NMT 5.0%, Standard
ci analogd
Any unspecified
impurity
1.21

—
1.5

1.0
0.15

0.10
solution Total impurities — — 0.50
Signal-to-noise ratio: NLT 10, Sensitivity solution
ffi
Analysis a The relative response factor is calculated against fentanyl related compound G.
Samples: Standard solution and Sample solution b 1-Phenethyl-4-(N-phenylpropionamido)piperidine 1-oxide.
Calculate the percentage of fentanyl related compound E c 2-Oxo-N-(1-phenethylpiperidin-4-yl)-N-phenylpropanamide.
in the portion of Fentanyl taken: d N-(1-Phenethylpiperidin-4-yl)-N-phenylbutyramide.

Result = (rU/rS) × (CS/CU) × 100 SPECIFIC TESTS

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• LOSS ON DRYING á731ñ

rU = peak response of fentanyl related compound E Analysis: Dry under vacuum at 60° for 2 h.
from the Sample solution Acceptance criteria: NMT 0.5%
rS = peak response of fentanyl related compound E
from the Standard solution ADDITIONAL REQUIREMENTS
CS = concentration of USP Fentanyl Related • PACKAGING AND STORAGE: Preserve in tightly closed,
Compound E RS in the Standard solution light-resistant containers. Store at 25°, excursions
(mg/mL) permitted between 15° and 30°.
CU = concentration of Fentanyl in the Sample solution • USP REFERENCE STANDARDS á11ñ
(mg/mL) USP Fentanyl RS
USP Fentanyl Related Compound E RS
Calculate the percentage of each other impurity in the 1-Phenethyl-N-phenylpiperidin-4-amine.
portion of Fentanyl taken: C19H24N2 280.41
USP Fentanyl Related Compound G RS
Result = (rU/rS) × (CS/CU) × (1/F) × 100 N-(1-Phenethylpiperidin-4-yl)-N-phenylacetamide.
C21H26N2O 322.44
rU = peak response of each other impurity from the
Sample solution

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