Printed on: Fri Jan 05 2024, 09:22:02 PM(EST) Status: Currently Official on 06-Jan-2024 DocId: GUID-C1E3DF2E-B4DA-4A71-9D65-176ADDD3F5FD_3_en-US

Printed by: USP NF Official Date: Official as of 01-Nov-2020 Document Type: USP @2024 USPC
Do Not Distribute DOI Ref: 8b8dj DOI: https://doi.org/10.31003/USPNF_M6130_03_01
1

Sample solution: Prepare as directed in the chapter using

Pregabalin ▲
1.0 g▲ (USP 1-Aug-2020) of Pregabalin.
Acceptance criteria: The turbidity produced in the Sample
solution is NMT that produced in the Standard solution
(0.1%).
C8H17NO2 159.23 Change to read:
(S)-3-(Aminomethyl)-5-methylhexanoic acid CAS RN®:
148553-50-8. • ORGANIC IMPURITIES
Buffer: 0.04 M dibasic ammonium phosphate. Adjust with
DEFINITION phosphoric acid to a pH of 6.5.
Pregabalin contains NLT 98.0% and NMT 102.0% of Solution A: Methanol and Buffer (20:80)
pregabalin (C8H17NO2), calculated on the dried basis. Solution B: Methanol and acetonitrile (10:90)
IDENTIFICATION Mobile phase: See Table 1.

Change to read: Table 1

Time Solution A Solution B
• A. SPECTROSCOPIC IDENTIFICATION TESTS á197ñ, Infrared (min) (%) (%)
Spectroscopy: á197Kñ ▲or á197Añ▲ (USP 1-Aug-2020)
• B. The retention time of the major peak of the Sample 0 98 2
solution corresponds to that of the Standard solution, as 5 98 2
obtained in the Assay.
16 80 20

al
ASSAY
33 80 20
• PROCEDURE
Mobile phase: Acetonitrile and water (5:95) 34 98 2
Standard solution: 2.0 mg/mL of USP Pregabalin RS in
45 98 2
Mobile phase
Sample solution: 2.0 mg/mL of Pregabalin in Mobile phase
Chromatographic system
(See Chromatography á621ñ, System Suitability.)
Mode: LC
ci ▲
Sensitivity solution: 0.005 mg/mL of USP Pregabalin RS
in water▲ (USP 1-Aug-2020)
Standard solution: 0.01 mg/mL of USP Mandelic Acid RS
Detector: UV 205 nm and 0.05 mg/mL each of USP Pregabalin RS and USP
Column: 4.6-mm × 25-cm; 5-µm packing L1 Pregabalin Related Compound C RS in water
ffi
Temperatures Sample solution: 10.0 mg/mL of Pregabalin in water
Autosampler: 10° Chromatographic system
Column: 25° (See Chromatography á621ñ, System Suitability.)
Flow rate: 1.0 mL/min Mode: LC
Injection volume: 20 µL Detector: UV 210 nm
Run time: NLT 2.5 times the retention time of pregabalin Column: 4.6-mm × 25-cm; 5-µm packing L1
O

System suitability Column temperature: 35°

Sample: Standard solution Flow rate: 1.0 mL/min
Suitability requirements Injection volume: 20 µL
Tailing factor: NMT 1.5 System suitability
Relative standard deviation: NMT 0.73% Samples: ▲Sensitivity solution and▲ (USP 1-Aug-2020) Standard
Analysis solution
Samples: Standard solution and Sample solution Suitability requirements
Calculate the percentage of pregabalin (C8H17NO2) in the Relative standard deviation: NMT 5% for mandelic
portion of Pregabalin taken: acid, pregabalin, and pregabalin related compound C,
▲
Standard solution
Result = (rU/rS) × (CS/CU) × 100 Signal-to-noise ratio: NLT 10, Sensitivity
solution▲ (USP 1-Aug-2020)
rU = peak response of pregabalin from the Sample Analysis
solution Samples: Standard solution and Sample solution
rS = peak response of pregabalin from the Standard Calculate the percentage of mandelic acid and pregabalin
solution related compound C in the portion of Pregabalin taken:
CS = concentration of USP Pregabalin RS in the
Standard solution (mg/mL) Result = (rU/rS) × (CS/CU) × 100
CU = concentration of Pregabalin in the Sample
solution (mg/mL) rU = peak response of mandelic acid or pregabalin
related compound C from the Sample solution
Acceptance criteria: 98.0%–102.0% on the dried basis rS = peak response of mandelic acid or pregabalin
related compound C from the Standard solution
IMPURITIES CS = concentration of USP Mandelic Acid RS or USP
• RESIDUE ON IGNITION á281ñ: NMT 0.1% Pregabalin Related Compound C RS in the
Standard solution (mg/mL)
Change to read: CU = concentration of Pregabalin in the Sample
• CHLORIDE AND SULFATE á221ñ, Chloride solution (mg/mL)
Standard solution: Prepare as directed in the chapter using
▲
1.4 mL.▲ (USP 1-Aug-2020)

https://online.uspnf.com/uspnf/document/1_GUID-C1E3DF2E-B4DA-4A71-9D65-176ADDD3F5FD_3_en-US 1/2
Printed on: Fri Jan 05 2024, 09:22:02 PM(EST) Status: Currently Official on 06-Jan-2024 DocId: GUID-C1E3DF2E-B4DA-4A71-9D65-176ADDD3F5FD_3_en-US
Printed by: USP NF Official Date: Official as of 01-Nov-2020 Document Type: USP @2024 USPC
Do Not Distribute DOI Ref: 8b8dj DOI: https://doi.org/10.31003/USPNF_M6130_03_01
2

Calculate the percentage of any individual unspecified bicarbonate, and heat at 55° for about 1 h. Allow to cool to
impurity in the portion of Pregabalin taken: room temperature and dilute with water to volume.
Sample stock solution: 1 mg/mL of Pregabalin prepared as
Result = (rU/rS) × (CS/CU) × (1/F) × 100 follows. Transfer an appropriate amount of Pregabalin to a
suitable volumetric flask. Dissolve in 50% of the final
rU = peak response of each impurity from the Sample volume of water and dilute with acetone to volume.
solution Sample solution: Transfer 0.5 mL of the Sample stock
rS = peak response of pregabalin from the Standard solution to a 5.0-mL volumetric flask, add 0.3 mL of the
solution Derivatizing reagent solution and 50 µL of 1 M sodium
CS = concentration of USP Pregabalin RS in the bicarbonate, and heat at 55° for about 1 h. Allow to cool to
Standard solution (mg/mL) room temperature and dilute with water to volume.
CU = concentration of Pregabalin in the Sample Chromatographic system
solution (mg/mL) (See Chromatography á621ñ, System Suitability.)
F = relative response factor (see Table 2) Mode: LC
Detector: UV 340 nm
Acceptance criteria: See Table 2. The reporting threshold is Column: 4.6-mm × 25-cm; 5-µm packing L1
0.05%. Temperatures
Autosampler: 10°
Table 2 Column: 25°
Relative Relative Acceptance Flow rate: 2.0 mL/min
Retention Response Criteria, Injection volume: 20 µL
Name Time Factor NMT (%)
Run time: NLT 3.8 times the retention time of pregabalin

al
Mandelic acid 0.66 — 0.10 System suitability
Samples: ▲Sensitivity solution and▲ (USP 1-Aug-2020) Standard
Isobutylgluta-
ric acida 0.85 2.1 0.15 solution
▲
[NOTE—The relative retention times for pregabalin
Pregabalin 1.0 1.0 — and pregabalin related compound A are 1.0 and 1.2,
respectively.]▲ (USP 1-Aug-2020)
Isobutylglutar-
monoamideb
Pregabalin relat-
ed compound C
1.52

3.95
3.3

—
ci
0.15

0.15
Suitability requirements
Resolution: NLT 3.0 between pregabalin and pregabalin
related compound A, ▲Standard solution▲ (USP 1-Aug-2020)
Relative standard deviation: NMT 5.0% for pregabalin
Any unspecified related compound A, ▲Standard solution
—
ffi
impurity 1.0 0.10
Signal-to-noise ratio: NLT 10, Sensitivity
Total impurities — — 0.8 solution▲ (USP 1-Aug-2020)
a 3-Isobutylpentanedioic acid.
Analysis
b 3-(2-Amino-2-oxoethyl)-5-methylhexanoic acid.
Sample: Sample solution
Calculate the percentage of pregabalin related
compound A in the portion of Pregabalin taken:
O

Change to read:
Result = (rU/rT) × 100
• ENANTIOMERIC PURITY
Buffer: Triethylamine and water (7.2: 1000). Adjust with rU = peak response of pregabalin related compound A
50% (v/v) phosphoric acid to a pH of 3.0. from the Sample solution
Mobile phase: Acetonitrile and Buffer (38:62) rT = sum of the peak responses of pregabalin and
Derivatizing reagent solution: 3 mg/mL of Marfey’s pregabalin related compound A from the Sample
reagent in acetone solution
▲
Sensitivity stock solution: 0.5 µg/mL of USP Pregabalin
RS prepared as follows. Transfer an appropriate amount of Acceptance criteria: NMT 0.15%
USP Pregabalin RS to a suitable volumetric flask. Dissolve in
50% of the final volume of water and dilute with acetone SPECIFIC TESTS
to volume. • LOSS ON DRYING á731ñ
Sensitivity solution: Transfer 1.0 mL of the Sensitivity stock Analysis: Dry at 105° for 3 h.
solution to a 10.0-mL volumetric flask, add 0.6 mL of the Acceptance criteria: NMT 0.5%
Derivatizing reagent solution and 100 µL of 1 M sodium ADDITIONAL REQUIREMENTS
bicarbonate, and heat at 55° for about 1 h. Allow to cool to • PACKAGING AND STORAGE: Preserve in tight containers.
room temperature and dilute with water to Protect from light and store at NMT 25°.
volume.▲ (USP 1-Aug-2020) • USP REFERENCE STANDARDS á11ñ
Standard stock solution: 1 mg/mL of USP Pregabalin RS USP Mandelic Acid RS
and 0.05 mg/mL of USP Pregabalin Related Compound A USP Pregabalin RS
RS prepared as follows. Transfer appropriate amounts of USP Pregabalin Related Compound A RS
USP Pregabalin RS and USP Pregabalin Related (R)-3-(Aminomethyl)-5-methylhexanoic acid.
Compound A RS to a suitable volumetric flask. Dissolve in C8H17NO2 159.23
50% of the final volume of water and dilute with acetone USP Pregabalin Related Compound C RS
to volume. 4-Isobutylpyrrolidin-2-one.
Standard solution: Transfer 0.5 mL of the Standard stock C8H15NO 141.21
solution to a 5.0-mL volumetric flask, add 0.3 mL of the
Derivatizing reagent solution and 50 µL of 1 M sodium

https://online.uspnf.com/uspnf/document/1_GUID-C1E3DF2E-B4DA-4A71-9D65-176ADDD3F5FD_3_en-US 2/2