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Printed by: Nguyen Nhan Official Date: Official as of 01-May-2020 Document Type: USP @2021 USPC
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CS = concentration of USP Cephalexin RS in the

Cephalexin Standard solution (mg/mL)
CU = concentration of Cephalexin in the Sample
solution (mg/mL)
P = potency of cephalexin in USP Cephalexin RS
(µg/mg)

Acceptance criteria: 950–1030 µg/mg on the anhydrous

basis
IMPURITIES
ORGANIC IMPURITIES
• Procedure 1
Solution A: Dissolve 1 g of sodium 1-pentanesulfonate in a
C16H17N3O4S · H2O 365.40 mixture of 1000 mL of water and 15 mL of triethylamine.
C16H17N3O4S 347.40 Adjust with phosphoric acid to a pH of 2.5 ± 0.1.
5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7- Solution B: Dissolve 1 g of sodium 1-pentanesulfonate in a
[(aminophenylacetyl)amino]-3-methyl-8-oxo-, mixture of 300 mL of water and 15 mL of triethylamine.
monohydrate, [6R-[6α,7β (R*)]]-; Adjust with phosphoric acid to a pH of 2.5 ± 0.1, and add
(6R,7R)-7-[(R)-2-Amino-2-phenylacetamido]-3-methyl-8- 350 mL of acetonitrile and 350 mL of methanol.
oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid Mobile phase: See the gradient table below.
monohydrate [23325-78-2].
Anhydrous [15686-71-2].

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Time Solution A Solution B
(min) (%) (%)
DEFINITION
Cephalexin has a potency of NLT 950 µg/mg and NMT 1030 0 100 0
µg/mg of C16H17N3O4S, calculated on the anhydrous basis. 1 100 0

IDENTIFICATION
Change to read:
• A. ▲SPECTROSCOPIC IDENTIFICATION TESTS á197ñ, Infrared
ci 33.3

34.3
0

0
100

100

Diluent: 18 mg/mL of monobasic potassium phosphate in

Spectroscopy: 197K▲ (CN 1-May-2020) water
ffi
• B. The retention time of the major peak of the Sample Standard solutions: 0.08 mg/mL and 0.16 mg/mL of
solution corresponds to that of the Standard solution, as cephalexin (C16H17N3O4S) from USP Cephalexin RS in
obtained in the Assay. Diluent, taking into account the stated potency of the USP
ASSAY Cephalexin RS
• PROCEDURE Sample solution: 5 mg/mL of Cephalexin in Diluent
Mobile phase: 0.985 g/L of sodium-1-pentanesulfonate in a Chromatographic system
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mixture of acetonitrile, methanol, triethylamine, and water (See Chromatography á621ñ, System Suitability.)
(20:10:3:170), adjusted with phosphoric acid to a pH of 3.0 Mode: LC
± 0.1 Detector: UV 254 nm
Standard stock solution: 1 mg/mL of USP Cephalexin RS in Column: 4.6-mm × 25-cm; packing L1 of low acidity
water Flow rate: 1 mL/min
Standard solution: 0.4 mg/mL of cephalexin in Injection size: 20 µL
Mobile phase from Standard stock solution Analysis
Sample stock solution: 1 mg/mL of Cephalexin in water Samples: Standard solutions and Sample solution
Sample solution: 0.4 mg/mL of Cephalexin in Mobile phase Plot the responses of the cephalexin peaks from the
from Sample stock solution Standard solutions versus their concentrations, calculated
Chromatographic system on the anhydrous basis, in mg/mL, and draw a straight
(See Chromatography á621ñ, System Suitability.) line through the two points and zero. From the line and
Mode: LC the peak responses of the Sample solution, determine the
Detector: UV 254 nm concentration, I, in mg/mL, of each cephalexin-related
Column: 4.6-mm × 25-cm; packing L1 of low acidity substance of the Sample solution other than the
Flow rate: 1.5 mL/min cephalexin peak.
Injection size: 20 µL Calculate the percentage of each cephalexin-related
System suitability substance:
Sample: Standard solution
Suitability requirements Result = I/C × 100
Relative standard deviation: NMT 2.0%
I = concentration of each cephalexin-related
Analysis
substance in the Sample solution as determined
Samples: Standard solution and Sample solution
from the calibration curve (mg/mL)
Calculate the quantity, in µg, of cephalexin (C16H17N3O4S)
C = concentration of cephalexin from the Sample
per mg of the Cephalexin taken: solution (mg/mL)
Result = (rU/rS) × (CS/CU) × P Acceptance criteria
rU = peak response from the Sample solution Individual impurities: NMT 1.0% of any individual
cephalexin-related substance
rS = peak response from the Standard solution Total impurities: NMT 5.0%

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• Procedure 2: Dimethylaniline á223ñ: Meets the • CRYSTALLINITY á695ñ: Meets the requirements
requirement • PH á791ñ: 3.0–5.5, in an aqueous suspension containing
50 mg/mL
SPECIFIC TESTS • WATER DETERMINATION, Method I á921ñ: 4.0%–8.0%
• OPTICAL ROTATION, Specific Rotation á781Sñ: +149° to
+158° ADDITIONAL REQUIREMENTS
Sample solution: 5 mg/mL, in pH 4.4 neutralized phthalate • PACKAGING AND STORAGE: Preserve in tight containers.
buffer (see Reagents, Indicators, and Solutions—Buffer • USP REFERENCE STANDARDS á11ñ
Solutions) USP Cephalexin RS

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