Ceftiofur Sodium

This article is compiled based on the United States Pharmacopeia (USP) – 2025 Edition

Issued and maintained by the United States Pharmacopeial Convention (USP)

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C₁₉H₁₆N₅NaO₇S₃     545.54 

C₁₉H₁₆N₅NaO₇S₃.H₂O 563.55 

C₁₉H₁₆N₅NaO₇S₃.3H₂O 599.59 

5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[(2-amino-4-thiazolyl)(methoxyimino)acetyl]amino]-3-[[(2- furanylcarbonyl)thio]methyl]-8-oxo-, monosodium salt, [6R-[6α,7β(Z)]]-; 

Sodium (6R,7R)-7-[2-(2-amino-4-thiazolyl)glyoxylamido]-3-(mercaptomethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate,72-(Z)-(O methyloxime), 2-furoate (ester) CAS RN®: 104010-37-9. 

1 DEFINITION 

Ceftiofur Sodium is the sodium salt of ceftiofur. It is anhydrous or contains one or three molecules of water of hydration. It has a potency equivalent to NLT 862 µg/mg and NMT 970 µg/mg of ceftiofur (C₁₉H₁₇N₅O₇S₃), calculated on the anhydrous and solvent-free basis.  

2 IDENTIFICATION 

Change to read: 

A. Spectroscopic Identification Tests 〈197〉, Infrared Spectroscopy: 197M 

B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. 

3 ASSAY 

Procedure 

Solution A: Tetrabutylammonium hydroxide, 40% in water 

Solution B: Dissolve 3.85 g of ammonium acetate and 13.5 mL of Solution A in 700 mL of water. Adjust with glacial acetic acid to a pH of 6.7. Mobile phase: Mix 700 mL of Solution B with 200 mL of methanol and 110 mL of tetrahydrofuran. 

Diluent: 0.05 M ammonium acetate 

Standard solution: 0.16 mg/mL of USP Ceftiofur Sodium Trihydrate RS in Diluent 

Sample solution: 0.16 mg/mL of Ceftiofur Sodium in Diluent 

Chromatographic system 

(See Chromatography 〈621〉, System Suitability.) 

Mode: LC 

Detector: UV 254 nm 

Column: 4.6-mm × 25-cm; 5-µm packing L7 

Column temperature: 40° 

Flow rate: 1.5 mL/min 

Injection volume: 20 µL 

System suitability 

Sample: Standard solution 

Suitability requirements 

Relative standard deviation: NMT 2.0% 

Analysis 

Samples: Standard solution and Sample solution 

Calculate the quantity, in µg/mg, of ceftiofur (C₁₉H₁₇N₅O₇S₃) in the portion of Ceftiofur Sodium taken: 

Result = (r_U/r_S) × (C_S/C_U) × P

r_U = peak response from the Sample solution 

r_S = peak response from the Standard solution 

C_S = concentration of USP Ceftiofur Sodium Trihydrate RS in the Standard solution (mg/mL)

C_U = concentration of Ceftiofur Sodium in the Sample solution, corrected for water (mg/mL)

P = potency of ceftiofur in USP Ceftiofur Sodium Trihydrate RS (μg/mg)

Acceptance criteria: 862–970 µg/mg on the anhydrous and solvent-free basis 

4 IMPURITIES 

Low Molecular Weight Impurities 

Solution A: Water and trifluoroacetic acid (1000:1) 

Solution B: Acetonitrile and trifluoroacetic acid (1000:1) 

Solution C: Solution A and Solution B (950:50) 

Mobile phase: See Table 1. 

Table 1 

Buffer: 3.5 g/L of sodium phosphate dibasic dihydrate, adjusted with phosphoric acid to a pH of 8.0 

Diluent: Acetonitrile and Buffer (40:60) 

System suitability solution: 1 mg/mL of USP Ceftiofur System Suitability Mixture RS in Diluent. Sonicate as needed to dissolve. Peak identification solution: 15 µg/mL of USP Cefotaxime Sodium RS in Diluent 

Standard solution: 0.005 mg/mL of USP Ceftiofur Sodium Trihydrate RS in Diluent 

Sample solution: 1 mg/mL of Ceftiofur Sodium in Diluent. Inject within 20 min of preparation. 

Chromatographic system 

(See Chromatography 〈621〉, System Suitability.) 

Mode: LC 

Detector: UV 254 nm 

Column: 4.6-mm × 25-cm; 5-µm packing L1 

Flow rate: 1.2 mL/min 

Injection volume: 20 µL 

Run time: 3.5 times the retention time of ceftiofur 

System suitability 

Samples: System suitability solution and Standard solution 

[Note—The relative retention times for ceftiofur delta-3 isomer and ceftiofur are 0.93 and 1.0, respectively.] 

Suitability requirements 

Resolution: NLT 1.5 between ceftiofur delta-3 isomer and ceftiofur, System suitability solution 

Relative standard deviation: NMT 3.0%, Standard solution 

Analysis 

Samples: Sample solution and Peak identification solution 

[Note—The elution order of the N-deacyl ceftiofur and cefotaxime peaks may be reversed depending on the column used. Determine the location of the cefotaxime peak by using the Peak identification solution.] 

Calculate the percentage of each impurity in the portion of Ceftiofur Sodium taken: 

Result = (r_U/r_S) × (C_S/C_U) × P × (F₁/F₂) × 100 

r_U = peak response of each impurity from the Sample solution 

r_S = peak response of ceftiofur from the Standard solution 

C_S = concentration of USP Ceftiofur Sodium Trihydrate RS in the Standard solution (mg/mL) 

C_U = concentration of Ceftiofur Sodium in the Sample solution, corrected for water (mg/mL) 

P = potency of ceftiofur in USP Ceftiofur Sodium Trihydrate RS (µg/mg) 

F₁ = conversion factor, 0.001 mg/µg 

F₂ = relative response factor (see Table 2) 

Acceptance criteria: See Table 2. The reporting threshold is 0.1%. 

Table 2 

^a(Z)-2-(2-Aminothiazol-4-yl)-2-(methoxyimino)acetic acid. 

^b Furan-2-carboxylic acid. 

^c Furan-2-carbothioic acid. 

^d(Z)-2-(2-Aminothiazol-4-yl)-N-((5aR,6R)-1,7-dioxo-1,3,4,5a,6,7-hexahydroazeto[2,1-b]thieno[3,4-d][1,3]thiazin-6-yl)-2- (methoxyimino)acetamide. 

^e(6R,7R)-7-Amino-3-((furan-2-carbonylthio)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid. 

^f (6R,7R)-3-(Acetoxymethyl)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(methoxyimino)acetamido)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylic acid. 

^gIf possible from the manufacturing process. 

^h(6R,7R)-7-[(Z)-2-(2-Aminothiazol-4-yl)-2-(methoxyimino)acetamido]-3-(mercaptomethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylic acid. 

ⁱ (6R,6′R,7R,7′R,Z)-3,3′-[Dithiobis(methylene)]bis{7-[(Z)-2-(2-aminothiazol-4-yl)-2-(methoxyimino)acetamido]-8-oxo-5-thia-1- azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid}. 

^j (6R,7R)-7-((Z)-2-(2-Aminothiazol-4-yl)-2-(methoxyimino)acetamido)-3-((furan-2-carbonylthio)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-3- ene-2-carboxylic acid. 

^k(6R,7R)-7-((E)-2-(2-Aminothiazol-4-yl)-2-(methoxyimino)acetamido)-3-((furan-2-carbonylthio)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2- ene-2-carboxylic acid. 

^l (6R,7R)-7-{(Z)-2-(2-[(Z)-2-(2-Aminothiazol-4-yl)-2-(methoxyimino)acetamido]thiazol-4-yl)-2-(methoxyimino)acetamido}-3-{[(furan-2- carbonyl)thio]methyl}-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid. 

^m (6R,7R)-3-{[(Furan-2-carbonyl)thio]methyl}-7-{(Z)-2-[2-(furan-2-carboxamido)thiazol-4-yl]-2-(methoxyimino)acetamido}-8-oxo-5-thia-1- azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid. 

ⁿ Difuran-2-oyl disulfide. Change to read: 

High Molecular Weight Impurities (Ceftiofur Polymers) 

[Note—Perform this test if this impurity is possible from the manufacturing process.] 

To minimize leaching of plastic components, avoid contact between the Mobile phase and plastics during all steps including during preparation of the Mobile phase, the System suitability solution, and the Sample solution, and when lling the injection vial. Solution A: 0.68 g/L of monobasic potassium phosphate. Adjust with 45% potassium hydroxide solution to a pH of 7.5. Mobile phase: 10 g/L of electrophoresis grade sodium dodecyl sulfate in Solution A. Stir or slightly heat to dissolve. Blank: Use the Mobile phase. 

System suitability solution: 0.15 mg/mL each of USP Ceftiofur System Suitability Mixture RS in Mobile phase. Sonicate if necessary to dissolve. Inject within 20 min of preparation. 

Sample solution: 0.15 mg/mL of Ceftiofur Sodium in Mobile phase. Inject within 20 min of preparation. 

Chromatographic system 

(See Chromatography 〈621〉, System Suitability.) 

Mode: LC 

Detector: UV 254 nm 

Column: 4.0-mm × 25-cm; 5-µm packing L20 

Flow rate: 1 mL/min 

Injection volume: 20 µL 

Run time: NLT 2 times the retention time of the ceftiofur peak 

System suitability 

Sample: System suitability solution 

[Note—Adjust the chromatographic system so that the retention time of ceftiofur is NLT 2.5 min. The relative retention times of ceftiofur E isomer and ceftiofur are 0.9 and 1.0, respectively.] 

Suitability requirements 

Resolution: NLT 1.0 between ceftiofur and ceftiofur E-isomer 

Analysis 

Samples: Blank and Sample solution 

Calculate the percentage of high molecular weight impurities in the portion of Ceftiofur Sodium taken: 

Result = {100 × (r_U/F)/[(r_U/F) + r_C + r_A ]} − T 

r_U = sum of the responses of all peaks that elute prior to ceftiofur from the Sample solution, corrected for the Blank if necessary 

F = relative response factor, 0.8 

r_C = peak response of ceftiofur from the Sample solution 

r_A = sum of the responses of all peaks that elute after ceftiofur from the Sample solution  

T = total impurities from Table 2

Acceptance criteria 

Total high molecular weight impurities: NMT 11.0% 

Residual Solvents 〈467〉 

Acceptance criteria 

Tetrahydrofuran (if present): NMT 0.7% 

Acetone (if present): NMT 3.5% 

[Note—For the Acceptance criteria for any other residual solvents, see Residual Solvents 〈467〉.] 

5 SPECIFIC TESTS 

pH 〈791〉 

Sample solution: 50 mg/mL 

Acceptance criteria: 5.0–7.5 

Optical Rotation 〈781S〉, Procedures, Specific Rotation 

Sample solution: 10 mg/mL in water 

Acceptance criteria: −60.0° to −74.0° 

Bacterial Endotoxins Test 〈85〉: Where the label states that Ceftiofur Sodium is sterile or that it must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 1.0 USP Endotoxin Unit/mg of ceftiofur sodium. 

Water Determination 〈921〉, Method I 

Acceptance criteria 

Where it is labeled as anhydrous: NMT 2.0% 

Where it is labeled as the monohydrate: NMT 4.0% 

Where it is labeled as the trihydrate: NMT 10.0% 

6 ADDITIONAL REQUIREMENTS 

Packaging and Storage: Preserve in tight, light-resistant containers. Store at controlled room temperature except where it is labeled as monohydrate, in which case it should be stored in the freezer. 

Labeling: Label it to indicate that it is intended for veterinary use only and to indicate whether it is anhydrous, monohydrate, or trihydrate. Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. 

USP Reference Standards 〈11〉 

USP Cefotaxime Sodium RS 

USP Ceftiofur Sodium Trihydrate RS 

USP Ceftiofur System Suitability Mixture RS 

This is a mixture of ceftiofur, ceftiofur delta-3 isomer, ceftiofur E-isomer, and other impurities. 

Ceftiofur delta-3 isomer 

(6R,7R)-7-((Z)-2-(2-Aminothiazol-4-yl)-2-(methoxyimino)acetamido)-3-((furan-2-carbonylthio)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-3-ene-2- carboxylic acid. 

C₁₉H₁₇N₅O₇S₃ 523.56 

Ceftiofur E-isomer 

(6R,7R)-7-((E)-2-(2-Aminothiazol-4-yl)-2-(methoxyimino)acetamido)-3-((furan-2-carbonylthio)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylic acid. 

C₁₉H₁₇N₅O₇S₃  523.56 

USP Endotoxin RS