Tetracycline Hydrochloride

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

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

DOWNLOAD PDF HERE

1 DEFINITION

Tetracycline Hydrochloride has a potency of NLT 900 µg/mg of tetracycline hydrochloride (C H N O · HCl).

IDENTIFICATION

A. SPECTROSCOPIC IDENTIFICATION TESTS 〈197〉 , Infrared Spectroscopy: 197A or 197K. Do not dry the specimen.

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

C. IDENTIFICATION TESTS—GENERAL 〈191〉 , Chemical Identification Tests, Chloride Sample solution: 1 mg/mL of Tetracycline Hydrochloride in methanol Acceptance criteria: Meets the requirements

2 ASSAY

PROCEDURE

Solution A: Dilute 1 mL of phosphoric acid with water to 1 L.

Solution B: Acetonitrile

Mobile phase: See Table 1.

Table 1

System suitability solution: 25 µg/mL each of USP Anhy drotetracy cline Hy drochloride RS, USP Epitetracy cline Hy drochloride RS, USP 4- Epianhydrotetracycline Hydrochloride RS, and USP Tetracycline Hydrochloride RS in Solution A

Standard solution: 100 µg/mL of USP Tetracy cline Hy drochloride RS in Solution A

Sample solution: 100 µg/mL of Tetracycline Hydrochloride in Solution A Chromatographic system

(See Chromatography 〈621〉, System Suitability.)

Mode: LC

Detector: UV 280 nm

Column: 4.6-mm × 15-cm; 3-µm packing L60. [NOTE—Column with L1 packing is also suitable.]

Temperatures

Autosampler: 10°

Column: 50°

Flow rate: 1.0 mL/min

Injection volume: 10 µL

System suitability

Samples: System suitability solution and Standard solution

Suitability requirements

Resolution: NLT 2.5 between epitetracycline and tetracycline; NLT 2.5 between anhydrotetracycline and 4-epianhydrotetracycline, System suitability solution

Tailing factor: NMT 1.5, Standard solution

Relative standard deviation: NMT 2.0%, Standard solution

Analysis

Samples: Standard solution and Sample solution

Calculate the potency, in µg/mg, of tetracycline hydrochloride (C H N O · HCl) in the portion of Tetracycline Hydrochloride taken:

Result = (r /r ) × (C /C ) × P

= peak response of tetracycline from the Sample solution

= peak response of tetracycline from the Standard solution

= concentration of USP Tetracycline Hydrochloride RS in the Standard solution (µg/mL)

= concentration of Tetracycline Hydrochloride in the Sample solution (µg/mL)

P    = potency of USP Tetracycline Hydrochloride RS (µg/mg) Acceptance criteria: NLT 900 µg/mg

3 IMPURITIES

ORGANIC IMPURITIES

Solution A, Solution B, Mobile phase, System suitability solution, Sample solution, and Chromatographic system: Proceed as directed in the Assay.

Standard solution 1: 0.5 µg/mL of USP Anhy drotetracy cline Hy drochloride RS and 3 µg/mL of USP Epitetracy cline Hy drochloride RS in

Solution A

Standard solution 2: 2 µg/mL of USP 4-Epianhydrotetracycline Hydrochloride RS in Solution A Standard solution 3: 0.1 µg/mL of USP Tetracy cline Hy drochloride RS in Solution A

System suitability

Sample: System suitability solution Suitability requirements

Resolution: NLT 2.5 between epitetracycline and tetracycline; NLT 2.5 between anhydrotetracycline and 4-epianhydrotetracycline Relative standard deviation: NMT 2%

Analysis

Samples: Standard solution 1, Standard solution 2, Standard solution 3, and Sample solution

Calculate the percentage of anhydrotetracycline hydrochloride and epitetracycline hydrochloride in the portion of Tetracycline Hydrochloride taken:

Result = (r /r ) × (C /C ) × 100

= peak response of anhydrotetracycline or epitetracycline from the Sample solution

= peak response of the corresponding USP Reference Standard from Standard solution 1

= concentration of the corresponding USP Reference Standard in Standard solution 1 (µg/mL)

= concentration of Tetracycline Hydrochloride in the Sample solution (µg/mL)

Calculate the percentage of 4-epianhydrotetracycline hydrochloride in the portion of Tetracycline Hydrochloride taken:

Result = (rU/rS) × (CS/CU) × 100

= peak response of 4-epianhydrotetracycline from the Sample solution

= peak response of 4-epianhydrotetracycline from Standard solution 2

= concentration of USP 4-Epianhydrotetracycline Hydrochloride RS in Standard solution 2 (µg/mL)

= concentration of Tetracycline Hydrochloride in the Sample solution (µg/mL)

Calculate the percentage of 2-acetyl analog or any unspecified impurity in the portion of Tetracycline Hydrochloride taken:

Result = (r /r ) × (C /C ) × 100

= peak response of 2-acetyl analog or any unspecified impurity from the Sample solution

= peak response of tetracycline from Standard solution 3

= concentration of USP Tetracy cline Hy drochloride RS in Standard solution 3 (µg/mL)

= concentration of Tetracycline Hydrochloride in the Sample solution (µg/mL)

Acceptance criteria: See Table 2. The reporting threshold is 0.05%.

Table 2

^a 2-Acetyl-2-decarbamoyltetracycline; (4S,4aS,5aS,6S,12aS)-2-Acetyl-4-(dimethylamino)-3,6,10,12,12a-pentahydroxy-6-methyl-4a,5a,6,12a- tetrahydrotetracene-1,11(4H,5H)-dione.

4 SPECIFIC TESTS

PTICAL ROTATION 〈781S〉 , Procedures, Specific Rotation

Sample solution: 5 mg/mL of Tetracycline Hydrochloride in 0.1 N hy drochloric acid Acceptance criteria: −240° to −255° on the dried basis

CRYSTALLINITY 〈695〉: Meets the requirements

PH 〈791〉

Sample solution: Prepare in a solution (1 in 100). Acceptance criteria: 1.8–2.8

LOSS ON DRYING 〈731〉

Sample: 100 mg of Tetracycline Hydrochloride

Analysis: Dry the Sample in a capillary-stoppered bottle under vacuum at a pressure not exceeding 5 mm of mercury at 60° for 3 h. Acceptance criteria: NMT 2.0%

STERILITY TESTS 〈71〉: Where the label states that Tetracycline Hydrochloride is sterile, it meets the requirements.

Change to read:

BACTERIAL ENDOTOXINS TEST 〈85〉: Where the label states Tetracycline Hydrochloride (ERR 1-Nov-2022) must be subjected to further processing during the preparation of injectable dosage forms, the level of bacterial endotoxins are such that the requirement under the relevant dosage form monograph(s) in which Tetracycline Hydrochloride (ERR 1-Nov-2022) is used can be met.

5 ADDITIONAL REQUIREMENTS

 PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Change to read:

LABELING: Where Tetracycline Hydrochloride (ERR 1-Nov-2022) must be sterile or subjected to further processing during the preparation of injectable dosage forms to ensure acceptable levels of bacterial endotoxins, it is so labeled.

USP REFERENCE STANDARDS 〈11〉

USP Anhydrotetracycline Hydrochloride RS

(4S,4aS,12aS)-4-(Dimethylamino)-3,10,11,12a-tetrahydroxy-6-methyl-1,12-dioxo-1,4,4a,5,12,12a-hexahydrotetracene-2-carboxamide monohydrochloride.

C H N O · HCl    462.88

USP 4-Epianhydrotetracycline Hydrochloride RS

(4R,4aS,12aS)-4-(Dimethylamino)-3,10,11,12a-tetrahydroxy-6-methyl-1,12-dioxo-1,4,4a,5,12,12a-hexahydrotetracene-2-carboxamide hydrochloride.

C H N O · HCl    462.88

USP Epitetracycline Hydrochloride RS

(4R,4aS,5aS,6S,12aS)-4-(Dimethylamino)-3,6,10,12,12a-pentahydroxy-6-methyl-1,11-dioxo-1,4,4a,5,5a,6,11,12a-octahydrotetracene-2-    carboxamide monohydrochloride.

C H N O · HCl    480.90

USP Tetracycline Hydrochloride RS