Tetracycline Hydrochloride
C22H24N2O8·HCl 480.90

2-Naphthacenecarboxamide,4-(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,6,10,12,12a-pentahydroxy-6-methyl-1,11-dioxo-,monohydrochloride,[4S-(4a,4aa,5aa,6b,12aa)]-.
(4S,4aS,5aS,6S,12aS)-4-(Dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,6,10,12,12a-pentahydroxy-6-methyl-1,11-dioxo-2-naphthacenecarboxamide monohydrochloride [64-75-5].
»Tetracycline Hydrochloride has a potency of not less than 900µg of C22H24N2O8·HCl per mg.
Packaging and storage— Preserve in tight,light-resistant containers.
Labeling— Where it is intended for use in preparing injectable or other sterile dosage forms,the label states that it is sterile or must be subjected to further processing during the preparation of injectable or other sterile dosage forms.
Identification—
A: Infrared Absorption á197Kñ—Do not dry specimen.
B: Ultraviolet Absorption á197Uñ—
Solution: 20µg per mL.
Medium: 0.25Nsodium hydroxide.
Absorptivity 6minutes after preparation,calculated on the dried basis,at 380nm is between 96.0%and 104.0%of that of USP Tetracycline Hydrochloride RS,the potency of the Reference Standard being taken into account.
C: The chromatogram of the Assay preparationobtained as directed in the Assayexhibits a major peak for tetracycline,the retention time of which corresponds to that exhibited in the chromatogram of the Standard preparationobtained as directed in the Assay.
D: To 0.5mg add 2mLof sulfuric acid:a purplish red color is produced.Add the solution to 1mLof water:the color becomes yellow.
E: Prepare a Test Solutionin methanol containing 1mg per mL,and proceed as directed for Method IIunder Identification—Tetracyclines á193ñ.
F: It responds to the silver nitrate test for Chloride á191ñ.
Crystallinity á695ñ: meets the requirements.
pHá791ñ: between 1.8and 2.8,in a solution containing 10mg per mL.
Specific rotation á781ñ: between -240and -255,calculated on the dried basis.
Test solution: 5mg per mL,in 0.1Nhydrochloric acid.
Loss on drying á731ñ— Dry about 100mg,accurately weighed,in a capillary-stoppered bottle in vacuum at a pressure not exceeding 5mm of mercury at 60for 3hours:it loses not more than 2.0%of its weight.
Limit of 4-epianhydrotetracycline— Using the Diluting solvent,Chromatographic system,and Procedureset forth in the Assay,chromatograph a Standard solution prepared by dissolving an accurately weighed quantity of USP4-Epianhydrotetracycline Hydrochloride RSin Diluting solventto obtain a solution having a known concentration of about 10µg per mL.Using the chromatogram so obtained and the chromatogram of the Assay preparationobtained as directed in the Assay,calculate the percentage of 4-epianhydrotetracycline hydrochloride in the Tetracycline Hydrochloride taken by the formula:
10(CE/W)(rU/rS),
in which CEis the concentration,in µg per mL,of USP4-Epianhydrotetracycline Hydrochloride RSin the Standard solution;Wis the weight,in mg,of Tetracycline Hydrochloride taken to prepare the Assay preparation;and rUand rSare the 4-epianhydrotetracycline peak responses obtained from the Assay preparationand the Standard solution,respectively:not more than 2.0%is found.
Other requirements— Where the label states that Tetracycline Hydrochloride is sterile,it meets the requirements for Sterilityand Bacterial endotoxinsunder Tetracycline Hydrochloride for Injection.Where the label states that Tetracycline Hydrochloride must be subjected to further processing during the preparation of injectable dosage forms,it meets the requirements for Bacterial endotoxinsunder Tetracycline Hydrochloride for Injection.Where it is intended for use in preparing nonparenteral sterile dosage forms,it is exempt from the requirements for Bacterial endotoxins.
Assay—
Diluting solvent— Mix 680mLof 0.1Mammonium oxalate and 270mLof dimethylformamide.
Mobile phase— Mix 680mLof 0.1Mammonium oxalate,270mLof dimethylformamide,and 50mLof 0.2Mdibasic ammonium phosphate.Adjust,if necessary,with 3Nammonium hydroxide or 3Nphosphoric acid to a pHof 7.6to 7.7.Make any other necessary adjustments (see System Suitabilityunder Chromatography á621ñ).Filter through a membrane filter of 0.5-µm or finer porosity.
Standard preparation— Dissolve an accurately weighed quantity of USP Tetracycline Hydrochloride RSin Diluting solvent,and dilute quantitatively with Diluting solventto obtain a solution having a known concentration of about 0.5mg per mL.
Assay preparation— Transfer about 50mg of Tetracycline Hydrochloride,accurately weighed,to a 100-mLvolumetric flask,dissolve in Diluting solvent,dilute with the same solvent to volume,and mix.
Resolution solution— Prepare a solution in Diluting solventcontaining about 100µg of tetracycline hydrochloride and 25µg of USP4-Epianhydrotetracycline Hydrochloride RSper mL.
Chromatographic system (see Chromatography á621ñ)—The liquid chromatograph is equipped with a 280-nm detector,a 4.6-mm ×3-cm guard column that contains 10-µm packing L7,and a 4.6-mm ×25-cm analytical column that contains 5-to 10-µm packing L7.The flow rate is about 2mLper minute.Chromatograph the Resolution solution,and record the peak responses as directed for Procedure:the relative retention times are about 0.9for 4-epianhydrotetracycline and 1.0for tetracycline;and the resolution,R,between the 4-epianhydrotetracycline and tetracycline peaks is not less than 1.2.Chromatograph the Standard preparation,and record the peak responses as directed for Procedure:the relative standard deviation for replicate injections is not more than 2.0%.
Procedure— Separately inject equal volumes (about 20µL)of the Standard preparationand the Assay preparationinto the chromatograph,record the chromatograms,and measure the responses for the major peaks.Calculate the quantity,in µg,of C22H24N2O8·HCl in each mg of the Tetracycline Hydrochloride taken by the formula:
100(CP/W)(rU/rS),
in which Cis the concentration,in mg per mL,of USP Tetracycline Hydrochloride RSin the Standard preparation;Pis the potency,in µg per mg,of USP Tetracycline Hydrochloride RS;Wis the weight,in mg,of Tetracycline Hydrochloride taken to prepare the Assay preparation;and rUand rSare the peak responses obtained from the Assay preparationand the Standard preparation,respectively.
Auxiliary Information— Staff Liaison:William W.Wright,Ph.D.,Scientific Fellow
Expert Committee:(PA7)Pharmaceutical Analysis 7
USP28–NF23Page 1885
Phone Number:1-301-816-8335