Revision Bulletin

Official October 1, 2010 Tetracycline 1

Acceptance criteria: 90.0%–125.0%

Tetracycline Hydrochloride Capsules
.

PERFORMANCE TESTS
DEFINITION
Tetracycline Hydrochloride Capsules contain NLT 90.0% and Change to read:
NMT 125.0% of the labeled amount of C22H24N2O8 · HCl.
• DISSOLUTION 〈711〉
IDENTIFICATION •Test 1• (RB 1-Oct-2010)
• The retention time of the major peak of the Sample solution Medium: Water; 900 mL
corresponds to that of the Standard solution, as obtained in Apparatus 2: 75 rpm; maintain a distance of 45 ± 5 mm
the Assay. between the blade and the inside bottom of the vessel.
Time: 60 min; 90 min for 500-mg Capsules
ASSAY Detector: UV 276 nm
• PROCEDURE Standard solution: USP Tetracycline Hydrochloride RS in
Diluent: Dimethylformamide and 0.1 M ammonium oxalate Medium
(27:68) Sample solution: Pass a portion of the solution under test
Mobile phase: Dimethylformamide, 0.1 M ammonium oxa- through a suitable filter.
late, and 0.2 M dibasic ammonium phosphate (27:68:5). Analysis: Determine the amount of C22H24N2O8 · HCl dis-
Adjust, if necessary, with 3 N ammonium hydroxide or 3 N solved by using UV absorption on portions of the Sample
phosphoric acid to a pH of 7.6–7.7. solution suitably diluted with Medium, if necessary, in com-
Standard solution: 0.5 mg/mL of USP Tetracycline Hydro- parison with a Standard solution having a known concentra-
chloride RS in Diluent tion of USP Tetracycline Hydrochloride RS.
Sample solution: Equivalent to 0.5 mg/mL of tetracycline Tolerances: NLT 80% (Q) of C22H24N2O8 · HCl is dissolved.
hydrochloride in Diluent from NLT 20 Capsules. [NOTE—Soni- •Test 2: If the product complies with this test, the labeling
cate if necessary to dissolve.] indicates that the product meets USP Dissolution Test 2.
System suitability solution: 100 µg/mL of tetracycline hy- Medium: Water; 900 mL, deaerated.
drochloride and 25 µg/mL of USP 4-Epianhydrotetracycline Apparatus 2: 75 rpm; maintain a distance of 45 ± 5 mm
Hydrochloride RS between the blade and the inside bottom of the vessel.
Chromatographic system Time
(See Chromatography 〈621〉, System Suitability.) For Capsules labeled to contain 250 mg: 30 and 60 min
Mode: LC For Capsules labeled to contain 500 mg: 30, 60, and 90
Detector: UV 280 nm min
Guard column: 4.6-mm × 3-cm; 10-µm packing L7 Detector: UV 276 nm
Column: 4.6-mm × 25-cm; 5- to 10-µm packing L7 Standard solution: USP Tetracycline Hydrochloride RS in
Flow rate: 2 mL/min Medium
System suitability Sample solution: At the times specified, withdraw a known
Samples: Standard solution and System suitability solution volume of the solution under test, and pass it through a
[NOTE—The relative retention times of 4-epianhydrotetra- suitable filter. Dilute with Medium, if necessary. Replace the
cycline and tetracycline are 0.9 and 1.0, respectively.] volume withdrawn with the same amount of Medium, pre-
Suitability requirements heated at 37.0 ± 0.5°.
Resolution: NLT 1.2 between the 4-epianhydrotetra- Blank: Medium
cycline and tetracycline peaks, System suitability solution Tolerances: See Tables 1 and 2.
Relative standard deviation: NMT 2.0%, Standard
solution
Analysis Table 1
Samples: Standard solution and Sample solution For Capsules Labeled to Contain 250 mg
Calculate the percentage of tetracycline hydrochloride Time (min) Amount Dissolved
(C22H24N2O8 · HCl) in the portion of Capsules taken: 30 NLT 60% (Q)
Result = (rU/rS) × (CS/CU) × P × F × 100 60 NLT 85% (Q)

rU = peak response of tetracycline from the Sample

solution Table 2
rS = peak response of tetracycline from the Standard For Capsules Labeled to Contain 500 mg
solution
CS = concentration of USP Tetracycline Hydrochloride Time (min) Amount Dissolved
RS in the Standard solution (mg/mL) 30 NLT 50% (Q)
CU = nominal concentration of the Sample solution 60 NLT 70% (Q)
(mg/mL) 90 NLT 85% (Q)
P = potency of USP Tetracycline Hydrochloride RS
(µg/mg) • (RB 1-Oct-2010)
F = conversion factor, 0.001 mg/µg • UNIFORMITY OF DOSAGE UNITS 〈905〉: Meet the requirements
IMPURITIES
Organic Impurities
• PROCEDURE: LIMIT OF 4-EPIANHYDROTETRACYCLINE
Diluent, Mobile phase, Sample solution, Chromato-
graphic system, and System suitability: Proceed as di-
rected in the Assay.

2010 The United States Pharmacopeial Convention All Rights Reserved.

 Revision Bulletin
2 Tetracycline Official October 1, 2010

Standard solution: 10 µg/mL of USP 4-Epianhydrotetra- ADDITIONAL REQUIREMENTS

cycline Hydrochloride RS in Diluent • PACKAGING AND STORAGE: Preserve in tight, light-resistant
Analysis containers.
Samples: Standard solution and Sample solution
Calculate the percentage of 4-epianhydrotetracycline hy-
drochloride in the portion of Capsules taken: Add the following:

Result = (rU/rS) × (CS/CU) × F × 100 •• LABELING: When more than one Dissolution test is
given, the labeling states the test used only if Test 1 is not
rU = peak response of 4-epianhydrotetracycline from used.• (RB 1-Oct-2010)
the Sample solution • USP REFERENCE STANDARDS 〈11〉
rS = peak response of 4-epianhydrotetracycline from USP 4-Epianhydrotetracycline Hydrochloride RS
the Standard solution 2-Naphthacenecarboxamide, 4-(dimethylamino)-
CS = concentration of USP 4-Epianhydrotetracycline 1,4,4a,5,12,12a-hexahydro-3,10,11,12a-tetrahydroxy-6-
Hydrochloride RS in the Standard solution methyl-1,12-dioxo, monohydrochloride, [4R-(4α, 4aβ,
(µg/mL) 12aβ)]
CU = nominal concentration of tetracycline hydro- C22H22N2O7 · HCl 462.89
chloride in the Sample solution (mg/mL) USP Tetracycline Hydrochloride RS
F = conversion factor, 0.001 mg/µg (4S,4aS,5aS,6S,12aS)-4-(Dimethylamino)-
Acceptance criteria: NMT 3.0% 1,4,4a,5,5a,6,11,12a-octahydro-3,6,10,12,12a-
pentahydroxy- 6-methyl-1,11-dioxo-2-naphthacenecarbox-
SPECIFIC TESTS amide monohydrochloride.
• LOSS ON DRYING 〈731〉: Dry 100 mg of Capsule contents in a C22H24N2O8 · HCl 480.90
capillary-stoppered bottle in vacuum at a pressure not ex-
ceeding 5 mm of mercury at 60° for 3 h: it loses NMT 4.0%
of its weight.

2010 The United States Pharmacopeial Convention All Rights Reserved.