Vancomycin Hydrochloride for Injection
Vancomycin Hydrochloride for Injection is a sterile dry mixture of Vancomycin Hydrochloride and may contain a suitable stabilizing agent. It contains NLT 90.0% and NMT 115.0% of the labeled amount of vancomycin.
• AntibioticsMicrobial Assays 81
Sample solution 1 (where it is represented as being in a single-dose container): Constitute a container of Vancomycin Hydrochloride for Injection in water corresponding to the volume of diluent specified in the labeling. Withdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe. Dilute to an equivalent of 1 mg/mL of vancomycin with water.
Sample solution 2 (where it is packaged for dispensing): Dissolve the contents of 1 container of Vancomycin Hydrochloride for Injection in water, and dilute with water to obtain a solution having a concentration of 1 mg/mL of vancomycin.
Sample solution 3 (where the label states the quantity of vancomycin in a given volume of constituted solution): Constitute a container of Vancomycin Hydrochloride for Injection in water corresponding to the volume of diluent specified in the labeling. Dilute a portion to obtain a final concentration equivalent to 1 mg/mL of vancomycin in water.
Analysis: Proceed as directed in AntibioticsMicrobial Assays 81. [NoteUse a measured volume of the appropriate Sample solution, diluted quantitatively with Buffer No. 4 to yield a Test Dilution having a concentration assumed to be equal to the median dose level of the Standard. ]
Acceptance criteria: 90.0%115.0%
• Uniformity of Dosage Units 905: Meets the requirements
• Particulate Matter in Injections 788: Meets the requirements under small-volume injections
• Bacterial Endotoxins Test 85: NMT 0.33 USP Endotoxin Unit/mg of vancomycin
• Sterility Tests 71: It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration, except to dissolve the specimen in water instead of in Fluid A.
• pH 791: 2.54.5, 50 mg/mL in water
• Water Determination, Method I 921: NMT 5.0%
• Injections, Constituted Solutions 1: Meets the requirements at the time of use.
• Content of Vancomycin
Solution A: Triethylamine and water (1:500). Adjust with phosphoric acid to a pH of 3.2.
Solution B: Acetonitrile, tetrahydrofuran, and Solution A (7:1:92)
Solution C: Acetonitrile, tetrahydrofuran, and Solution A (29:1:70)
Mobile phase: See the gradient table below.
[NoteMake adjustments if necessary, changing the acetonitrile proportion in Solution B to obtain a retention time of 7.510.5 min for the main vancomycin peak. ]
System suitability solution: 0.5 mg/mL of USP Vancomycin Hydrochloride RS. Heat at 65 for 48 h, and allow to cool.
Sample solution 1: Equivalent to 10 mg/mL of vancomycin from Vancomycin Hydrochloride for Injection in Solution B
Sample solution 2: 0.4 mg/mL of vancomycin from Sample solution 1 in Solution B
Detector: UV 280 nm
Column: 4.6-mm × 25-cm; 5-µm packing L1
Flow rate: 2 mL/min
Injection size: 20 µL
Sample: System suitability solution
[NoteThe elution order is compound 1, vancomycin B, and compound 2. Compound 2 elutes 36 min after the start of the period when the percentage of Solution C increases from 0% to 100%. ]
Resolution: NLT 3.0 between compound 1 and vancomycin B
Column efficiency: NLT 1500 theoretical plates, calculated from the vancomycin B peak
Samples: Sample solution 1 and Sample solution 2
[NoteWhere baseline separation is not achieved, peak areas are defined by vertical lines extended from the valleys between peaks to the baseline. The main component peak may include a fronting shoulder, which is attributed to monodechlorovancomycin. This shoulder should not be integrated separately. ]
[NoteCorrect any peak observed in the chromatograms obtained from Sample solution 1 and Sample solution 2 by subtracting the area response of any peak observed in the chromatogram of Solution B at the corresponding elution time. ]
Measure the area responses for all of the peaks.
Calculate the percentage of vancomycin B in the specimen tested:
Result = [D × rB/((D × rB) + rA)] × 100
Calculate the percentage of each other peak taken:
Result = [rI/(D × rB) + rA] × 100
Acceptance criteria: NLT 80.0% of vancomycin B; NMT 9.0% of any peak other than the main peak
• Other Requirements: It meets the requirements under Injections 1.
• Packaging and Storage: Preserve as described in Injections 1, Containers for Sterile Solids.
• Labeling: Meets the requirements under Injections 1
• USP Reference Standards 11
USP Endotoxin RS
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USP35NF30 Page 5004Pharmacopeial Forum: Volume No. 35(4) Page 881