Vancomycin Injection
DEFINITION
Vancomycin Injection is a sterile isoosmotic solution of Vancomycin Hydrochloride in Water for Injection. It contains NLT 90.0% and NMT 115.0% of the labeled amount of vancomycin (C66H75Cl2N9O24). It contains a suitable tonicity-adjusting agent.
IDENTIFICATION
• A.
The retention time of the main vancomycin peak of the Sample stock solution, obtained as directed in Composition of Vancomycin, corresponds to that of a similarly prepared solution of USP Vancomycin Hydrochloride RS.
ASSAY
• Antibiotics—Microbial Assays 
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Sample solution:
Allow a container of Injection to thaw, and mix the solution. Dilute a portion of this solution with Buffer No. 4 to yield a Test dilution having a concentration assumed to be equal to that of the median dose of the standard.
Analysis:
Proceed as directed for vancomycin in the chapter.
Acceptance criteria:
90.0%–115.0%
SPECIFIC TESTS
• Composition of Vancomycin
Buffer:
Triethylamine and water (1:500). Adjust with phosphoric acid to a pH of 3.2.
Solution A:
Acetonitrile, tetrahydrofuran, and Buffer (7:1:92)
Solution B:
Acetonitrile, tetrahydrofuran, and Buffer (29:1:70)
Mobile phase:
See Table 1. Make adjustments if necessary, changing the acetonitrile proportion in Solution A to obtain a retention time of 7.5–10.5 min for the main vancomycin peak.
Table 1
| Time (min) |
Solution A (%) |
Solution B (%) |
|---|---|---|
| 0 | 100 | 0 |
| 12 | 100 | 0 |
| 20 | 0 | 100 |
| 22 | 0 | 100 |
| 23 | 100 | 0 |
| 30 | 100 | 0 |
System suitability solution:
Allow a container of Injection to thaw, and mix the solution. Dilute a portion of the Injection with water to obtain a solution containing 0.5 mg/mL of vancomycin. Heat at 65
for 24 h, and allow to cool.
for 24 h, and allow to cool.
Sample stock solution:
Allow a container of Injection to thaw, and mix the solution.
Sample solution:
Equivalent to 0.4 mg/mL of vancomycin hydrochloride from the Sample stock solution in Solution A
Chromatographic system
Mode:
LC
Detector:
UV 280 nm
Column:
4.6-mm × 25-cm; 5-µm packing L1
Flow rate:
2 mL/min
Injection size:
20 µL
System suitability
Sample:
System suitability solution
Suitability requirements
[Note—The elution order is compound 1, vancomycin B, and compound 2. Compound 2 elutes 3–6 min after the start of the period, when the percentage of Solution B is increasing from 0% to 100%. ]
Resolution:
NLT 3.0 between compound 1 and vancomycin B
Column efficiency:
NLT 1500 theoretical plates for the vancomycin B peak
Analysis
Samples:
Sample stock solution and Sample solution
Where 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.
Correct any peak observed in the chromatograms from Sample stock solution and Sample solution by subtracting the area response of any peak observed in the chromatogram of Solution A at the corresponding retention time.
Calculate the percentage of vancomycin B in the portion of Injection taken:
Result = {(D × rB)/[(D × rB) + rA]} × 100
| D | = | = dilution factor, Sample stock solution to Sample solution, 25 |
| rB | = | = corrected peak area response of the main peak from the Sample solution |
| rA | = | = sum of the corrected peak area responses of all the peaks, other than the main peak, from the Sample stock solution |
Calculate the percentage of any individual peak, other than the main peak, in the portion of Injection taken:
Result = {ri/[(D × rB) + rA]} × 100
| ri | = | = corrected peak area response of any individual peak, other than the main peak, from the Sample stock solution |
| D | = | = dilution factor, Sample stock solution to Sample solution, 25 |
| rB | = | = corrected peak area response of the main peak from the Sample solution |
| rA | = | = sum of the corrected peak area responses of all the peaks, other than the main peak, from the Sample stock solution |
Acceptance criteria:
NLT 88% of vancomycin B; NMT 4% of any individual peak other than the main peak
• pH 791:
3.0–5.0
791:
3.0–5.0
• Bacterial Endotoxins Test 85:
NMT 0.33 USP Endotoxin Unit/mg of vancomycin
85:
NMT 0.33 USP Endotoxin Unit/mg of vancomycin
• Particulate Matter in Injections 788:
Meets the requirements for small-volume injections
788:
Meets the requirements for small-volume injections
• Sterility Tests 71:
Meets the requirements when tested as directed in Test for Sterility of the Product to Be Examined, Membrane Filtration, except use water instead of Diluting Fluid A
71:
Meets the requirements when tested as directed in Test for Sterility of the Product to Be Examined, Membrane Filtration, except use water instead of Diluting Fluid A
• Other Requirements:
Meets the requirements in Injections 1
1
ADDITIONAL REQUIREMENTS
• Packaging and Storage:
Preserve as described in Injections 1, Containers for Injections. Maintain in a frozen state.
1, Containers for Injections. Maintain in a frozen state.
• Labeling:
It meets the requirements in Injections 1, Labeling. The label states that it is to be thawed just before use, describes conditions for proper storage of the resultant solution, and directs that the solution is not to be refrozen.
1, Labeling. The label states that it is to be thawed just before use, describes conditions for proper storage of the resultant solution, and directs that the solution is not to be refrozen.
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Auxiliary Information—
Please check for your question in the FAQs before contacting USP.
| Topic/Question | Contact | Expert Committee |
|---|---|---|
| Monograph | Ahalya Wise, M.S.
Senior Scientific Liaison 1-301-816-8161 |
(SM12010) Monographs - Small Molecules 1 |
| 85 |
Radhakrishna S Tirumalai, Ph.D.
Principal Scientific Liaison 1-301-816-8339 |
(GCM2010) General Chapters - Microbiology |
| 71 |
Radhakrishna S Tirumalai, Ph.D.
Principal Scientific Liaison 1-301-816-8339 |
(GCM2010) General Chapters - Microbiology |
| Reference Standards | RS Technical Services 1-301-816-8129 rstech@usp.org |
USP35–NF30 Page 5003
Pharmacopeial Forum: Volume No. 27(4) Page 2784