Vancomycin Hydrochloride for Injection
DEFINITION
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.
IDENTIFICATION
ASSAY
• Antibiotics—Microbial Assays 
81
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 Antibiotics—Microbial Assays 81. [Note—Use 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. ]
81. [Note—Use 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%
PERFORMANCE TESTS
• Uniformity of Dosage Units 905:
Meets the requirements
905:
Meets the requirements
SPECIFIC TESTS
• Particulate Matter in Injections 788:
Meets the requirements under small-volume injections
788:
Meets the requirements under small-volume injections
• Bacterial Endotoxins Test 85:
NMT 0.33 USP Endotoxin Unit/mg of vancomycin
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.
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:
2.5–4.5, 50 mg/mL in water
791:
2.5–4.5, 50 mg/mL in water
• Water Determination, Method I 921:
NMT 5.0%
921:
NMT 5.0%
• Injections, Constituted Solutions 1:
Meets the requirements at the time of use.
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.
[Note—Make adjustments if necessary, changing the acetonitrile proportion in Solution B to obtain a retention time of 7.5–10.5 min for the main vancomycin peak. ]
| Time (min) |
Solution B (%) |
Solution C (%) |
|---|---|---|
| 0 | 100 | 0 |
| 12 | 100 | 0 |
| 20 | 0 | 100 |
| 22 | 0 | 100 |
| 23 | 100 | 0 |
| 30 | 100 | 0 |
System suitability solution:
0.5 mg/mL of USP Vancomycin Hydrochloride RS. Heat at 65
for 48 h, and allow to cool.
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
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
[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 C increases from 0% to 100%. ]
Suitability requirements
Resolution:
NLT 3.0 between compound 1 and vancomycin B
Column efficiency:
NLT 1500 theoretical plates, calculated from the vancomycin B peak
Analysis
Samples:
Sample solution 1 and Sample solution 2
[Note—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. ]
[Note—Correct 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
| D | = | = dilution factor, Sample solution 1 to Sample solution 2, 25 |
| rB | = | = corrected area response of the main peak from Sample solution 2 |
| rA | = | = sum of the corrected area responses of all the peaks, other than the main peak, from Sample solution 1 |
Calculate the percentage of each other peak taken:
Result = [rI/(D × rB) + rA] × 100
| rI | = | = corrected area response of any individual peak, other than the main peak, from Sample solution 1 |
| D | = | = dilution factor, Sample solution 1 to Sample solution 2, 25 |
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.
1.
ADDITIONAL REQUIREMENTS
• Packaging and Storage:
Preserve as described in Injections 1, Containers for Sterile Solids.
1, Containers for Sterile Solids.
• Labeling:
Meets the requirements under Injections 1
1
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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 5004
Pharmacopeial Forum: Volume No. 35(4) Page 881