Oxaliplatin Injection
DEFINITION
Oxaliplatin Injection is a sterile solution of Oxaliplatin in Water for Injection. It contains NLT 90.0% and NMT 110.0% of the labeled amount of oxaliplatin (C8H14N2O4Pt).
IDENTIFICATION
•  A. Ultraviolet Absorption 197U
Sample solution:  100 µg/mL
Medium:  Water
•  B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.
ASSAY
[Note—All HPLC autosampler vials should be made of polypropylene. ]
•  Procedure
Acidified water:  Adjust with phosphoric acid to a pH of 3.0.
Mobile phase:  Acetonitrile and Acidified water (1:99)
System suitability solution:  0.1 mg/mL of USP Oxaliplatin RS and 0.1 mg/mL of USP Oxaliplatin System Suitability RS in water. [Note—USP Oxaliplatin System Suitability RS is [SP-4-2-(1R-trans)]-(1,2-cyclohexanediamine-N,N') dichloridoplatinum(II). ]
Standard solution:  0.1 mg/mL of USP Oxaliplatin RS in water
Sample solution:  0.1 mg/mL of oxaliplatin in water, from the combined contents of NLT three vials of Injection
Chromatographic system 
Mode:  LC
Detector:  UV 210 nm
Column:  4.6-mm × 25-cm; 5-µm packing L1
Column temperature:  40
Flow rate:  1.2 mL/min
Injection size:  20 µL
System suitability 
Sample:  System suitability solution
[Note—The relative retention times for USP Oxaliplatin System Suitability RS and oxaliplatin are 0.9 and 1.0, respectively. ]
Suitability requirements 
Resolution:  NLT 2.0 between USP Oxaliplatin System Suitability RS and oxaliplatin
Tailing factor:  NMT 2.0, oxaliplatin peak
Relative standard deviation:  NMT 1.0%, oxaliplatin peak
Analysis 
Samples:  Standard solution and Sample solution
Calculate the percentage of C8H14N2O4Pt in the portion of Injection taken:
Result = (rU/rS) × (CS/CU) × 100
rU== peak response from the Sample solution
rS== peak response from the Standard solution
CS== concentration of USP Oxaliplatin RS in the Standard solution (mg/mL)
CU== nominal concentration of oxaliplatin in the Sample solution (mg/mL)
Acceptance criteria:  90.0%–110.0%
IMPURITIES
Organic Impurities 
•  Procedure 1: Limit of Oxalic Acid [Note—All HPLC autosampler vials should be made of polypropylene. ]
Solution A:  Dissolve 1.36 g of monobasic potassium phosphate in 10 mL of 10% tetrabutylammonium hydroxide, dilute with water to 1 L, and adjust with phosphoric acid to a pH of 6.0.
Mobile phase:  Acetonitrile and Solution A (1:4)
Standard solution:  35 µg/mL of USP Oxaliplatin Related Compound A RS in water. [Note—USP Oxaliplatin Related Compound A RS is available as dihydrate oxalic acid. ]
System suitability solution:  0.1 mg/mL of succinic acid in Standard solution
Sensitivity solution:  3.5 µg/mL of USP Oxaliplatin Related Compound A RS in water, from the Standard solution
Sample solution:  Combined contents of NLT three vials of Injection
Chromatographic system 
Mode:  LC
Detector:  UV 210 nm
Column:  4.6-mm × 25-cm; 5-µm packing L1
Column temperature:  40
Flow rate:  2 mL/min
Injection size:  10 µL
System suitability 
Samples:  Standard solution, System suitability solution, and Sensitivity solution
[Note—The relative retention times for succinic acid and oxalic acid are 0.8 and 1.0, respectively. ]
Suitability requirements 
Resolution:  NLT 2.0 between succinic acid and oxalic acid, System suitability solution
Tailing factor:  Between 0.5 and 2.0, oxalic acid peak, System suitability solution
Signal-to-noise ratio:  NLT 10, Sensitivity solution
Relative standard deviation:  NMT 3.0%, Standard solution
Analysis 
Sample:  Standard solution and Sample solution
Calculate the percentage of each impurity in the portion of Injection taken:
Result = (rU/rS) × (CS/CU) × (Mr1/Mr2) × 100
rU== peak response of oxalic acid from the Sample solution
rS== peak response of oxalic acid from the Standard solution
CS== concentration of USP Oxaliplatin Related Compound A RS in the Standard solution (mg/mL)
CU== concentration of oxaliplatin in the Sample solution (mg/mL)
Mr1== molecular weight of anhydrous oxalic acid, 90.03
Mr2== molecular weight of oxaliplatin related compound A, 126.07
Acceptance criteria 
Oxalic acid:  NMT 0.6%
•  Procedure 2: Limit of (SP-4-2)-Diaqua[(1R,2R)-cyclohexane-1,2-diamine-N,N']platinum and Unspecified Impurities
[Note—All HPLC autosampler vials should be made of polypropylene. ]
Solution A:  Dissolve 1.36 g of monobasic potassium phosphate and 0.55 g of sodium heptanesulfonate in 1 L of water. Adjust with phosphoric acid to a pH of 3.0.
Solution B:  Methanol and Solution A (19:81)
Solution C:  Methanol and Solution A (50.5:49.5)
Mobile phase:   See the gradient table below.
Time
(min)
Solution B
(%)
Solution C
(%)
0 100 0
45.0 0 100
45.5 100 0
53.0 100 0
System suitability solution:  2 mg/mL of USP Oxaliplatin RS in 0.005 M sodium hydroxide. Allow this solution to stand at room temperature for at least 5 days. Transfer 5 mL of this solution into a 50-mL volumetric flask, and dilute with water to volume. [Note—The preparation of the System suitability solution forms (SP-4-2)-diaqua[(1R,2R)-cyclohexane-1,2-diamine-N,N']platinum and diaquodiaminocyclohexaneplatinum dimer. ]
Standard stock solution:  Transfer a weighed quantity of USP Oxaliplatin Related Compound B RS into a suitable volumetric flask, add a volume of methanol equivalent to about 25% of the final volume, and sonicate for approximately 2 min to disperse the solids. Add a volume of 0.01 M nitric acid equivalent to about 65% of the final volume, and sonicate for approximately 30 min to dissolve. Allow to cool if necessary, and dilute with 0.01 M nitric acid to volume to obtain a solution of concentration of 0.125 mg/mL.
Standard solution:  31.25 µg/mL of USP Oxaliplatin Related Compound B RS in 0.01 M nitric acid, from the Standard stock solution. [Note—USP Oxaliplatin Related Compound B RS is converted to (SP-4-2)-diaqua[(1R,2R)-cyclohexane-1,2-diamine-N,N']platinum in the Standard solution preparation. ]
Sample solution:  Combined contents of NLT three vials of Injection
Chromatographic system 
Mode:  LC
Detector:  UV 210 nm
Column:  4.6-mm × 7.5-cm; 3-µm packing L1
Column temperature:  10
Flow rate:  1 mL/min
Injection size:  20 µL
System suitability 
Samples:  System suitability solution and Standard solution
Suitability requirements 
Resolution:  NLT 8.0 between the peaks of (SP-4-2)-diaqua[(1R,2R)-cyclohexane-1,2-diamine-N,N']platinum and diaquodiaminocyclohexaneplatinum dimer, System suitability solution.
Tailing factor:  NMT 2.0, (SP-4-2)-diaqua[(1R,2R)-cyclohexane-1,2-diamine-N,N']platinum peak, System suitability solution
Relative standard deviation:  NMT 3.0%, Standard solution
Analysis 
Samples:  Standard solution and Sample solution
Calculate the percentage of each impurity in the portion of Injection taken:
Result = (rU/rS) × (CS/CU) × (Mr1/Mr2) × (1/F) × 100
rU== peak response of each impurity from the Sample solution
rS== peak response of (SP-4-2)-diaqua[(1R,2R)-cyclohexane-1,2-diamine-N,N']platinum from the Standard solution
CS== concentration of USP Oxaliplatin Related Compound B RS in the Standard solution (mg/mL)
CU== concentration of oxaliplatin in the Sample solution (mg/mL)
Mr1== molecular weight of (SP-4-2)-diaqua[(1R,2R)-cyclohexane-1,2-diamine-N,N']platinum, 345.30
Mr2== molecular weight of oxaliplatin related compound B, 433.28
F== relative response factor for each individual impurity (See Impurity Table 1.)
Acceptance criteria 
Individual impurities:  See Impurity Table 1.
Total impurities:  NMT 2.45%, from Limit of oxalic acid and Limit of (SP-4-2)-Diaqua[(1R,2R)-cyclohexane-1,2-diamine-N,N']platinum and Unspecified Impurities
Impurity Table 1
Name Relative
Retention
Time
Relative
Response
Factor
Acceptance
Criteria,
NMT (%)
(SP-4-2)-Diaqua[(1R,2R)-cyclohexane-1,2-diamine-N,N']platinum 1.0 1.0 0.65
Diaquodiaminocyclohexaneplatinum dimera 1.4 2.5 0.50
Any individual unspecified impurity 4.0 0.20
a   (SP-4-2)-di-µ-Oxobis[(1R,2R)-cyclohexane-1,2-diamine-kN,kN']diplatinum.
SPECIFIC TESTS
•  Bacterial Endotoxins Test 85: It contains NMT 1.0 USP Endotoxin Units/mg of oxaliplatin.
•  Sterility Tests 71: It meets the requirements when tested as directed for Membrane Filtration under Test for Sterility of the Product to Be Examined.
•  pH 791: Between 4.0 and 7.0 using a polymer combination electrode
•  Particulate Matter in Injections 788: It meets the requirements for small-volume injections.
•  Other Requirements: It meets the requirements under Injections 1.
ADDITIONAL REQUIREMENTS
•  Packaging and Storage: Preserve in single-dose or multiple-dose containers, preferably of Type I glass, protected from light. Store at controlled room temperature. Do not freeze.
•  Labeling: Label it to indicate that it is to be diluted with a 5% dextrose solution. Oxaliplatin Injection must not be diluted in sodium chloride solutions or in chloride-containing solutions.
•  USP Reference Standards 11
USP Endotoxin RS
USP Oxaliplatin RS Click to View Structure
USP Oxaliplatin Related Compound A RS
Oxalic acid dihydrate.
    C2H2O4·2H2O        126.07
USP Oxaliplatin Related Compound B RS
[SP-4-2-(1R-trans)]-(1,2-Cyclohexanediamine-N,N¢) dinitratoplatinum(II).
    C6H14N4O6Pt         433.28
USP Oxaliplatin System Suitability RS
[SP-4-2-(1R-trans)]-(1,2-Cyclohexanediamine-N,N¢) dichloridoplatinum (II).
    C6H14Cl2N2Pt        380.17
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