Alteplase for Injection
»Alteplase for Injection is a sterile lyophilized preparation of Alteplase.Its biological activity is not less than 90percent and not more than 115percent of that stated on the label in USP Alteplase Units.It contains not less than 95percent and not more than 111percent of the total protein content stated on the label.
Packaging and storage— Preserve in hermetic,light-resistant containers,and store in a refrigerator.
Labeling— Label it to state the biological activity in USP Alteplase Units per vial and the amount of protein per vial.
Constituted solution— At the time of use,it meets the requirements for Constituted Solutionsunder Injections á1ñ.
Identification— It responds to the tests for Identificationand Peptide mappingunder Alteplase.
Bacterial endotoxins á85ñ— It contains less than 1USP Endotoxin Unit per mg.
Safety— It meets the requirements for biologics as set forth for Safety Tests—Biologicalsunder Biological Reactivity Tests,In Vivo á88ñ.
Sterility á71ñ— It meets the requirements when tested as directed for Membrane Filtrationunder Test for Sterility of the Product to be Examined.
Uniformity of dosage units á905ñ: meets the requirements for Content Uniformity.
pHá791ñ: between 7.1and 7.5,in the solution constituted as directed in the labeling.
Water,Method Iá921ñ: not more than 4.0%.
Single-chain content— When constituted with water,it meets the requirements for Single-chain contentunder Alteplase.
Percent monomer—
Mobile phase— Dissolve 34.84g of arginine,158.56g of ammonium sulfate,and 100mLof isopropyl alcohol in water,and dilute with water to 1000mL.Adjust with phosphoric acid to a pHof 7.3,degas,and pass through a 0.45-µm porosity filter.Make adjustments if necessary (see System Suitabilityunder Chromatography á621ñ).
Standard solution— Dissolve an accurately weighed quantity of USP Alteplase RSin water to obtain a solution having a known concentration of about 1mg per mL.
Test solution— Dissolve an accurately weighed quantity of Alteplase for Injection in water to obtain a solution having a concentration of about 1mg per mL.
Resolution solution— Prepare a solution containing 1mg per mLof each of chicken ovalbumin and bovine gamma globulin.
Chromatographic system (see Chromatography á621ñ)— The liquid chromatograph is equipped with a 280-nm detector and a 7.5-mm ×30-cm column that contains packing L25.The flow rate is between 0.5and 1.0mLper minute.Chromatograph the Resolution solution,and record the peak responses as directed for Procedure:the resolution,R,between gamma globulin and ovalbumin is not less than 1.6.Chromatograph the Standard solution,and record the peak responses as directed for Procedure:the column efficiency determined from the alteplase peak is not less than 1200theoretical plates.
Procedure— Separately inject equal volumes (about 50µL)of the Standard solutionand the Test solutioninto the chromatograph,record the chromatograms,and measure the peak responses.Calculate the percent monomer in the portion of Alteplase for Injection taken by the formula:
100(rm/rS),
in which rmis the peak response for the alteplase monomer,and rSis the sum of the responses of all of the alteplase related peaks:not less than 95.0%is found.
Protein content— Proceed as directed for Protein contentunder Alteplase.
Assay for biological potency— When constituted with water,Alteplase for Injection meets the requirements for Assay for biological potencyunder Alteplase.
Auxiliary Information— Staff Liaison:Larry N.Callahan,Ph.D.,Scientist
Expert Committee:(BNT)Biotechnology and Natural Therapeutics/Diagnostics
USP28–NF23Page 75
Pharmacopeial Forum:Volume No.27(3)Page 2514
Phone Number:1-301-816-8385