U.S. PHARMACOPEIA

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Alteplase for Injection
» Alteplase for Injection is a sterile lyophilized preparation of Alteplase. Its biological activity is not less than 90 percent and not more than 115 percent of that stated on the label in USP Alteplase Units. It contains not less than 95 percent and not more than 111 percent 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 Solutions under Injections 1.
Identification— It responds to the tests for Identification and Peptide mapping under Alteplase.
Bacterial endotoxins 85— It contains less than 1 USP Endotoxin Unit per mg.
Safety— It meets the requirements for biologics as set forth for Safety Tests—Biologicals under Biological Reactivity Tests, In Vivo 88.
Sterility 71— It meets the requirements when tested as directed for Membrane Filtration under Test for Sterility of the Product to be Examined.
Uniformity of dosage units 905: meets the requirements for Content Uniformity.
pH 791: between 7.1 and 7.5, in the solution constituted as directed in the labeling.
Water, Method I 921: not more than 4.0%.
Single-chain content— When constituted with water, it meets the requirements for Single-chain content under Alteplase.
Percent monomer—
Mobile phase— Dissolve 34.84 g of arginine, 158.56 g of ammonium sulfate, and 100 mL of isopropyl alcohol in water, and dilute with water to 1000 mL. Adjust with phosphoric acid to a pH of 7.3, degas, and pass through a 0.45-µm porosity filter. Make adjustments if necessary (see System Suitability under Chromatography 621).
Standard solution— Dissolve an accurately weighed quantity of USP Alteplase RS in water to obtain a solution having a known concentration of about 1 mg per mL.
Test solution— Dissolve an accurately weighed quantity of Alteplase for Injection in water to obtain a solution having a concentration of about 1 mg per mL.
Resolution solution— Prepare a solution containing 1 mg per mL of each of chicken ovalbumin and bovine gamma globulin.
Chromatographic system (see Chromatography 621)— 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 0.5 and 1.0 mL per 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 1200 theoretical plates.
Procedure— Separately inject equal volumes (about 50 µL) of the Standard solution and the Test solution into 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 rm is the peak response for the alteplase monomer, and rS is 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 content under Alteplase.
Assay for biological potency— When constituted with water, Alteplase for Injection meets the requirements for Assay for biological potency under Alteplase.
Auxiliary Information— Staff Liaison : Larry N. Callahan, Ph.D., Scientist
Expert Committee : (BBPP05) Biologics and Biotechnology - Proteins and Polysaccharides
USP29–NF24 Page 83
Pharmacopeial Forum : Volume No. 27(3) Page 2514
Phone Number : 1-301-816-8385