Insulin Injection
»Insulin Injection is an isotonic,sterile solution of Insulin.It has a potency of not less than 95.0percent and not more than 105.0percent of the potency stated on the label,expressed in USP Insulin Units.
Packaging and storage— Preserve in the unopened multiple-dose container provided by the manufacturer.Do not repackage.Store in a refrigerator,protect from sunlight,and avoid freezing.
Labeling— Label it to indicate the one or more animal species to which it is related,as pork,as beef,or as a mixture of pork and beef.If the Insulin Injection is made from Insulin that is purified,label it as such.Label it to state that it is to be stored in a refrigerator and that freezing is to be avoided.The label states the potency in USP Insulin Units per mL.
Identification— The retention time of the insulin peak in the chromatogram of the Assay preparationcorresponds to the retention time of the appropriate species in the chromatogram of the Identification preparation,as obtained in the Assay.[NOTE—It may be necessary to inject a mixture of Assay preparation and Identification preparation.]
Bacterial endotoxins á85ñ— It contains not more than 80USP Endotoxin Units for each 100USP Insulin Units.
Sterility á71ñ— It meets the requirements when tested as directed for Membrane Filtrationunder Test for Sterility of the Product to be Examined.
pHá791ñ: between 7.0and 7.8,determined potentiometrically.
Particulate matter á788ñ: meets the requirements for small-volume injections.
Zinc content á591ñ: between 10and 40µg for each 100USP Insulin Units of appropriate species.
Limit of high molecular weight proteins—
Arginine solution ,Mobile phase,System suitability solution,and Chromatographic system—Proceed as directed in the test for Limit of high molecular weight proteinsunder Insulin.
Test solution— Quantitatively add 4µLof 6Nhydrochloric acid per mLof an accurately measured volume of Injection,and mix.
Procedure— Proceed as directed for Procedurein the test for Limit of high molecular weight proteinsunder Insulin.Not more than 2.0%is found.
Other requirements— It meets the requirements under Injections á1ñ.
Assay—
Mobile phase,Identification preparation,Standard preparation,System suitability solution,and Chromatographic system— Proceed as directed in the Assayunder Insulin.
NOTE—The Identification preparation,Standard preparation,and Assay preparationmay be stored at room temperature for up to 12hours or in a refrigerator for up to 48hours.
Assay preparation 1 (for Injection labeled as containing 40USP Insulin Units per mL)—Add 2.5µLof 9.6Nhydrochloric acid per mLof an accurately measured volume of Injection.Allow the suspension,if present,to clarify,and mix.
Assay preparation 2 (for Injection labeled as containing 100USP Insulin Units per mL)—Add 2.5µLof 9.6Nhydrochloric acid per mLof an accurately measured volume of Injection.Allow the suspension,if present,to clarify,and mix.[NOTE—Pooling of several package units may be necessary to obtain sufficient volume of the test specimen.]Pipet 2mLof this solution into a 5-mLvolumetric flask,dilute with 0.01Nhydrochloric acid to volume,and mix.
Procedure— Separately inject equal volumes (about 20µL)of the appropriate Assay preparation,the Identification preparation,and the Standard preparationinto the chromatograph,record the chromatograms,and measure the peak responses for insulin and A-21desamido insulin,using the chromatogram of the Identification preparationto identify the insulin peaks.For Insulin Injection prepared from a single species,calculate the potency,in USP Insulin Units per mL,of the Injection taken by the formula:
(CD)(SrU/SrS),
in which Cis the concentration,in USP Insulin Units per mL,of USP Insulin RSin the Standard preparation;Dis the dilution factor;and SrUand SrSare the sums of the areas of the insulin and A-21desamido insulin peaks obtained from the chromatograms of the Assay preparationand the Standard preparation,respectively.For Injection prepared from a mixture of beef and pork insulins,calculate the total potency as the sum of the potencies of both beef and pork insulins,determined as directed above.
Auxiliary Information— Staff Liaison:Larry N.Callahan,Ph.D.,Scientist
Expert Committee:(BNT)Biotechnology and Natural Therapeutics/Diagnostics
USP28–NF23Page 1021
Pharmacopeial Forum:Volume No.27(2)Page 2168
Phone Number:1-301-816-8385