» Insulin Injection is an isotonic, sterile solution of Insulin. It has a potency of not less than 95.0 percent and not more than 105.0 percent 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.
USP Reference standards 11
USP Endotoxin RS
Identification The retention time of the insulin peak in the chromatogram of the Assay preparation corresponds to the retention time of the appropriate species in the chromatogram of the Identification preparation, as obtained in the Assay. [noteIt may be necessary to inject a mixture of Assay preparation and Identification preparation. ]
Bacterial endotoxins 85 It contains not more than 80 USP Endotoxin Units for each 100 USP Insulin Units.
Sterility 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 7.0 and 7.8, determined potentiometrically.
Particulate matter 788: meets the requirements for small-volume injections.
Zinc content 591: between 10 and 40 µg for each 100 USP 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 proteins under Insulin.
Test solution Quantitatively add 4 µL of 6 N hydrochloric acid per mL of an accurately measured volume of Injection, and mix.
Procedure Proceed as directed for Procedure in the test for Limit of high molecular weight proteins under Insulin. Not more than 2.0% is found.
Other requirements It meets the requirements under Injections 1.
Mobile phase, Identification preparation, Standard preparation, System suitability solution, and Chromatographic system Proceed as directed in the Assay under Insulin.
[noteThe Identification preparation, Standard preparation, and Assay preparation may be stored at room temperature for up to 12 hours or in a refrigerator for up to 48 hours. ]
Assay preparation 1 (for Injection labeled as containing 40 USP Insulin Units per mL)Add 2.5 µL of 9.6 N hydrochloric acid per mL of an accurately measured volume of Injection. Allow the suspension, if present, to clarify, and mix.
Assay preparation 2 (for Injection labeled as containing 100 USP Insulin Units per mL)Add 2.5 µL of 9.6 N hydrochloric acid per mL of an accurately measured volume of Injection. Allow the suspension, if present, to clarify, and mix. [notePooling of several package units may be necessary to obtain sufficient volume of the test specimen. ] Pipet 2 mL of this solution into a 5-mL volumetric flask, dilute with 0.01 N hydrochloric acid to volume, and mix.
Procedure Separately inject equal volumes (about 20 µL) of the appropriate Assay preparation, the Identification preparation, and the Standard preparation into the chromatograph, record the chromatograms, and measure the peak responses for insulin and A-21 desamido insulin, using the chromatogram of the Identification preparation to 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 C is the concentration, in USP Insulin Units per mL, of USP Insulin RS in the Standard preparation; D is the dilution factor; and SrU and SrS are the sums of the areas of the insulin and A-21 desamido insulin peaks obtained from the chromatograms of the Assay preparation and 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.
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USP35NF30 Page 3505Pharmacopeial Forum: Volume No. 27(2) Page 2168