INTRODUCTION
Glucagon is a peptide hormone that increases blood glucose levels via release of liver glycogen stores. A robust and precise physicochemical chromatographic procedure is used in the glucagon assay to assign potency on a mass basis. Bioidentity is still required in Glucagon for Injection, and two procedure options are presented here: an in vivo procedure based on release of glucose from freshly prepared rat liver cells (hepatocytes) stimulated with glucagon ex vivo or production of cyclic adenosine monophosphate (cAMP) in vitro in response to glucagon stimulation of the glucagon receptor cell line. To meet the acceptance criteria of the bioidentity test, only one of these bioidentity tests is required.
PROCEDURE
Change to read:
•A. Primary Liver Cell Bioidentity Test
[Note—All buffers are oxygenated, prepared with either Sterile Water for Injection or Sterile Water for Irrigation, warmed to 37°, and adjusted to a final pH of 7.4 unless otherwise indicated. At least two independent assays (replicates) must be performed utilizing two rat livers for each lot of glucagon. Figure 1 demonstrates the process used to generate one replicate value. A minimum of two replicates are combined according to the Calculations section. The concentration range of the Standard preparations and Assay preparations may be modified to fall within the linear range of the Assay, and the calculations can be adjusted accordingly. Alternatively, full curve analysis using validated nonlinear statistical methods can be used, provided that similarity is demonstrated when analysts compare the responses of the Standard preparations and Assay preparations.]
Hepatocyte preparation
Calcium-free perfusion buffer with dextrose:Prepare a solution containing 7.92 g/L of sodium chloride, 0.35 g/L of potassium chloride, 1.80 g/L of dextrose, 0.19 g/L of edetic acid (EDTA), and 2.38 g/L of 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES). Oxygenate before use.
Source from USP:https://online.uspnf.com/uspnf/document/2_GUID-1E931FF5-3C40-47BE-9249-1653566221F5_60101_en-US
Copyright © Shijiazhuang Xinfuda Medical Packaging Co., Ltd. All Rights
MAKE AN ENQUIRY
