{"id":999,"date":"2025-09-17T10:38:56","date_gmt":"2025-09-17T03:38:56","guid":{"rendered":"https:\/\/nhathuocngocanh.com\/bp\/?p=999"},"modified":"2025-10-01T17:46:24","modified_gmt":"2025-10-01T10:46:24","slug":"human-glucagon","status":"publish","type":"post","link":"https:\/\/nhathuocngocanh.com\/bp\/human-glucagon\/","title":{"rendered":"Human Glucagon"},"content":{"rendered":"<p><em>(Glucagon, Human, Ph. Eur. monograph 1635)<\/em><\/p>\n<p>C<sub>153<\/sub>H<sub>225<\/sub>N<sub>43<\/sub>O<sub>49<\/sub>S\u00a0 \u00a03483<\/p>\n<p><strong>Action and use<\/strong><\/p>\n<p>Hormone; treatment of hypoglycaemia.<\/p>\n<p><strong>Preparation<\/strong><\/p>\n<p>Human Glucagon Injection<\/p>\n<h2>DEFINITION<\/h2>\n<p>Polypeptide having the same structure (29 amino acids) as the hormone produced by the \u03b1-cells of the human pancreas, which increases the blood-glucose concentration by promoting rapid breakdown of liver glycogen.<\/p>\n<h3>Content<\/h3>\n<p>92.5 per cent to 105.0 per cent (anhydrous substance).<\/p>\n<h2>PRODUCTION<\/h2>\n<p>Human glucagon is produced by a method based on recombinant DNA (rDNA) technology. During the course of product development it must be demonstrated that the manufacturing process produces a product having a biological activity of not less than 1 IU\/mg using a suitable validated bioassay.<\/p>\n<h3>Host-cell-derived proteins<\/h3>\n<p>The limit is approved by the competent authority.<\/p>\n<h3>Host-cell- and vector-derived DNA<\/h3>\n<p>The limit is approved by the competent authority.<\/p>\n<h2>CHARACTERS<\/h2>\n<h3>Appearance<\/h3>\n<p>White or almost white powder.<\/p>\n<h3>Solubility<\/h3>\n<p>Practically insoluble in water and in most organic solvents. It is soluble in dilute mineral acids and in dilute solutions of alkali hydroxides.<\/p>\n<h2>IDENTIFICATION<\/h2>\n<p>A. Peptide mapping. Liquid chromatography (2.2.29).<\/p>\n<p><em>Test solution<\/em>\u00a0 Prepare a 5 mg\/mL solution of the substance to be examined in 0.01 M hydrochloric acid. Mix 200 \u03bcL of this solution with 800 \u03bcL of 0.1 M ammonium carbonate buffer solution pH 10.3 R (diluted stock solution). Prepare a 2 mg\/mL solution of \u03b1-chymotrypsin for peptide mapping R in 0.1 M ammonium carbonate buffer solution pH 10.3 R and add 25 \u03bcL of this solution to the diluted stock solution. Place the solution in a closed vial at 37 \u00b0C for 2 h. Remove the vial and stop the reaction immediately by adding 120 \u03bcL of glacial acetic acid R.<\/p>\n<p><em>Reference solution\u00a0<\/em> Prepare a 1 mg\/mL solution of human glucagon CRS in 0.1 M ammonium carbonate buffer solution pH 10.3 R (diluted stock solution) and continue as described for the test solution.<\/p>\n<p><em>Column:<\/em><\/p>\n<p><em>\u2014 size:<\/em> l = 0.05 m, \u00d8 = 4 mm;<\/p>\n<p><em>\u2014 stationary phase:<\/em> octadecylsilyl silica gel for chromatography R (5 \u03bcm).<\/p>\n<p><em>Mobile phase:<\/em><\/p>\n<p><em>\u2014 mobile phase A:<\/em> mix 500 \u03bcL of trifluoroacetic acid R and 1000 mL of water R;<\/p>\n<p><em>\u2014 mobile phase B:<\/em> mix 500 \u03bcL of trifluoroacetic acid R with 600 mL of anhydrous ethanol R and add 400 mL of<br \/>\nwater R;<\/p>\n<table style=\"border-collapse: collapse; width: 100%; height: 42px;\">\n<tbody>\n<tr style=\"height: 21px;\">\n<td style=\"width: 33.3333%; height: 21px; text-align: center;\"><strong>Time<\/strong><\/p>\n<p><strong>(min)<\/strong><\/td>\n<td style=\"width: 33.3333%; height: 21px; text-align: center;\"><strong>Mobile phase A<\/strong><\/p>\n<p><strong>(per cent V\/V)<\/strong><\/td>\n<td style=\"width: 33.3333%; height: 21px; text-align: center;\"><strong>Mobile phase B<\/strong><\/p>\n<p><strong>(per cent V\/V)<\/strong><\/td>\n<\/tr>\n<tr style=\"height: 21px;\">\n<td style=\"width: 33.3333%; height: 21px; text-align: center;\">0 &#8211; 35<\/p>\n<p>35 &#8211; 45<\/p>\n<p>45 &#8211; 46<\/p>\n<p>46 &#8211; 75<\/td>\n<td style=\"width: 33.3333%; height: 21px; text-align: center;\">100 \u2192 53<\/p>\n<p>53 \u2192 0<\/p>\n<p>0 \u2192 100<\/p>\n<p>100<\/td>\n<td style=\"width: 33.3333%; height: 21px; text-align: center;\">0 \u2192 47<\/p>\n<p>47 \u2192 100<\/p>\n<p>100 \u2192 0<\/p>\n<p>0<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p><em>Flow rate\u00a0<\/em> 1.0 mL\/min.<\/p>\n<p><em>Detection<\/em>\u00a0 Spectrophotometer at 215 nm.<\/p>\n<p><em>Equilibration<\/em>\u00a0 With mobile phase A for at least 15 min.<\/p>\n<p><em>Injection<\/em>\u00a0 20 \u03bcL.<\/p>\n<p><em>System suitability<\/em>\u00a0 The chromatogram obtained with the reference solution is qualitatively similar to the chromatogram supplied with human glucagon CRS.<\/p>\n<p><em>Results<\/em>\u00a0 The profile of the chromatogram obtained with the test solution corresponds to that of the chromatogram obtained with the reference solution.<\/p>\n<p>B. Examine the chromatograms obtained in the assay.<\/p>\n<p><em>Results\u00a0<\/em> The principal peak in the chromatogram obtained with the test solution is similar in retention time to the principal peak in the chromatogram obtained with reference solution (a).<\/p>\n<h2>TESTS<\/h2>\n<h3>Related proteins and deamidated forms<\/h3>\n<p>Liquid chromatography (2.2.29): use the normalisation procedure.<\/p>\n<p><em>Test solution\u00a0<\/em> Dissolve the substance to be examined in 0.01 M hydrochloric acid to obtain a concentration of 0.5 mg\/mL. Maintain the solution at 2-8 \u00b0C.<\/p>\n<p><em>Reference solution (a)<\/em> Dissolve the contents of a vial of human glucagon CRS in 0.01 M hydrochloric acid to obtain a concentration of 0.5 mg\/mL. Maintain the solution at 2-8 \u00b0C.<\/p>\n<p><em>Reference solution (b)\u00a0<\/em> Dissolve the substance to be examined in 0.01 M hydrochloric acid to obtain a concentration of about 0.5 mg\/mL. Heat at 50 \u00b0C for 48 h (in situ preparation of all 4 deamidated forms of glucagon at a total concentration of not less than 7 per cent).<\/p>\n<p><em>Column:<\/em><\/p>\n<p><em>\u2014 size:<\/em> l = 0.15 m, \u00d8 = 3 mm;<\/p>\n<p><em>\u2014 stationary phase:<\/em> octadecylsilyl silica gel for chromatography R (3 \u03bcm);<\/p>\n<p><em>\u2014 temperature:<\/em> 45 \u00b0C.<\/p>\n<p><em>Mobile phase:<\/em><\/p>\n<p><em>\u2014 mobile phase A:<\/em> dissolve 16.3 g of potassium dihydrogen phosphate R in 800 mL of water R, adjust to pH 2.7 with phosphoric acid R and add 200 mL of acetonitrile for chromatography R;<\/p>\n<p><em>\u2014 mobile phase B:<\/em> acetonitrile for chromatography R, water R (40:60 V\/V);<\/p>\n<table style=\"border-collapse: collapse; width: 100%; height: 42px;\">\n<tbody>\n<tr style=\"height: 21px;\">\n<td style=\"width: 33.3333%; height: 21px; text-align: center;\"><strong>Time<\/strong><\/p>\n<p><strong>(min)<\/strong><\/td>\n<td style=\"width: 33.3333%; height: 21px; text-align: center;\"><strong>Mobile phase A<\/strong><\/p>\n<p><strong>(per cent V\/V)<\/strong><\/td>\n<td style=\"width: 33.3333%; height: 21px; text-align: center;\"><strong>Mobile phase B<\/strong><\/p>\n<p><strong>(per cent V\/V)<\/strong><\/td>\n<\/tr>\n<tr style=\"height: 21px;\">\n<td style=\"width: 33.3333%; height: 21px; text-align: center;\">0 &#8211; 25<\/p>\n<p>25 &#8211; 29<\/p>\n<p>29 &#8211; 30<\/p>\n<p>30 &#8211; 31<\/td>\n<td style=\"width: 33.3333%; height: 21px; text-align: center;\">61<\/p>\n<p>61 \u2192 12<\/p>\n<p>12<\/p>\n<p>12 \u2192 61<\/td>\n<td style=\"width: 33.3333%; height: 21px; text-align: center;\">39<\/p>\n<p>39 \u2192 88<\/p>\n<p>88<\/p>\n<p>88 \u2192 39<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p><em>NOTE<\/em>\u00a0 The end time of the isocratic elution may be adjusted so that the gradient begins after elution of the peak due to deamidated glucagon 4 (see relative retention below).<\/p>\n<p><em>Flow rate\u00a0<\/em> 0.5 mL\/min.<\/p>\n<p><em>Detection<\/em>\u00a0 Spectrophotometer at 214 nm.<\/p>\n<p><em>Injection<\/em>\u00a0 15 \u03bcL.<\/p>\n<p><em>Relative retention<\/em>\u00a0 With reference to glucagon (retention time = about 21 min): deamidated glucagon 1 = about 1.1; deamidated glucagon 4 = about 1.4.<\/p>\n<p><em>System suitability:<\/em><\/p>\n<p><em>\u2014 resolution:<\/em> minimum 1.5 between the peaks due to glucagon and deamidated glucagon 1 in the chromatogram obtained with reference solution (b);<\/p>\n<p><em>\u2014 symmetry factor:<\/em> maximum 1.8 for the peak due to glucagon in the chromatogram obtained with reference solution (a);<\/p>\n<p><em>\u2014 repeatability:<\/em> maximum relative standard deviation of 2.0 per cent after 5 injections of reference solution (a);<\/p>\n<p>\u2014 4 peaks eluting after the principal peak, that correspond to the deamidated forms, are clearly visible in the<br \/>\nchromatogram obtained with reference solution (b).<\/p>\n<p><em>Limits:<\/em><\/p>\n<p><em>\u2014 deamidated forms:<\/em> maximum 0.8 per cent;<\/p>\n<p><em>\u2014 total:<\/em> maximum 3.0 per cent.<\/p>\n<h3>Water (2.5.32)<\/h3>\n<p>Maximum 10 per cent, determined on 50 mg.<\/p>\n<h3>Bacterial endotoxins (2.6.14)<\/h3>\n<p>Less than 10 IU\/mg.<\/p>\n<h2>ASSAY<\/h2>\n<p>Liquid chromatography (2.2.29) as described in the test for related proteins and deamidated forms with the following modification.<\/p>\n<p><em>Injection\u00a0<\/em> Test solution and reference solution (a).<\/p>\n<p>Calculate the percentage content of human glucagon (C153H225N43O49S) taking into account the assigned content of C153H225N43O49S in human glucagon CRS.<\/p>\n<h2>STORAGE<\/h2>\n<p>In an airtight container, protected from light, at a temperature lower than -15 \u00b0C.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>(Glucagon, Human, Ph. Eur. monograph 1635) C153H225N43O49S\u00a0 \u00a03483 Action and use Hormone; treatment of hypoglycaemia. Preparation Human Glucagon Injection DEFINITION Polypeptide having the same structure (29 amino acids) as the hormone produced by the \u03b1-cells of the human pancreas, which increases the blood-glucose concentration by promoting rapid breakdown of liver glycogen. Content 92.5 per cent&#8230;<\/p>\n","protected":false},"author":4,"featured_media":2065,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[174],"tags":[],"class_list":["post-999","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-medicinal-substances"],"acf":[],"_links":{"self":[{"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/posts\/999","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/users\/4"}],"replies":[{"embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/comments?post=999"}],"version-history":[{"count":6,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/posts\/999\/revisions"}],"predecessor-version":[{"id":7370,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/posts\/999\/revisions\/7370"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/media\/2065"}],"wp:attachment":[{"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/media?parent=999"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/categories?post=999"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/tags?post=999"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}