(Ph. Eur. monograph 2829)<\/p>\n
C181<\/sub>H291<\/sub>N55<\/sub>O51<\/sub>S2<\/sub> 4118 52232-67-4<\/p>\n Action and use<\/strong><\/p>\n Parathyroid hormone analogue; treatment of osteoporosis.<\/p>\n Tetratriacontapeptide in which the sequence of amino acids is the same as that of the 1-34 N-terminal fragment of endogeneous human parathyroid hormone (rhPTH).<\/p>\n Content<\/strong><\/p>\n 95.0 per cent to 105.0 per cent (anhydrous, acetic acid- and chloride-free substance).<\/p>\n Teriparatide 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 biologically active protein using a suitable bioassay as approved by the competent authority.<\/p>\n Prior to release, the following tests are carried out on each batch of teriparatide, unless exemption has been granted by the competent authority.<\/p>\n The limit is approved by the competent authority.<\/p>\n The limit is approved by the competent authority.<\/p>\n White or almost white, very hygroscopic powder.<\/p>\n Freely soluble in water and in methanol, practically insoluble in acetonitrile.<\/p>\n A. Peptide mapping (2.2.55).<\/p>\n Solution A<\/em> Dissolve 230 mg of anhydrous disodium hydrogen phosphate R and 60 mg of sodium dihydrogen phosphate monohydrate R in 100 mL of water R and adjust to pH 7.8 with sodium hydroxide solution R.<\/p>\n Test solution<\/em> Dissolve the substance to be examined in solution A to obtain a concentration of 1.5 mg\/mL and transfer 150 \u03bcL of the solution to a clean tube. Add 90 \u03bcL of a 0.25 mg\/mL solution of glutamyl endopeptidase for peptide mapping R in solution A. Mix and incubate at 37 \u00b0C for 18-24 h. Stop the reaction by adding 660 \u03bcL of mobile phase A to reach a final digested protein concentration of about 0.25 mg\/mL.<\/p>\n NOTE: if a teriparatide concentration of 1.5 mg\/mL is not obtainable, a similar ratio of micrograms of endopeptidase per milligram of teriparatide may be used.<\/p>\n Reference solution<\/em> Prepare at the same time and in the same manner as for the test solution but using teriparatide CRS instead of the substance to be examined.<\/p>\n Blank solution<\/em> Prepare at the same time and in the same manner as for the test solution but omitting the substance to be examined.<\/p>\n Liquid chromatography (2.2.29). Store the solutions at 2-8 \u00b0C and use them within 72 h.<\/p>\n Column:<\/p>\n \u2014 size: l = 0.15 m, \u00d8 = 4.6 mm;<\/p>\n \u2014 stationary phase: octadecylsilyl silica gel for chromatography R (3.5 \u03bcm) with a pore size of 30 nm;<\/p>\n \u2014 temperature: 40 \u00b0C.<\/p>\n Mobile phase:<\/p>\n \u2014 mobile phase A: mix 1 mL of trifluoroacetic acid R and 1000 mL of water R; filter and degas;<\/p>\n \u2014 mobile phase B: mix 1 mL of trifluoroacetic acid R, 400 mL of water R and 600 mL of acetonitrile for chromatography R; filter and degas;<\/p>\n Flow rate<\/em> 1 mL\/min.<\/p>\n Detection<\/em> Spectrophotometer at 214 nm.<\/p>\n Autosampler<\/em> Set at 2-8 \u00b0C.<\/p>\n Injection<\/em> 20 \u03bcL.<\/p>\n System suitability:<\/em><\/p>\n \u2014 the chromatogram obtained with the reference solution is qualitatively similar to the chromatogram of teriparatide digest supplied with teriparatide CRS;<\/p>\n \u2014 in the chromatogram obtained with the reference solution, identify the peaks due to digest fragments I, II, III, IV and V:<\/p>\n symmetry<\/em> factor Maximum 2.3 for the peak due to fragment IV;<\/p>\n resolution<\/em> Minimum 1.5 between the peaks due to fragments I and III.<\/p>\n Results<\/em> The profile of the chromatogram obtained with the test solution corresponds to that of the chromatogram obtained with the reference solution.<\/p>\n B. Examine the chromatograms obtained in the assay.<\/p>\n Results<\/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 the reference solution.<\/p>\n Size-exclusion chromatography (2.2.30): use the normalisation procedure. Store the solutions at 2-8 \u00b0C and use them within 72 h.<\/p>\n Test solution<\/em> Dissolve the substance to be examined in water R to obtain a concentration of 1 mg\/mL.<\/p>\n Reference solution<\/em> Dissolve the contents of a vial of teriparatide CRS in water R to obtain a concentration of 1 mg\/mL.<\/p>\n Blank solution<\/em> water R.<\/p>\n Resolution solution<\/em> Incubate a vial of teriparatide CRS at 75 \u00b0C for 16-24 h. After incubation, dissolve the contents of the vial in water R to obtain a concentration of 1 mg\/mL of degraded teriparatide.<\/p>\n Column:<\/p>\n \u2014 size: l = 0.30 m, \u00d8 = 7.8 mm;<\/p>\n \u2014 stationary phase: hydrophilic silica gel for chromatography R (5-10 \u03bcm) with a pore size of 12.5 nm, of a grade suitable for fractionation of globular proteins of relative molecular mass up to 150 000.<\/p>\n Mobile phase<\/em> Add 1 mL of trifluoroacetic acid R to 750 mL of water R, mix with 250 mL of acetonitrile for chromatography R and degas.<\/p>\n Flow rate<\/em> 0.5 mL\/min.<\/p>\n Detection<\/em> Spectrophotometer at 214 nm.<\/p>\n Autosampler<\/em> Set at 2-8 \u00b0C.<\/p>\n Injection<\/em> 20 \u03bcL.<\/p>\n Run time<\/em> 1.5 times the retention time of teriparatide monomer.<\/p>\n Retention time<\/em> Teriparatide monomer = about 17 min.<\/p>\n System suitability:<\/p>\n \u2014 the chromatogram obtained with the reference solution is similar to the chromatogram supplied with teriparatide CRS;<\/p>\n \u2014 resolution: minimum 2.0 between the peaks due to teriparatide dimer and monomer in the chromatogram obtained with the resolution solution.<\/p>\n Limit:<\/p>\n \u2014 sum of the peaks eluted before the principal peak: maximum 0.3 per cent; disregard any peak with a retention time greater than that of the peak due to teriparatide monomer.<\/p>\n Liquid chromatography (2.2.29): use the normalisation procedure. Store the solutions at 2-8 \u00b0C and use them within 48 h.<\/p>\n Buffer solution<\/em> Dissolve 28.4 g of anhydrous sodium sulfate R in 900 mL of water R and adjust to pH 2.3 with phosphoric acid R. Dilute to 1000 mL with water for chromatography R and filter.<\/p>\n Test solution<\/em> Dissolve the substance to be examined in mobile phase A to obtain a concentration of 0.7 mg\/mL.<\/p>\n Reference solution<\/em> Dissolve the contents of a vial of teriparatide CRS in mobile phase A to obtain a concentration of 0.7 mg\/mL.<\/p>\n Blank solution<\/em> Mobile phase A.<\/p>\n Resolution solution<\/em> Dissolve the contents of a vial of teriparatide for system suitability CRS in mobile phase A to obtain a concentration of 1 mg\/mL.<\/p>\n Column:<\/p>\n \u2014 size: l = 0.15 m, \u00d8 = 4.6 mm;<\/p>\n \u2014 stationary phase: octadecylsilyl silica gel for chromatography R (3.5 \u03bcm) with a pore size of 30 nm;<\/p>\n \u2014 temperature: 40 \u00b0C.<\/p>\n Mobile phase:<\/p>\n \u2014 mobile phase A: mix 10 volumes of acetonitrile for chromatography R and 90 volumes of the buffer solution and degas; apply mild heating at 20 25 \u00b0C while stirring continuously during analysis;<\/p>\n \u2014 mobile phase B: mix equal volumes of acetonitrile for chromatography R and the buffer solution and degas; apply mild heating at 20-25 \u00b0C while stirring continuously during analysis;<\/p>\n Flow rate<\/em> 1.0 mL\/min.<\/p>\n Detection<\/em> Spectrophotometer at 214 nm.<\/p>\n Autosampler<\/em> Set at 2-8 \u00b0C.<\/p>\n Injection<\/em> 20 \u03bcL.<\/p>\n Relative retention<\/em> With reference to teriparatide (retention time = 20-25 min): related protein A ([MetO ,MetO]teriparatide) = about 0.40; related protein B ([MetO ]teriparatide) = about 0.49; related protein C ([MetO]teriparatide) = about 0.57; related protein D = about 1.06; related protein E = about 1.14.<\/p>\n System suitability<\/em> Resolution solution:<\/p>\n \u2014 the chromatogram obtained is similar to the chromatogram supplied with teriparatide for system suitability CRS;<\/p>\n \u2014 resolution: minimum 1.5 between the peaks due to teriparatide and related protein D;<\/p>\n \u2014 symmetry factor: 0.8 to 2.0 for the peak due to teriparatide;<\/p>\n Results:<\/p>\n \u2014 the profile of the chromatogram obtained with the test solution corresponds to that of the chromatogram obtained with the reference solution.<\/p>\n Limits:<\/p>\n \u2014 sum of related proteins A, B and C: maximum 0.5 per cent;<\/p>\n \u2014 any other related protein: maximum 0.5 per cent;<\/p>\n \u2014 total: maximum 2.0 per cent;<\/p>\n \u2014 reporting threshold: 0.05 per cent.<\/p>\n Maximum 7.0 per cent, determined on 10 mg using the evaporation technique:<\/p>\n \u2014 temperature: 100 \u00b0C;<\/p>\n \u2014 heating time: 8 min.<\/p>\n Less than 50 IU\/mg.<\/p>\n Liquid chromatography (2.2.29): use the normalisation procedure. Store the solutions at 2-8 \u00b0C and use them within 72 h.<\/p>\n Test solution<\/em> Dissolve the substance to be examined in the mobile phase to obtain a concentration of 5 mg\/mL.<\/p>\n Reference solutions<\/em> Dissolve separately 100 mg, 200 mg and 300 mg of anhydrous sodium acetate R in the mobile phase and dilute to 100 mL with the mobile phase. Further dilute 1.0 mL of each solution to 10.0 mL with the mobile phase to prepare a standard curve with acetate concentrations in the range of 0.072-0.216 mg\/mL.<\/p>\n Plot peak areas versus injected acetate content and perform linear regression to create a standard curve.<\/p>\n Column:<\/p>\n \u2014 size: l = 0.25 m, \u00d8 = 9.0 mm;<\/p>\n \u2014 stationary phase: ion-exclusion resin for chromatography R (7.5 \u03bcm).<\/p>\n Mobile phase<\/em> 0.5 per cent V\/V solution of dilute sulfuric acid R.<\/p>\n Flow rate<\/em> 1.0 mL\/min.<\/p>\n Detection<\/em> Spectrophotometer at 210 nm.<\/p>\n Autosampler<\/em> Set at 2-8 \u00b0C.<\/p>\n Injection<\/em> 100 \u03bcL.<\/p>\n Run time<\/em> 1.5 times the retention time of acetate.<\/p>\n Retention time<\/em> Acetate = about 10 min.<\/p>\n System suitability:<\/p>\n \u2014 repeatability: maximum relative standard deviation of 1.25 per cent for the area of the principal peak, determined on 3 injections of the middle reference solution;<\/p>\n \u2014 the correlation coefficient (r) calculated for the standard curve is not less than 0.999.<\/p>\n Calculate the acetate content using the standard curve and the area of the peak due to acetate in the chromatogram obtained with the test solution.<\/p>\n Liquid chromatography (2.2.29): use the normalisation procedure. Use the solutions within 72 h.<\/p>\n Test solution<\/em> Dissolve the substance to be examined in water R to obtain a concentration of 1 mg\/mL.<\/p>\n Reference solution (a)<\/em> Dissolve 165.9 mg of sodium chloride R, previously dried at 105 \u00b0C for 30 min, in water R and dilute to 100 mL with the same solvent.<\/p>\n Reference solution (b)<\/em> Dissolve 150 mg of sodium nitrite R in water R and dilute to 100 mL with the same solvent. Mix 1.0 mL of the solution and 2.5 mL of reference solution (a) and dilute to 100 mL with water R.<\/p>\n Reference solutions<\/em> Dilute reference solution (a) with water R to prepare a standard curve with at least 4 concentrations in the range of 10-40 \u03bcg\/mL.<\/p>\n Plot peak areas versus injected chloride content and perform linear regression to create a standard curve.<\/p>\n Precolumn:<\/p>\n \u2014 size: l = 0.05 m, \u00d8 = 4.0 mm;<\/p>\n \u2014 stationary phase: strongly basic anion-exchange resin for chromatography R (15 \u03bcm).<\/p>\n Column:<\/p>\n \u2014 size: l = 0.25 m, \u00d8 = 4.0 mm;<\/p>\n \u2014 stationary phase: strongly basic anion-exchange resin for chromatography R (15 \u03bcm).<\/p>\n Mobile phase<\/em> Dissolve 285.7 mg of sodium hydrogen carbonate R and 381.6 mg of anhydrous sodium carbonate R in water R and dilute to 2000 mL with the same solvent.<\/p>\n Flow rate<\/em> 2.0 mL\/min.<\/p>\n Detection<\/em> Conductivity detector; use a self-regenerating anion suppressor at 100 mA.<\/p>\n Injection<\/em> 50 \u03bcL.<\/p>\n Run time<\/em> 6 times the retention time of chloride.<\/p>\n Retention time<\/em> Chloride = about 1.6 min; nitrite = about 1.8 min.<\/p>\n System suitability<\/em> Reference solution (b):<\/p>\n \u2014 resolution: minimum 1.5 between the peaks due to chloride and nitrite;<\/p>\n \u2014 symmetry factor: maximum 2.0 for the peaks due to chloride and nitrite;<\/p>\n \u2014 repeatability: maximum relative standard deviation of 2.0 per cent for the areas of the peaks due to chloride and nitrite, determined on 5 injections;<\/p>\n \u2014 the correlation coefficient (r) calculated for the standard curve is not less than 0.999.<\/p>\n Calculate the chloride content using the standard curve and the area of the peak due to chloride in the chromatogram obtained with the test solution.<\/p>\n Liquid chromatography (2.2.29) as described in the test for related proteins with the following modifications.<\/p>\n Test solution<\/em> Dissolve the substance to be examined in the mobile phase to obtain a concentration of 0.25 mg\/mL.<\/p>\n Reference solution<\/em> Dissolve the contents of a vial of teriparatide CRS in the mobile phase to obtain a concentration of 0.25 mg\/mL.<\/p>\n Mobile phase<\/em> Mobile phase A, mobile phase B (63:37 V\/V); apply mild heating at 20-25 \u00b0C while stirring continuously during analysis.<\/p>\n Run time<\/em> 1.5 times the retention time of teriparatide.<\/p>\n Retention time<\/em> Teriparatide = about 10 min.<\/p>\n System suitability<\/em> Reference solution:<\/p>\n \u2014 symmetry factor: 0.8 to 1.5 for the peak due to teriparatide;<\/p>\n \u2014 repeatability: maximum relative standard deviation of 1.25 per cent for the area of the peak due to teriparatide, determined on 3 injections.<\/p>\n Calculate the percentage content of teriparatide (C181<\/sub>H291<\/sub>N55<\/sub>O51<\/sub>S2<\/sub>) taking into account the assigned content of teriparatide CRS.<\/p>\n In an airtight container, protected from light, at -10 \u00b0C or below.<\/p>\n","protected":false},"excerpt":{"rendered":" (Ph. Eur. monograph 2829) C181H291N55O51S2 4118 52232-67-4 Action and use Parathyroid hormone analogue; treatment of osteoporosis. DEFINITION Tetratriacontapeptide in which the sequence of amino acids is the same as that of the 1-34 N-terminal fragment of endogeneous human parathyroid hormone (rhPTH). Content 95.0 per cent to 105.0 per cent (anhydrous, acetic acid- and chloride-free substance)….<\/p>\n","protected":false},"author":5,"featured_media":30431,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[174],"tags":[],"class_list":["post-30409","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\/30409","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\/5"}],"replies":[{"embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/comments?post=30409"}],"version-history":[{"count":2,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/posts\/30409\/revisions"}],"predecessor-version":[{"id":30434,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/posts\/30409\/revisions\/30434"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/media\/30431"}],"wp:attachment":[{"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/media?parent=30409"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/categories?post=30409"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/tags?post=30409"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}DEFINITION<\/h2>\n
PRODUCTION<\/h2>\n
Host-cell-derived proteins<\/h3>\n
Host-cell- and vector-derived DNA<\/h3>\n
CHARACTERS<\/h2>\n
Appearance<\/h3>\n
Solubility<\/h3>\n
IDENTIFICATION<\/h2>\n
SELECTIVE CLEAVAGE OF THE PEPTIDE BONDS<\/h2>\n
CHROMATOGRAPHIC SEPARATION<\/h2>\n
\n\n
\n Time (min)<\/strong><\/td>\n Mobile phase A (per cent V\/V)<\/strong><\/td>\n Mobile phase B\u00a0(per cent V\/V)<\/strong><\/td>\n<\/tr>\n \n 0 – 6<\/td>\n 96<\/td>\n 4<\/td>\n<\/tr>\n \n 6 – 20<\/td>\n 96 \u2192 45<\/td>\n 4 \u2192 55<\/td>\n<\/tr>\n \n 20 – 25<\/td>\n 45 \u2192 0<\/td>\n 55 \u2192 100<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n TESTS<\/h2>\n
Impurities with molecular masses greater than that of teriparatide<\/h3>\n
Related proteins<\/h3>\n
\n\n
\n Time (min)<\/strong><\/td>\n Mobile phase A (per cent V\/V)<\/strong><\/td>\n Mobile phase B\u00a0(per cent V\/V)<\/strong><\/td>\n<\/tr>\n \n 0 – 5<\/td>\n 100 \u2192 65<\/td>\n 0 \u2192 35<\/td>\n<\/tr>\n \n 5 – 35<\/td>\n 65 \u2192 60<\/td>\n 35 \u2192 40<\/td>\n<\/tr>\n \n 35 – 45<\/td>\n 60 \u2192 0<\/td>\n 40 \u2192 100<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n Water (2.5.32)<\/h3>\n
Bacterial endotoxins (2.6.14)<\/h3>\n
ASSAY<\/h2>\n
Acetate<\/h3>\n
Chloride<\/h3>\n
Content<\/h3>\n
STORAGE<\/h2>\n