{"id":5081,"date":"2025-09-29T10:19:19","date_gmt":"2025-09-29T03:19:19","guid":{"rendered":"https:\/\/nhathuocngocanh.com\/bp\/?p=5081"},"modified":"2025-10-03T09:06:18","modified_gmt":"2025-10-03T02:06:18","slug":"calcitonin-salmon","status":"publish","type":"post","link":"https:\/\/nhathuocngocanh.com\/bp\/calcitonin-salmon\/","title":{"rendered":"Calcitonin (Salmon)"},"content":{"rendered":"

(Ph. Eur. monograph 0471)<\/p>\n

C_{145<\/sub>H_{240<\/sub>N_{44<\/sub>O_{48<\/sub>S_{2<\/sub>\u00a0 \u00a03432<\/p>\n}}}}}

Action and use<\/strong><\/p>\n

Hormone.<\/p>\n

Preparation<\/strong><\/p>\n

Calcitonin (Salmon) Injection<\/p>\n

DEFINITION<\/h2>\n
Polypeptide having the structure determined for salmon calcitonin I. It lowers the calcium concentration in plasma of mammals by diminishing the rate of bone resorption. It is obtained by chemical synthesis or by a method based on recombinant DNA (rDNA) technology. It is available as an acetate.<\/p>\n
Content<\/h3>\n
90.0 per cent to 105.0 per cent of the peptide C_{145<\/sub>H_{240<\/sub>N_{44<\/sub>O_{48<\/sub>S_{2 <\/sub>(anhydrous and acetic acid-free substance).<\/p>\n}}}}}
By convention, for the purpose of labelling calcitonin (salmon) preparations, 1 mg of calcitonin (salmon) (C_{145<\/sub>H_{240<\/sub>N_{44<\/sub>O_{48<\/sub>S_{2<\/sub>) is equivalent to 6000 IU of biological activity.<\/p>\n}}}}}
PRODUCTION<\/h2>\n
The following requirements apply only to calcitonin (salmon) produced by a method based on rDNA technology.<\/p>\n
Prior to release, the following tests are carried out on each batch of calcitonin (salmon), unless exemption has been granted by the competent authority.<\/p>\n
Host-cell-derived proteins<\/h3>\n
The limit is approved by the competent authority.<\/p>\n
Host-cell or vector-derived DNA<\/h3>\n
The limit is approved by the competent authority.<\/p>\n
CHARACTERS<\/h2>\n
Appearance<\/h3>\n
White or almost white powder.<\/p>\n
Solubility<\/h3>\n
Freely soluble in water.<\/p>\n
IDENTIFICATION<\/h2>\n
A. Examine the chromatograms obtained in the assay.<\/p>\n
Results: The principal peak in the chromatogram obtained with the test solution is similar in retention time and size to the principal peak in the chromatogram obtained with the reference solution.<\/p>\n
The following requirement applies only to calcitonin (salmon) obtained by chemical synthesis.<\/p>\n
B. Amino acid analysis (2.2.56).<\/p>\n
Express the content of each amino acid in moles. Calculate the relative proportions of the amino acids taking as equivalent to 1 the sum, divided by 20, of the number of moles of aspartic acid, glutamic acid, proline, glycine, valine, leucine, histidine, arginine and lysine. The values fall within the following limits: aspartic acid: 1.8 to 2.2; glutamic acid: 2.7 to 3.3; proline: 1.7 to 2.3; glycine: 2.7 to 3.3; valine: 0.9 to 1.1; leucine: 4.5 to 5.3; histidine: 0.9 to 1.1; arginine: 0.9 to 1.1; lysine: 1.8 to 2.2; serine: 3.2 to 4.2; threonine: 4.2 to 5.2; tyrosine: 0.7 to 1.1; half-cystine: 1.4 to 2.1. The following requirement applies only to calcitonin (salmon) produced by a method based on rDNA technology.<\/p>\n
C. Peptide mapping (2.2.55).<\/p>\n
SELECTIVE CLEAVAGE OF THE PEPTIDE BONDS<\/em><\/p>\n
Test solution: Prepare a 1 mg\/mL solution of the substance to be examined. Transfer 1.0 mL to a clean tube. Add 100 \u03bcL of 1 M tris-hydrochloride buffer solution pH 8.0 R and 20 \u03bcL of a freshly prepared 1.0 mg\/mL solution of trypsin for peptide mapping R. Allow to stand at 2-8 \u00b0C for 16-20 h. Stop the reaction by adding 10 \u03bcL of a 50 per cent V\/V solution of trifluoroacetic acid R. Cap the vial and mix. Centrifuge the vials to remove air bubbles.<\/p>\n
Reference solution: Prepare at the same time and in the same manner as for the test solution but using calcitonin (salmon) CRS instead of the substance to be examined.<\/p>\n
CHROMATOGRAPHIC SEPARATION <\/em><\/p>\n
Liquid chromatography (2.2.29).<\/p>\n
Column:<\/p>\n
\u2014 size: l = 0.25 m, \u00d8 = 4.6 mm;<\/p>\n
\u2014 stationary phase: octadecylsilyl silica gel for chromatography R (5 \u03bcm) with a pore size of 30 nm.<\/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 0.850 mL of trifluoroacetic acid R, 200 mL of water R and 800 mL of acetonitrile for
\nchromatography R; filter and degas;<\/p>\n\n\n\n\n\n\n\n\n\n
Time<\/strong><\/p>\n
(min)<\/strong><\/td>\n
Mobile phase A<\/strong><\/p>\n
(per cent V\/V)<\/strong><\/td>\n
Mobile phase B<\/strong><\/p>\n
(per cent V\/V)<\/strong><\/td>\n<\/tr>\n
0 – 50<\/td>\n 100 \u2192 65<\/td>\n 0 \u2192 35<\/td>\n<\/tr>\n
50 – 60<\/td>\n 65 \u2192 40<\/td>\n 35 \u2192 60<\/td>\n<\/tr>\n
60 – 60.1<\/td>\n 40 \u2192 0<\/td>\n 60 \u2192 100<\/td>\n<\/tr>\n
60.1 – 65.1<\/td>\n 0<\/td>\n 100<\/td>\n<\/tr>\n
65.1 – 65.2<\/td>\n 0 \u2192 100<\/td>\n 100 \u2192 0<\/td>\n<\/tr>\n
65.2 – 80.2<\/td>\n 100<\/td>\n 0<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n
Flow rate: 1.2 mL\/min.<\/p>\n
Detection: Spectrophotometer at 214 nm.<\/p>\n
Equilibration: At initial conditions for at least 15 min. Carry out a blank run using the above-mentioned gradient.<\/p>\n
Injection: 20 \u03bcL.<\/p>\n
System suitability: The chromatogram obtained with the reference solution is qualitatively similar to the chromatogram of calcitonin (salmon) digest supplied with calcitonin (salmon) CRS.<\/p>\n
Results: The profile of the chromatogram obtained with the test solution corresponds to that of the chromatogram obtained with the reference solution: the retention times of the fragment peaks in the chromatogram obtained with the test solution are within 5 per cent of the retention times of the fragments obtained with the reference solution; the peak area ratios of the fragment peaks in the chromatogram obtained with the test solution, normalised to the area of peak T2, are within 5 per cent of the corresponding peak ratios in the chromatogram obtained with the reference solution.<\/p>\n
TESTS<\/h2>\n
Acetic acid (2.5.34)<\/h3>\n
4.0 per cent to 15.0 per cent.<\/p>\n
Test solution: Dissolve 10.0 mg of the substance to be examined in a mixture of 5 volumes of mobile phase B and 95 volumes of mobile phase A and dilute to 10.0 mL with the same mixture of mobile phases.<\/p>\n
Related substances<\/h3>\n
Liquid chromatography (2.2.29): use the normalisation procedure.<\/p>\n
The following requirement applies to calcitonin (salmon), whether obtained by chemical synthesis or by a method based on rDNA technology.<\/p>\n
A. Test solution. Prepare a 1.0 mg\/mL solution of the substance to be examined in mobile phase A.<\/p>\n
Reference solution: Dissolve the contents of a vial of calcitonin (salmon) CRS in mobile phase A to obtain a concentration of 1.0 mg\/mL.<\/p>\n
Resolution solution: Dissolve the contents of a vial of N-acetyl-Cys -calcitonin CRS in 400 \u03bcL of mobile phase A and add 100 \u03bcL of the test solution.<\/p>\n
Column:<\/p>\n
\u2014 size: l = 0.25 m, \u00d8 = 4.6 mm;<\/p>\n
\u2014 stationary phase: octadecylsilyl silica gel for chromatography R (5 \u03bcm);<\/p>\n
\u2014 temperature: 65 \u00b0C.<\/p>\n
Mobile phase:<\/p>\n
\u2014 mobile phase A: dissolve 3.26 g of tetramethylammonium hydroxide R in 900 mL of water R, adjust to pH 2.5 with phosphoric acid R and mix with 100 mL of acetonitrile for chromatography R; filter and degas;<\/p>\n
<\/p>\n
\u2014 mobile phase B: dissolve 1.45 g of tetramethylammonium hydroxide R in 400 mL of water R, adjust to pH 2.5 with phosphoric acid R and mix with 600 mL of acetonitrile for chromatography R; filter and degas;<\/p>\n\n\n\n\n\n\n
Time<\/strong><\/p>\n
(min)<\/strong><\/td>\n
Mobile phase A<\/strong><\/p>\n
(per cent V\/V)<\/strong><\/td>\n
Mobile phase B<\/strong><\/p>\n
(per cent V\/V)<\/strong><\/td>\n<\/tr>\n
0 – 30<\/td>\n 72 \u2192 48<\/td>\n 28 \u2192 52<\/td>\n<\/tr>\n
30 – 32<\/td>\n 48 \u2192 72<\/td>\n 35 \u2192 60<\/td>\n<\/tr>\n
32 – 55<\/td>\n 72<\/td>\n 28<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n
Flow rate: 1.0 mL\/min.<\/p>\n
Detection: Spectrophotometer at 220 nm.<\/p>\n
Injection: 20 \u03bcL.<\/p>\n
Relative retention: With reference to calcitonin (salmon) (retention time = about 20 min): impurity B = about 0.8; impurity C = about 0.9; impurity D = about 1.05; impurity A = about 1.15.<\/p>\n
System suitability Resolution solution:<\/p>\n
\u2014 resolution: minimum 5.0 between the peaks due to calcitonin (salmon) and impurity A,<\/p>\n
\u2014 symmetry factor: maximum 2.5 for the peak due to impurity A.<\/p>\n
Limits:<\/p>\n
\u2014 impurities A, B, C, D: for each impurity, maximum 3.0 per cent; other unidentified, specified impurities may occur that co-elute with impurities A, B, C and D; the acceptance criterion applies irrespective of whether these impurities co-elute;<\/p>\n
\u2014 total: maximum 5.0 per cent;<\/p>\n
\u2014 disregard limit: 0.1 per cent.<\/p>\n
The following requirement applies only to calcitonin (salmon) produced by a method based on rDNA technology.<\/p>\n
B. Test solution. Prepare a 0.5 mg\/mL solution of the substance to be examined. To 1.0 mL of this solution add 100 \u03bcL of 0.25 M citrate buffer solution pH 3.0 R.<\/p>\n
Resolution solution: Prepare a 1 mg\/mL solution of the substance to be examined. Mix 1 volume of the solution and 1 volume of calcitonin-Gly CRS. To 1.0 mL of this mixture add 100 \u03bcL of 0.25 M citrate buffer solution pH 3.0 R.<\/p>\n
Column:<\/p>\n
\u2014 size: l = 0.20 m, \u00d8 = 4.6 mm;<\/p>\n
\u2014 stationary phase: a suitable polysulfoethylaspartamide ion-exchange gel (5 \u03bcm).<\/p>\n
Mobile phase:<\/p>\n
\u2014 mobile phase A: mix 15 volumes of acetonitrile for chromatography R and 85 volumes of a 2.72 g\/L solution of potassium dihydrogen phosphate R adjusted to pH 5.0 with a 600 g\/L solution of potassium hydroxide R;<\/p>\n
\u2014 mobile phase B: mix 15 volumes of acetonitrile for chromatography R and 85 volumes of a solution containing 2.72 g\/L of potassium dihydrogen phosphate R and 29.22 g\/L of sodium chloride R adjusted to pH 4.6 with a 600 g\/L solution of potassium hydroxide R;<\/p>\n\n\n\n\n\n\n\n
Time<\/strong><\/p>\n
(min)<\/strong><\/td>\n
Mobile phase A<\/strong><\/p>\n
(per cent V\/V)<\/strong><\/td>\n
Mobile phase B<\/strong><\/p>\n
(per cent V\/V)<\/strong><\/td>\n<\/tr>\n
0 – 10<\/td>\n 100 \u2192 0<\/td>\n 0 \u2192 100<\/td>\n<\/tr>\n
10 – 15<\/td>\n 0<\/td>\n 100<\/td>\n<\/tr>\n
15 – 15.1<\/td>\n 0 \u2192 100<\/td>\n 100 \u2192 0<\/td>\n<\/tr>\n
15.1 – 22.1<\/td>\n 100<\/td>\n 0<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n
Flow rate: 1.2 mL\/min.<\/p>\n
Detection: Spectrophotometer at 220 nm.<\/p>\n
Injection: 50 \u03bcL; rinse the injector with a 40 per cent V\/V solution of acetonitrile for chromatography R.<\/p>\n
Relative retention: With reference to calcitonin (salmon) (retention time = about 9 min): impurity G = about 0.4; impurity F = about 0.6; impurity E = about 0.9.<\/p>\n
System suitability: Resolution solution:<\/p>\n
\u2014 resolution: minimum 3.0 between the peaks due to impurity E and calcitonin (salmon).<\/p>\n
Limits:<\/p>\n
\u2014 impurity E: maximum 0.6 per cent;<\/p>\n
\u2014 impurities F, G: for each impurity, maximum 0.2 per cent.<\/p>\n
Water (2.5.32)<\/h3>\n
Maximum 10.0 per cent.<\/p>\n
Acetic acid and water<\/h3>\n
Maximum 20 per cent, calculated by adding together the percentage contents of acetic acid and water determined by the methods described above.<\/p>\n
Bacterial endotoxins (2.6.14)<\/h3>\n
Less than 25 IU\/mg, if intended for use in the manufacture of parenteral preparations without a further appropriate procedure for the removal of bacterial endotoxins.<\/p>\n
ASSAY<\/h2>\n
Liquid chromatography (2.2.29) as described in the test for related substances. Use method A for calcitonin (salmon) obtained by chemical synthesis and method B for calcitonin (salmon) obtained by a method based on rDNA technology.<\/p>\n
Calculate the content of calcitonin (salmon) (C_{145<\/sub>H_{240<\/sub>N_{44<\/sub>O_{48<\/sub>S_{2<\/sub>) from the area of the principal peak in each of the chromatograms obtained with the test solution and the reference solution and the declared content of C_{145<\/sub>H_{240<\/sub>N_{44<\/sub>O_{48<\/sub>S_{2 <\/sub>in calcitonin (salmon) CRS. Proceed with tangential integration of the peak areas.<\/p>\n}}}}}}}}}}
STORAGE<\/h2>\n
Protected from light at a temperature between 2 \u00b0C and 8 \u00b0C. If the substance is sterile, store in a sterile, airtight, tamper-evident container.<\/p>\n
LABELLING<\/h2>\n
The label states:<\/p>\n
\u2014 the calcitonin peptide content (C_{145<\/sub>H_{240<\/sub>N_{44<\/sub>O_{48<\/sub>S_{2<\/sub>);
\n\u2014 the origin: synthetic or rDNA technology.<\/p>\n}}}}}
IMPURITIES<\/h2>\n
Specified impurities A, B, C, D, E, F, G.<\/p>\n
$\"Calcitonin$ <\/p>\n
A. acetylcalcitonin (salmon),<\/p>\n
$\"Calcitonin$ <\/p>\n
B. [9-D-leucine]calcitonin (salmon),<\/p>\n
$\"Calcitonin$ <\/p>\n
C. des-22-tyrosine-calcitonin (salmon),<\/p>\n
D. O-acetylated calcitonin (salmon),<\/p>\n
$\"Calcitonin$ <\/p>\n
E. salmon calcitoninylglycine,<\/p>\n
$\"Calcitonin$ <\/p>\n
F. [1,7-bis(3-sulfo-L-alanine)]calcitonin (salmon),<\/p>\n
$\"Calcitonin$ <\/p>\n
G. [1,7-bis(3-sulfo-L-alanine)]calcitoninylglycine (salmon).<\/p>\n","protected":false},"excerpt":{"rendered":"
(Ph. Eur. monograph 0471) C145H240N44O48S2\u00a0 \u00a03432 Action and use Hormone. Preparation Calcitonin (Salmon) Injection DEFINITION Polypeptide having the structure determined for salmon calcitonin I. It lowers the calcium concentration in plasma of mammals by diminishing the rate of bone resorption. It is obtained by chemical synthesis or by a method based on recombinant DNA (rDNA)…<\/p>\n","protected":false},"author":4,"featured_media":5159,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[174],"tags":[],"class_list":["post-5081","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\/5081","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=5081"}],"version-history":[{"count":2,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/posts\/5081\/revisions"}],"predecessor-version":[{"id":5861,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/posts\/5081\/revisions\/5861"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/media\/5159"}],"wp:attachment":[{"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/media?parent=5081"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/categories?post=5081"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/tags?post=5081"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}

PRODUCTION<\/h2>\nThe following requirements apply only to calcitonin (salmon) produced by a method based on rDNA technology.<\/p>\nPrior to release, the following tests are carried out on each batch of calcitonin (salmon), unless exemption has been granted by the competent authority.<\/p>\n

Host-cell-derived proteins<\/h3>\nThe limit is approved by the competent authority.<\/p>\n

Host-cell or vector-derived DNA<\/h3>\nThe limit is approved by the competent authority.<\/p>\n

CHARACTERS<\/h2>\n

Appearance<\/h3>\nWhite or almost white powder.<\/p>\n

Solubility<\/h3>\nFreely soluble in water.<\/p>\n

TESTS<\/h2>\n

Acetic acid (2.5.34)<\/h3>\n4.0 per cent to 15.0 per cent.<\/p>\nTest solution: Dissolve 10.0 mg of the substance to be examined in a mixture of 5 volumes of mobile phase B and 95 volumes of mobile phase A and dilute to 10.0 mL with the same mixture of mobile phases.<\/p>\n

Water (2.5.32)<\/h3>\nMaximum 10.0 per cent.<\/p>\n

Acetic acid and water<\/h3>\nMaximum 20 per cent, calculated by adding together the percentage contents of acetic acid and water determined by the methods described above.<\/p>\n

Bacterial endotoxins (2.6.14)<\/h3>\nLess than 25 IU\/mg, if intended for use in the manufacture of parenteral preparations without a further appropriate procedure for the removal of bacterial endotoxins.<\/p>\n

STORAGE<\/h2>\nProtected from light at a temperature between 2 \u00b0C and 8 \u00b0C. If the substance is sterile, store in a sterile, airtight, tamper-evident container.<\/p>\n

LABELLING<\/h2>\nThe label states:<\/p>\n\u2014 the calcitonin peptide content (C145<\/sub>H240<\/sub>N44<\/sub>O48<\/sub>S2<\/sub>);\n\u2014 the origin: synthetic or rDNA technology.<\/p>\n

PRODUCTION<\/h2>\n
The following requirements apply only to calcitonin (salmon) produced by a method based on rDNA technology.<\/p>\n
Prior to release, the following tests are carried out on each batch of calcitonin (salmon), unless exemption has been granted by the competent authority.<\/p>\n

Host-cell-derived proteins<\/h3>\n
The limit is approved by the competent authority.<\/p>\n

Host-cell or vector-derived DNA<\/h3>\n
The limit is approved by the competent authority.<\/p>\n

Appearance<\/h3>\n
White or almost white powder.<\/p>\n

Solubility<\/h3>\n
Freely soluble in water.<\/p>\n

Acetic acid (2.5.34)<\/h3>\n
4.0 per cent to 15.0 per cent.<\/p>\n
Test solution: Dissolve 10.0 mg of the substance to be examined in a mixture of 5 volumes of mobile phase B and 95 volumes of mobile phase A and dilute to 10.0 mL with the same mixture of mobile phases.<\/p>\n

Water (2.5.32)<\/h3>\n
Maximum 10.0 per cent.<\/p>\n

Acetic acid and water<\/h3>\n
Maximum 20 per cent, calculated by adding together the percentage contents of acetic acid and water determined by the methods described above.<\/p>\n

Bacterial endotoxins (2.6.14)<\/h3>\n
Less than 25 IU\/mg, if intended for use in the manufacture of parenteral preparations without a further appropriate procedure for the removal of bacterial endotoxins.<\/p>\n

STORAGE<\/h2>\n
Protected from light at a temperature between 2 \u00b0C and 8 \u00b0C. If the substance is sterile, store in a sterile, airtight, tamper-evident container.<\/p>\n

LABELLING<\/h2>\n
The label states:<\/p>\n
\u2014 the calcitonin peptide content (C_{145<\/sub>H_{240<\/sub>N_{44<\/sub>O_{48<\/sub>S_{2<\/sub>);
\n\u2014 the origin: synthetic or rDNA technology.<\/p>\n}}}}}