{"id":13229,"date":"2025-10-13T11:40:32","date_gmt":"2025-10-13T04:40:32","guid":{"rendered":"https:\/\/nhathuocngocanh.com\/bp\/?p=13229"},"modified":"2025-10-13T11:40:32","modified_gmt":"2025-10-13T04:40:32","slug":"fluorescein-sodium","status":"publish","type":"post","link":"https:\/\/nhathuocngocanh.com\/bp\/fluorescein-sodium\/","title":{"rendered":"Fluorescein Sodium"},"content":{"rendered":"

Soluble Fluorescein<\/p>\n

(Ph. Eur. monograph 1213)<\/p>\n

C_{20<\/sub>H_{10<\/sub>Na_{2<\/sub>O_{5<\/sub>\u00a0 \u00a0 \u00a0 376.3\u00a0 \u00a0 \u00a0 \u00a0518-47-8<\/p>\n}}}}

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

Detection of corneal lesions, retinal angiography and pancreatic function testing.<\/p>\n

Preparations<\/strong><\/p>\n

Fluorescein Eye Drops<\/p>\n

Fluorescein Injection<\/p>\n

DEFINITION<\/h2>\n
Disodium 2-(6-oxido-3-oxo-3H-xanthen-9-yl)benzoate.<\/p>\n
Content<\/h3>\n
95.0 per cent to 103.0 per cent (dried substance).<\/p>\n
CHARACTERS<\/h2>\n
Appearance<\/h3>\n
Orange-red, fine powder, hygroscopic.<\/p>\n
Solubility<\/h3>\n
Freely soluble in water, soluble in ethanol (96 per cent), practically insoluble in hexane and in methylene chloride.<\/p>\n
IDENTIFICATION<\/h2>\n
First identification: B, D.<\/p>\n
Second identification: A, C, D.<\/p>\n
A. Dilute 0.1 mL of solution S (see Tests) to 10 mL with water R. The solution shows yellowish-green fluorescence. The fluorescence disappears on addition of 0.1 mL of dilute hydrochloric acid R and reappears on addition of 0.2 mL of dilute sodium hydroxide solution R.<\/p>\n
B. Infrared absorption spectrophotometry (2.2.24).<\/p>\n
Preparation: Discs.<\/p>\n
Comparison: Ph. Eur. reference spectrum of fluorescein sodium.<\/p>\n
C. The absorption by a piece of filter paper of 0.05 mL of the solution prepared for identification A (before the addition of dilute hydrochloric acid R) colours the paper yellow. On exposing the moist paper to bromine vapour for 1 min and then to ammonia vapour, the colour becomes deep pink.<\/p>\n
D. Ignite 0.1 g in a porcelain crucible. Dissolve the residue in 5 mL of water R and filter. 2 mL of the filtrate gives reaction (a) of sodium (2.3.1).<\/p>\n
TESTS<\/h2>\n
Solution S<\/h3>\n
Dissolve 1.0 g in carbon dioxide-free water R prepared from distilled water R and dilute to 50 mL with the same solvent.<\/p>\n
Appearance of solution<\/h3>\n
Solution S is clear (2.2.1) and orange-yellow with yellowish-green fluorescence.<\/p>\n
pH (2.2.3)<\/h4>\n
7.0 to 9.0 for solution S.<\/p>\n
Related substances<\/h3>\n
Liquid chromatography (2.2.29).<\/p>\n
Test solution (a): Dissolve 0.100 g of the substance to be examined in a mixture of 30 volumes of acetonitrile R and 70 volumes of mobile phase A and dilute to 100.0 mL with the same mixture of solvents.<\/p>\n
Test solution (b): Dilute 5.0 mL of test solution (a) to 250.0 mL with a mixture of 30 volumes of acetonitrile R and 70 volumes of mobile phase A.<\/p>\n
Reference solution (a): Dissolve 55.0 mg of diacetylfluorescein CRS in a mixture of 1 mL of 2.5 M sodium hydroxide and 5 mL of ethanol (96 per cent) R, heat on a water-bath for 20 min mixing frequently, cool and dilute to 50.0 mL with water R. Dilute 5.0 mL of the solution to 250.0 mL with a mixture of 30 volumes of acetonitrile R and 70 volumes of mobile phase A.<\/p>\n
Reference solution (b): Dissolve 10.0 mg of phthalic acid R (impurity B) and 10.0 mg of resorcinol R (impurity A) in a mixture of 30 volumes of acetonitrile R and 70 volumes of mobile phase A and dilute to 100.0 mL with the same mixture of solvents. Dilute 5.0 mL of the solution to 100.0 mL with a mixture of 30 volumes of acetonitrile R and 70 volumes of mobile phase A.<\/p>\n
Reference solution (c): Dilute 5.0 mL of test solution (b) to 20.0 mL with a mixture of 30 volumes of acetonitrile R and 70 volumes of mobile phase A.<\/p>\n
Column:<\/p>\n
\u2014 size: l = 0.25 m, \u00d8 = 4.6 mm;<\/p>\n
\u2014 stationary phase: octylsilyl silica gel for chromatography R (5 \u03bcm);<\/p>\n
\u2014 temperature: 35 \u00b0C.<\/p>\n
Mobile phase:<\/p>\n
\u2014 mobile phase A: dissolve 0.610 g of potassium dihydrogen phosphate R in water R and dilute to 1000 mL with the same solvent; adjust to pH 2.0 with phosphoric acid R;<\/p>\n
\u2014 mobile phase B: acetonitrile for chromatography R;<\/p>\n\n\n\n\n\n\n\n
Time<\/strong>
\n(min)<\/strong><\/td>\n Mobile phase A<\/strong>
\n(per cent V\/V)<\/strong><\/td>\n Mobile phase B<\/strong>
\n(per cent V\/V)<\/strong><\/td>\n<\/tr>\n
0 – 20<\/td>\n 85 \u2192 20<\/td>\n 15 \u2192 80<\/td>\n<\/tr>\n
20 – 29<\/td>\n 20<\/td>\n 80<\/td>\n<\/tr>\n
29 – 30<\/td>\n 20 \u2192 85<\/td>\n 80 \u2192 15<\/td>\n<\/tr>\n
30 – 35<\/td>\n 85<\/td>\n 15<\/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 of test solution (a) and reference solutions (b) and (c).<\/p>\n
Relative retention: With reference to fluorescein (retention time = about 15 min): impurity A = about 0.4; impurity B = about 0.5; impurity C = about 0.9.<\/p>\n
System suitability: Reference solution (b):<\/p>\n
\u2014 resolution: minimum 1.5 between the peaks due to impurity A and impurity B.<\/p>\n
Limits:<\/p>\n
\u2014 correction factor: for the calculation of content, multiply the peak area of impurity C by 1.6;<\/p>\n
\u2014 impurities A, B: for each impurity, not more than the area of the corresponding peak in the chromatogram obtained with reference solution (b) (0.5 per cent);<\/p>\n
\u2014 impurity C: not more than the area of the principal peak in the chromatogram obtained with reference solution (c)(0.5 per cent);<\/p>\n
\u2014 unspecified impurities: for each impurity, not more than 0.2 times the area of the principal peak in the chromatogram obtained with reference solution (c) (0.10 per cent);<\/p>\n
\u2014 sum of impurities other than A, B, C: not more than the area of the principal peak in the chromatogram obtained with reference solution (c) (0.5 per cent);<\/p>\n
\u2014 disregard limit: 0.1 times the area of the principal peak in the chromatogram obtained with reference solution (c) (0.05 per cent).<\/p>\n
Chlorides (2.4.4)<\/h3>\n
Maximum 0.25 per cent.<\/p>\n
To 10 mL of solution S add 90 mL of water R and 1 mL of dilute nitric acid R, wait for at least 10 min and filter. Dilute 10 mL of the filtrate to 15 mL with water R.<\/p>\n
Sulfates (2.4.13)<\/h3>\n
Maximum 1.0 per cent.<\/p>\n
To 5 mL of solution S add 90 mL of distilled water R, 2.5 mL of dilute hydrochloric acid R and dilute to 100 mL with distilled water R. Filter.<\/p>\n
Zinc<\/h4>\n
Dilute 5 mL of solution S to 10 mL with water R. Add 2 mL of hydrochloric acid R1, filter and add 0.1 mL of potassium ferrocyanide solution R. No turbidity or precipitate is formed immediately.<\/p>\n
Loss on drying (2.2.32)<\/h4>\n
Maximum 10.0 per cent, determined on 1.000 g by drying in an oven at 105 \u00b0C.<\/p>\n
ASSAY<\/h2>\n
Liquid chromatography (2.2.29) as described in the test for related substances with the following modification.<\/p>\n
Injection Test solution (b) and reference solution (a).<\/p>\n
Calculate the percentage content of C_{20<\/sub>H_{10<\/sub>Na_{2<\/sub>O_{5<\/sub> using the chromatogram obtained with reference solution (a) and the declared content of diacetylfluorescein CRS.<\/p>\n}}}}
1 mg of diacetylfluorescein CRS is equivalent to 0.9037 mg of C_{20<\/sub>H_{10<\/sub>Na_{2<\/sub>O_{5<\/sub>.<\/p>\n}}}}
STORAGE<\/h2>\n
In an airtight container, protected from light.<\/p>\n
IMPURITIES<\/h2>\n
Specified impurities A, B, C.<\/p>\n
$\"Fluorescein$ <\/p>\n
A. benzene-1,3-diol (resorcinol),<\/p>\n
$\"Fluorescein$ <\/p>\n
B. benzene-1,2-dicarboxylic acid (phthalic acid),<\/p>\n
$\"Fluorescein$ <\/p>\n
C. 2-(2,4-dihydroxybenzoyl)benzoic acid.<\/p>\n","protected":false},"excerpt":{"rendered":"
Soluble Fluorescein (Ph. Eur. monograph 1213) C20H10Na2O5\u00a0 \u00a0 \u00a0 376.3\u00a0 \u00a0 \u00a0 \u00a0518-47-8 Action and use Detection of corneal lesions, retinal angiography and pancreatic function testing. Preparations Fluorescein Eye Drops Fluorescein Injection DEFINITION Disodium 2-(6-oxido-3-oxo-3H-xanthen-9-yl)benzoate. Content 95.0 per cent to 103.0 per cent (dried substance). CHARACTERS Appearance Orange-red, fine powder, hygroscopic. Solubility Freely soluble in…<\/p>\n","protected":false},"author":2,"featured_media":13238,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[174],"tags":[],"class_list":["post-13229","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\/13229","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\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/comments?post=13229"}],"version-history":[{"count":2,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/posts\/13229\/revisions"}],"predecessor-version":[{"id":13246,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/posts\/13229\/revisions\/13246"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/media\/13238"}],"wp:attachment":[{"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/media?parent=13229"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/categories?post=13229"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/tags?post=13229"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}

Time<\/strong> \n(min)<\/strong><\/td>\n	Mobile phase A<\/strong> \n(per cent V\/V)<\/strong><\/td>\n	Mobile phase B<\/strong> \n(per cent V\/V)<\/strong><\/td>\n<\/tr>\n
0 – 20<\/td>\n	85 \u2192 20<\/td>\n	15 \u2192 80<\/td>\n<\/tr>\n
20 – 29<\/td>\n	20<\/td>\n	80<\/td>\n<\/tr>\n
29 – 30<\/td>\n	20 \u2192 85<\/td>\n	80 \u2192 15<\/td>\n<\/tr>\n
30 – 35<\/td>\n	85<\/td>\n	15<\/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 of test solution (a) and reference solutions (b) and (c).<\/p>\n Relative retention: With reference to fluorescein (retention time = about 15 min): impurity A = about 0.4; impurity B = about 0.5; impurity C = about 0.9.<\/p>\n System suitability: Reference solution (b):<\/p>\n \u2014 resolution: minimum 1.5 between the peaks due to impurity A and impurity B.<\/p>\n Limits:<\/p>\n \u2014 correction factor: for the calculation of content, multiply the peak area of impurity C by 1.6;<\/p>\n \u2014 impurities A, B: for each impurity, not more than the area of the corresponding peak in the chromatogram obtained with reference solution (b) (0.5 per cent);<\/p>\n \u2014 impurity C: not more than the area of the principal peak in the chromatogram obtained with reference solution (c)(0.5 per cent);<\/p>\n \u2014 unspecified impurities: for each impurity, not more than 0.2 times the area of the principal peak in the chromatogram obtained with reference solution (c) (0.10 per cent);<\/p>\n \u2014 sum of impurities other than A, B, C: not more than the area of the principal peak in the chromatogram obtained with reference solution (c) (0.5 per cent);<\/p>\n \u2014 disregard limit: 0.1 times the area of the principal peak in the chromatogram obtained with reference solution (c) (0.05 per cent).<\/p>\n Chlorides (2.4.4)<\/h3>\n Maximum 0.25 per cent.<\/p>\n To 10 mL of solution S add 90 mL of water R and 1 mL of dilute nitric acid R, wait for at least 10 min and filter. Dilute 10 mL of the filtrate to 15 mL with water R.<\/p>\n Sulfates (2.4.13)<\/h3>\n Maximum 1.0 per cent.<\/p>\n To 5 mL of solution S add 90 mL of distilled water R, 2.5 mL of dilute hydrochloric acid R and dilute to 100 mL with distilled water R. Filter.<\/p>\n Zinc<\/h4>\n Dilute 5 mL of solution S to 10 mL with water R. Add 2 mL of hydrochloric acid R1, filter and add 0.1 mL of potassium ferrocyanide solution R. No turbidity or precipitate is formed immediately.<\/p>\n Loss on drying (2.2.32)<\/h4>\n Maximum 10.0 per cent, determined on 1.000 g by drying in an oven at 105 \u00b0C.<\/p>\n ASSAY<\/h2>\n Liquid chromatography (2.2.29) as described in the test for related substances with the following modification.<\/p>\n Injection Test solution (b) and reference solution (a).<\/p>\n Calculate the percentage content of C_{20<\/sub>H_{10<\/sub>Na_{2<\/sub>O_{5<\/sub> using the chromatogram obtained with reference solution (a) and the declared content of diacetylfluorescein CRS.<\/p>\n}}}} 1 mg of diacetylfluorescein CRS is equivalent to 0.9037 mg of C_{20<\/sub>H_{10<\/sub>Na_{2<\/sub>O_{5<\/sub>.<\/p>\n}}}} STORAGE<\/h2>\n In an airtight container, protected from light.<\/p>\n IMPURITIES<\/h2>\n Specified impurities A, B, C.<\/p>\n $\"Fluorescein$ <\/p>\n A. benzene-1,3-diol (resorcinol),<\/p>\n $\"Fluorescein$ <\/p>\n B. benzene-1,2-dicarboxylic acid (phthalic acid),<\/p>\n $\"Fluorescein$ <\/p>\n C. 2-(2,4-dihydroxybenzoyl)benzoic acid.<\/p>\n","protected":false},"excerpt":{"rendered":" Soluble Fluorescein (Ph. Eur. monograph 1213) C20H10Na2O5\u00a0 \u00a0 \u00a0 376.3\u00a0 \u00a0 \u00a0 \u00a0518-47-8 Action and use Detection of corneal lesions, retinal angiography and pancreatic function testing. Preparations Fluorescein Eye Drops Fluorescein Injection DEFINITION Disodium 2-(6-oxido-3-oxo-3H-xanthen-9-yl)benzoate. Content 95.0 per cent to 103.0 per cent (dried substance). CHARACTERS Appearance Orange-red, fine powder, hygroscopic. Solubility Freely soluble in…<\/p>\n","protected":false},"author":2,"featured_media":13238,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[174],"tags":[],"class_list":["post-13229","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\/13229","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\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/comments?post=13229"}],"version-history":[{"count":2,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/posts\/13229\/revisions"}],"predecessor-version":[{"id":13246,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/posts\/13229\/revisions\/13246"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/media\/13238"}],"wp:attachment":[{"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/media?parent=13229"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/categories?post=13229"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/nhathuocngocanh.com\/bp\/wp-json\/wp\/v2\/tags?post=13229"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}

Content<\/h3>\n95.0 per cent to 103.0 per cent (dried substance).<\/p>\n

CHARACTERS<\/h2>\n

Appearance<\/h3>\nOrange-red, fine powder, hygroscopic.<\/p>\n

Solubility<\/h3>\nFreely soluble in water, soluble in ethanol (96 per cent), practically insoluble in hexane and in methylene chloride.<\/p>\n

TESTS<\/h2>\n

Solution S<\/h3>\nDissolve 1.0 g in carbon dioxide-free water R prepared from distilled water R and dilute to 50 mL with the same solvent.<\/p>\n

Appearance of solution<\/h3>\nSolution S is clear (2.2.1) and orange-yellow with yellowish-green fluorescence.<\/p>\n

pH (2.2.3)<\/h4>\n7.0 to 9.0 for solution S.<\/p>\n

Chlorides (2.4.4)<\/h3>\nMaximum 0.25 per cent.<\/p>\nTo 10 mL of solution S add 90 mL of water R and 1 mL of dilute nitric acid R, wait for at least 10 min and filter. Dilute 10 mL of the filtrate to 15 mL with water R.<\/p>\n

Sulfates (2.4.13)<\/h3>\nMaximum 1.0 per cent.<\/p>\nTo 5 mL of solution S add 90 mL of distilled water R, 2.5 mL of dilute hydrochloric acid R and dilute to 100 mL with distilled water R. Filter.<\/p>\n

Zinc<\/h4>\nDilute 5 mL of solution S to 10 mL with water R. Add 2 mL of hydrochloric acid R1, filter and add 0.1 mL of potassium ferrocyanide solution R. No turbidity or precipitate is formed immediately.<\/p>\n

Loss on drying (2.2.32)<\/h4>\nMaximum 10.0 per cent, determined on 1.000 g by drying in an oven at 105 \u00b0C.<\/p>\n

STORAGE<\/h2>\nIn an airtight container, protected from light.<\/p>\n

Content<\/h3>\n
95.0 per cent to 103.0 per cent (dried substance).<\/p>\n

Appearance<\/h3>\n
Orange-red, fine powder, hygroscopic.<\/p>\n

Solubility<\/h3>\n
Freely soluble in water, soluble in ethanol (96 per cent), practically insoluble in hexane and in methylene chloride.<\/p>\n

Solution S<\/h3>\n
Dissolve 1.0 g in carbon dioxide-free water R prepared from distilled water R and dilute to 50 mL with the same solvent.<\/p>\n

Appearance of solution<\/h3>\n
Solution S is clear (2.2.1) and orange-yellow with yellowish-green fluorescence.<\/p>\n

pH (2.2.3)<\/h4>\n
7.0 to 9.0 for solution S.<\/p>\n

Chlorides (2.4.4)<\/h3>\n
Maximum 0.25 per cent.<\/p>\n
To 10 mL of solution S add 90 mL of water R and 1 mL of dilute nitric acid R, wait for at least 10 min and filter. Dilute 10 mL of the filtrate to 15 mL with water R.<\/p>\n

Sulfates (2.4.13)<\/h3>\n
Maximum 1.0 per cent.<\/p>\n
To 5 mL of solution S add 90 mL of distilled water R, 2.5 mL of dilute hydrochloric acid R and dilute to 100 mL with distilled water R. Filter.<\/p>\n

Zinc<\/h4>\n
Dilute 5 mL of solution S to 10 mL with water R. Add 2 mL of hydrochloric acid R1, filter and add 0.1 mL of potassium ferrocyanide solution R. No turbidity or precipitate is formed immediately.<\/p>\n

Loss on drying (2.2.32)<\/h4>\n
Maximum 10.0 per cent, determined on 1.000 g by drying in an oven at 105 \u00b0C.<\/p>\n

STORAGE<\/h2>\n
In an airtight container, protected from light.<\/p>\n