LectureLecture № №77Vitamins as medicinesVitamins as medicines: : general general

characteristiccharacteristic, , methods of obtainingmethods of obtaining,, classificationclassification, , analysisanalysis, , storagestorage,,

precondition of their usage in medical precondition of their usage in medical practicepractice. . Vitamins of the aliphatic and Vitamins of the aliphatic and

aromatic rowsaromatic rows. .

prepared ass.prepared ass. MedvidMedvid II..II..,, ass. Kozachok S.S.ass. Kozachok S.S.

VitaminsVitamins - - are organic compounds of are organic compounds of different chemical structures, which in small different chemical structures, which in small amounts required for the normal organism livingamounts required for the normal organism living. . They are a part of enzyme systems, which are They are a part of enzyme systems, which are biological catalysts of the chemical reactions in biological catalysts of the chemical reactions in living cells and are involved in metabolismliving cells and are involved in metabolism. . The The human and animals can not synthesize vitamins or human and animals can not synthesize vitamins or synthesize them in insufficient quantities (nicotinic synthesize them in insufficient quantities (nicotinic acid) and therefore must obtain them from foodacid) and therefore must obtain them from food. . In In some cases vitamins are formed in the tissues as a some cases vitamins are formed in the tissues as a result of chemical transformation of substances result of chemical transformation of substances that are their precursors (provitamins). that are their precursors (provitamins).

InIn 1912 1912 yy. . polish scientist K. Funkpolish scientist K. Funk proposed proposed term “vitamins” that means “amines necessary for term “vitamins” that means “amines necessary for life”life”..

ClassificationClassification of of vitaminsvitamins1.1. By the By the solubilitysolubility::• fat solublefat soluble ( (A, D, E, F, K A, D, E, F, K ))• Water-solubleWater-soluble ( (groupgroup В, С, В, С, synthetic synthetic analogsanalogs of of

К, РР, Р)К, РР, Р)2.2. By lettersBy letters::• by by the disease that occurs the disease that occurs at theat the insufficient insufficient

amount of this vitaminamount of this vitamin (С – (С – scurvyscurvy ((antiscorbuticantiscorbutic), А –), А –antixerophthalmicantixerophthalmic, В – , В – beriberi--beriberi, Е – , Е – one that one that ensuresensures procreation.procreation.

• additional figures additional figures at theat the expanding expanding of of the groupthe group of of vitamins vitamins (В(В11 – В – В1515, В, Всс))

3.3. Chemical classification.Chemical classification.

Chemical classification of vitaminsChemical classification of vitamins I.I. Vitamins Vitamins of the of the aliphatic aliphatic rowrow: : а) а) Derivatives of the unsaturated polyoxyDerivatives of the unsaturated polyoxy--- - lactoneslactones ( (ascorbic acid ascorbic acid

(vitamin C)(vitamin C)););

bb) ) Derivatives of gluconic acidDerivatives of gluconic acid esters esters ((pangamic pangamic acid acid ((vitaminvitamin В В1515));));

cc) ) Derivatives ofDerivatives of - - aminoacidsaminoacids ( (pantothenic acid (vitaminpantothenic acid (vitamin ВВ33)).)).

II.II. Vitamins of the alicyclic rowVitamins of the alicyclic row: : а) а) cyclohexyl isoprenoidscyclohexyl isoprenoids((retinols retinols (vitamin(vitamins of A groups of A group));));bb) ) Cyclohexanolethylenehydrindanoic Cyclohexanolethylenehydrindanoic ((calciferols calciferols (vitamin(vitamins of groups of group

DD)).)).III.III. Vitamins of the aromatic rowVitamins of the aromatic row ( (naphthoquinone derivativesnaphthoquinone derivatives): ):

cc) ) natural vitaminnatural vitamins ofs of К К group group (phylloquinone(phylloquinone– К– К11, , menaquinonemenaquinone – К – К22););

dd) ) ssynthetic analogues of vitaminynthetic analogues of vitamins ofs of К К group group ((menadionmenadione, ve, vikasolikasol).).

IV.IV. VitaminsVitamins of the of the heterocyclic heterocyclic rowrow: :

1) 1) Chromane derivativesChromane derivatives

а) а) tocopherolstocopherols ( (vitamins ofvitamins of Е Е group group););

bb) ) bioflavonoidsbioflavonoids ( (vitamins ofvitamins of Р Р group group).).

2) 2) Pyridine derivativesPyridine derivatives

а) а) nicotinic acid and its amidenicotinic acid and its amide ( (vitamine of vitamine of РРРР group group););

bb) ) oxymethylpyridine vitamines oxymethylpyridine vitamines ((vitamins of vitamins of ВВ66 group) group)..

3) 3) pyrimidine derivativespyrimidine derivatives ( (thiaminthiamin ( (vitaminvitamin В В11).).

4) 4) Pterine derivatives Pterine derivatives ((folic acidfolic acid ( (vitamin vitamin Вс).Вс).

5) 5) Isoalloxazine derivatives or flavin vitamins Isoalloxazine derivatives or flavin vitamins ((riboflavinriboflavin ((vitaminvitamin В В22))))

6) 6) Corrine derivativesCorrine derivatives – – cobalamins cobalamins ((vitamins ofvitamins of В В1212 group) group)..

Obtaining of vitaminsObtaining of vitamins• Synthetic and semi-synthetic methodsSynthetic and semi-synthetic methods (С, А, Е, (С, А, Е, DD, В, В11, В, В22, РР , РР

and othersand others).).• From From plant and animal raw materials plant and animal raw materials , , microorganismsmicroorganisms (С - (С -

fruit-bloomfruit-bloom, Р –, Р –waste of the tea industrywaste of the tea industry, , DD – –natural sterinesnatural sterines, , А –А –fish oilfish oil, Е –, Е –vegetable fatsvegetable fats, В12 –, В12 –product of product of microbiological synthesis of antibioticsmicrobiological synthesis of antibiotics,, effluents effluents).).

PropertiesProperties White crystals or crystalline powder White crystals or crystalline powder ((rutin - a rutin - a

green- yellow, folic acid - a yellow or yellow-green- yellow, folic acid - a yellow or yellow-orange, riboflavin - yellow-orange, orange, riboflavin - yellow-orange, cyanocobalamin - dark redcyanocobalamin - dark red).). Tocopherol acetate- Tocopherol acetate-light yellow, transparent, dense, oily liquid with light yellow, transparent, dense, oily liquid with slight odorslight odor..

Vitamins are unstable under the action of light, air Vitamins are unstable under the action of light, air and heat (some of them).and heat (some of them).

Identification of vitaminsIdentification of vitamins

• There are not general reactionsThere are not general reactions,, except nitrogen except nitrogen determination by the Kjeldahl methoddetermination by the Kjeldahl method..

• Often the group reactionOften the group reaction((on the pyridine cycle etc.on the pyridine cycle etc.))

• More often - reactions that depend on the individual More often - reactions that depend on the individual characteristics of the chemical structurecharacteristics of the chemical structure. .

• According to theAccording to the SPhU requirementsSPhU requirements – – 1010 preparations preparations ((DD, С, РР, , С, РР, nicotinamidenicotinamide В В66, В, В22, В, В11

((hydrobromidehydrobromide), В), В11 ( (hydrochloridehydrochloride), В), Всс, В, В1212: :

ІІRR-, -, UVUV-- spectroscopy spectroscopy, , chromatographychromatography..

Assay of vitaminsAssay of vitamins• Biological methods - determining of the biological Biological methods - determining of the biological

activity of vitaminsactivity of vitamins. . Studies conducted on rats, Studies conducted on rats, pigeons, guinea pigs, which are moved on a diet pigeons, guinea pigs, which are moved on a diet ((exclude the study vitamin from the food)exclude the study vitamin from the food).. Then Then determines which of the study vitamin can cure or determines which of the study vitamin can cure or save the animal from avitaminosissave the animal from avitaminosis. . Parallel conduct Parallel conduct similar studies with the standard drugsimilar studies with the standard drug. . Activity in IU Activity in IU (international unit) - the notional amount of the (international unit) - the notional amount of the standard drug in mg or mcg standard drug in mg or mcg (().). For one unit is For one unit is considered to be the minimum amount of vitamin considered to be the minimum amount of vitamin that cures or prevents the animal from avitaminosisthat cures or prevents the animal from avitaminosis. . 1 1 IU is different in different vitamins (1 IU of IU is different in different vitamins (1 IU of vitamin A corresponds to 0,344 vitamin A corresponds to 0,344 g g of tocopherol of tocopherol acetate, vitamin D - 0,25 acetate, vitamin D - 0,25 g g of ergocalciferolof ergocalciferol).).

• Currently, the most frequently for quantification use Currently, the most frequently for quantification use physical, physico-chemical and chemical methodsphysical, physico-chemical and chemical methods..

USAGEUSAGE• Parts of biocatalysts (enzymes) that cause different Parts of biocatalysts (enzymes) that cause different

functions in the body metabolismfunctions in the body metabolism.. Hypo-and Hypo-and avitaminosis of the some vitaminavitaminosis of the some vitamin.. In addition In addition, , vitamins ofvitamins of В В group group – –CNS diseases, heart diseasesCNS diseases, heart diseases; ; ВВ12 12 –– anemia anemia; РР –; РР – a violation of the peripheral a violation of the peripheral circulation;circulation; Е – Е – for the pregnancy keeping for the pregnancy keeping; ; DD – – rachitisrachitis; А –; А – skin and eye disease skin and eye disease. . MultivitaminsMultivitamins––support during the cold season, reduce the risk of support during the cold season, reduce the risk of heart diseases and othersheart diseases and others..

• Side effects - allergic reactionsSide effects - allergic reactions ( (Multi-tabs, Kinder-Multi-tabs, Kinder-biovitalbiovital), ), dyspepticdyspeptic disordersdisorders. . Supervitaminosis Supervitaminosis – – vitaminesvitamines А, А, DD, К; , К; essentiale essentiale Н (Н (vitamins of vitamins of ВВgroupgroup) – ) – at the expense of long receptionat the expense of long reception..

Vitamins of the aliphaticVitamins of the aliphatic rowrow

• Ascorbic acid belongs to the derivatives of polyoxyAscorbic acid belongs to the derivatives of polyoxy-γ--γ-lactones oflactones of the unsaturated carboxylic acidsthe unsaturated carboxylic acids,, On the other On the other hand it can be attributed to the vitamins of heterocyclic hand it can be attributed to the vitamins of heterocyclic series as furan derivativeseries as furan derivative. . It is widely distributed in nature. It is widely distributed in nature. Especially rich in it is floraEspecially rich in it is flora:: fresh vegetables, fruits, etc fresh vegetables, fruits, etc.. In In industry, ascorbic acid synthesized from D-glucoseindustry, ascorbic acid synthesized from D-glucose..

• To the vTo the vitamins itamins ofofaliphatic aliphatic rowrow, , derivatives ofderivatives of gluconic gluconic acid esteracid esterss, , pangamic acid belongs pangamic acid belongs ((vitaminvitamin В В1515). ). In In medical practice is usedmedical practice is used its calcium salt its calcium salt. . Pangamic acid is Pangamic acid is a part of the rice bran, yeast, blood, livera part of the rice bran, yeast, blood, liver. .

• To the vTo the vitamins itamins ofofaliphatic aliphatic rowrow, , dertivatives ofdertivatives of β- β-aminoacidsaminoacids,, pantothenic acid belongs pantothenic acid belongs. . Yeast, eggs, liver Yeast, eggs, liver and egg yolk are rich on pentothenic acidand egg yolk are rich on pentothenic acid.. In In medical medical practice is usedpractice is used its calcium salt its calcium salt..

Ascorbic acidAscorbic acid ( (Acidum Acidum ascorbicumascorbicum) ) VitaminVitamin С ( С (SPhUSPhU))

((RR)-5-[()-5-[(SS)-1,2-)-1,2- Dihydroxyethyl Dihydroxyethyl]-3,4-]-3,4-dihydroxydihydroxy-5-5HH--furanfuran-2--2-onon

--lactonelactone-2,3--2,3-dehydrodehydro--LL--gulonic acidgulonic acid

OH OH

O

O

CH2OH

OHH

Extraction of ascorbic acidExtraction of ascorbic acid Available in fresh vegetables - cabbage, beets, lettuce, tomatoes, Available in fresh vegetables - cabbage, beets, lettuce, tomatoes,

potatoes, berries - strawberries, currents, fruits - lemon, orangepotatoes, berries - strawberries, currents, fruits - lemon, orange.. t is t is also found in milk, eggs, rose hips, fennel, etc.also found in milk, eggs, rose hips, fennel, etc.. . For the extraction For the extraction of ascorbic acid from the rose hips produced water of ascorbic acid from the rose hips produced water extract, which is thickened in vacuum to the thicken of extract, which is thickened in vacuum to the thicken of syrupsyrup. . From the remainder precipitated ballast substances with From the remainder precipitated ballast substances with alcohol and ether, and the filtrate evaporated to the dry statealcohol and ether, and the filtrate evaporated to the dry state.. Residue Residue is clearing by crystallization or chromatographic methodis clearing by crystallization or chromatographic method..

The basic amount of ascorbic acid is now extracted synthetically, The basic amount of ascorbic acid is now extracted synthetically, based on D-glucosebased on D-glucose, , that can be transformed by the reduction in the that can be transformed by the reduction in the D-sorbitol and D-sorbitol by enzymatic oxidation - in the D-sorbitol and D-sorbitol by enzymatic oxidation - in the LL--sorbosesorbose

To protect the alcohol group of sorbose, at first it is condensed with To protect the alcohol group of sorbose, at first it is condensed with two molecules of acetone, and then oxidize diacetone sorbose by two molecules of acetone, and then oxidize diacetone sorbose by ККMnOMnO44. . Obtained diacetone ketoglutaric acid is saponificated to Obtained diacetone ketoglutaric acid is saponificated to ketogulonic acidketogulonic acid..

Properties of ascorbic acidProperties of ascorbic acid • White or almost white crystalline powder of or White or almost white crystalline powder of or

colorless crystals that change color under the colorless crystals that change color under the action of air and moistureaction of air and moisture.. Easily soluble in water, Easily soluble in water, soluble in 96% alcohol, practically indissoluble in soluble in 96% alcohol, practically indissoluble in etherether.. Melts at about 190 ° C with decomposition Melts at about 190 ° C with decomposition..

• By endiol group ascorbic acid shows both acidic By endiol group ascorbic acid shows both acidic and restorative propertiesand restorative properties..

• Acidic nature is cased by the mobility of hydrogen Acidic nature is cased by the mobility of hydrogen atom in hydroxyl group in position 3; at the alkali atom in hydroxyl group in position 3; at the alkali titration ascorbic acid behaves like a monobasic titration ascorbic acid behaves like a monobasic acidacid..

Ascorbic acid oxidizes in two stages :

1. reversible oxidation to dehydroascorbic acid (keto-form);

2. irreversible process of oxidation, which leads to the formation of furfural:

CH2OH

O

OH OH

OHH

O[O]

[H]

OOHH

CH2OH

O

OO

[O] OC

H

O

Identification of ascorbic acid 1. By the physico-chemical constants : UV- and IR-

spectroscopy, determination of рН and the specific optical rotation.

2. Add to the solution of ascorbic acid diluted nitrate acid and silver nitrate solution – silver gray metallic precipitate falls:

OH OH

O

O

CH2OH

OHH

2Ag O O

O

O

CH2OH

OHH

+ 2AgNO3 +

2HNO3+ +OH OH

O

O

CH2OH

OHH

2Ag O O

O

O

CH2OH

OHH

+ 2AgNO3 +

2HNO3+ +

3. Unpharmacopoeial reactions:

a) To 1 ml of obtained solution add 2 drops of iron (III) chloride R2 solution and after1 min. 1 drop of of potassium ferricyanide R; blue color appeares:

b) To 1 ml of obtained solution add 5-7 drops of 0,05 М iodine solution; solution becomes transparent:

OH OH

O

O

CH2OH

OHH

2FeCl2

O O

O

O

CH2OH

OHH

+ 2FeCl3 + 2HCl ;+

3FeCl2 + 2K3[Fe(CN)6] → Fe3[Fe(CN)6]2 + 6KCl.

OH OH

O

O

CH2OH

OHH

O O

O

O

CH2OH

OHH

+ I2 + 2HI

c) To 1 ml of obtained solution add 1 drop of CuSO4 solution and 2-3 drops of 1 % NH4CNS solution; white precipitate forms:

d) To the aqueous solution of the preparation add NaHCO3 and FeSO4, shake and leave to stand; a dark purple color appears which disappears at the adding of H2SO4 dil.:

OH OH

O

O

CH2OH

OHH

2CuCNS (NH4)2SO4

O O

O

O

CH2OH

OHH

+ 2CuSO4 + 2NH4CNS +

H2SO4+ + +

OH OH

O

O

CH2OH

OHH

O- OH

O

O

CH2OH

OHH

Fe

+ FeSO4 + 2NaHCO3 +

+

Na2SO4 + 2CO2 + 2H2O

2

2

e) at the adding by drops 2,6- dichlorophenolinedophenolate solution to the solution of ascorbic acid blue color disappears:

f) Ascorbic acid is easily oxidized at the interaction with : • Phosphoric-molybdenum acid (the products of blue color

form);

• methylene blue (discoloration occurs due to formation of leucobase);

• Potassium permanganate(discoloration;

• Fehling reagent (red color)

Assay of ascorbic acid

Iodometry, direct titration (SPhU). Indicator – starch. E m .= М. m./2

Alkalimetry, direct titration . Indicator – phenolphthalein).

E.m. = М. m.

OH OH

O

O

CH2OH

OHH

O O

O

O

CH2OH

OHH

+ I2 + 2HI .

OH OH

O

O

CH2OH

OHH

ONa OH

O

O

CH2OH

OHH

+ NaOH + H2O .

• Iodatometry, direct titration in acidic medium in the presence of potassium iodide, indicator – starch. At the point of equivalence excess of potassium iodate causes the blue color of solution. E.m. = М. m./2.

• Titration by sodium 2,6-dichlorophenolindophenolate solution (for the determination of ascorbic acid in row material). E.m. = М. m./2

• Other redox methods (Iodochlorometry, cerimetry and others).

Ascorbic acid storageAscorbic acid storage• In nonmetallicIn nonmetallic airtight containersairtight containers,, in the dark place in the dark place..

Usage of ascorbic acidUsage of ascorbic acid• Takes part in oxidative-reduction processes, in Takes part in oxidative-reduction processes, in

carbohydrates metabolism and tissue regeneration, carbohydrates metabolism and tissue regeneration, the formation of steroid hormonesthe formation of steroid hormones. . Daily demand of Daily demand of a healthy person - 80-100 mga healthy person - 80-100 mg. .

• In preventive and therapeutic purposes at the In preventive and therapeutic purposes at the scorbutusscorbutus ( (scurvyscurvy),), bleeding of various etiologies, bleeding of various etiologies, infectious diseases and poisonings, liver and infectious diseases and poisonings, liver and kidneys diseaseskidneys diseases. .

• IssueIssue: : tabletstablets, , dragee bydragee by 0,025; 0,05; 0,1; 0,5 0,025; 0,05; 0,1; 0,5 gg; 5% ; 5% andand 10% 10% solutions for injectionssolutions for injections. . Soluble Soluble tablets bytablets by 1,0 1,0 gg..

Calcium pangamateCalcium pangamate((Calcii pangamasCalcii pangamas), ), vitvit. В. В1515, , KalhamKalham

By the chemical structure pangamic acid is an ester ofBy the chemical structure pangamic acid is an ester of DD--gluconic and dimethylaminoaceticgluconic and dimethylaminoacetic acidsacids ( (dimethylglycinedimethylglycine). ).

Substance, except for calcium pangamate contains calcium Substance, except for calcium pangamate contains calcium gluconate (25%) and calcium chloride (6%)gluconate (25%) and calcium chloride (6%)..

Extraction of pangamic acidExtraction of pangamic acid• Isolated from rice bran, yeast, blood, liverIsolated from rice bran, yeast, blood, liver..• Synthesized from D-glucoseSynthesized from D-glucose::

• PropertiesProperties.. White, sometimes yellowish crystalline White, sometimes yellowish crystalline powder with characteristic odorpowder with characteristic odor.. Hygroscopic. Easily Hygroscopic. Easily soluble in water and hardly soluble in organic soluble in water and hardly soluble in organic solventssolvents..

Identification of calcium pangamateIdentification of calcium pangamate

• By theBy the physico-chemical constants: IR physico-chemical constants: IR spectroscopyspectroscopy..

• Substance gives reactions on calcium (4 Substance gives reactions on calcium (4 pharmacopoeial reaction)pharmacopoeial reaction)..

• Residue of gluconic is confirmed by the reaction Residue of gluconic is confirmed by the reaction with iron (III) salts by the formation of light with iron (III) salts by the formation of light green colorgreen color..

• Alkaline hydrolysisAlkaline hydrolysis. . After the heating of substance After the heating of substance with solution of sodium hydroxide smell of with solution of sodium hydroxide smell of amines appears.amines appears.

• The reaction of formation of colored iron The reaction of formation of colored iron hydroxamate (esteric group)hydroxamate (esteric group)::

Assay of calcium pengamateAssay of calcium pengamate• Given that the substance except calcium Given that the substance except calcium

pangamate, also includes calcium gluconate and pangamate, also includes calcium gluconate and calcium chloride, quantitatively determine the calcium chloride, quantitatively determine the content ofcontent of::

1.1. NitrogenNitrogen (3,6 - 4,2%) - (3,6 - 4,2%) - aacidimetry in non-cidimetry in non-aqueous mediumaqueous medium; ;

2.2. CalciumCalcium (5,8 - 7,4%) – (5,8 - 7,4%) – complexonometry complexonometry methodmethod;;

3.3. ChloridesChlorides ( (not more thannot more than 2,2%) - 2,2%) - a method a method ofof reverse reverse argentometry byargentometry by Fol Folggardard;;

4.4. The amount of carboxyl groupsThe amount of carboxyl groups (11 - 15%) -(11 - 15%) - by by ion exchange chromatographyion exchange chromatography..

STORAGESTORAGE

• InIn glassglass airtight containersairtight containers, , in a dry placein a dry place..

Usage of calcium pangamateUsage of calcium pangamate• Activates the transfer of oxygen to the cells of tissues Activates the transfer of oxygen to the cells of tissues

and takes part in the methylation of biosubstrates, and takes part in the methylation of biosubstrates, improves lipid metabolismimproves lipid metabolism. .

• Similar to vitamins preparationSimilar to vitamins preparation. . Calcium pangamate Calcium pangamate is used for the treatment of various forms of is used for the treatment of various forms of atherosclerosis, cirrhosis, alcoholism and other atherosclerosis, cirrhosis, alcoholism and other diseasesdiseases..

• Issue Issue : : tablets bytablets by 0,05 0,05 gg..

Calcium pantothenateCalcium pantothenate ((Calcii pantothenasCalcii pantothenas), ), vitvit. В. В33

Calcium salt ofCalcium salt of DD-(+)-α,γ--(+)-α,γ-dioxydioxy-β,β--β,β-dimethylbutyryldimethylbutyryl--NN--amideamide-β'--β'-aminopropionic acidaminopropionic acid (β- (β-alaninealanine).).

Properties.Properties. White fine crystalline powder without smell White fine crystalline powder without smell.. Easily soluble in water, very little soluble in organic Easily soluble in water, very little soluble in organic solventssolvents..

Extraction of calcium Extraction of calcium pantothenatepantothenate• In large quantities contained in caviar, yeast, In large quantities contained in caviar, yeast,

liver, egg yolk, riceliver, egg yolk, rice..

• Synthetically is extracted by the following Synthetically is extracted by the following scheme:scheme:

Identification of calcium pantothenateIdentification of calcium pantothenate

1.1. By the physico-chemical constants: the specific By the physico-chemical constants: the specific rotation fromrotation from +25 +25 toto +28° (5 % +28° (5 % aqueous solution) aqueous solution)

2.2. Substance gives reactions on calciumSubstance gives reactions on calcium..

3.3. With the solution of copper (II) sulfate in an With the solution of copper (II) sulfate in an alkaline medium substance forms a blue complex alkaline medium substance forms a blue complex ((ββ-alanine)-alanine)::

4.4. Residue ofResidue of α,γ - α,γ -dioxydioxy- β,β –- β,β –dimethylbutyric aciddimethylbutyric acid determine after alkaline hydrolysisdetermine after alkaline hydrolysis. . . Substance is boiled . Substance is boiled with sodium hydroxide solution, after cooling acidified by with sodium hydroxide solution, after cooling acidified by acid chloride solution and added iron (III) chloride - acid chloride solution and added iron (III) chloride - yellow color appearsyellow color appears::

5.5. The reaction of colored iron hydroxamate The reaction of colored iron hydroxamate formation formation :: γγ-butyrolactone-butyrolactone

Assay of calcium pantothenateAssay of calcium pantothenate

Determine the content ofDetermine the content of:: NitrogenNitrogen (5,7 – 6,0%) – (5,7 – 6,0%) – Kjeldahl methodKjeldahl method; ; CalciumCalcium (8,2 – 8,6%) - (8,2 – 8,6%) - complexonometry methodcomplexonometry method..

Test on purityTest on purity Specific impuritSpecific impurityy. . Pantoyl lactonePantoyl lactone – –is determined is determined

spectrophotometricaly in the visible part of the spectrophotometricaly in the visible part of the spectrum by the reaction of iron hydroxammate spectrum by the reaction of iron hydroxammate formationformation..

StorageStorage• In glass airtight containersIn glass airtight containers,, in a dry place at the room in a dry place at the room

temperaturetemperature..

Usage of calcium pantothenateUsage of calcium pantothenate• Participates in hydrocarbon and fat metabolism, Participates in hydrocarbon and fat metabolism,

stimulates the formation of corticosteroids and stimulates the formation of corticosteroids and is part of coenzyme Ais part of coenzyme A..

• The daily demand - 10-12 mgThe daily demand - 10-12 mg.. Since it is formed Since it is formed during the vital functions of Escherichia coli, its during the vital functions of Escherichia coli, its avitaminosis is not observedavitaminosis is not observed. .

• To treat neuralgia, eczema, allergies, To treat neuralgia, eczema, allergies, polyneuritis, and other diseases related with polyneuritis, and other diseases related with violation of metabolism and at the violation of metabolism and at the inflammatory processesinflammatory processes..

• IssueIssue: : tablets bytablets by 0,1 0,1 gg. .

Vitamins of the alicyclic rowVitamins of the alicyclic row • Vitamins of the alyciclic row include retinolsVitamins of the alyciclic row include retinols ( (vitamins of vitamins of АА group group) )

and calciferols and calciferols ((vitamins ofvitamins of D groupD group). ). • In the core of the retinol molecule is trimethylcyclohexanoic cycle In the core of the retinol molecule is trimethylcyclohexanoic cycle

associated with tetraenol conjugate chain, in the end of which is associated with tetraenol conjugate chain, in the end of which is hydroxyl or aldehyde grouphydroxyl or aldehyde group.. Retinol was obtained from the liver of Retinol was obtained from the liver of fish in 1909 yearfish in 1909 year. . InIn 1928 1928 yy.., Euler found that some plants are , Euler found that some plants are substances that have provitamin activity, scilicet are the precursors of substances that have provitamin activity, scilicet are the precursors of vitaminsvitamins.. Provitamins of vitamin A are Provitamins of vitamin A are αα-, -, ββ-and -and γγ-carotene-carotene..

• . . To vitamins of D group, which is called calciferols or antirachitis To vitamins of D group, which is called calciferols or antirachitis include derivatives of cyclohexanol ethylene-hydrindaneinclude derivatives of cyclohexanol ethylene-hydrindane.. Is opened Is opened few vitamins of D group: D1-D7 similar by the chemical structure, few vitamins of D group: D1-D7 similar by the chemical structure, physico-chemical properties and pharmacological actionphysico-chemical properties and pharmacological action.. The The practical usage hame the following vitamins: vitamin D2 practical usage hame the following vitamins: vitamin D2 (ergocalciferol) and vitamin D3 (cholecalciferol)(ergocalciferol) and vitamin D3 (cholecalciferol)..

• Vitamins D2 and D3 are in egg yolks, caviar, butter, milk. A Vitamins D2 and D3 are in egg yolks, caviar, butter, milk. A significant number of them accompanied by retinol in liver and significant number of them accompanied by retinol in liver and adipose tissue of fish and marine animalsadipose tissue of fish and marine animals..

• In medicine is used retinol acetate and ergocalciferolIn medicine is used retinol acetate and ergocalciferol..

Retinol acetateRetinol acetate((Retinoli acetasRetinoli acetas) ) vvitit. А. А

TransTrans--9,13-9,13-DimethylDimethyl-7-( 1,1,5--7-( 1,1,5-trimethylcyclohexenetrimethylcyclohexene5-5-yl-yl-6)-6)-nonatatraenenonatatraene-7,9,11,13--7,9,11,13-olol-15 -15 acetateacetate

Extraction of retinolExtraction of retinol Hydrolysis of fish liver by Hydrolysis of fish liver by 15 % КОН 15 % КОН solution in the solution in the

atmosphere of inert gas is used for the vitamin A atmosphere of inert gas is used for the vitamin A obtainingobtaining..

Main quantity of vitamin A is obtained synthetically Main quantity of vitamin A is obtained synthetically from citral by the following schemefrom citral by the following scheme:: ββ-ionone-ionone

Properties of the retinol acetateProperties of the retinol acetate White or pale yellow crystals with a weak odorWhite or pale yellow crystals with a weak odor..

Extremely unstable under the action of air Extremely unstable under the action of air oxygen and light (easily oxidizes by the oxygen and light (easily oxidizes by the atmospheric oxygen, especially at elevated atmospheric oxygen, especially at elevated temperatures and light, with the formation of temperatures and light, with the formation of heronic acid)heronic acid).. Practically insoluble in water, Practically insoluble in water, soluble in 95% alcohol, chloroform, ether and soluble in 95% alcohol, chloroform, ether and oilsoils..

Identification of retinol acetateIdentification of retinol acetate1.1. The reaction with antimony (III) chloride in The reaction with antimony (III) chloride in

chloroform environment - blue color appears. chloroform environment - blue color appears. 2.2. The reaction with aluminum chloride - blue The reaction with aluminum chloride - blue

color appears. color appears. 3.3. Reactions on the unsaturated bonds Reactions on the unsaturated bonds

(discoloration of iodine, bromine water, (discoloration of iodine, bromine water, reduction of phosphorwolframic acid, AuCl3)reduction of phosphorwolframic acid, AuCl3)..

Assay of retinol acetateAssay of retinol acetate• UVUV--spectrophotometryspectrophotometry

• Photocolorimetry by the reaction withPhotocolorimetry by the reaction with SbClSbCl33

Storige of retinol acetateStorige of retinol acetateDue to the fact that the substance is easily oxidizes, Due to the fact that the substance is easily oxidizes,

store it in sealed ampoules in a stream of nitrogen, store it in sealed ampoules in a stream of nitrogen, which keep from the action of light, at the which keep from the action of light, at the temperatures no more than temperatures no more than +5°. +5°. Oil solutions of Oil solutions of retinol acetate are stored in chock-filled, well retinol acetate are stored in chock-filled, well corked cups of dark glasses at the temperatures no corked cups of dark glasses at the temperatures no more than more than +10 °С.+10 °С.

Usage of retinol acetateUsage of retinol acetate• In the treatment of avitaminosis, diseases and lesions of the In the treatment of avitaminosis, diseases and lesions of the

skin, eye diseasesskin, eye diseases.. Prescribed as pills, granules, oil solutions Prescribed as pills, granules, oil solutions orally, intramuscularly and topicallyorally, intramuscularly and topically.. During the treatment is During the treatment is necessary to consider the possibility of hypervitaminosisnecessary to consider the possibility of hypervitaminosis..

• Daily dose for a healthy person - 1 mg treatment dose - up Daily dose for a healthy person - 1 mg treatment dose - up to 10 mg (33000 UA), but not more than 30 mg (100 000 to 10 mg (33000 UA), but not more than 30 mg (100 000 UA)UA). .

• 1 1 IUIU = = 0,344 microgram or 100000 IU = 0,0344 g of retinol 0,344 microgram or 100000 IU = 0,0344 g of retinol acetateacetate..

• There are two forms of retinol: acetate and palmytate (more There are two forms of retinol: acetate and palmytate (more stable then acetate)stable then acetate)..

• IssueIssue. . VitaminVitaminА А gel. capsgel. caps. . By By 2500, 8000, 12000, 50000, 2500, 8000, 12000, 50000, 100000 100000 IUIU..

• CapsCaps. . AevitAevit – – vitvit. А + Е – . А + Е – capscaps. №24 (. №24 (retinol retinol 30000 30000 IUIU, , tocopheroltocopherol 70 70 IUIU). ).

VVit. it. A are carotenes, which contained in fruits, carrots, A are carotenes, which contained in fruits, carrots, red peppers and others.red peppers and others. --CCarotene in the human arotene in the human body decomposes under the influence of liver body decomposes under the influence of liver enzymes into two molecules of vitamin enzymes into two molecules of vitamin А, А, - - andand --carotenescarotenes form only one molecule of vitamin form only one molecule of vitamin А. А.ββ αα γγ

ErgocalciferolErgocalciferol ( (ErgocalciferolumErgocalciferolum), ), vitvit. . DD22 ( (SPhUSPhU))

(5(5ZZ,7,7EE,22,22EE)-9,10-)-9,10-SecoergostaSecoergosta-5,7,-5,7,1010(19),22-(19),22-tetraenetetraene-3β--3β-olol

CyclopentanophenantreneCyclopentanophenantrene General formula General formula of calciferolsof calciferols

Extraction of ergocalciferolExtraction of ergocalciferol VitaminVitamin DD22 by its structure is similar to steroidsby its structure is similar to steroids. . This This

vitamin is obtained by the UV-radiation of ergosterinevitamin is obtained by the UV-radiation of ergosterine,, which contains inwhich contains in yeast, the uterine horn etc.yeast, the uterine horn etc.. . Process of Process of ergosterine transformation in ergocalciferol conducts ergosterine transformation in ergocalciferol conducts through the formation of lumisterine and tachysterinethrough the formation of lumisterine and tachysterine::

Properties of ergocalciferolProperties of ergocalciferol • White or slightly yellowish crystalline powder or White or slightly yellowish crystalline powder or

white or almost white crystalswhite or almost white crystals.. Practically insoluble Practically insoluble in water, easily soluble in 96%alcohol, soluble in in water, easily soluble in 96%alcohol, soluble in fatty oils. Sensitive to air, heat and lightfatty oils. Sensitive to air, heat and light.. Solutions Solutions in volatile solvents are volatile and must been used in volatile solvents are volatile and must been used immediately after preparationimmediately after preparation.. In the solutions is In the solutions is possible the depending on temperature and time possible the depending on temperature and time reversible isomerization in pre-ergocalciferolreversible isomerization in pre-ergocalciferol.. Activity of the substance is caused by both Activity of the substance is caused by both componentscomponents..

• Solution of ergocalciferol in oil - is a transparent Solution of ergocalciferol in oil - is a transparent oily liquid from light yellow to dark yellowoily liquid from light yellow to dark yellow..

Identification of ergocalciferolIdentification of ergocalciferol1.1. By the physico-chemical constants By the physico-chemical constants : : IRIR--spectroscopyspectroscopy..2.2. Steroid part of molecule cases Liberman reactionSteroid part of molecule cases Liberman reaction

((chloroform solution of the drug at the shakingchloroform solution of the drug at the shaking with acetic with acetic anhydride and sulfate acid becomes redanhydride and sulfate acid becomes red,, color transforms color transforms to purpleto purple, , then to blue and finally to green)then to blue and finally to green)..

3.3. At the interaction with antimony (III) chloride solution in At the interaction with antimony (III) chloride solution in the presence of acetylchloride formed orange-pink colorthe presence of acetylchloride formed orange-pink color.. This reaction is also used for determination of impurities This reaction is also used for determination of impurities by TLC and assay by the photocolorimetry methodby TLC and assay by the photocolorimetry method..

4.4. Sobel-MeyerSobel-Meyer’s reaction’s reaction (with glycerin-1 ,3-d (with glycerin-1 ,3-dichlohydrineichlohydrine) ) - green color- green colorationation..

5.5. Shaltegger’s reaction (drug is boiled with benzene Shaltegger’s reaction (drug is boiled with benzene solutions of aldehydes (vanillin, furfural, anisic, etc.),then solutions of aldehydes (vanillin, furfural, anisic, etc.),then add perchlorate acid) - red color appearsadd perchlorate acid) - red color appears..

Assay fo ergocalciferolAssay fo ergocalciferol• Liquid chromatographyLiquid chromatography (SPhU(SPhU).).

• PhotocolorimetryPhotocolorimetry

Storage of ergocalciferolStorage of ergocalciferol Ergocalciferol is Ergocalciferol is stored in an airtight container stored in an airtight container

under nitrogen in a dark place at a temperature under nitrogen in a dark place at a temperature from from 2 °С 2 °С toto 8 °С. 8 °С. The contents of opened containers The contents of opened containers should be used immediatelyshould be used immediately. . Medical forms of Medical forms of ergocalciferol are stored in chock-filled, well corked ergocalciferol are stored in chock-filled, well corked cups of dark glasses, because it easily oxidizes by cups of dark glasses, because it easily oxidizes by the air oxygen, under the action of light gradually the air oxygen, under the action of light gradually decomposes and forms toxic productsdecomposes and forms toxic products..

CholecalciferolCholecalciferol ((CholecalciferolumCholecalciferolum), ), vitvit. . DD33

(5(5ZZ,7,7EE)-9,10-)-9,10-SecocholestaSecocholesta-5,7,-5,7,1010(19)-(19)-trienetriene-3β--3β-olol

At first was obtained from fish oilAt first was obtained from fish oil. . Provitamin of vitamin Provitamin of vitamin DD33 is is 7-7-dihydrocholesteroldihydrocholesterol

Because of the presence of cholesterol and 7-dehydrocholesterol Because of the presence of cholesterol and 7-dehydrocholesterol in the humane skin lipids content it is possible to synthesize vitamin in the humane skin lipids content it is possible to synthesize vitamin DD33 under the action of sun radiation or UV-radiation on the surface of under the action of sun radiation or UV-radiation on the surface of the humane body.the humane body.

Usage of calciferolsUsage of calciferols • In the medical practice use alcoholic In the medical practice use alcoholic (0,5 %(0,5 %)) and oiland oil

(0,125 %(0,125 %)) solutions of vitamin Dsolutions of vitamin D22 to prevent and to prevent and

treat rachitis, but also at the bone diseases treat rachitis, but also at the bone diseases associated with violations of calcium metabolismassociated with violations of calcium metabolism. . Vitamins ofVitamins of DD group group areare effective for treatment of effective for treatment of all forms of erythematosus and other skin diseasesall forms of erythematosus and other skin diseases. .

• Daily demand for a healthy manDaily demand for a healthy man– 1000 – 1000 IUIU..

• VitaminVitamin DD33 – – cholecalciferol. Tabletscholecalciferol. Tablets VideineVideine (2000 (2000

ііandand5000 5000 IUIU). ). VitVit. А+. А+DD3 3 – – aqueousaqueous solution solution bottles bottles

byby 10 10 mlml..

• Fish OilFish Oil..

Aromatic vitaminsAromatic vitamins • 2-2-methylmethyl-1,4--1,4-naphthoquinonenaphthoquinone ((vitamin ofvitamin of К К group group))

derivativesderivatives belong to the aromatic vitamin’s rawbelong to the aromatic vitamin’s raw. . They have antiThey have antihemorragic actionhemorragic action and and participate participate in the formation of prothrombinin the formation of prothrombin..

• VitaminVitamin К К11 ( (phylloquinonephylloquinone) ) present in the plantspresent in the plants ( (lucerlucernne e , ,

spinach spinach , , cabbagecabbage), ), vitaminvitamin К К22 ( (pharnoquinonepharnoquinone)) is is in in animal products and is produced by intestinal animal products and is produced by intestinal microfloramicroflora. . VitaminVitamin К К33 ( (menadionemenadione) ) is called is called 2-2-

methylmethyl-1,4--1,4-naphtoquinonenaphtoquinone..

• In medical practice, using a synthetic In medical practice, using a synthetic analogue of vitamin K - vikasol.analogue of vitamin K - vikasol.

Coagulation effect of vitamin K is very specific, since the Coagulation effect of vitamin K is very specific, since the small changes in their molecule lead to a significant change small changes in their molecule lead to a significant change in their activity. The discovery of Ansbaher and Ferngolts in their activity. The discovery of Ansbaher and Ferngolts (1939) was a great achievement, that the 2-(1939) was a great achievement, that the 2-methylnaphthoquinone (it was called vitamin Kmethylnaphthoquinone (it was called vitamin K33) is by three ) is by three times more active than vitamin Ktimes more active than vitamin K11. Insolubility in water of 3-. Insolubility in water of 3-methylnaphthoquinone has led to the synthesis of a number methylnaphthoquinone has led to the synthesis of a number of its water-soluble derivatives (by O.V. Palladin), including of its water-soluble derivatives (by O.V. Palladin), including vikasol.vikasol.

VikasolVikasol (Vikasolum) (Vikasolum) (Menadione of sodium (Menadione of sodium bisulfitebisulfite))

SodiumSodium 2,3- 2,3-dihydrodihydro-2--2-methylmethyl-1,4--1,4-naphthoquinonenaphthoquinone-2--2-sulfonatesulfonate threehydratethreehydrate

• CHARACTERS. CHARACTERS. A pale-A pale-yyellow, crystalline powder, ellow, crystalline powder, freely freely soluble soluble in water, in water, very slightlyvery slightly soluble in soluble in ether, hard soluble in alcohol. ether, hard soluble in alcohol.

CH3

SO3Na

O

O

* 3 H2O

Vikasol obtainingVikasol obtaining Precursor for the synthesis of the drug is a β-Precursor for the synthesis of the drug is a β-

methylnaphthalene, which is extracted from the methylnaphthalene, which is extracted from the waste cokecoal industry. Methylnaphthalene is waste cokecoal industry. Methylnaphthalene is oxidized in acetate acid by chromic anhydride oxidized in acetate acid by chromic anhydride to methylnaphthoquinone, which is heated with to methylnaphthoquinone, which is heated with an aqueous solution of sodium hydrosulfite:an aqueous solution of sodium hydrosulfite:

IIdentification reaction of vikasol is based on its dentification reaction of vikasol is based on its lability in alkaline and acid solutions.lability in alkaline and acid solutions.

IIdentification of vikasoldentification of vikasol 1.1. It gives reaction of a sodiumIt gives reaction of a sodium..

2.2. At the interaction with a sodium hydroxide solution there is At the interaction with a sodium hydroxide solution there is settled down a yellow crystalline precipitate of 2-methyl-1 ,4-settled down a yellow crystalline precipitate of 2-methyl-1 ,4-naphthoquinone, which is extracted by chloroform, purified from naphthoquinone, which is extracted by chloroform, purified from impurities and determine the melting point (104-107 ° C):impurities and determine the melting point (104-107 ° C):

Sodium sulfite is determined after the removal of the Sodium sulfite is determined after the removal of the excess of alkali solution by a iodine solution excess of alkali solution by a iodine solution according to the iodine discoloration reaction. according to the iodine discoloration reaction. Vikasol itself does not react with iodine.Vikasol itself does not react with iodine.

NaNa22SOSO33 + I + I22 + H + H22O O → Na→ Na22SOSO44 + 2HI + 2HI

3.3. At the interaction of vikasol with a At the interaction of vikasol with a concentrated sulfuric acid there is concentrated sulfuric acid there is sulfurdioxide small:sulfurdioxide small:

4.4. SubstanceSubstance + + ethanolethanol + + HClHCl concconc. . red colourred colour..

5.5. Aqua solution of the substanceAqua solution of the substance + + sodium ethylatesodium ethylate

Red-brown colorRed-brown color, , according to the formation of 2-oxi-3-methyl-according to the formation of 2-oxi-3-methyl-naphthoquinonenaphthoquinone ( (phtyoxolphtyoxol).).

ImpuritiesImpurities• Sodium bisulfite and 2-methyl-1,4-naphthohydroquinone-3-Sodium bisulfite and 2-methyl-1,4-naphthohydroquinone-3-

sulfonate are the specific impurities in vikasolsulfonate are the specific impurities in vikasol..

• Sodium bisulfiteSodium bisulfite NaHSONaHSO3 3 is determined by iodometric titration is determined by iodometric titration methodmethod ( (less thanless than 2 %). 2 %).

• 2-2-methylmethyl-1,4--1,4-naphthohydroquinonenaphthohydroquinone-3--3-sulfonatesulfonate is determined is determined by the adding ofby the adding of о- о-phenanthrolinephenanthroline - - hould not form the hould not form the precipitate (the impurity is not allowed.precipitate (the impurity is not allowed.

AssayAssay1.1. CerimetriCerimetri. Direct titration, the indicator is o-. Direct titration, the indicator is o-

phenanthroline. By the interaction with a sodium phenanthroline. By the interaction with a sodium hydroxide solution 2-methyl-1 ,4-naphthoquinone is hydroxide solution 2-methyl-1 ,4-naphthoquinone is precipitated, which is extracted with chloroform. After precipitated, which is extracted with chloroform. After removal of chloroform, it is reduced in an acidic removal of chloroform, it is reduced in an acidic medium to 2-methyl-1 ,4-dioxinaphthalene, then it is medium to 2-methyl-1 ,4-dioxinaphthalene, then it is titrated by a solution of cerium (IV) sulfate until the titrated by a solution of cerium (IV) sulfate until the green color:green color:

2.2. GGravimetryravimetry. (. (Precipitation formPrecipitation form – 2- – 2-methylmethyl-1,4--1,4-naphthoquinonenaphthoquinone).).

StorageStorage• Store protected from light.Store protected from light.

ApplicationApplication To increase the clotting of blood at various To increase the clotting of blood at various

bleedings. Water-soluble synthetic bleedings. Water-soluble synthetic substitute of K group vitamins, which take substitute of K group vitamins, which take part in the formation of liver prothrombin part in the formation of liver prothrombin

and promotes the normal blood and promotes the normal blood coagulation. At Haemophilia it does not coagulation. At Haemophilia it does not

act. It acts during 12-18 hours after act. It acts during 12-18 hours after injection.injection.

ProducedProduced: : powderpowder, , tabletstablets. о. оnn 0,015 0,015 gg, 1% , 1% solution for solution for injectioninjection..

• Per oral: the highest day dosePer oral: the highest day dose – 60 – 60 mg,mg, intraintra//muscularmuscular – 30 – 30 mgmg..

Thank you for attention!