LectureLecture № №33Alkaloids, indole derivatives and Alkaloids, indole derivatives and purine alkaloids and their salts, purine alkaloids and their salts, and some synthetic analogs by and some synthetic analogs by the biological action of the biological action of substances like drugs and substances like drugs and components of dosage forms.components of dosage forms.

Ass. Medvid I. I.Ass. Medvid I. I.

Indole - condensed system of pyrrole and Indole - condensed system of pyrrole and benzene cycles which have two share benzene cycles which have two share

atoms atoms ::

Indole is a structural basis Indole is a structural basis of physostigmine, of physostigmine, strychnine, reserpine alkaloidsstrychnine, reserpine alkaloids..

Group reaction on indole derivatives Group reaction on indole derivatives – – Van-Urk’s reactionVan-Urk’s reaction

In the base of reaction is the process of electrophyllic In the base of reaction is the process of electrophyllic substitution. Reagent p-dimethylaminobenzaldehydesubstitution. Reagent p-dimethylaminobenzaldehyde. . Reaction conducts at the presence of conc. HReaction conducts at the presence of conc. H22SOSO44 and and

FeClFeCl33 as oxidantas oxidant..

Derivatives of indole which have free 2 and 3 positions Derivatives of indole which have free 2 and 3 positions give this reactiongive this reaction. . Reserpine gives this reaction by the Reserpine gives this reaction by the opening of ring C in the presence of acidsopening of ring C in the presence of acids, , as a result as a result position 2 becomes freeposition 2 becomes free..

Product of reaction can exist in 2 formsProduct of reaction can exist in 2 forms. . Color of the Color of the reaction product depends on the chemical structure of reaction product depends on the chemical structure of primer compounds conditions of the reactionprimer compounds conditions of the reaction. . Van-Urk’s Van-Urk’s reaction can be hold with another aldehydereaction can be hold with another aldehyde. . So, for So, for reserpine solution of vanillin in chloride acid is usedreserpine solution of vanillin in chloride acid is used..

Physostigmine salicylate (Physostigmini salicylas)

Eserini salicylas

Salicylate of ester of methylcarbaminic acid and eseroline or 5-methylcarbaminoiloxy-1,3,1’-trimethyl-2,3,2’,3’-

tetrahydropyrrole indole

N N

OCH3

CH3 CH3

C

O

NH

H3C COOH

OH

*

PhysostigminePhysostigmine– – the the main alkaloid main alkaloid of of

calabaric calabaric beans beans ((Faba Faba calabarica) – calabarica) –

poisonous seeds of poisonous seeds of West AfricanWest African plant plant

Physostigma Physostigma venenosus , Fabaceae venenosus , Fabaceae

Physical propertiesPhysical properties Physostigmine salicylate Physostigmine salicylate -- brilliant colorless brilliant colorless or or almost colourless almost colourless prismaticprismatic crystalscrystals. . Soluble in water, Soluble in water, easilyeasily soluble in alcohol, practically soluble in alcohol, practically insoluble in ether insoluble in ether.. Aqueous solutions are unstable. It melts at about 182 °C, Aqueous solutions are unstable. It melts at about 182 °C, with decompositionwith decomposition Optic active compoundOptic active compound. . When heated When heated with water easy hydrolyze and therefore its solutions for with water easy hydrolyze and therefore its solutions for parenteral usage produce in aseptic introductionsparenteral usage produce in aseptic introductions. . On the On the air and light product paints in the red color – air and light product paints in the red color – pharmacological inactive rubrezerine formedpharmacological inactive rubrezerine formed..

PharmacologicalPharmacological action cased by the methylurethane groupaction cased by the methylurethane group.. ProserineProserine - - white crystalline powder with bitter tastewhite crystalline powder with bitter taste. . Very Very

easily soluble in water, easilyeasily soluble in water, easily soluble in alcohol and soluble in alcohol and chloroformchloroform, , ether ether. . HygroscopicHygroscopic. . Becomes pink on the Becomes pink on the lightlight..

Identification of Identification of Physostigmine salicylatePhysostigmine salicylate 1.1. Melting point, the specific rotationMelting point, the specific rotation..

2.2. Substance gives reaction to salicylates (2 reactions in Substance gives reaction to salicylates (2 reactions in SPU).SPU).

3.3. Total Pharmacopeial reaction Total Pharmacopeial reaction on on alkaloids (withalkaloids (with Dragendorff's reagentDragendorff's reagent))

4.4. After evaporation of the preparation with ammonium After evaporation of the preparation with ammonium forms blue residue forms blue residue ((physostigmine base)physostigmine base),, which is which is dissolved in ethanol with formation of a blue solution dissolved in ethanol with formation of a blue solution which after the acidificationwhich after the acidification by by СНСН33СООН СООН becomes redbecomes red..

5.5. Drug solution in HDrug solution in H22SOSO44 conc conc. . gradually becomes yellowgradually becomes yellow. .

6.6. Erdman and Frede reagents with medication give reddish-Erdman and Frede reagents with medication give reddish-yellow coloryellow color, , withwith HNOHNO33 concconc. – . – yellow coloryellow color..

7. 7. When heated with alkalis (and gradually when heated with When heated with alkalis (and gradually when heated with water)water) physostigmine salycilate hydrolyzed and appears physostigmine salycilate hydrolyzed and appears character odor methylaminecharacter odor methylamine::

8. 8. At the heating with At the heating with 0,1 % 0,1 % ninhydrine solution in conc. ninhydrine solution in conc. HH22SOSO44 on the water bath at on the water bath at 60 60 00С С duringduring 10 10 minmin. . And than And than

after cooling solution have green fluorescenceafter cooling solution have green fluorescence. . Proserine Proserine gives blue fluorescence at this conditionsgives blue fluorescence at this conditions. .

9. 9. At the gradually adding to the solution boric acidAt the gradually adding to the solution boric acid, 0,1 М , 0,1 М solution of nitrate acid and sodium nitritesolution of nitrate acid and sodium nitrite,, after 1 min. add after 1 min. add sodium hydroxide solution, violet color appearssodium hydroxide solution, violet color appears..

Assay Assay

Physostigmine salicylatePhysostigmine salicylate 1.1. Alkalimetriya, direct titrationAlkalimetriya, direct titration . . The drug is dissolved in The drug is dissolved in

a mixture of ethanol and chloroform anda mixture of ethanol and chloroform and titrated by 0,1 М 0,1 М NNаОН аОН to the pink color to the pink color ((indicatorindicator – – phenolphthaleinphenolphthalein). Е = М.). Е = М.mm. .

2.2. Acidimetry in non-aqueous mediumAcidimetry in non-aqueous medium.. The drug is dissolved in a The drug is dissolved in a mixture of chloroform and concmixture of chloroform and conc. . CHCH33COOHCOOH,, titrated by titrated by 0,1 М 0,1 М ННClClОО44.. For determination of the end-point use potentiometry. For determination of the end-point use potentiometry. Equivalent point is fixed by potentiometric methodEquivalent point is fixed by potentiometric method. Е = М.. Е = М.m./m./22. .

3.3. Complexonometry, reverse titrationComplexonometry, reverse titration.. As a stable titrant acetic-As a stable titrant acetic-acidic solution of bismuth nitrate is used in the presence of acidic solution of bismuth nitrate is used in the presence of potassium iodidepotassium iodide. . Scheme of reactionScheme of reaction::

Product of the interaction is filtrated and an excess of reagent is Product of the interaction is filtrated and an excess of reagent is titrated by titrated by 0,1 М 0,1 М sodium EDTA solutionsodium EDTA solution. Е = М.. Е = М.mm. .

STORAGESTORAGE and USAGE of Physostigmine USAGE of Physostigmine salicylatesalicylate

InIn an airtight containers of dark glass, protected from an airtight containers of dark glass, protected from

lightlight. . Poison compoundPoison compound. .

Cholinesterase inhibitorCholinesterase inhibitor, , myotic mean myotic mean ((atropine atropine antagonistantagonist). ). Used for the treatment of glaucoma as Used for the treatment of glaucoma as 0,25-1% 0,25-1% eye dropseye drops. . IntroduceIntroduce subcutaneoussubcutaneous 0,1 0,1% % --1,0 1,0 solution the neuromuscular solution the neuromuscular diseases (Alzheimer's diseases (Alzheimer's

disease)disease). H. dH. d. – 0,0005 . – 0,0005 gg, , H. d. dH. d. d. – 0,001 . – 0,001 gg..

Synthetic substitute of physostigmine

Proserine (Proserinum) Neostigmine methylsulfate*

N-(m-dimethylcarbamoiloxiphenyl)-N,N,N-trimethylammonium methylsulfate

OC

O

NH3C

H3CN CH3

CH3

CH3 CH3SO4-

+

Identification of proserineIdentification of proserine1.1. Reaction toReaction to methylsulfate-ion. . If after heating If after heating the the

drug withdrug with HNOHNO3 3 concconc. . add solution ofadd solution of BaClBaCl22, ,

white precipitatewhite precipitate falls ((BaSOBaSO44))..

2.2. With a solution of iodine preparation forms brown With a solution of iodine preparation forms brown sedimentsediment of periodide..

3.3. At the heating of drug with alkali dissolution of At the heating of drug with alkali dissolution of urethane groupurethane group takes place ((mm--dimethylaminophenol formeddimethylaminophenol formed, , which is detected which is detected by the condensation with diazotative sulfanylic by the condensation with diazotative sulfanylic acid acid – – cherry-red color (azo-dyescherry-red color (azo-dyes)):)):

AssayAssay The modified K'yeldal methodThe modified K'yeldal method.. The drug is boiled in a The drug is boiled in a

K'yeldalK'yeldal flask flask with NaOHNaOH. . DimethylamineDimethylamine, , whichwhich evaporates, , distilled with water vapor in the receiver distilled with water vapor in the receiver with a solution of boric acidwith a solution of boric acid. . Metaborate and tetraborate Metaborate and tetraborate of dimethylamine formedof dimethylamine formed, , which are titrated by which are titrated by 0,1 М 0,1 М solution of HCl (mixed indicatorsolution of HCl (mixed indicator). Е = М.). Е = М.mm. .

STORAGESTORAGE and USAGE of proserine and USAGE of proserine In an airtight containers of dark glass, protected from In an airtight containers of dark glass, protected from

lightlight. . Poison compoundPoison compound.. Substitute of physostigmineSubstitute of physostigmine. .

Anticholinesterase, antimyasthenic meanAnticholinesterase, antimyasthenic mean. . Curare Curare antagonist drugsantagonist drugs.. Used for treatment of Used for treatment of myasthenia, paralysis, neuritis, atony of intestine myasthenia, paralysis, neuritis, atony of intestine and urinary bladder, glaucoma, for stimulating and urinary bladder, glaucoma, for stimulating labor activity as labor activity as 0,25-1% 0,25-1% as eye dropsas eye drops. .

IssueIssue – – tabletstablets 0,015 0,015 gg, , amp.amp. 0,05%-1,0. 0,05%-1,0. H. dH. d..,, internallyinternally – 0,015 – 0,015 gg, , H. d. d.,H. d. d., internally internally – –

0,05 0,05 gg; ; H. dH. d. . subcutaneoussubcutaneous – 0,002 – 0,002 gg, , H. d. dH. d. d. . s/c s/c – – 0,006 0,006 gg..

Strychnine nitrate (Strychnini nitras)

Cycles АВ, BD, ED – derivatives of indole.

Cycle А – aromatic and strychnine can be nitrated and halogenated.

N19 – tertiary atom, has a base character and gives salts with acids.

N9 – is in the lactam group, which may be disclosed by the interaction with alcohol solution of KOH with formation of carboxyl and secondary amino-groups.

N

O

N

O

* HNO3

A B

C

D

E

F

G

12

34

5

6 7

8

9

1011

12

13

14

1516

17 18

19

20

21

2223

Strychnine can be found with brucine in the

seeds of tropical plant Strychnos

Nux Vomica (emetic nut )

Reserpine (Reserpinum)

11,17-dimethoxy-16-carbmethoxy-18(3’,4’,5’,-trimethoxibenzoyloxy)-alloyohimban

NH

NH3CO

H3COOC

OCH3

O C

O

OCH3

OCH3

OCH3

Reserpine molecule contains indole (АВ), dihydroquinolysidine (СD),

partially hydrogenated 3-carbolynic (АВС), hydrogenated isoquinoline (ED) cycles.

NH

NA C

D

E

12

34

56

789

10

1112

13

14

15

1617

18

19

20

21

B

Àëî é î õ³ì áàí

AlloyohibanAlloyohiban

Reserpine contains in the

roots of the plant

Rauwolfia serpentina

Benth

Physical propertiesPhysical properties

Strychnine nitrateStrychnine nitrate - a - a colorless colorless brilliantbrilliant crystals crystals with with very very bitter tastebitter taste. . Difficultly soluble in water and alcohol, easily Difficultly soluble in water and alcohol, easily soluble in boiling watersoluble in boiling water, , practically insoluble in etherpractically insoluble in ether. .

ReserpineReserpine - - colorless, white or slightly yellow, small colorless, white or slightly yellow, small crystals or crystalline powdercrystals or crystalline powder with melting point melting point 261- 261-265265°°С. С. Insoluble in water, soluble in chloroform, acetone, Insoluble in water, soluble in chloroform, acetone, pyridine and ether, darkening slowly on the exposure topyridine and ether, darkening slowly on the exposure to

llightight. Optic active compound. Optic active compound. . At the heating with acids or At the heating with acids or alkalis hydrolysisalkalis hydrolysis takes place (reserpinic acid, methanol, trimethoxybenzoic acid form).).

Identification of strychnine nitrateIdentification of strychnine nitrate

1. Pharmacopeial reaction on alkaloids.

2. Solution of the drug in H2SO4 conc. + crystal of K2Cr2O7 – formed the blue-violet strips which pass into the red and lilac-green.

3. Vitali-Moren’s reaction. At the interaction with HNO3

conc. drug becomes yellow (as opposed to brucine, which becomes blood-red) by nitration of benzene cycle А; after the evaporation of reaction product the residue gives with alcohol solution of КОН formed red-violet color.

4. Van-Urk’s reaction (on indole cycles). With 1% vanillin in glycerol in the presence of H2SO4 dil. Pink-violet color appears.

5. Reaction on nitrate ions NO3–-:

а) SPU. The interaction with nitrobenzene in the presence of sulfate acid

Quantity of substance, listed in a separate article, add to the mixture of 0,1 ml of nitrobenzol R and 0,2 ml of sulfate acid R and after 5 min. cooled in ice water. Continuing to cool slowly and while stirring add 5 ml of water R, 5 ml of sodium hydroxide concentrated solution R NaOH, 5 ml of acetone R, shake and put for standing; the apper layer becomes dark purple.

b) SPU, N. Not discolors potassium permanganate The solution of the substance, acidified by acid sulfate diluted R

H2SO4, not discolors solution of 1 g/l potassium permanganate R (difference of nitrite ).

с) Not pharmacopeial reaction. Interaction with iron (ІІ) sulfate FeSO4 in the medium of conc. H2SO4; brown ring is formed (FeSO4NO) (on the clock glass ):2 Strychnine•НNO3 + 6FeSO4 + 4H2SO4 = 2NO + 3Fe2(SO4)3 +

(Strychnine)2•Н2SO4 + 4H2ONO + Fe2+ + SO4

2– [Fe(NO)]SO4

d) Unpharmacopeial reaction. Interaction with diphenylamine in acidic medium. (conc. H2SO4), formed an bright blue organic dye:

diphenylbenzidine

Sulfimmonium salt of diphenylbenzidine (blue dye)

H2SO4NH NHNH

NO2

H2SO4

2+

N N

H

HSO4

+

Identification of reserpineIdentification of reserpine1.1. Specific optical rotation Specific optical rotation ((6 assymetric carbon 6 assymetric carbon

atomsatoms).).2.2. UV-spectroscopy UV-spectroscopy ((chromophor groups chromophor groups – – indole indole

andand trimethoxybenzoatic acid – 2 maximum of trimethoxybenzoatic acid – 2 maximum of absorption on UV-spectrumabsorption on UV-spectrum).).

3.3. Reactions of indole cycleReactions of indole cycle.. With chloric water With chloric water ––purple, withpurple, with KMnOKMnO44 – – dark lilac, with vanillin in dark lilac, with vanillin in the presence of HCl - pink, with the presence of HCl - pink, with НН22ОО22 – – yellow-yellow-lilac colorlilac color..

4.4. Water solutions of reserpine in UV-light Water solutions of reserpine in UV-light – – blue blue fluorescencefluorescence..

5.5. Alcohol solution of the preparation Alcohol solution of the preparation + + HH22SOSO44 + + NaNONaNO22 – – green fluorescencegreen fluorescence..

6.6. With Frede reagentWith Frede reagent – – red color, which goes to the red color, which goes to the blueblue..

7.7. On ester groupsOn ester groups:: а) а) alkali hydrolysisalkali hydrolysis;; б)б) hydroxame sample hydroxame sample..8.8. Van-Urk’s reactionVan-Urk’s reaction. . With p- With p-

dimethylaminobenzaldehydedimethylaminobenzaldehyde + + HH22SOSO44 ++

СНСН33СООН –СООН – green coloring that goes into the green coloring that goes into the

redred. . With vanillin in chloride acid With vanillin in chloride acid – – pink colorpink color..

AssayAssayStrychnine nitrate Strychnine nitrate – – AlkalimetryAlkalimetry, , direct titrationdirect titration. .

Titration of the drug in alcohol-chloroform solution Titration of the drug in alcohol-chloroform solution of of 0,1 М 0,1 М NaOHNaOH ( (phenolphthalein indicatorphenolphthalein indicator). ).

Е = М.Е = М.mm..Specific additive - brucineSpecific additive - brucine. .

ReserpineReserpine – – Acidimetry in non-aqueous medium.Acidimetry in non-aqueous medium. Hatch is titrated in the medium of anhydrous Hatch is titrated in the medium of anhydrous СНСН33СООН СООН by by 0,1 М 0,1 М solutionsolution HClOHClO44 ( (indicator indicator crystal violet) to the appearance of green colorcrystal violet) to the appearance of green color. Е = . Е = М.М.mm..

StorageStorage, , usageusage Strychnine nitrate Strychnine nitrate

In airtight containersIn airtight containers. . Poison compoundPoison compound. .

CNS stimulant, tonic CNS stimulant, tonic meanmean. .

Issue - amp. Issue - amp. 0,1%-1,0.0,1%-1,0. H. dH. d. . ss//cc – 0,002 – 0,002 gg; ;

H. d. dH. d. d. . ss//cc – 0,005 – 0,005 gg..

ReserpineReserpine In airtight containersIn airtight containers , in a dark , in a dark

placeplace.. Powder Powder -- poisonous poisonous substancesubstance. .

Neuroleptic Neuroleptic ,, treatment of treatment of hypertensionhypertension. . Included in Included in tablets: Adelphane (0.1 mg of tablets: Adelphane (0.1 mg of reserpine,reserpine,10 10 mgmg of of dihydralasinedihydralasine), ), Adelphan Adelphan –– esidrex esidrex ((TriresidTriresid)) (Adelfane (Adelfane + + 10 10 mg ofmg of dichlorothiazidedichlorothiazide), ), Adelphane Adelphane –– esidrex esidrex -К -К ((TriresidTriresid К)(0,6 К)(0,6 g ofg of КС КСll), ), CrystepinCrystepin, , NeocrystepinNeocrystepin. . RaunatineRaunatine––the amount of the amount of Rauwolfia alkaloidsRauwolfia alkaloids..

Alkaloids, purine derivatives Alkaloids, purine derivatives PurinePurine – –condensed system condensed system of pyrimidine and imidazolof pyrimidine and imidazol

If in the core of purine hydrogen atoms in the pyrimidine If in the core of purine hydrogen atoms in the pyrimidine nucleus replaced by hydroxyl groupsnucleus replaced by hydroxyl groups, we will get xantine :

Caffeine is contained in coffee beans Caffeine is contained in coffee beans ((Coffea Coffea arabicaarabica), ), tea leaves tea leaves ((Thea sinensisThea sinensis), ), cola cola ((Cola Cola

acuminataacuminata) )

PharmacologyPharmacology

Caffeine stimulates the central nervous system Caffeine stimulates the central nervous system first at the higher levels, resulting in increased alertness first at the higher levels, resulting in increased alertness and wakefulness, faster and clearer flow of thought, and wakefulness, faster and clearer flow of thought, increased focus, and better general body coordination, increased focus, and better general body coordination, and later at the spinal cord level at higher doses. Once and later at the spinal cord level at higher doses. Once inside the body, it has a complex chemistry, and acts inside the body, it has a complex chemistry, and acts through several mechanisms as described below.through several mechanisms as described below.

Metabolism and half-lifeMetabolism and half-life

Theophylline first Theophylline first was isolated from was isolated from tea leaves tea leaves ((Thea Thea

sinensissinensis))

Theobromine is extracted Theobromine is extracted from cocoa beans from cocoa beans

((Theobroma cacaoTheobroma cacao))

Three natural alkaloids, derivatives Three natural alkaloids, derivatives xantine: caffeine, theophylline, xantine: caffeine, theophylline,

theobromine theobromine ::

Extracted from semi-synthetic uric acid, guanine, urea.Extracted from semi-synthetic uric acid, guanine, urea.

Very convenient is the method of extraction of caffeine Very convenient is the method of extraction of caffeine and theobromine from xantine, which can be extracted and theobromine from xantine, which can be extracted from uric acid from uric acid ((waste poultry farms) and guanine (fish waste poultry farms) and guanine (fish flakes, waste of paper) flakes, waste of paper) ::

Synthesis of caffeine and theophyllin by method Synthesis of caffeine and theophyllin by method Hmelevskiy - AbramovaHmelevskiy - Abramova( firs was synthesed by Traube

in1900 year)

Caffeine Medications• Caffeine (Соffеіnuт) ,

Caffeine monohydrate (Соffеіnuт

monohydricum) (SPU)

1,3,7-Trimethyl-3,7-dihydro- 1H-purine-2,6-dione

1,3,7-trimethylxantine

• Caffeine-sodium benzoate (Coffeinum-

natrii benzoas)

N

N N

N

O

O

H3C

CH3

CH3

N

N N

N

O

O

H3C

CH3

CH3

C

O

ONa

*

Physical propertiesPhysical properties Caffeine - White or almost White or almost white, crystalline powder or white, crystalline powder or silky crystals, sublimes readilysilky crystals, sublimes readily. Moderatly solubleModeratly soluble

in water, freely soluble in in water, freely soluble in boiling water, slightly soluble boiling water, slightly soluble in ethanol and ether. Soluble in in ethanol and ether. Soluble in the concentrated solutions of the concentrated solutions of alkali benzoate or salicylatesalkali benzoate or salicylates. Very weak base, forms Very weak base, forms unstable salts with acids by unstable salts with acids by nitrogen in position 9nitrogen in position 9.

Caffeine-sodiume benzoate –white powder, white powder, odorless, bitter tasteodorless, bitter taste.

Easily soluble in water, Easily soluble in water, slightly soluble in ethanol. slightly soluble in ethanol. Contains 38-40% caffeine.Contains 38-40% caffeine.

Extracted by the mixing and Extracted by the mixing and evaporation to the dry state evaporation to the dry state of aqueous solutions of aqueous solutions containing equimolar containing equimolar quantity of caffeine and quantity of caffeine and sodium benzoatesodium benzoate.

General Pharmacopeial reaction - a reaction Pharmacopeial reaction - a reaction on xantines on xantines (Murexide reaction or reaction on

purine alkaloids):

Identification of caffeineIdentification of caffeine1.1. By the By the physicphysicoo-chemical constants: melting point, IR-chemical constants: melting point, IR--

spectroscopyspectroscopy..2.2. With potassium iodide in iodine in the presence of HCl dilWith potassium iodide in iodine in the presence of HCl dil.- .-

brown precipitate formed(periodidebrown precipitate formed(periodide С С88НН1010NN44ОО22•J•J22•HJ), •HJ), which which dissolves in NaOH dil. solution at the neutralizationdissolves in NaOH dil. solution at the neutralization..

3.3. Murexide sampleMurexide sample..4.4. Unpharmacopoeial reaction Unpharmacopoeial reaction - - with 1% solution of tannin – white

precipitate dissolved in excess of reagent. .

5.5. With HgClWith HgCl22 – white crystalline sediment, which is a complex – white crystalline sediment, which is a complex compound with the following content Ccompound with the following content C55HH1010NN44OO22· HgCl· HgCl22..

6.6. WithWith acetylacetone and dimethylaminobenzaldehydeacetylacetone and dimethylaminobenzaldehyde. . Solution of Solution of the substance in a mixture of acetylacetone and dil. NaOH heated the substance in a mixture of acetylacetone and dil. NaOH heated in a water bath, cooledin a water bath, cooled, , than add solution of than add solution of dimethylaminobenzaldehyde and heat again, cool and add water - dimethylaminobenzaldehyde and heat again, cool and add water - appears an intense blue colorappears an intense blue color::

Caffeine monohydrate gives all reactions on Caffeine monohydrate gives all reactions on caffeine after a preliminary drying at 100-105 ° C. caffeine after a preliminary drying at 100-105 ° C.

Identification of caffeine-sodium Identification of caffeine-sodium benzoatebenzoate

1. 1. Caffeine identify byCaffeine identify by: : aa) ) melting temperature (234-237 ° C) after extraction melting temperature (234-237 ° C) after extraction

by chloroform from alkaline solutionby chloroform from alkaline solution;;bb) ) Murexide sampleMurexide sample;;cc) ) reaction reaction with 1% solution of tannin;;dd)) reaction with iodine solution reaction with iodine solution;; 2. 2. Sodium benzoate identify bySodium benzoate identify by::ee) ) the reaction with solution of iron (III) chloride - the reaction with solution of iron (III) chloride -

pink-yellow sedimentpink-yellow sediment;;ff)) the sodium cation paints the flame in yellow color the sodium cation paints the flame in yellow color..

AssayAssayCaffeine

1.1. Acidimetry in non-aqueous Acidimetry in non-aqueous medium medium in a mixture of acetic in a mixture of acetic acidacid anhydrous anhydrous, acetic , acetic anhydride and toluene, a direct anhydride and toluene, a direct titration. Potentiometric titration. Potentiometric indication, the control indication, the control experiment experiment (Е=М.(Е=М.mm).).

2.2. IodometryIodometry,, reverse titration reverse titration, , indicatorindicator - - starchstarch (Е=М.(Е=М.mm/4)./4).

3.3. CerimetryCerimetry,, reverce titration reverce titration, , with iodometric endingwith iodometric ending. . An An excess of cerium is neutralized excess of cerium is neutralized by potassium iodide solutionby potassium iodide solution. . Sodium thiosulfate used as Sodium thiosulfate used as titranttitrant. (Е=М.. (Е=М.mm/4)./4).

Caffeine-sodium benzoate

1. Caffeine content is determined by iodometric method (Е=М.m/4). ). ). In the dry matter it should be In the dry matter it should be 38,0 - 40,0 %.

2.2. Sodium is Sodium is determined in the determined in the presence of mixed indicator presence of mixed indicator (methyl orange solution and (methyl orange solution and methylene blue at a ratio of methylene blue at a ratio of 1:1) and ether (for the 1:1) and ether (for the extraction of benzoic acid, extraction of benzoic acid, available in the titration) available in the titration) (Е=М.(Е=М.mm). ). Sodium benzoate Sodium benzoate in in the dry matter must be not less the dry matter must be not less than than 58,0 % 58,0 % and not more thanand not more than 62,0 %.62,0 %.

Cerimetric determination of caffeine

2 Ce(SO4)2 + 2 KI I2 + K2SO4 + Ce2(SO4)3

I2 + 2 Na2S2O3 2 NaI + Na2S4O6

N

N N

N

O

O

H3C

CH3

CH3

+ 4 Ce(SO4)2 + 3 H2O H2N C NH

O

CH3

N

N

H3C

CH3

O

O

OO

+ 2 Ce2(SO4)3 + 2 H2SO4

+

+

StorageStorage, , UsageUsageCaffeine

In a dry, dark In a dry, dark placeplace.. Central nervous system stimulantCentral nervous system stimulant,,

cardiotonic meancardiotonic mean, , at the at the angiospasmsangiospasms;; enuresis in enuresis in children; stimulant of mental children; stimulant of mental and physical disability, and physical disability, poisoning with drugspoisoning with drugs.. Produced Produced as powderas powder. .

Caffeine monohydrate is part of Caffeine monohydrate is part of the tabletsthe tablets: : TheophedrineTheophedrine, , CytramoneCytramone, , CytropakCytropak, , AskofeneAskofene, , CofficyllCofficyll, , Cophetamine,Cophetamine, BenalginBenalgin, , ColdrexColdrex, , SolpadeinSolpadein, , PanadolPanadol--екстра. екстра. Apply in doses Apply in doses by by 0,05-0,1 0,05-0,1 gg as CNS stimulantas CNS stimulant..

Caffeine-sodium benzoate

In a dry, dark In a dry, dark placeplace.. Central nervous system Central nervous system

stimulantstimulant,, cardiotonic cardiotonic meanmean. . Thanks to the Thanks to the solubility in water used in solubility in water used in the form of injection the form of injection solutionssolutions. . Issue - powderIssue - powder, , tabletstablets 0,1 0,1 andand 0,2 0,2 gg, 0,075 , 0,075 gg ( (for childrenfor children); 10% і 20% ); 10% і 20% solutions in amp. bysolutions in amp. by 1,0- 1,0-2,0 2,0 mlml. . Included in the Included in the tabletstablets: : AnapryllineAnaprylline, , PentalginPentalgin..

• Theobromine Theobrominum

(SPU)

3,7-Dimethyl-3,7- dihydro-1Н-purine-2,6-dione, or 3,7- dimethylxantine

• Theophylline monohydrate

Theophyllinum monohydricum (SPU)

1,3-Dimethyl-3,7- dihydro-1Н-purine-2,6-dione monohydrate,

or monohydrate of 1,3- dimethylxantine

N

N N

NH

O

O

H3C

CH3

* H2O

HN

N N

N

O

O

CH3

CH3

Properties

Theobromine

White crystalline powder, with bitter taste. very slightly soluble in water, ethanol ,ether and chloroform; slightly soluble in hot water; easily in dil. Solutions of alkalis and acids.

Theophylline

White crystalline powder. Sightly soluble in water, ethanol and chloroform; easily soluble in hot water; soluble in dil. Solutions of acids and alkalis.

Theobromine and theophylline - amphoteric Theobromine and theophylline - amphoteric compounds with a predominance of acidic compounds with a predominance of acidic properties (by moving the hydrogen atom at the properties (by moving the hydrogen atom at the nitrogen atom in position 1 or 7)nitrogen atom in position 1 or 7)

Identification of theobromine Identification of theobromine 1.1. IR-spectrophotometryIR-spectrophotometry..

2.2. Dissolve the substance in ammonium solution at the heatingDissolve the substance in ammonium solution at the heating. . After cooling add silver nitrate solution After cooling add silver nitrate solution – – solution must still be solution must still be colorlesscolorless. . After the boiling during few minutes white precipitate After the boiling during few minutes white precipitate formedformed..

3.3. Reaction on xantinesReaction on xantines ( (murexide samplemurexide sample).). At the oxidation of At the oxidation of theobromine 3-methylalloxane and methylurea formedtheobromine 3-methylalloxane and methylurea formed. . Ammonium salt of dimethylpuepuric acid formed as a result of Ammonium salt of dimethylpuepuric acid formed as a result of murexide reactionmurexide reaction::

Unpharmacopoeial reactionsUnpharmacopoeial reactions

4.4. Reaction of the sodium salt of theobromine, obtained by the Reaction of the sodium salt of theobromine, obtained by the interaction of alkali with an excess of preparationinteraction of alkali with an excess of preparation, , with cobalt with cobalt ((IIII))chloride solutionchloride solution – – intense violet color appearsintense violet color appears,, which quickly which quickly disappearsdisappears, , grey-blue precipitate: grey-blue precipitate:

5.5. The reaction of theobromine sodium salt with silver nitrate The reaction of theobromine sodium salt with silver nitrate - - formed a dense gelatin mass formed a dense gelatin mass ((silver saltsilver salt), ), which is thinning which is thinning by heating to by heating to 80° С80° С and solidifies again at the cooling and solidifies again at the cooling..

6.6. WithWith HgClHgCl22 – white crystalline precipitate – white crystalline precipitate..

Identification of theophylline1.1. Determination of the melting temperature alter the dryingDetermination of the melting temperature alter the drying..

2.2. IR spectrophotometryIR spectrophotometry..

3.3. Theophylline in the alkali medium decomposes to teophyllidine, Theophylline in the alkali medium decomposes to teophyllidine, which can be identified by the reaction of azojoining with which can be identified by the reaction of azojoining with diazonium salts, red color azo-dye formeddiazonium salts, red color azo-dye formed..

4.4. Determination of the water by semimicromethod Determination of the water by semimicromethod (К. (К. FisherFisher) ) (8-9,5 %).(8-9,5 %).

5.5. Reaction on xantinesReaction on xantines ( (murexide samplemurexide sample). ). At the oxidation of At the oxidation of theophylline theophylline 1,3-1,3-dimethylalloxane and ureadimethylalloxane and urea. . Ammonium salt of Ammonium salt of tetramethylpuepuric acid forme as a result of murexide reactiontetramethylpuepuric acid forme as a result of murexide reaction::

2.2. The reaction of theobromine sodium salt obtained by the The reaction of theobromine sodium salt obtained by the interaction of alkali with an excess of drug, with a solution of interaction of alkali with an excess of drug, with a solution of cobalt (II) chloride - formed white with pink tinge precipitate of cobalt (II) chloride - formed white with pink tinge precipitate of cobalt saltcobalt salt..

3.3. WithWith HgClHgCl22 –white crystalline precipitate –white crystalline precipitate..

4.4. With alkali solution of sodium nitroprussideWith alkali solution of sodium nitroprusside green color formed green color formed dissappearsdissappears at the adding an excess of acid.at the adding an excess of acid.

5.5. The reaction of theobromine sodium salt with silver nirate The reaction of theobromine sodium salt with silver nirate -- formed a dense gelatin massformed a dense gelatin mass..

6.6. Theophylline withTheophylline with 2,6- 2,6-dichloroqiunonechloroimide in borate dichloroqiunonechloroimide in borate

buffer solutionbuffer solution

(рН 8,5) (рН 8,5) gives intense blue gives intense blue

merocyanic dyemerocyanic dye::

AssayTheophylline and Theobromine

Alkalimetry by the substituentsubstituent ((indirect indirect alkalimetryalkalimetry).).IndicatorIndicator – – phenolphthalein phenolphthalein ( (theobromine) ortheobromine) or Bromothymol Bromothymol

blue blue ((theophyllinetheophylline). Е = М.). Е = М.mm..

StorageStorage, , usageusageTheobromine

In airtight containersIn airtight containers,, place place protected from lightprotected from light..

Stimulates the activity of the Stimulates the activity of the heart, somewhat expands heart, somewhat expands the coronary vessels and the coronary vessels and bronchi, shows diuretic bronchi, shows diuretic effecteffect. . Issue - powder and Issue - powder and tablets bytablets by 0,25 0,25 gg. .

Included in tabletsIncluded in tablets: : Theminal Theminal ((with amidopyrinewith amidopyrine and and PhenobarbitalPhenobarbital), ), Theodibaverine Theodibaverine ((withwith papaverine and dibazolepapaverine and dibazole), ), TheoephedrineTheoephedrine..

Theophylline In airtight containersIn airtight containers,, place place

protected from lightprotected from light..

Non-selective phosphodiesterase Non-selective phosphodiesterase inhibitor (xanthine); treatment of inhibitor (xanthine); treatment of reversible airways obstruction. reversible airways obstruction. BroncholiticBroncholitic, , cardiotonic and cardiotonic and diuretic mean with moderate diuretic mean with moderate influence on the stagnation influence on the stagnation phenomena in the cardiac and phenomena in the cardiac and renal origin organsrenal origin organs.. Issue – Issue – powder and tablets bypowder and tablets by 0,1 0,1 andand 0,3 0,3 gg; ; amp.amp. 2%-5,0; 2%-5,0; candles by candles by 0,2 0,2 gg. . TeopekTeopek, , TheotardTheotard, , NeophyllineNeophylline, , EuphyllineEuphylline.. Included in tablets: Included in tablets: TheoephedrineTheoephedrine..

Pentoxifylline (Pentoxifyllinium)Agapurine, Pentyline, Trental

A synthetic analogue of theobromine

3,7-dimethyl-1-(5’-oxohexyl)-3,7-dihydro-7Н-purine-2,6-dion or 1-(5’-oxohexyl)-theobromine

N

N N

NCH2

CH3

CH3

O

O

CH2

CH2

CH2

C

O

H3C

Identification of pentoxifyllineIdentification of pentoxifylline

Temperature of meltingTemperature of meltingIR-spectroscopy IR-spectroscopy TLCTLCReaction on xantinesReaction on xantines

Formation of azo-dye Formation of azo-dye ((look theophyllinelook theophylline))AssayAssay

pentoxifyllinepentoxifylline

Acidimetry in non-aqueous media in a mixture of Acidimetry in non-aqueous media in a mixture of anhydrous acetic acid and acetic anhydrideanhydrous acetic acid and acetic anhydride,, direct direct titration. Potentiometric indication. titration. Potentiometric indication. (Е=М.(Е=М.mm).).

Usage of pentoxifyllineUsage of pentoxifyllineHas a vasodilator effect, improves tissue oxygen Has a vasodilator effect, improves tissue oxygen

supplysupply, decreases thrombosis aggregation and decreases thrombosis aggregation and reduces blood viscosityreduces blood viscosity. Apply at the peripheral Apply at the peripheral circulatory disorders, atherosclerotic disorders, circulatory disorders, atherosclerotic disorders, ischemic condition after heart attack, in ischemic condition after heart attack, in ophthalmology, at the hearing disordersophthalmology, at the hearing disorders. Issue – tablets of 0,1 g, ampoules of ampoules of 2%-5,0. Adopt Adopt inside by inside by 0,2 g 3 times daily after meals daily after meals. In the In the acute disorders of peripheral or cerebral acute disorders of peripheral or cerebral circulation injected circulation injected 0,1 g intravenousintravenous in 250-500 ml of NaCl or 5% glucose solution.

Synthetic analogues of theophylline

Theophylline-ethylenediamine (Theophyllinum et ethylenediaminum) (SPU),

Euphylline (Euphyllinum), Aminophilline

Theophylline withTheophylline with 1,2- 1,2-ethylenediamineethylenediamineWhite, sometimes with yellowish crystalline powder with slight White, sometimes with yellowish crystalline powder with slight ammonia odorammonia odor. . On tOn the air absorbs carbon dioxide, thus he air absorbs carbon dioxide, thus decreasing its solubilitydecreasing its solubility.. Soluble in water, aqueous solutions Soluble in water, aqueous solutions have an alkaline reactionhave an alkaline reaction..

N

N N

NHH3C

CH3

O

O

H2C

H2C

NH2

NH2

*

Identification of euphylline

1. Theophylline is identified after the separation by HCl of ethylenediamine according to SPU:

а) By the melting temperature of theophylline (269 - 274°С) after HCl acidification to рН 4-5;

Б) IR-spectroscopy;

в) Reaction of azojoining (look theophylline);

г) murexide sample.

2. Ethylenediamine can be confirmed by the following reactions:

а) determination of the melting temperature of the product of reaction with benzoyl in alkali medium (dibenzoylethylenediamine) (SPU):

б) with a solution of copper (II) sulfate a bright purple color forms :

в) with 2,4-dinitrochlorobenzene –yellow precipitate falls.

H2C NH2

H2C NH2

CuH2C NH2

H2C NH2

SO4+ CuSO4

2

2

H2C

H2C

NH2

NH2

+ 2C Cl

OC N

HO

CH2

CH2

NH

C

O- 2 HCl

AssayEuphylline

1. Ethylenediamine can be determined by acidimetry, indicator –methyl orange. Е = М.m./2.

Ethylenediamine in euphylline should be 13,5—15 % in the dry matter.

2. Theophylline can be determined by alkalimetry by substituent 1 after drying in a drying cabinet at 25-130 °С to the disappearance of amine odor.

The content of waterless theophylline in euphylline should be 84- 87,4 %.

H2C NH2

H2C NH2

+ 2HClH2C NH2

H2C NH2

* 2 HCl

Storage, usage of euphylline

Given the ability to absorb carbon dioxide, stored in a well corked filled to the end container, protected from the effects of light and moisture.

Antispasmodic, bronchodilatin, diuretic mean. At the bronchial asthma and bronchospasm, hypertension, cardiac asthma, to improve blood circulation to the brain, decreasing the intraperitoneal pressure and brain edema in ischemic stroke.

Used oral by 0,15g after food, i/v (2,4% solutions by 5,0) and i/m (24 % solution by 1 ml).

Diprophylline (Diprophyllinum)

7-(2',3'-Dioxipropyl)-theophylline

Less toxic than theophylline. Used for treatment of coronary spasm, cardiac and bronchial asthma, hypertension. Issue – tablets by 0,2 g; amp. 10%-5,0; candles by 0,5 g.

N

N N

NH3C

CH3

O

O

CH2

CH

CH2OH

OH

Xantinole nicotinate (Xantinoli nicotinas) Complamine, Theonicol

7-[2’-oxi-3’-(N'-methyl-β-oxiethylamino)-propyl]-theophylline nicotinate

Used to improve peripheral and cerebral circulation

Issue – tablets by 0,15 g; amp. 15%-2,0 і 10,0.

N

N N

NH3C

CH3

O

O

CH2

CH

CH2

OH

N CH3

CH2CH2OHN

COOH

*

Thank you for Thank you for attention!attention!