Caffeine
Caffeine C8H10N4O2 = 194.2 (58-08-2)
General reagent grade of commerce.
Melting point, about 236°.
Anhydrous Caffeine
(Ph Eur monograph 0267)
C8H10N4O2 194.2 56-08-2
Action and use
Central nervous stimulant.
Preparation
Aspirin and Caffeine Tablets
Ph Eur
DEFINITION
Caffeine contains not less than 98.5 per cent and not more than the equivalent of 101.5 per cent of
1,3,7-trimethyl-3,7-dihydro-1H-purine-2,6-dione, calculated with reference to the dried substance.
CHARACTERS
A white, crystalline powder or silky, white crystals, sublimes readily, sparingly soluble in water, freely
soluble in boiling water, slightly soluble in ethanol. It dissolves in concentrated solutions of alkali
benzoates or salicylates.
IDENTIFICATION
First identification A, B, E.
Second identification A, C, D, E, F.
A. Melting point (2.2.14): 234 °C to 239 °C.
B. Examine by infrared absorption spectrophotometry (2.2.24), comparing with the spectrum obtained
with caffeine CRS.
C. To 2 ml of a saturated solution add 0.05 ml of iodinated potassium iodide solution R. The solution
remains clear. Add 0.1 ml of dilute hydrochloric acid R. A brown precipitate is formed. Neutralise with
dilute sodium hydroxide solution R. The precipitate dissolves.
D. In a glass-stoppered tube, dissolve about 10 mg in 0.25 ml of a mixture of 0.5 ml of acetylacetone
R and 5 ml of dilute sodium hydroxide solution R. Heat in a water-bath at 80 °C for 7 min. Cool and add
0.5 ml of dimethylaminobenzaldehyde solution R2. Heat again in a water-bath at 80 °C for 7 min. Allow to
cool and add 10 ml of water R. An intense blue colour develops.
E. It complies with the test for loss on drying (see Tests).
F. It gives the reaction of xanthines (2.3.1).
TESTS
Solution S
Dissolve 0.5 g with heating in 50 ml of carbon dioxide-free water R prepared from distilled water R, cool
and dilute to 50 ml with the same solvent.
Appearance of solution
Solution S is clear (2.2.1) and colourless (2.2.2, Method II).
Acidity
To 10 ml of solution S add 0.05 ml of bromothymol blue solution R1. The solution is green or yellow. Not
more than 0.2 ml of 0.01 M sodium hydroxide is required to change the colour of the indicator to blue.
Related substances
Examine by thin-layer chromatography (2.2.27) using silica gel GF254 R as the coating substance.
Test solution Dissolve 0.2 g of the substance to be examined in a mixture of 4 volumes of methanol R
and 6 volumes of methylene chloride R and dilute to 10 ml with the same mixture of solvents.
Reference solution Dilute 0.5 ml of the test solution to 100 ml with a mixture of 4 volumes of methanol
R and 6 volumes of methylene chloride R.
Apply to the plate 10 µl of each solution. Develop over a path of 15 cm using a mixture of 10 volumes of
concentrated ammonia R, 30 volumes of acetone R, 30 volumes of methylene chloride R and 40
volumes of butanol R. Allow the plate to dry in air and examine in ultraviolet light at 254 nm. Any spot in
the chromatogram obtained with the test solution, apart from the principal spot, is not more intense than
the spot in the chromatogram obtained with the reference solution (0.5 per cent).
Sulphates (2.4.13)
15 ml of solution S complies with the limit test for sulphates (500 ppm). Prepare the standard using a
mixture of 7.5 ml of sulphate standard solution (10 ppm SO4) R and 7.5 ml of distilled water R.
Heavy metals (2.4.8)
1.0 g complies with limit test C for heavy metals (20 ppm). Prepare the standard using 2 ml of lead
standard solution (10 ppm Pb) R.
Loss on drying (2.2.32)
Not more than 0.5 per cent, determined on 1.000 g by drying in an oven at 100-105 °C for 1 h.
Sulphated ash (2.4.14)
Not more than 0.1 per cent, determined on 1.0 g.
ASSAY
Dissolve 0.170 g with heating in 5 ml of anhydrous acetic acid R. Allow to cool, add 10 ml of acetic
anhydride R and 20 ml of toluene R. Titrate with 0.1 M perchloric acid, determining the end-point
potentiometrically (2.2.20).
1 ml of 0.1 M perchloric acid is equivalent to 19.42 mg of C8H10N4O2.
IMPURITIES
Specified impurities A.
Other detectable impurities B, C.
A. theophylline,
B. N-[6-amino-1,3-dimethyl-2,4(1H,3H)-dioxopyrimidin-5-yl]formamide,
C. 1,3,9-trimethyl-3,9-dihydro-1H-purine-2,6-dione (isocaffeine).
Ph Eur
Caffeine Hydrate
(Caffeine Monohydrate, Ph Eur monograph 0268)
C8H10N4O2,H2O 212.2 5743-12-4
Action and use
Central nervous stimulant.
Ph Eur
DEFINITION
Caffeine monohydrate contains not less than 98.5 per cent and not more than the equivalent of 101.5
per cent of 1,3,7-trimethyl-3,7-dihydro-1H-purine-2,6-dione, calculated with reference to the dried
substance.
CHARACTERS
A white, crystalline powder or silky, white crystals, sublimes readily, sparingly soluble in water, freely
soluble in boiling water, slightly soluble in ethanol. It dissolves in concentrated solutions of alkali
benzoates or salicylates.
IDENTIFICATION
First identification A, B, E.
Second identification A, C, D, E, F.
A. Melting point (2.2.14): 234 °C to 239 °C, determined after drying at 100-105 °C.
B. Dry the substance to be examined at 100-105 °C. Examine by infrared absorption
spectrophotometry (2.2.24), comparing with the spectrum obtained with caffeine CRS.
C. To 2 ml of a saturated solution add 0.05 ml of iodinated potassium iodide solution R. The solution
remains clear. Add 0.1 ml of dilute hydrochloric acid R. A brown precipitate is formed. Neutralise with
dilute sodium hydroxide solution R. The precipitate dissolves.
D. In a glass-stoppered tube, dissolve about 10 mg in 0.25 ml of a mixture of 0.5 ml of acetylacetone
R and 5 ml of dilute sodium hydroxide solution R. Heat in a water-bath at 80 °C for 7 min. Cool and add
0.5 ml of dimethylaminobenzaldehyde solution R2. Heat again in a water-bath at 80 °C for 7 min. Allow to
cool and add 10 ml of water R. An intense blue colour develops.
E. It complies with the test for loss on drying (see Tests).
F. It gives the reaction of xanthines (2.3.1).
TESTS
Solution S
Dissolve 0.5 g with heating in 50 ml of carbon dioxide-free water R prepared from distilled water R, cool
and dilute to 50 ml with the same solvent.
Appearance of solution
Solution S is clear (2.2.1) and colourless (2.2.2, Method II).
Acidity
To 10 ml of solution S add 0.05 ml of bromothymol blue solution R1. The solution is green or yellow. Not
more than 0.2 ml of 0.01 M sodium hydroxide is required to change the colour of the indicator to blue.
Related substances
Examine by thin-layer chromatography (2.2.27) using silica gel GF254 R as the coating substance.
Test solution Dissolve 0.2 g of the substance to be examined in a mixture of 4 volumes of methanol R
and 6 volumes of methylene chloride R and dilute to 10 ml with the same mixture of solvents.
Reference solution Dilute 0.5 ml of the test solution to 100 ml with a mixture of 4 volumes of methanol
R and 6 volumes of methylene chloride R.
Apply to the plate 10 µl of each solution. Develop over a path of 15 cm using a mixture of 10 volumes of
concentrated ammonia R, 30 volumes of acetone R, 30 volumes of methylene chloride R and 40
volumes of butanol R. Allow the plate to dry in air and examine in ultraviolet light at 254 nm. Any spot in
the chromatogram obtained with the test solution, apart from the principal spot, is not more intense than
the spot in the chromatogram obtained with the reference solution (0.5 per cent).
Sulphates (2.4.13)
15 ml of solution S complies with the limit test for sulphates (500 ppm). Prepare the standard using a
mixture of 7.5 ml of sulphate standard solution (10 ppm SO4) R and 7.5 ml of distilled water R.
Heavy metals (2.4.8)
1.0 g complies with limit test C for heavy metals (20 ppm). Prepare the standard using 2 ml of lead
standard solution (10 ppm Pb) R.
Loss on drying (2.2.32)
5.0 per cent to 9.0 per cent, determined on 1.000 g by drying in an oven at 100-105 °C for 1 h.
Sulphated ash (2.4.14)
Not more than 0.1 per cent, determined on 1.0 g.
ASSAY
Dissolve 0.170 g, previously dried at 100-105 °C, with heating in 5 ml of anhydrous acetic acid R. Allow
to cool, add 10 ml of acetic anhydride R and 20 ml of toluene R. Titrate with 0.1 M perchloric acid,
determining the end-point potentiometrically (2.2.20).
1 ml of 0.1 M perchloric acid is equivalent to 19.42 mg of C8H10N4O2.
IMPURITIES
Specified impurities A.
Other detectable impurities B, C.
A. theophylline,
B. N-[6-amino-1,3-dimethyl-2,4(1H, 3H)-dioxopyrimidin-5-yl]formamide,
C. 1,3,9-trimethyl-3,9-dihydro-1H-purine-2,6-dione (isocaffeine).
Ph Eur
Aspirin and Caffeine Tablets
DEFINITION
Aspirin and Caffeine Tablets contain, in each, 350 mg of Aspirin and 30 mg of Caffeine.
The tablets comply with the requirements stated under Tablets and with the following requirements.
Content of aspirin, C9H8O4
330 to 370 mg.
Content of caffeine, C8H10N4O2
27.5 to 32.5 mg.
IDENTIFICATION
A. Boil 1 g of the powdered tablets with 10 ml of 1 M sodium hydroxide, cool and filter. Acidify the
filtrate with 1M sulphuric acid; a white precipitate is produced. To a solution of the precipitate add iron(III)
chloride solution R1; a deep violet colour is produced.
B. Shake 0.5 g of the powdered tablets with 10 ml of water for 5 minutes, filter and add 10 ml of 1M
sodium hydroxide. Extract with three 30 ml quantities of dichloromethane, washing each extract with the
same 10 ml of water . Filter the combined extracts through absorbent cotton and evaporate the filtrate to
dryness. Reserve a quantity of the residue for test C. Dissolve 10 mg of the residue in 1 ml of
hydrochloric acid, add 0.1 g of potassium chlorate and evaporate to dryness in a porcelain dish. A
reddish residue remains, which becomes purple on exposure to ammonia vapour.
C. The light absorption, Appendix II B, in the range 240 to 350 nm of a 0.001% w/v solution of the
residue reserved in test B exhibits a maximum at 273 nm.
TESTS
Salicylic acid
To a quantity of the powdered tablets containing 0.50 g of Aspirin add 50 ml of chloroform and 10 ml of
water , shake well and allow to separate. Filter the chloroform layer through a dry filter paper and
evaporate 10 ml of the filtrate to dryness at room temperature using a rotary evaporator. To the residue
add 4 ml of ethanol (96%), stir well, dilute to 100 ml with water at a temperature not exceeding 10°, filter
immediately, rapidly transfer 50 ml to a Nessler cylinder, add 1 ml of freshly prepared ammonium iron(III)
sulphate solution R1, mix and allow to stand for 1 minute. Any violet colour produced is not more intense
than that obtained by adding 1 ml of freshly prepared ammonium iron(III) sulphate solution R1 to a
mixture of 3 ml of a freshly prepared 0.050% w/v solution of salicylic acid, 2 ml of ethanol (96%) and
sufficient water to produce 50 ml contained in a second Nessler cylinder (3.0%).
Dissolution
For aspirin
Comply with the dissolution test for tablets and capsules, Appendix XII D, using Apparatus II. Use as the
medium 500 ml of a pH 4.5 buffer prepared by mixing 29.9 g of sodium acetate and 16.6 ml of glacial
acetic acid with sufficient water to produce 10 litres and rotate the paddle at 50 revolutions per minute.
Withdraw a sample of 20 ml of the medium and filter. Immediately measure the absorbance of the filtrate,
Appendix II B, diluted with the dissolution medium if necessary, at 265 nm using dissolution medium in
the reference cell. Measure the absorbance of a suitable solution of aspirin BPCRS in the dissolution
medium and calculate the total content of aspirin, C 9H8O4, in the medium using the declared content of
C9H8O4 in aspirin BPCRS.
ASSAY
Weigh and powder 20 tablets.
For aspirin
To a quantity of the powder containing 0.7 g of Aspirin add 20 ml of water and 2 g of sodium citrate and
boil under a reflux condenser for 30 minutes. Cool, wash the condenser with 30 ml of warm water and
titrate with 0.5M sodium hydroxide VS using phenolphthalein solution R1 as indicator. Each ml of 0.5M
sodium hydroxide VS is equivalent to 45.04 mg of C9H8O4.
For caffeine
To a quantity of the powder containing 30 mg of Caffeine add 200 ml of water and shake for 30 minutes.
Add sufficient water to produce 250 ml and filter. To 10 ml of the filtrate add 10 ml of 1 M sodium
hydroxide and extract immediately with five 30 ml quantities of chloroform, washing each extract with the
same 10 ml of water . Filter the combined chloroform extracts, if necessary, through absorbent cotton
previously moistened with chloroform. Evaporate the solution to dryness and dissolve the residue as
completely as possible in water , warming gently if necessary. Cool, add sufficient water to produce 100
ml, mix and filter if necessary. Measure the absorbance of the resulting solution at the maximum at 273
nm, Appendix II B. Calculate the content of C 8H10N4O2 taking 504 as the value of A(1%, 1 cm) at the
maximum at 273 nm.
LABELLING
The label states that the tablets contain Aspirin, unless this word appears in the name of the tablets. This
requirement does not apply in countries where exclusive proprietary rights in the name Aspirin are
claimed.
