Frangula bark dry extract, standardised EUROPEAN PHARMACOPOEIA 5.

Loss on drying (2.2.32). Not more than 10.0 per cent, calculated with reference to the dried extract. The measured
determined on 1.000 g of the powdered drug (355) by drying content does not deviate from that stated on the label by
in an oven at 100-105 °C for 2 h. more than ± 10 per cent.
Total ash (2.4.16). Not more than 6.0 per cent. PRODUCTION
ASSAY The extract is produced from the drug and ethanol (50 to
Carry out the assay protected from bright light. 80 per cent V/V) by an appropriate procedure.
In a tared, round-bottomed flask with a ground-glass neck, CHARACTERS
weigh 0.250 g of powdered drug (180). Add 25.0 ml of a A yellowish-brown, fine powder.
70 per cent V/V solution of methanol R ; mix and weigh
again. Heat in a water-bath under a reflux condenser for IDENTIFICATION
15 min. Allow to cool, weigh and adjust to the original mass A. Examine by thin-layer chromatography (2.2.27), using a
with a 70 per cent V/V solution of methanol R. Filter and suitable silica gel as the coating substance.
transfer 5.0 ml of the filtrate to a separating funnel. Add
50 ml of water R and 0.1 ml of hydrochloric acid R. Shake Test solution. To 0.05 g add 5 ml of alcohol (70 per
with five quantities, each of 20 ml, of light petroleum R. cent V/V) R and heat to boiling. Cool and centrifuge.
Allow the layers to separate and transfer the aqueous layer Decant the supernatant solution immediately and use
to a 100 ml volumetric flask. Combine the light petroleum within 30 min.
layers and wash with two quantities, each of 15 ml, of Reference solution. Dissolve 20 mg of barbaloin R in
water R. Use this water for washing the separating funnel alcohol (70 per cent V/V) R and dilute to 10 ml with the
and add it to the aqueous solution in the volumetric flask. same solvent.
Add 5 ml of a 50 g/l solution of sodium carbonate R and Apply separately to the plate, as bands, 10 µl of each
dilute to 100.0 ml with water R. Discard the light petroleum solution. Develop over a path of 10 cm using a mixture
layer. Transfer 40.0 ml of the aqueous solution to a 200 ml of 13 volumes of water R, 17 volumes of methanol R and
round-bottomed flask with a ground-glass neck. Add 20 ml 100 volumes of ethyl acetate R. Allow the plate to dry for
of a 200 g/l solution of ferric chloride R and heat under 5 min, then spray with a 50 g/l solution of potassium
a reflux condenser for 20 min in a water-bath with the hydroxide R in alcohol (50 per cent V/V) R and heat
water level above that of the liquid in the flask. Add 2 ml at 100-105 °C for 15 min. Examine immediately after
of hydrochloric acid R and continue heating for 20 min, heating. The chromatogram obtained with the reference
shaking frequently, until the precipitate is dissolved. Allow solution shows a reddish-brown zone in the median third
to cool, transfer the mixture to a separating funnel and shake corresponding to barbaloin. The chromatogram obtained
with three quantities, each of 25 ml, of ether R, previously with the test solution shows two orange-brown zones
used to rinse the flask. Combine the ether extracts and wash (glucofrangulins) in the lower third and two to four red
with two quantities, each of 15 ml, of water R. Transfer the zones (frangulins, not always clearly separated, and above
ether layer to a volumetric flask and dilute to 100.0 ml with them frangula-emodin) in the upper third.
ether R. Evaporate 20.0 ml carefully to dryness and dissolve B. To about 25 mg add 25 ml of dilute hydrochloric acid R
the residue in 10.0 ml of a 5 g/l solution of magnesium and heat the mixture on a water-bath for 15 min. Allow
acetate R in methanol R. Measure the absorbance (2.2.25) to cool, shake with 20 ml of ether R and discard the
at 515 nm using methanol R as the compensation liquid. aqueous layer. Shake the ether layer with 10 ml of dilute
Calculate the percentage of glucofrangulins, expressed as ammonia R1. The aqueous layer becomes reddish-violet.
glucofrangulin A, from the expression :
TESTS
Loss on drying (2.8.17) : maximum 5.0 per cent.
Microbial contamination. Total viable aerobic count (2.6.12)
i.e. taking the specific absorbance of glucofrangulin A to not more than 104 per gram of which not more than 102
be 204. fungi per gram, determined by plate count. It complies with
A = absorbance at 515 nm, the test for Escherichia coli and Salmonella (2.6.13).
m = mass of the sample, in grams. ASSAY
Carry out the assay protected from bright light.
STORAGE
In a tared round-bottomed flask with a ground-glass neck,
Store protected from light. weigh 0.100 g of the preparation to be examined. Add
25.0 ml of a 70 per cent V/V solution of methanol R, mix and
weigh again. Heat the flask in a water-bath under a reflux
01/2005:1214 condenser at 70 °C for 15 min. Allow to cool, weigh and
adjust to the original mass with a 70 per cent V/V solution
FRANGULA BARK DRY EXTRACT, of methanol R. Filter and transfer 5.0 ml of the filtrate to
STANDARDISED a separating funnel. Add 50 ml of water R and 0.1 ml of
hydrochloric acid R. Shake with five quantities, each of
20 ml, of light petroleum R1. Allow the layers to separate
Frangulae corticis extractum siccum and transfer the aqueous layer to a 100 ml volumetric flask.
normatum Combine the light petroleum layers and wash with two
quantities, each of 15 ml, of water R. Use this water for
DEFINITION washing the separating funnel and add it to the aqueous
Standardised frangula bark dry extract is produced from solution in the volumetric flask. Add 5 ml of a 50 g/l solution
Frangula bark (0025). It contains not less than 15.0 per of sodium carbonate R and dilute to 100.0 ml with water R.
cent and not more than 30.0 per cent of glucofrangulins, Discard the light petroleum layer. Transfer 40.0 ml of the
expressed as glucofrangulin A (C27H30O14 ; Mr 578.5) and aqueous solution to a 200 ml round-bottomed flask with a

1642 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 5.0
ground-glass neck. Add 20 ml of a 200 g/l solution of ferric
chloride R and heat under a reflux condenser for 20 min in a
water-bath with the water level above that of the liquid in the
flask. Add 2 ml of hydrochloric acid R and continue heating
for 20 min, shaking frequently, until the precipitate is
dissolved. Allow to cool, transfer the mixture to a separating
funnel and shake with three quantities, each of 25 ml, of
ether R, previously used to rinse the flask. Combine the
ether extracts and wash with two quantities, each of 15 ml,
of water R. Transfer the ether layer to a volumetric flask and
dilute to 100.0 ml with ether R. Evaporate 20.0 ml carefully
to dryness and dissolve the residue in 10.0 ml of a 5 g/l
solution of magnesium acetate R in methanol R. Measure
the absorbance (2.2.25) at 515 nm using methanol R as the
compensation liquid.
Calculate the percentage of glucofrangulins, expressed as
glucofrangulin A from the expression :
i.e. taking the specific absorbance of glucofrangulin A to
be 204, calculated on the basis of the specific absorbance
of barbaloin,
A = absorbance at 515 nm,
m = mass of the preparation to be examined, in grams.
STORAGE
Store in an airtight container, protected from light.
LABELLING
The label states the content of glucofrangulins.
01/2005:0188
FRUCTOSE
Fructosum
C6H12O6 Mr 180.2 of the indicator to pink.
DEFINITION
Fructose is (-)-D-arabino-hex-2-ulopyranose. The substance
described in this monograph is not necessarily suitable for
parenteral use.
CHARACTERS
A white, crystalline powder, with a very sweet taste, very
soluble in water, soluble in alcohol.
IDENTIFICATION
A. Examine by thin-layer chromatography (2.2.27), using
silica gel G R as the coating substance.
Test solution. Dissolve 10 mg of the substance to be
examined in a mixture of 2 volumes of water R and
3 volumes of methanol R and dilute to 20 ml with the
same mixture of solvents.
Reference solution (a). Dissolve 10 mg of fructose CRS
in a mixture of 2 volumes of water R and 3 volumes of
methanol R and dilute to 20 ml with the same mixture
of solvents.
General Notices (1) apply to all monographs and other texts
Fructose
Reference solution (b). Dissolve 10 mg each of
fructose CRS, glucose CRS, lactose CRS and
sucrose CRS in a mixture of 2 volumes of water R and
3 volumes of methanol R and dilute to 20 ml with the
same mixture of solvents.
Apply separately to the plate 2 µl of each solution and
thoroughly dry the starting points. Develop over a path
of 15 cm using a mixture of 10 volumes of water R,
15 volumes of methanol R, 25 volumes of anhydrous
acetic acid R and 50 volumes of ethylene chloride R. The
solvents should be measured accurately since a slight
excess of water produces cloudiness. Dry the plate in a
current of warm air. Repeat the development immediately,
after renewing the mobile phase. Dry the plate in a
current of warm air and spray evenly with a solution of
0.5 g of thymol R in a mixture of 5 ml of sulphuric acid R
and 95 ml of alcohol R. Heat at 130 °C for 10 min. The
principal spot in the chromatogram obtained with the
test solution is similar in position, colour and size to
the principal spot in the chromatogram obtained with
reference solution (a). The test is not valid unless the
chromatogram obtained with reference solution (b) shows
four clearly separated spots.
B. Dissolve 0.1 g in 10 ml of water R. Add 3 ml of
cupri-tartaric solution R and heat. A red precipitate is
formed.
C. To 1 ml of solution S (see Tests) add 9 ml of water R. To
1 ml of the solution add 5 ml of hydrochloric acid R and
heat to 70 °C. A brown colour develops.
D. Dissolve 5 g in water R and dilute to 10 ml with the
same solvent. To 0.5 ml of the solution add 0.2 g of
resorcinol R and 9 ml of dilute hydrochloric acid R and
heat on a water-bath for 2 min. A red colour develops.
TESTS
Solution S. Dissolve 10.0 g in distilled water R and dilute to
100 ml with the same solvent.
Appearance of solution. Dissolve 5.0 g in water R and
dilute to 10 ml with the same solvent. The solution is clear
(2.2.1). Add 10 ml of water R. The solution is colourless
(2.2.2, Method II).
Acidity or alkalinity. Dissolve 6.0 g in 25 ml of carbon
dioxide-free water R and add 0.3 ml of phenolphthalein
solution R. The solution is colourless. Not more than 0.15 ml
of 0.1 M sodium hydroxide is required to change the colour
Specific optical rotation (2.2.7). Dissolve 10.0 g in 80 ml of
water R, add 0.2 ml of dilute ammonia R1, allow to stand
for 30 min and dilute to 100.0 ml with water R. The specific
optical rotation is − 91.0 to − 93.5, calculated with reference
to the anhydrous substance.
Foreign sugars. Dissolve 5.0 g in water R and dilute to
10 ml with the same solvent. To 1 ml of the solution add
9 ml of alcohol R. Any opalescence in the solution is not
more intense than that in a mixture of 1 ml of the initial
solution and 9 ml of water R.
5-Hydroxymethylfurfural and related compounds. To 5 ml
of solution S add 5 ml of water R. The absorbance (2.2.25)
measured at 284 nm is not greater than 0.32.
Barium. To 10 ml of solution S add 1 ml of dilute sulphuric
acid R. When examined immediately and after 1 h, any
opalescence in the solution is not more intense than that in a
mixture of 1 ml of distilled water R and 10 ml of solution S.
Lead in sugars (2.4.10). It complies with the limit test for
lead in sugars (0.5 ppm).
1643