EUROPEAN PHARMACOPOEIA 10.
0 Bearberry leaf
Results : see below the sequence of zones present in the
chromatograms obtained with the reference solution and
the test solution. Furthermore, other faint zones may
be present in the chromatogram obtained with the test
solution.
Top of the plate
Scopoletin : a bright blue A blue fluorescent zone
fluorescent zone (scopoletin)
A blue fluorescent zone
_______
2 blue fluorescent zones
Rutoside : an orange fluorescent An orange fluorescent zone
zone (rutoside)
3-4 blue fluorescent zones
_______
An orange fluorescent zone
Reference solution Test solution
TESTS
Loss on drying (2.2.32) : maximum 11.0 per cent, determined
on 2.000 g of the powdered herbal drug (355) (2.9.12) by
drying in an oven at 105 °C for 2 h.
Total ash (2.4.16) : maximum 5.0 per cent.
Extractable matter : minimum 55.0 per cent.
To 2.00 g of the powdered herbal drug (355) (2.9.12) add
50.0 g of water R, boil under reflux for 1 h, compensate the
loss of water and filter. Evaporate 25.0 g of the filtrate to
dryness on a water-bath and dry in an oven at 105 °C for 3 h.
The residue weighs a minimum of 0.55 g.
07/2013:1054
BEARBERRY LEAF
Uvae ursi folium
DEFINITION
Whole or fragmented, dried leaf of Arctostaphylos uva-ursi (L.)
Spreng.
Content : minimum 7.0 per cent of anhydrous arbutin
(C12H16O7 ; Mr 272.3) (dried drug).
IDENTIFICATION
A. The leaf, shiny and dark green on the adaxial surface,
lighter on the abaxial surface, is generally 7-30 mm long
and 5-12 mm wide. The entire leaf is obovate with smooth
margins, somewhat reflexed downwards, narrowing at the
base into a short petiole. The leaf is obtuse or retuse at its
apex. The lamina is thick and coriaceous. The venation,
pinnate and finely reticulate, is clearly visible on both
surfaces. The adaxial surface is marked with sunken
veinlets, giving it a characteristic grainy appearance. Only
the young leaf has ciliated margins. Old leaves are glabrous.
B. Microscopic examination (2.8.23). The powder is green,
greenish-grey or yellowish-green. Examine under a
microscope using chloral hydrate solution R. The powder
shows the following diagnostic characters (Figure 1054.-1) :
fragments of adaxial epidermis (surface view [A]) showing
thick and irregularly pitted polygonal cells [Aa] usually
accompanied by palisade parenchyma [Ab] ; fragments
of adaxial epidermis (transverse section [G]), showing
straight-walled cells [Ga] covered by a thick smooth cuticle
[Gb], and accompanied by palisade parenchyma [Gc]
consisting of 3 or 4 layers of cells of unequal lengths, some
General Notices (1) apply to all monographs and other texts
of which contain numerous prisms of calcium oxalate
[Gd] ; fragments of abaxial epidermis (surface view [B, E]),
showing anomocytic stomata (2.8.3) [Ba] surrounded
by 5-11 subsidiary cells, scars of hair bases [Ea], and
accompanied by spongy parenchyma [Eb] ; groups of
lignified fibres from the pericycle [D] ; fragments of the
vascular system [F] consisting of pitted vessels [Fa] and
fibres [Fb] accompanied by rows of cells containing prisms
of calcium oxalate [Fc] ; oil droplets are present in the
parenchymatous cells ; occasional fragments of conical,
_______ unicellular covering trichomes [C].
_______
Figure 1054.-1. – Illustration for identification test B of
powdered herbal drug of bearberry leaf
C. Thin-layer chromatography (2.2.27).
Test solution. To 0.5 g of the powdered herbal drug
(355) (2.9.12) add 5 mL of a mixture of equal volumes of
methanol R and water R, and heat under a reflux condenser
for 10 min. Filter whilst hot. Wash the flask and the filter
with a mixture of equal volumes of methanol R and water R
and dilute to 5 mL with the same mixture of solvents.
Reference solution. Dissolve 50 mg of arbutin R and 25 mg
of gallic acid R in methanol R and dilute to 20.0 mL with
the same solvent.
Plate : TLC silica gel plate R (5-40 μm) [or TLC silica gel
plate R (2-10 μm)].
Mobile phase : anhydrous formic acid R, water R, ethyl
acetate R (6:6:88 V/V/V).
Application : 10 μL [or 2 μL] as bands of 15 mm [or 8 mm].
Development : over a path of 15 cm [or 6 cm].
Drying : at 105-110 °C until the mobile phase has
evaporated.
Detection : treat with a 10 g/L solution of
dichloroquinonechlorimide R in methanol R, then
treat with a 20 g/L solution of anhydrous sodium
carbonate R.
Results : see below the sequence of zones present in the
chromatograms obtained with the reference solution and
the test solution. Furthermore, other blue or brown zones
may be present in the chromatogram obtained with the
test solution.
1333
Belamcanda chinensis rhizome
Top of the plate
Gallic acid : a brownish zone A brownish zone
_______
A brown zone
_______
Arbutin : a blue zone An intense blue zone (arbutin)
Reference solution Test solution
TESTS
Foreign matter (2.8.2): maximum 5 per cent of stems and
maximum 3 per cent of other foreign matter.
Leaves of different colour : maximum 10 per cent, determined
in the same manner as foreign matter (2.8.2).
Loss on drying (2.2.32) : maximum 10.0 per cent, determined
on 1.000 g of the powdered herbal drug (355) (2.9.12) by
drying in an oven at 105 °C for 2 h.
Total ash (2.4.16) : maximum 5.0 per cent.
ASSAY
Liquid chromatography (2.2.29).
Test solution. In a 100 mL flask with a ground-glass neck,
place 0.800 g of the powdered herbal drug (250) (2.9.12).
Add 20 mL of water R and heat under a reflux condenser on
a water-bath for 30 min. Allow to cool and filter the liquid
through a plug of absorbent cotton. Add the absorbent cotton
to the residue in the 100 mL flask and extract with 20 mL of
water R under a reflux condenser on a water-bath for 30 min.
Allow to cool and filter through a paper filter. Combine the
filtrates and dilute to 50.0 mL with water R. Filter through a
paper filter. Discard the first 10 mL of the filtrate.
Reference solution (a). Dissolve 50.0 mg of arbutin CRS in the
mobile phase and dilute to 50.0 mL with the mobile phase.
Reference solution (b). Dissolve 2.5 mg of hydroquinone R in
the mobile phase and dilute to 10.0 mL with the mobile phase.
To 5.0 mL of the solution, add 2.5 mL of reference solution (a) B. Microscopic examination (2.8.23). The powder is
and dilute to 10.0 mL with the mobile phase.
Column :
– size : l = 0.25 m, Ø = 4 mm ;
– stationary phase : base-deactivated octadecylsilyl silica gel for
chromatography R (5 μm).
Mobile phase : methanol R, water R (10:90 V/V).
Flow rate : 1.2 mL/min.
Detection : spectrophotometer at 280 nm.
Injection : 20 μL.
System suitability : reference solution (b) :
– resolution : minimum 4.0 between the peaks due to arbutin
and hydroquinone.
Calculate the percentage content of arbutin using the following
expression :
F1 ´ m 2 ´ p
F2 ´ m1
1334
EUROPEAN PHARMACOPOEIA 10.0
F1 = area of the peak due to arbutin in the chromatogram
obtained with the test solution ;
F2 = area of the peak due to arbutin in the chromatogram
obtained with reference solution (a) ;
m1 = mass of the herbal drug to be examined used to
_______ prepare the test solution, in grams ;
m2 = mass of arbutin CRS used to prepare reference
solution (a), in grams ;
p = percentage content of arbutin in arbutin CRS.
_______
01/2014:2561
BELAMCANDA CHINENSIS RHIZOME
Belamcandae chinensis rhizoma
DEFINITION
Dried, whole or fragmented rhizome of Iris domestica (L.)
Goldblatt et Mabb. (syn. Belamcanda chinensis (L.) DC.),
collected in early spring while the plant is budding or in late
autumn while the aerial part is withering, with roots removed.
Content : minimum 0.10 per cent of irisflorentin (C20H18O8 ;
Mr 386.4) (dried drug).
IDENTIFICATION
A. The whole rhizome is nodular, rounded, about 3-10 cm
long and 1-2 cm in diameter, irregular, more or less
branched, with numerous annular striations ; there are
crateriform, annular stem scars on the upper surface and
small roots about 2-3 mm in diameter on the lower surface.
The longitudinally fragmented rhizome is found as pieces
about 2-8 cm long, 2 cm wide and 1 cm thick ; stem scars
and root fragments are present. The orange-brown or dark
brown outer surface is the same colour as the fracture. The
central parenchyma has a pitted appearance due to the
numerous primary vascular bundles. The texture is hard.
The fracture is granular.
orange-brown or dark brown. Examine under a microscope
using chloral hydrate solution R. The powder shows the
following diagnostic characters : rare fragments of brown
cork with superimposed polyhedral cells ; numerous,
somewhat rounded parenchyma cells with irregularly
thickened and pitted walls, granular and oily contents, with
some of the cells containing a very large calcium oxalate
prism up to 250 μm long and about 50 μm in diameter ;
very numerous free calcium oxalate prisms, usually broken ;
reticulate or pitted lignified vessels. Examine under a
microscope using a 50 per cent V/V solution of glycerol R.
The powder shows very numerous rounded or ovoid starch
granules, 3-15 μm in diameter, simple or rarely compound
with 2-5 components. The punctiform hilum is sometimes
visible. The starch granules are free and very often included
in the parenchymatous cells.
C. Examine the chromatograms obtained in the test for Iris
tectorum Maxim.
Results A : see below the sequence of zones present in the
chromatograms obtained with the reference solution and
the test solution. Furthermore, other faint zones may
be present in the chromatogram obtained with the test
solution.
See the information section on general monographs (cover pages)