EUROPEAN PHARMACOPOEIA 11.

0 Wild thyme

in a water-bath at 60 °C for 10 min, shaking frequently. Allow B. Microscopic examination (2.8.23). The powder is
to cool and filter into the same 100 mL volumetric flask as
used previously. Rinse the 200 mL flask with a further quantity
of alcohol (60 per cent V/V) R, filter and transfer to the same
100 mL volumetric flask. Dilute to volume with alcohol (60 per
cent V/V) R and filter.
Test solution. Introduce 5.0 mL of the stock solution into
a round-bottomed flask and evaporate to dryness under
reduced pressure. Take up the residue with 8 mL of a mixture
of 10 volumes of methanol R and 100 volumes of glacial acetic
acid R and transfer into a 25 mL volumetric flask. Rinse the
round-bottomed flask with 3 mL of a mixture of 10 volumes
of methanol R and 100 volumes of glacial acetic acid R
and transfer into the same 25 mL volumetric flask as used
previously. Add 10.0 mL of a solution containing 25.0 g/L of
boric acid R and 20.0 g/L of oxalic acid R in anhydrous formic
acid R and dilute to 25.0 mL with anhydrous acetic acid R.
Compensation liquid. Introduce 5.0 mL of the stock solution
into a round-bottomed flask and evaporate to dryness under
reduced pressure. Take up the residue with 8 mL of a mixture
of 10 volumes of methanol R and 100 volumes of glacial
acetic acid R and transfer into a 25 mL volumetric flask.
Rinse the round-bottomed flask with 3 mL of a mixture of
10 volumes of methanol R and 100 volumes of glacial acetic
acid R and transfer into the same 25 mL volumetric flask as
used previously. Add 10.0 mL of anhydrous formic acid R and
dilute to 25.0 mL with anhydrous acetic acid R.
Measure the absorbance (2.2.25) of the test solution at 405 nm
after 30 min.
Calculate the percentage content of total flavonoids, expressed
as violanthin from the expression :
A ´ 1.25
m
taking the specific absorbance of violanthin to be 400.
A = measured absorbance at 405 nm,
m = mass of the herbal drug to be examined, in grams.
greyish-green or greenish-brown. Examine under a
microscope using chloral hydrate solution R. The powder
shows the following diagnostic characters (Figure 1891.-1) :
fragments of the leaf epidermises [A, B, F] covered by a
finely striated cuticle and consisting of cells with sinuous
anticlinal walls [Aa, Ba, Fa] and diacytic stomata (2.8.3)
[Ab, Bb, Fb] ; cells of the adaxial leaf epidermis [B] with
wavy, irregularly thickened anticlinal walls [Ba] ; numerous
covering trichomes on both epidermises and the leaf
margins, with some of the cells containing very small
crystals of calcium oxalate [Af, Ca, Fd], the majority are
short, conical, unicellular, with thickened and warty walls
(surface view [Bc], side view [Fc]) ; fewer multicellular
covering trichomes, long, tapering to a point, composed
of up to 8 cells, slightly swollen at the joints, with finely
pitted walls, on an epidermis [Ae] or fragmented [C] ;
abundant glandular trichomes, mostly multicellular of
the lamiaceous type [Ac] with a unicellular stalk and a
glandular head consisting of 12 inconspicuous cells, others
with a unicellular stalk and a unicellular globular or ovoid
head [Ad] ; purplish-violet fragments of the corolla whose
inner epidermis consists of cells with rounded papillae [D]
and whose outer epidermis [E], with a striated cuticle,
consists of cells with lobed walls [Ea], unicellular [Eb] or
multicellular [Ec] uniseriate covering trichomes, glandular
trichomes with a unicellular head and a unicellular stalk
[Ed] and glandular trichomes of the lamiaceous type ;
relatively rare pollen grains, spherical, about 30 μm in
diameter, with a finely pitted exine and 6 germinal pores
[G].

07/2014:1891

WILD THYME
Serpylli herba
DEFINITION
Whole or cut, dried, flowering aerial parts of Thymus
serpyllum L.
Content : minimum 3.0 mL/kg of essential oil (dried drug).
IDENTIFICATION
A. The stem is much branched, up to about 1.5 mm in
diameter, cylindrical or indistinctly quadrangular, green,
reddish or purplish, the older stems brown and woody, the
younger stems pubescent. The leaves are opposite, 3-12 mm
long and up to 4 mm wide, elliptical to ovate-lanceolate
with an obtuse apex, cuneate and shortly petiolate at the
base ; the margin is entire and markedly ciliate, especially
near the base ; both surfaces are more or less glabrous but
distinctly punctate. The inflorescence is composed of about Figure 1891.-1.– Illustration for identification test B of
6-12 flowers in rounded to ovoid, terminal heads. The calyx powdered herbal drug of wild thyme
is tubular, 2-lipped with the upper lip dividing to form
3 teeth, the lower lip with 2 teeth, edged with long hairs ; C. Thin-layer chromatography (2.2.27).
the inner surfaces are strongly pubescent, the hairs forming Test solution. To 0.5 g of the powdered herbal drug (355)
a closed tube after flowering. The corolla is purplish-violet (2.9.12) add 5 mL of methanol R. Sonicate for 10 min.
or red, 2-lipped, the lower lip with 3 lobes and the upper Centrifuge or filter ; use the supernatant or the filtrate.
lip notched, the inner surface is strongly pubescent ; 4 Reference solution. Dissolve 1 mg of rutoside trihydrate R
epipetalous stamens project from the corolla tube. and 1 mg of rosmarinic acid R in 5 mL of methanol R.

General Notices (1) apply to all monographs and other texts 1777
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Willow bark EUROPEAN PHARMACOPOEIA 11.0

Plate : TLC silica gel F254 plate R (5-40 μm) [or TLC silica 01/2013:1583
gel F254 plate R (2-10 μm)].

Mobile phase : anhydrous formic acid R, water R, ethyl

acetate R (1:1:15 V/V/V).

Application : 20 μL [or 5 μL] as bands of 20 mm [or 8 mm]. WILLOW BARK

Development : over a path of 15 cm [or 6 cm]. Salicis cortex
Drying : in air. DEFINITION
Whole or fragmented dried bark of young branches or
Detection : heat at 100 °C for 3 min ; treat the still-hot plate whole dried pieces of current-year twigs of various species of
with a 5 g/L solution of diphenylboric acid aminoethyl genus Salix including S. purpurea L., S. daphnoides Vill. and
ester R in ethyl acetate R, then treat with a 50 g/L solution S. fragilis L.
of macrogol 400 R in methylene chloride R ; examine in
Content : minimum 1.5 per cent of total salicylic derivatives,
ultraviolet light at 365 nm.
expressed as salicin (C13H18O7 ; Mr 286.3) (dried drug).
Results : see below the sequence of zones present in the IDENTIFICATION
chromatograms obtained with the reference solution and
A. The bark is 1-2 mm thick and occurs in flexible, elongated,
the test solution. Furthermore, other faint fluorescent
quilled or curved pieces. The outer surface is smooth
zones may be present in the chromatogram obtained with
or slightly wrinkled longitudinally and greenish-yellow
the test solution.
or brownish-grey. The inner surface is smooth or
finely striated longitudinally and white, pale yellow or
Top of the plate reddish-brown, depending on the species. The fracture is
A red fluorescent zone short in the outer part and coarsely fibrous in the inner
region. The diameter of current-year twigs is not greater
than 10 mm. The wood is white or pale yellow.
Rosmarinic acid : a blue A blue fluorescent zone
fluorescent zone (rosmarinic acid)
_______ _______

1 or 2 blue fluorescent zones

_______ _______

A yellow or orange fluorescent

zone
A green or blue fluorescent zone
may be present
Rutoside : an orange-yellow
fluorescent zone
Reference solution Test solution

TESTS
Foreign matter (2.8.2) : maximum 3 per cent, determined on
30 g.
Thymus vulgaris L. or Thymus zygis L. Adulteration with
T. vulgaris L. or T. zygis L. is indicated by the presence of
acicular to linear-lanceolate leaves with a strongly bent margin,
the adaxial surface showing covering trichomes shaped as
pointed teeth with warty walls, the abaxial surface showing
many types of warty covering trichomes : unicellular, straight
or slightly curved, bicellular or tricellular, often elbow-shaped,
and bicellular or tricellular, more or less straight.
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.
Figure 1583.-1. – Illustration for identification test B of
Total ash (2.4.16) : maximum 10.0 per cent. powdered herbal drug of willow bark
Ash insoluble in hydrochloric acid (2.8.1): maximum 3.0 per B. Microscopic examination (2.8.23). The powder is pale
cent. yellow, greenish-yellow or light brown. Examine under a
microscope using chloral hydrate solution R. The powder
shows the following diagnostic characters (Figure 1583.-1) :
bundles [B, C] of narrow fibres [Ba, Ca], up to about 600 μm
ASSAY
long, with very thick walls and surrounded by a crystal
Essential oil (2.8.12). Use 50.0 g of the cut herbal drug, a sheath containing prisms of calcium oxalate [Bb, Cb] ;
1000 mL round-bottomed flask and 500 mL of water R as parenchymatous cells of the cortex [D, J], with thick, pitted
the distillation liquid. Distil at a rate of 2-3 mL/min for 2 h and deeply beaded walls [Da], and containing large cluster
without xylene R in the graduated tube. crystals of calcium oxalate [Ga, Ja] ; some parenchyma

1778 See the information section on general monographs (cover pages)

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