Fenugreek EUROPEAN PHARMACOPOEIA 11.

transversely elongated cells 2-9 μm wide, having a Water (2.2.13): maximum 80 mL/kg, determined on 20.0 g of
parquetry arrangement, sometimes accompanied by the powdered herbal drug (710) (2.9.12).
the inner layer of the mesocarp [Aa] ; fragments of the Total ash (2.4.16) : maximum 10.0 per cent.
epicarp with stomata accompanied by oil droplets [C] ; very
numerous oil droplets [J].
ASSAY
C. Thin-layer chromatography (2.2.27).
Essential oil (2.8.12). Use 10.0 g of the herbal drug reduced
Test solution. Shake 0.3 g of the freshly powdered herbal to a powder (1400) (2.9.12) immediately before the assay,
drug (1400) (2.9.12) with 5.0 mL of methylene chloride R a 500 mL round-bottomed flask, 200 mL of water R as the
for 15 min. Filter and carefully evaporate the filtrate to distillation liquid, and 0.50 mL of xylene R in the graduated
dryness on a water-bath at 60 °C. Dissolve the residue in tube. Distil at a rate of 2-3 mL/min for 2 h.
0.5 mL of toluene R.
Anethole. Gas chromatography (2.2.28) as described in
Reference solution. Dissolve 60 μL of anethole R in 5.0 mL the test for estragole and fenchone with the following
of hexane R. modification.
Plate : TLC silica gel GF254 plate R. Reference solution. Dissolve 5 mg of anethole R in 0.5 mL of
Mobile phase : hexane R, toluene R (20:80 V/V). xylene R.
Application : 10 μL as bands of 20 mm by 3 mm.
STORAGE
Development : over a path of 10 cm.
Protected from moisture.
Drying : in air.
Detection A : examine in ultraviolet light at 254 nm.
Results A : the chromatograms show in the central part a
quenching zone due to anethole.
Detection B : spray with sulfuric acid R and heat at 140 °C 01/2019:1323
for 5 min ; examine in daylight.
Results B : the chromatograms show in the central part a
violet band due to anethole ; the chromatogram obtained
with the test solution also shows a reddish-brown zone in
the upper third (terpenes). FENUGREEK
TESTS
Estragole and fenchone. Gas chromatography (2.2.28): use Trigonellae foenugraeci semen
the normalisation procedure.
Test solution. Dilute the mixture of essential oil and xylene R DEFINITION
obtained in the assay of essential oil to 5.0 mL with xylene R, Dried, ripe seeds of Trigonella foenum-graecum L.
by rinsing the apparatus.
Reference solution. Dissolve 5 mg of estragole R and 5 mg of IDENTIFICATION
fenchone R in 0.5 mL of xylene R.
A. The seed is hard, flattened, brown or reddish-brown and
Column : more or less rhomboidal with rounded edges. It is 3-5 mm
– size : l = 30-60 m, Ø = 0.3 mm ; long, 2-3 mm wide and 1.5-2 mm thick. The widest
surfaces are marked by a groove that divides the seed into
– stationary phase : macrogol 20 000 R. 2 unequal parts. The smaller part contains the radicle ; the
Carrier gas : nitrogen for chromatography R. larger part contains the cotyledons.
Flow rate : 0.40 mL/min. B. Microscopic examination (2.8.23). The powder is
yellowish-brown. Examine under a microscope using
Split ratio : 1:200. chloral hydrate solution R. The powder shows the following
Time Temperature
diagnostic characters (Figure 1323.-1): fragments of the
(min) (°C)
testa (transverse section [B]) with lageniform epidermal
0-4 60
cells [Bb] covered by a thick cuticle [Ba], a hypodermis
Column
consisting of large cells, narrower at the upper end and
4 - 26 60 → 170 constricted in the middle, with bar-like thickenings of
26 - 41 170
the radial walls [Bc] and parenchyma with flattened
cells [Bd] ; fragments of the epidermis in surface view,
Injection port 220 yellowish-brown, consisting of small polygonal cells, either
Detector 270
with thick, channelled walls and a narrow lumen (view
from above [G]) or with smooth walls and a larger lumen
(view from below [C]) ; fragments of the hypodermis in
Detection : flame ionisation. surface view, with cells having either a circular outline and
Injection : 1 μL. thickened walls, closely beaded (view from above [H]), or
having a polyhedral outline whose bar-like thickenings
Limits : extend from the lower to the upper walls (view from
– estragole : maximum 10.0 per cent in the essential oil ; below [A]) ; parenchyma of the testa consisting of loosely
arranged cells leaving numerous spaces [E] ; fragments of
– fenchone : maximum 7.5 per cent in the essential oil. endosperm with cells that are rounded [F] or elongated [D]
Foreign matter (2.8.2) : maximum 1.5 per cent of peduncles depending on the orientation, associated with mucilage [Fa]
and maximum 1.5 per cent of other foreign matter. and sometimes with small spiral or annular vessels [J].

1510 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 11.0 Feverfew

Content : minimum 0.20 per cent of parthenolide (C15H20O3 ;

Mr 248.3) (dried drug).

CHARACTERS

Camphoraceous odour.

IDENTIFICATION

A. The leafy, more or less branched stem has a diameter

of up to 5 mm ; it is almost quadrangular, channelled
longitudinally and slightly pubescent. The leaves are ovate,
2-5 cm long, sometimes up to 10 cm, yellowish-green,
petiolate and alternate. They are pinnate or bipinnate,
deeply divided into 5-9 segments, each with a coarsely
crenate margin and an obtuse apex. Both surfaces are
somewhat pubescent and the midrib is prominent on the
Figure 1323.-1. – Illustration for identification test B of lower surface. When present, the flowering heads are
powdered herbal drug of fenugreek 12-22 mm in diameter with long pedicels ; they are clustered
C. Thin-layer chromatography (2.2.27). into broad corymbs consisting of 5-30 flower-heads. The
Test solution. Place 1.0 g of the powdered herbal drug hemispherical involucre is 6-8 mm wide and consists of
(710) (2.9.12) in a 25 mL conical flask and add 5.0 mL of many overlapping bracts, which are rather narrow, obtuse
methanol R. Heat in a water-bath at 65 °C for 5 min. Cool and scarious and have membranous margins. The central
and filter. flowers are yellow, hermaphrodite, tube-shaped with 5
Reference solution. Dissolve 3.0 mg of trigonelline teeth and have 5 stamens inserted in the corolla ; the
hydrochloride R in 1.0 mL of methanol R. filaments of the stamens are separate from each other but
Plate : TLC silica gel F254 plate R. the anthers are fused into a tube through which passes the
style, bearing 2 stigmatic branches. The peripheral flowers
Mobile phase : water R, methanol R (30:70 V/V). are female and have a white, three-toothed ligule, 2-7 mm
Application : 20 μL of the test solution and 10 μL of the long. The fruit is an achene, 1.2-1.5 mm long, brown when
reference solution, as bands. ripe, with 5-10 white longitudinal ribs. It is glandular and
Development : over a path of 10 cm. bears a short, crenate, membranous crown.
Drying : in air.
Detection A : examine in ultraviolet light at 254 nm. B. Microscopic examination (2.8.23). The powder is
Results A : the chromatogram obtained with the test yellowish-green. Examine under a microscope using
solution shows in its lower half a quenching zone similar in chloral hydrate solution R. The powder shows the following
position and fluorescence to the zone in the chromatogram diagnostic characters (Figure 1516.-1) : numerous isolated,
obtained with the reference solution. large, multicellular, uniseriate covering trichomes [J]
consisting of a rhomboidal basal cell, 3-5 smaller,
Detection B : spray with potassium iodobismuthate
thick-walled rectangular cells, and a very long, flat, slender
solution R2.
terminal cell, often curved at a right angle to the axis of
Results B : the chromatogram obtained with the test solution the basal cell ; isolated glandular trichomes [C, F] with a
shows an intense orange-red zone similar in position and short, biseriate, 2- to 4-celled stalk and a biseriate head
colour to the zone in the chromatogram obtained with the of 4-6 cells around which the cuticle forms a bladder-like
reference solution. It also shows in its upper half, a broad covering ; fragments of the epidermis of the leaves [A]
light brownish-yellow zone (triglycerides). consisting of cells with sinuous anticlinal walls, with a
TESTS striated cuticle [Aa], anomocytic stomata (2.8.3) [Ab],
covering trichomes [Ac] or their basal part [Ad] and
Loss on drying (2.2.32) : maximum 12.0 per cent, determined glandular trichomes ; numerous fragments of the stems
on 1.000 g of the powdered herbal drug by drying in an oven (longitudinal section [H, L]) showing pitted or reticulate
at 105 °C for 2 h. vessels [Ha] accompanied by cells of the pith with pitted
Total ash (2.4.16) : maximum 5.0 per cent. walls [Hb] and groups of fibres [La]. If flowers are present,
Swelling index (2.8.4) : minimum 6, determined on the the powder also shows : fragments of the outer epidermis of
powdered herbal drug (710) (2.9.12). the bracts of the involucre (surface view [M]), consisting,
on the margin, of thin-walled cells [Ma] and occasional
04/2021:1516 covering trichomes [Mb], and a central region composed of
elongated sclereids with strongly thickened and channelled
walls [Mc] ; fragments of the inner epidermis of the
corolla [K] consisting of polygonal cells, papillose, with
finely striated walls ; fragments of the outer epidermis
of the corolla (surface view [B]) consisting of sinuous
FEVERFEW cells, covered by a striated cuticle, bearing numerous
glandular trichomes [Ba] ; fragments of the ovary [E] with
Tanaceti parthenii herba parenchymatous cells containing small crystals of calcium
oxalate [Ea] occurring as cluster crystals or occasionally
DEFINITION as prisms [Eb], accompanied by small spiral vessels [Ec] ;
Dried, whole or fragmented aerial parts of Tanacetum spherical pollen grains about 25 μm in diameter, with
parthenium (L.) Sch. Bip. 3 pores and a spiny exine [D, G].

General Notices (1) apply to all monographs and other texts 1511