Yarrow
Mobile phase : acetone R, glacial acetic acid R, toluene R,
methylene chloride R (10:10:30:50 V/V/V/V).
Application : 10 μL as bands.
Development : over a path of 15 cm.
Drying : in air.
Detection A : spray with acetic anhydride - sulfuric acid
solution R and examine in daylight.
Results A : the chromatogram obtained with the test
solution shows a blue zone due to artabsin shortly above a
red zone due to methyl red in the chromatogram obtained
with the reference solution.
Detection B : examine in daylight while heating at
100-105 °C for 5 min.
Results B : the chromatogram obtained with the reference
solution shows in the middle third a red zone due to methyl
red and below it a light pink zone due to resorcinol. The
chromatogram obtained with the test solution shows an
intense red or brownish-red zone due to absinthin with a
similar RF value to that of the zone due to resorcinol in the
chromatogram obtained with the reference solution. Other
zones are visible, but less intense than that due to absinthin.
TESTS
Foreign matter (2.8.2) : maximum 5 per cent of stems with a
diameter greater than 4 mm and maximum 2 per cent of other
foreign matter.
Bitterness value (2.8.15) : minimum 10 000.
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 12.0 per cent.
Ash insoluble in hydrochloric acid (2.8.1): maximum 1.0 per
cent.
ASSAY
Essential oil (2.8.12). Use 50.0 g of the cut drug, a 1000 mL
round-bottomed flask and 500 mL of water R as the distillation
liquid. Add 0.5 mL of xylene R in the graduated tube. Distil at
a rate of 2-3 mL/min for not less than 3 h.
07/2014:1382
YARROW
Millefolii herba
DEFINITION
Whole or cut, dried flowering tops of Achillea millefolium L.
Content :
– essential oil : minimum 2 mL/kg (dried drug) ;
– proazulenes, expressed as chamazulene (C14H16 ; Mr 184.3) :
minimum 0.02 per cent (dried drug).
IDENTIFICATION
A. The leaves are green or greyish-green, faintly pubescent on
the upper surface and more pubescent on the lower surface, C. To 2.0 g of the powdered herbal drug (710) (2.9.12)
2-3 pinnately divided with linear lobes and a finely pointed
whitish tip. The capitula are arranged in a corymb at the
end of the stem. Each capitulum, 3-5 mm in diameter,
consists of the receptacle, usually 4-5 ligulate ray-florets and this solution add 2.5 mL of dimethylaminobenzaldehyde
3-20 tubular disk-florets. The involucre consists of 3 rows of
imbricate lanceolate, pubescent green bracts arranged with
a brownish or whitish, membranous margin. The receptacle
is slightly convex and, in the axillae of paleae, bears ligulate greenish-blue colour.
1674
EUROPEAN PHARMACOPOEIA 10.
ray-florets with a three-lobed, whitish or reddish ligule and
tubular disk-florets with a radial, five-lobed, yellowish or
light brownish corolla. The pubescent green, partly brown
or violet stems are longitudinally furrowed, up to 3 mm
thick with a light-coloured medulla.
B. Microscopic examination (2.8.23). The powder is green
or greyish-green. Examine under a microscope using
chloral hydrate solution R. The powder shows the following
diagnostic characters (Figure 1382.-1) : fragments of the
stem epidermis (surface view [K]), with cells having a
smooth cuticle and anomocytic stomata (2.8.3) ; fragments
of leaf and bract epidermises (surface view [B]), with
cells having wavy and irregularly thickened walls, a finely
striated cuticle and anomocytic stomata (2.8.3) ; very rare
glandular trichomes with a short stalk and a head formed of
2 rows of 3-5 cells enclosed in a bladder-like membrane [H] ;
uniseriate, whole or fragmented covering trichomes [A]
consisting of 4-6 small, more or less isodiametric cells at the
base and a thick-walled, often somewhat tortuous terminal
cell, about 400 μm to greater than 1000 μm long ; fragments
of the ligulate corolla with papillary epidermal cells [D] ;
fragments of the corolla tubes, with sinuous epidermal
cells, covered by a thin striated cuticle (surface view [F]);
small-celled parenchyma from the corolla tubes containing
cluster crystals of calcium oxalate [E] ; groups of lignified
and pitted cells from the bracts [G] ; spherical pollen grains,
about 30 μm in diameter, with 3 germinal pores and a spiny
exine [C] ; groups of sclerenchymatous fibres and small
vessels with spiral or annular thickening, from the stem [J].
Figure 1382.-1. – Illustration for identification test B of
powdered herbal drug of yarrow
add 25 mL of ethyl acetate R, shake for 5 min and filter.
Evaporate to dryness on a water-bath and dissolve the
residue in 0.5 mL of toluene R (solution A). To 0.1 mL of
solution R8 and heat on a water-bath for 2 min. Allow
to cool. Add 5 mL of light petroleum R and shake the
mixture vigorously. The aqueous layer shows a blue or
See the information section on general monographs (cover pages)
EUROPEAN PHARMACOPOEIA 10.0
D. Thin-layer chromatography (2.2.27).
Test solution. Use solution A prepared in identification
test C.
Reference solution. Dissolve 10 mg of cineole R and 10 mg
of guaiazulene R in 20 mL of toluene R.
Plate : TLC silica gel plate R.
Mobile phase : ethyl acetate R, toluene R (5:95 V/V).
Application : 20 μL as bands.
Development : over a path of 10 cm.
Drying : in air.
Detection : treat with anisaldehyde solution R, heat at
100-105 °C for 5-10 min and examine in daylight.
Results : the chromatogram obtained with the reference
solution shows in the upper part a red zone (guaiazulene)
and in the middle part a blue or greyish-blue zone (cineole).
The chromatogram obtained with the test solution shows a
violet zone a little above the zone due to guaiazulene in the
chromatogram obtained with the reference solution ; below
this zone a reddish-violet zone ; below which, 1-2 not clearly
separated greyish-violet or greyish zones (which changes to
greenish-grey after a few hours) and a reddish-violet zone a
little above the zone due to cineole in the chromatogram
obtained with the reference solution. Furthermore, other
faint zones may be present.
TESTS
Foreign matter (2.8.2) : maximum 5 per cent of stems with a
diameter greater than 3 mm and maximum 2 per cent of other
foreign matter.
Loss on drying (2.2.32) : maximum 12.0 per cent, determined
on 0.500 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 10.0 per cent.
Ash insoluble in hydrochloric acid (2.8.1): maximum 2.5 per
cent.
ASSAY
Essential oil (2.8.12). Use 20.0 g of cut herbal drug, a 1000 mL
round-bottomed flask and 500 mL of a mixture of 1 volume of
water R and 9 volumes of ethylene glycol R as the distillation
liquid. Add 0.50 mL of 1,2,4-trimethylbenzene R in the
graduated tube. Distil at a rate of 3-4 mL/min for 4 h.
Stop cooling at the end of distillation and continue distilling C. Thin-layer chromatography (2.2.27).
until the blue, steam-volatile components have reached the
lower end of the cooler. Immediately start cooling again, to
avoid warming the separation space. Stop the distillation after
10 min.
Proazulenes. To ensure that as little water as possible is
transferred, transfer the blue mixture of essential oil and
1,2,4-trimethylbenzene obtained in the assay of essential oil
into a 50 mL volumetric flask with the aid of small portions
of xylene R, rinsing the graduated tube of the apparatus
with xylene R, and dilute to 50.0 mL with the same solvent.
Measure the absorbance (2.2.25) at 608 nm using xylene R
as the compensation liquid.
Calculate the percentage content of proazulenes, expressed as
chamazulene, using the following expression :
A ´ 2.1
m
i.e. taking the specific absorbance of chamazulene to be 23.8.
A = absorbance at 608 nm ;
m zones may be present in the chromatogram obtained with
= mass of the herbal drug to be examined, in grams.
General Notices (1) apply to all monographs and other texts
Zanthoxylum bungeanum pericarp
01/2017:2656
ZANTHOXYLUM BUNGEANUM
PERICARP
Zanthoxyli bungeani pericarpium
DEFINITION
Dried pericarp of the ripe fruit, with seeds removed, of
Zanthoxylum bungeanum Maxim.
Content : minimum 15 mL/kg of essential oil (anhydrous
drug).
IDENTIFICATION
A. Single, dehiscent spherical capsules, split along the ventral
suture, 4-5 mm in diameter ; the outer surface is dark
red or brownish-red, with numerous convex, translucent
verrucose oil dots ; the inner surface is light yellow and
smooth ; the endocarp is mostly separated from the
mesocarp at the base and is rolled up.
B. Microscopic examination (2.8.23). The powder is
reddish-brown. Examine under a microscope using chloral
hydrate solution R. The powder shows the following
diagnostic characters : fragments of the epicarp, covered by
a striated cuticle, consisting of polygonal cells, with rigid
walls and anomocytic stomata (2.8.3) with 5-7 subsidiary
cells, some cells of the epicarp contain orange-yellow
granular contents ; fragments of the mesocarp consisting
of parenchymatous cells some of which contain cluster
crystals of calcium oxalate, elongated cells of the oil
glands containing droplets of essential oil, vascular
bundles with spiral vessels accompanied by fibres with
regularly thickened, pitted walls and pointed ends,
sometimes bifurcated, prisms of calcium oxalate, especially
in the area of the vascular bundles ; fragments of the
endocarp consisting of several layers of narrow, elongated,
thick-walled, pitted cells in a parquetry arrangement, with
the arrangements in the different layers oriented crosswise
to each other.
Test solution. To 0.5 g of the powdered herbal drug (355)
(2.9.12) add 5 mL of methanol R. Sonicate for 10 min.
Centrifuge or filter. Use the supernatant or filtrate.
Reference solution. Dissolve 1 mg of chlorogenic acid R and
1 mg of emodin R in 1 mL of methanol R.
Plate : TLC silica gel plate R (2-10 μm).
Mobile phase : water R, methanol R, ethyl acetate R
(10:20:80 V/V/V).
Application : 5 μL as bands of 8 mm.
Development : over a path of 6 cm.
Drying : in air.
Detection : examine in ultraviolet light at 366 nm.
Results : see below the sequence of zones present in the
chromatograms obtained with the reference solution and
the test solution. Furthermore, other faint fluorescent
the test solution.
1675