Wormwood

Wormwood EUROPEAN PHARMACOPOEIA 11.
PRODUCTION Mobile phase:

The extract is produced from the herbal drug by a suitable – mobile phase A : tetrahydrofuran R, 0.5 per cent V/V
procedure using either water or a hydroalcoholic solvent solution of phosphoric acid R (1.8:98.2 V/V) ;
equivalent in strength to a maximum of 80 per cent V/V – mobile phase B : tetrahydrofuran R ;
ethanol. Time Mobile phase A Mobile phase B
CHARACTERS (min) (per cent V/V) (per cent V/V)
0 - 15 100 0
Appearance : yellowish-brown amorphous powder.
15 - 17 100 → 90 0 → 10
IDENTIFICATION
17 - 23 90 10
Thin-layer chromatography (2.2.27).
23 - 25 90 → 100 10 → 0
Test solution (a). To 0.200 g of the extract to be examined add
5 mL of methanol R. Sonicate for 5 min, filter and dilute to 25 - 40 100 0
10 mL with methanol R.
Test solution (b). To 5.0 mL of test solution (a) add 1.0 mL of Flow rate : 1.0 mL/min.
a 50 g/L solution of anhydrous sodium carbonate R and heat Detection : spectrophotometer at 270 nm.
in a water-bath at about 60 °C for 10 min. Cool and filter if Injection : 10 μL.
necessary. Retention time : salicin = about 6.4 min ; pi-
Reference solution. Dissolve 2.0 mg of salicin R and 2.0 mg of cein = about 7.7 min.
chlorogenic acid R in 1.0 mL of methanol R. System suitability : reference solution :
Plate : TLC silica gel plate R (5-40 μm) [or TLC silica gel plate R – resolution : minimum 1.5 between the peaks due to salicin
(2-10 μm)]. and picein.
Mobile phase : water R, methanol R, ethyl acetate R Calculate the percentage content of total salicylic derivatives,
(8:15:77 V/V/V). expressed as salicin, from the following expression :
Application : 10 μL [or 2 μL] as bands. A1 ´ m 2 ´ p ´ 2
Development : over a path of 15 cm [or 6 cm]. A 2 ´ m1
Drying : in a current of warm air.
A1 = area of the peak due to salicin in the chromatogram
Detection : spray with a mixture of 5 volumes of sulfuric acid R
and 95 volumes of methanol R. Heat at 100-105 °C for 5 min obtained with the test solution ;
and examine in daylight. A2 = area of the peak due to salicin in the chromatogram
Results : see below the sequence of the zones present in the obtained with the reference solution ;
chromatograms obtained with the reference solution and m1 = mass of the extract to be examined used to prepare
test solutions (a) and (b). Furthermore, other zones may be the test solution, in grams ;
present in the chromatogram obtained with test solutions (a) m2 = mass of salicin CRS used to prepare the reference
and (b). solution, in grams ;
Top of the plate p = percentage content of salicin in salicin CRS.
_______ _______
04/2011:1380
Several reddish-violet
zones may be present
Salicin : a reddish-violet A weak reddish-violet A reddish-violet zone
zone zone (salicin) (salicin)
_______ _______
Chlorogenic acid : a WORMWOOD

brown zone
Reference solution Test solution (a) Test solution (b) Absinthii herba
DEFINITION
ASSAY
Basal leaves or slightly leafy, flowering tops, or mixture of
Liquid chromatography (2.2.29). these dried, whole or cut organs of Artemisia absinthium L.
Test solution. To 0.300 g of the extract to be examined add Content : minimum 2 mL/kg of essential oil (dried drug).
40 mL of methanol R and 40.0 mL of 0.1 M sodium hydroxide.
Heat in a water-bath at about 60 °C under a reflux condenser, IDENTIFICATION
with frequent shaking, for about 1 h. After cooling, add A. The leaves are greyish or greenish, densely tomentose on
4.0 mL of 1 M hydrochloric acid. Filter the suspension into a both surfaces. The basal leaves, with long petioles, have
100 mL volumetric flask, then wash and dilute to 100.0 mL triangular or oval bipinnatisect or tripinnatisect lamina,
with a mixture of equal volumes of water R and methanol R. with rounded or lanceolate segments. The cauline leaves
Filter through a membrane filter (nominal pore size 0.45 μm). are less segmented and the apical leaves are lanceolate.
Reference solution. Dissolve 5.0 mg of picein R in 25.0 mL The stem of the flower-bearing region is greenish-grey,
of a mixture of 20 volumes of water R and 80 volumes of tomentose, up to 2.5 mm in diameter and usually with
methanol R (solution A). Dissolve 15.0 mg of salicin CRS in 5 flattened longitudinal grooves. The capitula are arranged
25 mL of a mixture of 20 volumes of water R and 80 volumes as loose, axillary panicles, inserted at the level of the
of methanol R. Add 5.0 mL of solution A and dilute to 50.0 mL lanceolate or slightly pinnatisect leaves ; they are spherical
with water R. or flattened hemispherical, 2-4 mm in diameter and consist
Column: of a grey, tomentose involucre, the outer bracts linear, inner
layer ovate, blunt at the apices with scarious margins, a
– size : l = 0.10 m, Ø = 4.6 mm ; receptacle with very long paleae up to 1 mm or more long,
– stationary phase : octadecylsilyl silica gel for numerous yellow, tubular, hermaphroditic florets about
chromatography R (3 μm). 2 mm long and few yellow, ray florets.
1780 See the information section on general monographs (cover pages)

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EUROPEAN PHARMACOPOEIA 11.0 Yarrow
B. Microscopic examination (2.8.23). The powder is Mobile phase : acetone R, glacial acetic acid R, toluene R,
greenish-grey. Examine under a microscope using chloral methylene chloride R (10:10:30:50 V/V/V/V).
hydrate solution R. The powder shows the following Application : 10 μL as bands.
diagnostic characters (Figure 1380.-1.) : many T-shaped Development : over a path of 15 cm.
trichomes [A] with a short uniseriate stalk consisting of
1-5 small cells, perpendicularly capped by a very long, Drying : in air.
undulating terminal cell tapering at the ends ; fragments of Detection A : spray with acetic anhydride - sulfuric acid
epidermises (surface view [D]) with sinuous or wavy walls, solution R and examine in daylight.
anomocytic stomata (2.8.3) [Da], covering trichomes [Db] Results A : the chromatogram obtained with the test
and glandular trichomes containing oil [Dc] or not solution shows a blue zone due to artabsin shortly above a
containing oil [Dd], each with a short, biseriate, 2-celled red zone due to methyl red in the chromatogram obtained
stalk and a biseriate head with 2-4 cells ; free glandular with the reference solution.
trichomes (side view [C]) ; fragments of the corollas of Detection B : examine in daylight while heating at
the tubular and ray florets, some containing small cluster 100-105 °C for 5 min.
crystals of calcium oxalate [H] ; numerous paleae each Results B : the chromatogram obtained with the reference
composed of a small cell forming a stalk and a very long, solution shows in the middle third a red zone due to methyl
cylindrical and thin-walled terminal cell about 1-1.5 mm red and below it a light pink zone due to resorcinol. The
long, either whole [E] or limited to the distal part [B] ; chromatogram obtained with the test solution shows an
spheroidal pollen grains, about 30 μm in diameter, with intense red or brownish-red zone due to absinthin with a
3 pores and a finely warty exine [G] ; fragments of vascular similar RF value to that of the zone due to resorcinol in the
tissue from the leaves [F] or the stems [J] consisting of chromatogram obtained with the reference solution. Other
vessels with spiral or annular thickenings [Fa], or with zones are visible, but less intense than that due to absinthin.
bordered pits [Ja], fibres [Fb, Jb] and parenchymatous cells
with pitted, moderately thickened walls [Fc, Jc]. 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 :
Figure 1380.-1. – Illustration for identification test B of – essential oil : minimum 2 mL/kg (dried drug) ;
powdered herbal drug of wormwood – proazulenes, expressed as chamazulene (C14H16 ; Mr 184.3) :
C. Thin-layer chromatography (2.2.27). minimum 0.02 per cent (dried drug).
Test solution. Place 2 g of the powdered herbal drug (355) IDENTIFICATION
(2.9.12) in 50 mL of boiling water R and allow to stand A. The leaves are green or greyish-green, faintly pubescent
for 5 min, shaking the flask several times. After cooling, on the upper surface and more pubescent on the lower
add 5 mL of a 100 g/L solution of lead acetate R. Mix and surface, 2-3 pinnately divided with linear lobes and a
filter. Rinse the flask and the residue on the filter with finely pointed whitish tip. The capitula are arranged in a
20 mL of water R. Shake the filter with 50 mL of methylene corymb at the end of the stem. Each capitulum, 3-5 mm
chloride R. Separate the organic layer, dry over anhydrous in diameter, consists of the receptacle, usually 4-5 ligulate
sodium sulfate R, filter and evaporate the filtrate to dryness ray-florets and 3-20 tubular disk-florets. The involucre
on a water-bath. Dissolve the residue in 0.5 mL of ethanol consists of 3 rows of imbricate lanceolate, pubescent green
(96 per cent) R. bracts arranged with a brownish or whitish, membranous
Reference solution. Dissolve 2 mg of methyl red R and 2 mg margin. The receptacle is slightly convex and, in the axillae
of resorcinol R in 10.0 mL of methanol R. of paleae, bears ligulate ray-florets with a three-lobed,
Plate : TLC silica gel plate R. whitish or reddish ligule and tubular disk-florets with
General Notices (1) apply to all monographs and other texts 1781
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Wormwood EUROPEAN PHARMACOPOEIA 11.

PRODUCTION Mobile phase:

Chlorogenic acid : a WORMWOOD

1780 See the information section on general monographs (cover pages)

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