Matricaria flower

EUROPEAN PHARMACOPOEIA 5.0

C. Thin-layer chromatography (2.2.27).

Test solution. Dilute 50 l of essential oil obtained in the
assay of essential oil in 1 ml of xylene R.
Reference solution. Dissolve 2 l of chamazulene R, 5 l
of levomenol R and 10 mg of bornyl acetate R in 5 ml
of toluene R.
Plate : TLC silica gel plate R.
STORAGE
Mobile
phase : ethyl acetate R, toluene R (5:95 V/V).
Do not powder.
Application : 10 l, as bands.
Development : over a path of 10 cm.
01/2005:0404
Drying : in air.
MATRICARIA FLOWER
Detection : spray with anisaldehyde solution R and heat
at 100-105 C for 5-10 min. Examine immediately in
daylight.
Matricariae flos
Results : see below the sequence of the zones present in
DEFINITION
the chromatograms obtained with the reference solution
Dried capitula of Matricaria recutita L. (Chamomilla
and the test solution. Furthermore, other zones are
recutita (L.) Rauschert).
present in the chromatogram obtained with the test
solution.
Content :
blue essential oil : minimum 4 ml/kg (dried drug),
Top of the plate
total apigenin 7-glucoside (C21H20O10) : minimum 0.25 per
1 or 2 blue to bluish-violet zones
cent (dried drug).

ASSAY
Essential oil (2.8.12). Use a 500 ml round-bottomed flask
and 200 ml of water R as the distillation liquid. Reduce the
drug to a coarse powder (1400) and immediately use 20.0 g
for the determination. Introduce 0.50 ml of xylene R in the
graduated tube. Distil at a rate of 2-3 ml/min for 2 h.

CHARACTERS
Macroscopic and microscopic characters described under
identification tests A and B.
IDENTIFICATION
A. Capitula, when spread out, consisting of an involucre
made up of many bracts arranged in 1 to 3 rows ; an
elongated-conical receptacle, occasionally hemispherical
(young capitula) ; 12 to 20 marginal ligulate florets with
a white ligule ; several dozen yellow central tubular
florets. The involucre bracts are ovate to lanceolate,
with a brownish-grey scarious margin. The receptacle
is hollow, without paleae. The corolla of the ligulate
florets has a brownish-yellow tube at the base extending
to form a white, elongated-oval ligule. The inferior ovary
is dark brown, ovoid to spherical, and has a long style
and bifid stigma. The tubular florets are yellow and have
a five-toothed corolla tube, 5 syngenesious, epipetalous
stamens and a gynoecium similar to that of the ligulate
florets.
B. Separate the capitulum into its different parts. Examine
under a microscope using chloral hydrate solution R.
The bracts have a margin composed of thin-walled cells
and a central region composed of elongated sclereids
with occasional stomata. The inner epidermis of the
corolla of the ligulate florets, in surface view, consisting
of thin-walled, polygonal cells, slightly papillose, those
of the outer epidermis markedly sinuous and strongly
striated ; corolla of the tubular florets with longitudinally
elongated epidermal cells, and with small groups of
papillae near the apex of the lobes. Glandular trichomes
each consisting of a short stalk and a head of 2 to 3 tiers
of 2 cells each occur on the outer surfaces of the bracts
and on the corollas of both types of florets. The ovaries
have a sclerous ring at the base and the wall is composed
of vertical bands of thin-walled, longitudinally elongated
cells with numerous glandular trichomes, alternating
with fusiform groups of small, radially elongated cells
containing mucilage. The cells at the apex of the stigmas
are extended to form rounded papillae. Numerous small,
cluster crystals of calcium oxalate occur in the inner
tissues of the ovaries and the anther lobes. Pollen grains
spherical to triangular, about 30 m in diameter with
3 pores and a spiny exine.
1976

Chamazulene : a red to
reddish-violet zone
_______

A red to reddish-violet zone

(chamazulene)
_______

Bornyl acetate : a
yellowish-brown zone
A brown zone (en-ynedicycloether)
_______
Levomenol : a reddish-violet to
bluish-violet zone
Reference solution

_______
A reddish-violet to bluish-violet
zone (levomenol)
Test solution

TESTS
Broken drug : maximum 25 per cent, determined on 20.0 g,
passes through a sieve (710).
Foreign matter (2.8.2) : maximum 2 per cent m/m.
Loss on drying (2.2.32) : maximum 12.0 per cent, determined
on 1.000 g of the powdered drug (355) by drying in an oven
at 100-105 C for 2 h.
Total ash (2.4.16) : maximum 13.0 per cent.
ASSAY
Essential oil (2.8.12). Use 30 g of whole drug, a 1000 ml
flask, 300 ml of water R as distillation liquid and 0.50 ml of
xylene R in the graduated tube. Distil at a rate of 3-4 ml/min
for 4 h. Towards the end of this period, stop the flow of
water to the condenser assembly but continue distilling until
the blue, steam-volatile components have reached the lower
end of the condenser. Immediately re-start the flow of water
to the condenser assembly to avoid warming the separation
space. Stop the distillation after a further 10 min.
Total apigenin 7-glucoside. Liquid chromatography (2.2.29).
Test solution. Reduce 40 g of the drug to a powder
(500). Place 2.00 g of the powdered drug in a 500 ml
round-bottomed flask. Add 200 ml of alcohol R. Heat the
mixture under a reflux condenser on a water-bath for 15 min.
Cool and filter. Rinse the filter and the residue with a few
millilitres of alcohol R. To the filtrate add 10 ml of freshly
prepared dilute sodium hydroxide solution R and heat the
mixture under a reflux condenser on a water-bath for about
1 h. Cool. Dilute to 250.0 ml with alcohol R. To 50.0 ml of
the solution add 0.5 g of citric acid R. Shake for 5 min and

See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 5.0

filter. Dilute 5.0 ml to 10.0 ml with the mobile phase (initial

mixture).
Reference solution (a). Dissolve 10.0 mg of apigenin
7-glucoside R in 100.0 ml of methanol R. Dilute 25.0 ml
of this solution to 200 ml with the mobile phase (initial
mixture).
Reference solution (b). Dissolve 10.0 mg of
5,7-dihydroxy-4-methylcoumarin R in 100.0 ml of
methanol R. Dilute 25.0 ml of this solution to 100 ml of the
mobile phase (initial mixture). To 4.0 ml of this solution add
4.0 ml of reference solution (a) and dilute to 10.0 ml with the
mobile phase (initial mixture).
Precolumn :
size : l = 8 mm, = 4.6 mm,
stationary phase: octadecylsilyl silica gel for
chromatography R (5 m).
Column :
size : l = 0.25 m, = 4.6 mm,

Matricaria liquid extract

01/2005:1544

MATRICARIA LIQUID EXTRACT

Matricariae extractum fluidum
DEFINITION
Matricaria liquid extract is produced from Matricaria
flower (0404). It contains not less than 0.30 per cent of
blue residual oil.
PRODUCTION
The extract is produced from the drug and a mixture of
2.5 parts of a 10 per cent m/m solution of ammonia (NH3),
47.5 parts of water and 50 parts of alcohol with an
appropriate procedure for liquid extracts.
CHARACTERS
A brownish, clear liquid with an intense characteristic odour
and characteristic bitter taste ; miscible with water and with
alcohol with development of turbidity, soluble in alcohol
(50 per cent V/V).

stationary phase: octadecylsilyl silica gel for

chromatography R (5 m).

IDENTIFICATION
A. Examine by thin-layer chromatography (2.2.27), using a
Mobile phase :
TLC silica gel F254 plate R.
Test
solution. Place 10 ml of the extract in a separating
mobile phase A : phosphoric acid R, water R
funnel
and shake with 2 quantities, each of 10 ml, of
(0.5:99.5 V/V).
pentane R. Combine the pentane layers, dry over 2 g
mobile phase B : phosphoric acid R, acetonitrile R
of anhydrous sodium sulphate R and filter. Evaporate
(0.5:99.5 V/V).
the filtrate to dryness on a water-bath and dissolve the
residue in 0.5 ml of toluene R.
Time
Mobile phase A
Mobile phase B
Reference solution. Dissolve 4 mg of guaiazulene R,
(min)
(per cent V/V)
(per cent V/V)
20 mg of levomenol R and 20 mg of bornyl acetate R
0-9
75
25
in 10 ml of toluene R.
9 - 19
75 25
25 75
Apply to the plate as bands 10 l of each solution. Develop
over a path of 10 cm using a mixture of 5 volumes of ethyl
19 - 24
25
75
acetate R and 95 volumes of toluene R. Allow the plate
24 - 29
25 75
75 25
to dry in air and examine in ultraviolet light at 254 nm.
The chromatogram obtained with the test solution
29 - 30
75 90
25 10
shows several quenching zones, of which 2 main zones
are in the middle third (en-yne-dicycloether). Examine
Flow rate : 1 ml/min.
in ultraviolet light at 365 nm. The chromatogram
Detection : spectrophotometer at 340 nm.
obtained with the test solution shows in the middle part
an intense blue fluorescent zone (herniarin). Spray
Injection : 20 l.
the plate with anisaldehyde solution R. Examine in
daylight while heating at 100-105 C for 5-10 min. The
System suitability : reference solution (b) :
chromatogram obtained with the reference solution
resolution : minimum 1.8 between the peaks due to
shows in the lower third a reddish-violet to bluish-violet
apigenin 7-glucoside and 5,7-dihydroxy-4-methylcoumarin.
zone (levomenol), in the middle third a yellowish-brown
to greyish-green zone (bornyl acetate) and in the upper
Calculate the percentage content of total apigenin
third a red to reddish-violet zone (guaiazulene). The
7-glucoside from the expression :
chromatogram obtained with the test solution shows
in the lower third yellowish-brown to greenish-yellow
and violet zones and a reddish-violet to bluish-violet
zone corresponding to levomenol in the chromatogram
obtained with the reference solution ; a brownish zone
A1 = area of the peak due to apigenin 7-glucoside in the
(en-yne-dicycloether) similar in position to bornyl acetate
chromatogram obtained with the test solution,
in the chromatogram obtained with the reference
A2 = area of the peak due to apigenin 7-glucoside in
solution ; a red or reddish-violet zone (chamazulene)
the chromatogram obtained with the reference
corresponding to guaiazulene in the chromatogram
solution,
obtained with the reference solution and immediately
above it 1 or 2 blue to bluish-violet zones ; further weak
m1 = mass of the drug in the test solution, in grams,
zones may be present in the chromatogram obtained with
m2 = mass of apigenin 7-glucoside R in reference
the test solution.
solution (a), in grams,
B. Examine by thin-layer chromatography (2.2.27), using a
P
= percentage content of apigenin 7-glucoside in the
TLC silica gel plate R.
reagent.
Test solution. Use the extract.
General Notices (1) apply to all monographs and other texts

1977