Bitter-orange epicarp and mesocarp EUROPEAN PHARMACOPOEIA 5.

— cis-anethole : maximum 0.5 per cent, Results : see below the sequence of the zones present
— trans-anethole : 55.0 per cent to 75.0 per cent, in the chromatograms obtained with the reference and
— anisaldehyde : maximum 2.0 per cent. test solutions. Furthermore, other fluorescent zones
are present in the chromatogram obtained with the test
The ratio of α-pinene content to limonene content is greater solution.
than 1.0.
Top of the plate
STORAGE
A light blue fluorescent zone
In a well-filled, airtight container, protected from light and
at a temperature not exceeding 25 °C. A light blue fluorescent zone
Caffeic acid : a light blue
fluorescent zone
01/2005:1603
A light blue fluorescent zone

BITTER-ORANGE EPICARP AND A light blue fluorescent zone

MESOCARP Naringin : a dark green

fluorescent zone
A dark green fluorescent zone
(naringin)
A red fluorescent zone
Aurantii amari epicarpium et mesocarpium (neoeriocitrin)
DEFINITION An orange fluorescent zone
Dried epicarp and mesocarp of the ripe fruit of Citrus Reference solution Test solution
aurantium L. ssp. aurantium (C. aurantium L. ssp. amara
Engl.) partly freed from the white spongy tissue of the TESTS
mesocarp and endocarp.
Foreign matter (2.8.2) : it complies with the test for foreign
Content : minimum 20 ml/kg of essential oil (anhydrous matter.
drug).
Water (2.2.13) : maximum 10.0 per cent, determined on
CHARACTERS 20.0 g of powdered drug (355) by distillation.
Aromatic odour and spicy bitter taste. Total ash (2.4.16) : maximum 7.0 per cent
Macroscopic and microscopic characters described under Extractable matter : minimum 6.0 per cent.
identification A and B. To 2.000 g of the powdered drug (250) add a mixture of 3 ml
IDENTIFICATION of water R and 7 ml of alcohol R and extract for 2 h, shaking
frequently. Filter, evaporate 2.000 g of the filtrate to dryness
A. The drug consists of elliptical to irregular pieces 5 cm to on a water-bath and dry in an oven at 100-105 °C for 3 h.
8 cm long, 3 cm to 5 cm broad and about 3 mm thick. The Allow to cool in a dessicator over diphosphorus pentoxide R
outer surface is yellowish to reddish-brown and distinctly and weigh. The residue weighs a minimum of 120 mg.
punctate, the inner surface is yellowish to brownish-white.
B. Reduce to a powder (355). The powder is light brown. ASSAY
Examine under a microscope using chloral hydrate Carry out the determination of essential oil in vegetable
solution R. The powder shows small polygonal cells with drugs (2.8.12). Use a 500 ml round-bottomed flask, 200 ml
slightly thickened anticlinal walls, filled with orange-red of water R as the distillation liquid and 0.5 ml of xylene R
chromatophores, and very occasional anomocytic in the graduated tube. Reduce the drug to a powder (710)
stomata (2.8.3) ; fragments of the hypodermic showing and immediately use 15.0 g for the determination. Distil at a
collenchymatous thickening ; groups of parenchyma rate of 2-3 ml/min for 90 min.
with each cell containing a prism crystal of calcium
oxalate ; fragments of lysigenous oil glands ; parenchyma
01/2005:1604
containing crystals of hesperidin which dissolve in a
20 g/l potassium hydroxide R solution giving a yellow
colour. BITTER-ORANGE-EPICARP AND
C. Thin-layer chromatography (2.2.27). MESOCARP TINCTURE
Test solution. To 1.0 g of the powdered drug (710) add
10 ml of methanol R and heat in a water-bath at 65 °C for Aurantii amari epicarpii et mesocarpii
5 min shaking frequently. Allow to cool and filter. tinctura
Reference solution. Dissolve 1.0 mg of naringin R and
1.0 mg of caffeic acid R in 1 ml of methanol R. DEFINITION
Plate : TLC silica gel plate R. Tincture produced from Bitter-orange epicarp and
Mobile phase : water R, anhydrous formic acid R, ethyl mesocarp (1603).
acetate R (10:15:75 V/V/V). PRODUCTION
Application : 20 µl, as bands.
The tincture is produced from 1 part of the freshly powdered
Development : over a path of 10 cm. drug (2000) and 5 parts of alcohol (70 per cent V/V/V) by
Drying : in air, and heat in an oven at 110-120 °C for an appropriate procedure.
5 min.
Detection : spray the warm plate with a 10 g/l solution CHARACTERS
of diphenylboric acid aminoethyl ester R in methanol R Liquid with a bitter taste.
and then with a 50 g/l solution of macrogol 400 R in
methanol R. After at least 1 h, examine in ultraviolet light IDENTIFICATION
at 365 nm. Examine by thin-layer chromatography (2.2.27).

1110 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 5.0 Bitter-orange flower

Test solution. The tincture to be examined. peduncle which is about 5 mm to 10 mm long. The flower
Reference solution. Dissolve 1.0 mg of naringin R and buds contain at least 20 stamens with yellow anthers and
1.0 mg of caffeic acid R in 1 ml of methanol R. with filaments fused at the base into groups of 4 or 5 ; the
Plate : TLC silica gel plate R. ovary is superior, brownish-black and spherical, consists
of 8 to 10 multi-ovular loculi and is surrounded at the
Mobile phase : water R, anhydrous formic acid R, ethyl base by an annular granular hypogynous disc ; the thick,
acetate R (10:15:75 V/V/V). cylindrical style ends in a capitate stigma.
Application : 20 µl, as bands. B. Reduce to a powder (355). The powder is brownish-yellow.
Development : over a path of 10 cm. Examine under a microscope using chloral hydrate
Drying : in air, and heat in an oven at 110-120 °C for 5 min. solution R. The powder shows numerous spherical pollen
Detection : spray the warm plate with a 10 g/l solution grains, with a finely pitted exine and 3 to 5 germinal
of diphenylboric acid aminoethyl ester R in methanol R pores ; fragments of the epidermis of the sepals with
and then with a 50 g/l solution of macrogol 400 R in unicellular trichomes and with large prism crystals of
methanol R. After 1 h, examine in ultraviolet light at 365 nm. calcium oxalate in the underlying mesophyll ; fragments
of the epidermis of the petals with a distinctly striated
Results : see below the sequence of the zones present in cuticle ; fragments of large schizolysigenous oil glands
the chromatograms obtained with the reference and test which measure up to 100 µm in diameter, numerous
solutions. Furthermore, other zones are present in the anomocytic stomata (2.8.3). Examine under a microscope
chromatogram obtained with the test solution. using a 300 g/l solution of potassium hydroxide R. The
Top of the plate powder shows yellow crystals of hesperidin.
A light blue fluorescent zone C. Examine the chromatograms obtained in the test for
sweet-orange flower.
A light blue fluorescent zone
Results : see below the sequence of the zones present in
Caffeic acid : a light blue the chromatograms obtained with the reference solution
fluorescent zone
and the test solution.
A light blue fluorescent zone
Top of the plate
A light blue fluorescent zone
A weak yellow fluorescent zone
Naringin : a dark green fluorescent A dark green fluorescent zone
zone (naringin) A weak yellow fluorescent zone
A red fluorescent zone
(neoeriocitrin) Hesperidin : a greenish-yellow A greenish-yellow fluorescent
fluorescent zone zone (hesperidin)
An orange fluorescent zone
Naringin : a yellow fluorescent A yellow fluorescent zone
Reference solution Test solution zone (naringin)
A red fluorescent zone
TESTS (neoeriocitrin)
A yellow fluorescent zone
Ethanol content (2.9.10) : 63 per cent to 67 per cent V/V. (diosmin and neodiosmin)
Methanol and 2-propanol (2.9.11) : maximum 0.05 per Reference solution Test solution
cent V/V of methanol and maximum 0.05 per cent V/V of
2-propanol. TESTS
Dry residue : minimum 6.0 per cent m/m, determined on Sweet-orange flower. Thin-layer chromatography (2.2.27).
2.00 g of tincture to be examined.
Test solution. To 0.5 g of the powdered drug (355), add 5 ml
of methanol R. Heat with stirring at 40 °C for 10 min. Filter.
01/2005:1810 Reference solution. Dissolve 3.0 mg of naringin R and
3.0 mg of hesperidin R in 10 ml of methanol R.
BITTER-ORANGE FLOWER Plate : TLC silica gel plate R.
Mobile phase : water R, anhydrous formic acid R, ethyl
Aurantii amari flos acetate R (10:15:75 V/V/V).
Application : 10 µl, as bands.
DEFINITION
Development : over a path of 10 cm.
Whole, dried, unopened flower of Citrus aurantium L.
ssp. aurantium (C. aurantium L. ssp. amara Engl.). Drying : in air, then heat in an oven at 110-120 °C for 5 min.
Content : minimum 8.0 per cent of total flavonoids, expressed Detection : spray the hot plate with a 10 g/l solution of
as naringin (C27H32O14 ; Mr 580.5) (dried drug). diphenylboric acid aminoethyl ester R in methanol R
and then with a 50 g/l solution of macrogol 400 R in
CHARACTERS methanol R. After at least 1 h, examine in ultraviolet light
Macroscopic and microscopic characters described under at 365 nm.
identification tests A and B. Results : the chromatogram obtained with the test solution
shows a yellow zone similar in position to the zone of
IDENTIFICATION naringin in the chromatogram obtained with the reference
A. The flower buds are white or yellowish-white and may solution and immediately below it a red zone (neoeriocitrin).
reach up to 25 mm in length. The dialypetalous corolla Foreign matter (2.8.2) : maximum 2 per cent.
is composed of 5 thick, oblong and concave petals
dotted with oil glands visible under a hand lens ; the Loss on drying (2.2.32) : maximum 11.0 per cent, determined
short, yellowish-green persistent gamosepalous calyx has on 1.000 g of the powdered drug (355) by drying in an oven
5 spreading sepals, connate at the base and forming a at 100-105 °C.
star-shaped structure attached to the yellowish-green Total ash (2.4.16) : maximum 10.0 per cent.

General Notices (1) apply to all monographs and other texts 1111