Citronella oil
Readily carbonisable substances. To 1.0 g in a cleaned test
tube add 10 ml of sulphuric acid R and immediately heat the
mixture in a water-bath at 90 ± 1 °C for 60 min. Immediately
cool rapidly. The solution is not more intensely coloured
than a mixture of 1 ml of red primary solution and 9 ml of
yellow primary solution (2.2.2, Method I).
Oxalic acid. Dissolve 0.80 g in 4 ml of water R. Add 3 ml of
hydrochloric acid R and 1 g of zinc R in granules. Boil for
1 min. Allow to stand for 2 min. Transfer the supernatant
liquid to a test-tube containing 0.25 ml of a 10 g/l solution
of phenylhydrazine hydrochloride R and heat to boiling.
Cool rapidly, transfer to a graduated cylinder and add an
equal volume of hydrochloric acid R and 0.25 ml of a 50 g/l
solution of potassium ferricyanide R. Shake and allow to
stand for 30 min. Any pink colour in the solution is not more
intense than that in a standard prepared at the same time in
the same manner using 4 ml of a 0.1 g/l solution of oxalic
acid R (360 ppm, calculated as anhydrous oxalic acid).
Sulphates (2.4.13). Dissolve 2.0 g in distilled water R and
dilute to 30 ml with the same solvent. The solution complies
with the limit test for sulphates (150 ppm).
Aluminium (2.4.17). If intended for use in the manufacture
of dialysis solutions, it complies with the test for aluminium.
Dissolve 20 g in 100 ml of water R and add 10 ml of acetate
buffer solution pH 6.0 R. The solution complies with the
limit test for aluminium (0.2 ppm). Use as the reference
solution a mixture of 2 ml of aluminium standard solution
(2 ppm Al) R, 10 ml of acetate buffer solution pH 6.0 R and
98 ml of water R. To prepare the blank use a mixture of 10 ml
of acetate buffer solution pH 6.0 R and 100 ml of water R.
Heavy metals (2.4.8). Dissolve 5.0 g in several portions in
39 ml of dilute sodium hydroxide solution R and dilute to
50 ml with distilled water R. 12 ml complies with limit test A
for heavy metals (10 ppm). Prepare the standard using lead
standard solution (1 ppm Pb) R.
Water (2.5.12) : 7.5 per cent to 9.0 per cent, determined on
0.500 g by the semi-micro determination of water.
Sulphated ash (2.4.14). Not more than 0.1 per cent,
determined on 1.0 g.
Bacterial endotoxins (2.6.14) : less than 0.5 IU/mg, if
intended for use in the manufacture of parenteral dosage
forms without a further appropriate procedure for the
removal of bacterial endotoxins.
ASSAY
Dissolve 0.550 g in 50 ml of water R. Titrate with 1 M sodium
hydroxide, using 0.5 ml of phenolphthalein solution R as
indicator.
1 ml of 1 M sodium hydroxide is equivalent to 64.03 mg
of C6H8O7.
STORAGE
In an airtight container.
LABELLING
The label states :
— where applicable, that the substance is free from bacterial Column :
endotoxins,
— where applicable, that the substance is intended for use
in the manufacture of dialysis solutions.
1308 See the information section on general monographs (cover pages)
EUROPEAN PHARMACOPOEIA 5.
01/2005:1609
CITRONELLA OIL
Citronellae aetheroleum
DEFINITION
Oil obtained by steam distillation from the fresh or partially
dried aerial parts of Cymbopogon winterianus Jowitt.
CHARACTERS
Pale yellow to brown-yellow liquid, with a very strong odour
of citronellal.
IDENTIFICATION
First identification : B.
Second identification : A.
A. Thin-layer chromatography (2.2.27).
Test solution. Dilute 0.1 g of citronella oil in 10.0 ml of
alcohol R.
Reference solution. Dilute 20 µl of citronellal R in
10.0 ml of alcohol R.
Plate : TLC silica gel plate R.
Mobile phase : ethyl acetate R, toluene R (10:90 V/V).
Application : 5 µl, as bands.
Development : over a path of 15 cm.
Drying : in air.
Detection : spray with anisaldehyde solution R and heat
at 100-105 °C for 10 min. Examine in ultraviolet light at
365 nm.
Result : see below the sequence of the zones present in
the chromatograms obtained with the reference and test
solutions. Furthermore, other zones are present in the
chromatogram obtained with the test solution.
Top of the plate
Citronellal : a violet zone A zone similar in colour to the
citronellal zone
An orange zone (citronellol-
geraniol)
Reference solution Test solution
B. Examine the chromatograms obtained in the test for
chromatographic profile.
Results : the characteristic peaks in the chromatogram
obtained with the test solution are similar in retention
time to those in the chromatogram obtained with the
reference solution. Neral and geranial may be absent in
the chromatogram obtained with the test solution.
TESTS
Relative density (2.2.5) : 0.881 to 0.895.
Refractive index (2.2.6) : 1.463 to 1.475.
Optical rotation (2.2.7) : − 4° to + 1.5°.
Chromatographic profile. Gas chromatography (2.2.28) :
use the normalisation procedure.
Test solution. The substance to be examined.
Reference solution. Dilute 25 µl of limonene R, 100 µl of
citronellal R, 25 µl of citronellyl acetate R, 25 µl of citral R,
25 µl of geranyl acetate R, 25 µl of citronellol R and 100 µl
of geraniol R in 5 ml of hexane R.
— material : fused silica,
— size : l = 60 m, Ø = 0.25 mm,
— stationary phase : macrogol 20 000 R (0.2 µm).
EUROPEAN PHARMACOPOEIA 5.0 Clarithromycin

Carrier gas : helium for chromatography R. DEFINITION

Flow rate : 1.0 ml/min. (3R,4S,5S,6R,7R,9R,11R,12R,13S,14R)-4-[(2,6-Dideoxy-
Split ratio : 1:100. 3-C-methyl-3-O-methyl-α-L-ribo-hexopyranosyl)oxy]-
Temperature : 14-ethyl-12,13-dihydroxy-7-methoxy-3,5,7,9,11,13-
hexamethyl-6-[[3,4,6-trideoxy-3-(dimethylamino)-β-D-
Time Temperature xylo-hexopyranosyl]oxy]oxacyclotetradecane-2,10-dione
(min) (°C) (6-O-methylerythromycin A).
Column 0-2 80 Content : 96.0 per cent to 102.0 per cent (anhydrous
2 - 26 80 → 150 substance).
26 - 42 150 → 185 CHARACTERS
42 - 49 185 → 250 Appearance : white or almost white, crystalline powder.
Solubility : practically insoluble in water, soluble in acetone
Injection port 260
and in methylene chloride, slightly soluble in methanol.
Detector 260
IDENTIFICATION
Detection : flame ionisation. Infrared absorption spectrophotometry (2.2.24).
Injection : 1 µl of the reference solution, 0.2 µl of the test Comparison : clarithromycin CRS.
solution.
Elution order : the order indicated in the composition of TESTS
the reference solution. Record the retention times of these Solution S. Dissolve 0.500 g in methylene chloride R and
substances. dilute to 50.0 ml with the same solvent.
System suitability : reference solution : Appearance of solution. Solution S is clear or not more
— resolution : minimum of 1.2 between the peaks due to opalescent than reference suspension II (2.2.1) and not
geranyl acetate and citronellol. more intensely coloured than reference solution Y7 (2.2.2,
Using the retention times determined from the chromatogram Method II).
obtained with the reference solution, locate the components Specific optical rotation (2.2.7) : − 94 to − 102 (anhydrous
of the reference solution in the chromatogram obtained with substance), determined on solution S.
the test solution. Related substances. Liquid chromatography (2.2.29).
Determine the percentage content of each of these Test solution. Dissolve 75.0 mg of the substance to be
components. examined in 25 ml of acetonitrile R1 and dilute to 50.0 ml
The percentages are within the following values : with water R.
— limonene : 1.0 per cent to 5.0 per cent, Reference solution (a). Dissolve 75.0 mg of
— citronellal : 30.0 per cent to 45.0 per cent, clarithromycin CRS in 25 ml of acetonitrile R1 and
— citronellyl acetate : 2.0 per cent to 4.0 per cent, dilute to 50.0 ml with water R.
— neral: less than 2.0 per cent, Reference solution (b). Dilute 5.0 ml of reference
— geranial : less than 2.0 per cent, solution (a) to 100.0 ml with a mixture of equal volumes of
acetonitrile R1 and water R.
— geranyl acetate : 3.0 per cent to 8.0 per cent,
Reference solution (c). Dilute 1.0 ml of reference solution (b)
— citronellol : 9.0 per cent to 15.0 per cent, to 10.0 ml with a mixture of equal volumes of acetonitrile R1
— geraniol : 20.0 per cent to 25.0 per cent. and water R.
STORAGE Reference solution (d). Dissolve 15.0 mg of clarithromycin
for peak identification CRS in 5.0 ml of acetonitrile R1 and
In a well-filled container, protected from light. dilute to 10.0 ml with water R.
Blank solution. Dilute 25.0 ml of acetonitrile R1 to 50.0 ml
01/2005:1651 with water R and mix.
corrected
Column :
— size : l = 0.10 m, Ø = 4.6 mm,
CLARITHROMYCIN
— stationary phase : octadecylsilyl silica gel for
chromatography R (3.5 µm),
Clarithromycinum — temperature : 40 °C.
Mobile phase :
— mobile phase A : a 4.76 g/l solution of potassium
dihydrogen phosphate R adjusted to pH 4.4 with dilute
phosphoric acid R or a 45 g/l solution of potassium
hydroxide R, filtered through a C18 filtration kit,
— mobile phase B : acetonitrile R1,
Time Mobile phase A Mobile phase B
(min) (per cent V/V) (per cent V/V)
0 - 32 75 → 40 25 → 60
32 - 34 40 60
34 - 36 40 → 75 60 → 25
36 - 42 75 25
C38H69NO13 Mr 748

General Notices (1) apply to all monographs and other texts 1309