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Thyme oil, thymol type

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Thymol : a brownish-pink zone Carvacrol : a pale violet zone

A brownish-pink zone (thymol)

A pale violet zone (carvacrol)

A greyish-pink zone

A violet zone (cineole and linalol)

A greyish-brown zone (borneol)

A violet-blue zone

An intense violet zone

Reference solution

Test solution
D. Examine the chromatograms obtained in the assay for
thymol and carvacrol.

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 refere
solution.

TESTS

(tamed with the reterence

yaozn

Foreign matter (2.8.2): maximum 10 per cent of stems and

maximum 2 per cent of other foreign matter. Stems must not
be more than 1 mm in diameter and 15 mm in length. Leaves
with long trichomes at their base and with weakly pubescent
other parts ( Thymus serpyllum L.) are absent.

Water (2.2.13): maximum 100 mL/kg, determined on 20.0 g

of the powdered herbal drug (355) (2.9.12).
Total ash (2.4.16): maximum 15.0 per cent.

Ash insoluble in hydrochloric acid (2.8.1): maximum 3.0 per

cent.

ASSAY

Essential oil (2.8.12). Use 30.0 g of the drug, a 1000 mL

round-bottomed flask and 400 mL of water R as the distillation
liquid. Distil at a rate of 2-3 mL/min for 2 h without xylene R
in the graduated tube.

Thymol and carvacrol. Gas chromatography (2.2.28) : use

the normalisation procedure.

Test solution. Filter the essential oil obtained in the

determination of essential oil over a small amount of
anhydrous sodium sulfate R and dilute to 5.0 mL with hexane R
by rinsing the apparatus and the anhydrous sodium sulfate.

Reference solution. Dissolve 0.20 g of thymol R and 50 mg of

carvacrol R in hexane R and dilute to 5.0 mL with the same
solvent.

Column:
- material: fused silica;

- size : l = 30-60 m, 0 = 0.25 mm;

- stationary phase: macrogol 20 000 R (film thickness

0.25 pm).

Carrier gas: nitrogen for chromatography R or helium for

chromatography R.

Flow rate: 1-2 mL/min.

Split ratio: 1:100.

Temperature:

Time

(min)

Temperature

(°C)

Column

0 - 45
40 220

Injection port

190

Detector

210

Detection: flame ionisation.

Injection: 0.2 pL.

Elution order: order indicated in the composition of the

reference solution. Record the retention times of these
substances.

System suitability: reference solution:

- resolution: minimum 1.5 between the peaks due to thymol

and carvacrol.

Using the retention times determined from the chromatogram

obtained with the reference solution, locate the components
of the reference solution in the chromatogram obtained with
the test solution.

Determine the percentage content of thymol and carvacrol.

01/2012:1374

THYME OIL, THYMOL TYPE

Thymi typo thymolo aetheroleum

DEFINITION

cfi obtained by steam distillation from the fresh

lowt rin j er al Darts of Thymus vulgaris L., T. zygis L. or a
mixture ot both species.

CHARACTERS

Appearance: clear, yellow or very dark reddish-brown, mobile

liquid.

Odour reminiscent of thymol.

Solubility: miscible with anhydrous ethanol and with light

petroleum.

IDENTIFICATION
First identification: B.
Second identification: A.

A. Thin-layer chromatography (2.2.27).

Test solution. Dissolve 0.2 mL of the substance to be

examined in methylene chloride R and dilute to 10 mL with
the same solvent.

Reference solution. Dissolve 5 mg of thymol R and 10 pL of

carvacrol R in methylene chloride R and dilute to 10 mL
with the same solvent.

Plate: TLC silica gel plate R (5-40 pm) [or TLC silica gel
plate R (2-10 pm)].

Mobile phase: methylene chloride R.

Application: 10 pL [or 4 pL] as bands of 10 mm [or 8 mm].

Development: over a path of 12 cm [or 6 cm].

Drying: in air.

Detection: treat with anisaldehyde solution R and heat at

100-105 °C for 5-10 min; examine in daylight.

Results: see below the sequence of zones present in the

chromatograms obtained with the reference solution and
the test solution. Furthermore, other faint zones may
be present in the chromatogram obtained with the test
solution.

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A pink zone

Thymol : an orange-brown zone

Carvacrol : an orange-grey zone

An intense orange-brown zone

(thymol)

A faint orange -grey zone

(carvacrol) may be present

A pink zone

A violet zone

A brownish -grey zone

Reference solution

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

1405

Herbal drugs

Herbal drugs

Tolu balsam

EUROPEAN PHARMACOPOEIA 8.0

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 t