Valerian root, cut EUROPEAN PHARMACOPOEIA 11.

m1 = mass of the herbal drug to be examined used to

prepare the test solution, in grams ;
m2 = mass of valerian dry extract HRS used to prepare
the reference solution, in grams ;
p = percentage content of valerenic acid in valerian dry
extract HRS.

04/2017:2526

VALERIAN ROOT, CUT

Valerianae radix minutata

DEFINITION

Dried, cut underground parts of Valeriana officinalis L. s.l., Figure 2526.-1. – Illustration for identification test A of
including the rhizome, roots and stolons. powdered herbal drug of cut valerian root
It is produced from Valerian root (0453) for the purpose of B. Thin-layer chromatography (2.2.27).
being used in herbal teas.
Test solution. Suspend 1 g of the powdered herbal
drug (355) (2.9.12) in 10 mL of methanol R and sonicate for
Content : 10 min. Filter the supernatant through a membrane filter
(nominal pore size 0.45 μm). Use the filtrate.
– essential oil : minimum 3 mL/kg (dried drug) ;
Reference solution. Dissolve 5 mg of acetoxyvalerenic acid R
– sesquiterpenic acids : minimum 0.10 per cent m/m expressed and 5 mg of valerenic acid R in 20 mL of methanol R.
as valerenic acid (C15H22O2 ; Mr 234.3) (dried drug).
Plate : TLC silica gel plate R (5-40 μm) [or TLC silica gel
plate R (2-10 μm)].
Mobile phase : glacial acetic acid R, ethyl acetate R,
IDENTIFICATION cyclohexane R (2:38:60 V/V/V).
A. Microscopic examination (2.8.23). The powder is pale Application : 20 μL [or 5 μL] as bands of 10 mm [or 8 mm].
yellowish-grey or pale greyish-brown. Examine under a Development : over a path of 10 cm [or 6 cm].
microscope using chloral hydrate solution R. The powder
shows the following diagnostic characters (Figure 2526.-1) : Drying : in air.
occasional groups of rectangular sclereids with moderately
thickened walls and a large lumen, from the stem base [H] ; Detection : treat with anisaldehyde solution R and heat at
very numerous fragments of parenchyma with large ovoid 100-105 °C for 5-10 min ; examine in daylight.
cells (longitudinal section [K], transverse section [J]) ; Results : see below the sequence of zones present in the
spiral, reticulate or pitted lignified vessels, isolated or in chromatograms obtained with the reference solution and
small groups [D, G] ; thin-walled, elongated cells of the the test solution. Furthermore, other violet zones may
piliferous layer (surface view [A], transverse section [B]), be present in the chromatogram obtained with the test
some with root hairs [Aa, Ba] or their scars [Ab] ; the solution.
piliferous layer is usually accompanied by an underlying
layer of cells with slightly thickened and elongated Top of the plate
walls [Ac, Bb] ; fragments of dermal tissue from the _______ _______
rhizome composed of 1 or 2 layers of polygonal cells with
irregularly thickened walls [F] ; a few groups of sclereids Valerenic acid : a violet zone A violet zone (valerenic acid)
with thick walls and a narrow lumen [E] from the pith
Acetoxyvalerenic acid : a violet A violet zone (acetoxyvalerenic
of the rhizome. Examine under a microscope using a zone acid)
50 per cent V/V solution of glycerol R. The powder shows _______ _______
numerous starch granules, simple or 2- to 6-compound,
but frequently separated, rounded or irregular and up to 2 faint or very faint violet zones
about 15 μm in diameter ; most of the granules show a Reference solution Test solution
rather indistinct cleft or radiate hilum [C].

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EUROPEAN PHARMACOPOEIA 11.0 Valerian tincture

TESTS A1 = area of the peak due to acetoxyvalerenic acid in the

Foreign matter (2.8.2) : maximum 5 per cent of stem bases chromatogram obtained with the test solution ;
and maximum 2 per cent of other foreign matter, determined A2 = area of the peak due to valerenic acid in the
on the herbal drug prior to cutting. chromatogram obtained with the test solution ;
Loss on drying (2.2.32) : maximum 12.0 per cent, determined A3 = area of the peak due to valerenic acid in the
on 1.000 g of well-homogenised powdered herbal drug (355) chromatogram obtained with the reference
(2.9.12) by drying in an oven at 105 °C for 2 h. solution ;
Total ash (2.4.16) : maximum 12.0 per cent. m1 = mass of the herbal drug to be examined used to
prepare the test solution, in grams ;
Ash insoluble in hydrochloric acid (2.8.1): maximum 5.0 per
cent. m2 = mass of valerian dry extract HRS used to prepare
the reference solution, in grams ;
ASSAY p = percentage content of valerenic acid in valerian dry
Essential oil (2.8.12). Use 40.0 g of freshly powdered herbal extract HRS.
drug (500) (2.9.12), a 2000 mL flask, 500 mL of water R as the
distillation liquid and 0.50 mL of 1,2,4-trimethylbenzene R in
the graduated tube. Distil at a rate of 3-4 mL/min for 4 h. 07/2010:1899
Sesquiterpenic acids. Liquid chromatography (2.2.29).
Test solution. Place 1.50 g of the powdered herbal drug
(710) (2.9.12) in a 100 mL round-bottomed flask with a
ground-glass neck. Add 20 mL of methanol R1. Mix and heat
on a water-bath under a reflux condenser for 30 min. Allow to VALERIAN TINCTURE
cool and filter. Place the filter with the residue in the 100 mL
round-bottomed flask. Add 20 mL of methanol R1 and heat
on a water-bath under the reflux condenser for 15 min. Allow Valerianae tinctura
to cool and filter. Combine the filtrates and dilute to 50.0 mL DEFINITION
with methanol R1, rinsing the round-bottomed flask and the
filter. Tincture produced from Valerian root (0453).
Reference solution. Dissolve an amount of valerian dry Content : minimum 0.015 per cent m/m of sesquiterpenic
extract HRS corresponding to 1.0 mg of valerenic acid in acids, expressed as valerenic acid (C15H22O2 ; Mr 234.3).
methanol R1 and dilute to 10.0 mL with the same solvent. PRODUCTION
Sonicate for 10 min and filter through a membrane filter
(nominal pore size 0.45 μm). The tincture is produced from 1 part of the drug and 5 parts of
ethanol (60 to 80 per cent V/V) by an appropriate procedure.
Column :
CHARACTERS
– size : l = 0.25 m, Ø = 4.6 mm ;
Appearance : brown liquid.
– stationary phase : octadecylsilyl silica gel for
chromatography R (5 μm). IDENTIFICATION
Mobile phase : Thin-layer chromatography (2.2.27).
– mobile phase A : acetonitrile R1, 5 g/L solution of phosphoric Test solution. Dilute 5 mL of the tincture to be examined with
acid R (20:80 V/V) ; 5 mL of ethanol (70 per cent V/V) R.
– mobile phase B : 5 g/L solution of phosphoric acid R, Reference solution. Dissolve 5 mg of acetoxyvalerenic acid R
acetonitrile R1 (20:80 V/V); and 5 mg of valerenic acid R in 20 mL of methanol R.
Plate : TLC silica gel plate R (5-40 μm) [or TLC silica gel plate R
Time Mobile phase A Mobile phase B (2-10 μm)].
(min) (per cent V/V) (per cent V/V)
Mobile phase : glacial acetic acid R, ethyl acetate R,
0-5 55 45 cyclohexane R (2:38:60 V/V/V).
5 - 18 55 → 20 45 → 80 Application : 20 μL [or 5 μL] as bands of 10 mm [or 8 mm].
18 - 22 20 80 Development : over a path of 10 cm [or 6 cm].
Drying : in air.
Flow rate : 1.5 mL/min. Detection : spray with anisaldehyde solution R and heat at
Detection : spectrophotometer at 220 nm. 100-105 °C for 5-10 min ; examine in daylight.
Injection : 20 μL. Results : see below the sequence of zones present in the
chromatograms obtained with the reference solution and the
Peak identification : use the chromatogram supplied with test solution. Furthermore, other violet zones may be present
valerian dry extract HRS and the chromatogram obtained in the chromatogram obtained with the test solution.
with the reference solution to identify the peaks due to
acetoxyvalerenic acid and valerenic acid. Top of the plate
_______ _______
System suitability : reference solution :
– relative retention with reference to valerenic acid (retention Valerenic acid : a violet zone A violet zone (valerenic acid)
time = about 19 min) : acetoxyvalerenic acid = about 0.5. Acetoxyvalerenic acid : a violet zone A violet zone (acetoxyvalerenic
Calculate the percentage content of sesquiterpenic acids, acid)
_______ _______
expressed as valerenic acid, using the following expression :
2 faint or very faint violet zones
( A1 + A 2 ) ´ m 2 ´ p ´ 5
Reference solution Test solution
A3 ´ m1

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