Goldenrod
cartridge. Wash the cartridge with 20 mL of water R followed
by 15 mL of a 30 per cent V/V solution of methanol R. Discard
the eluates after confirming that no ginsenosides are present,
otherwise repeat the preparation of the solution with another
brand of cartridge where no ginsenosides are eluted with a
30 per cent V/V solution of methanol R. Elute the cartridge
with 20 mL of methanol R ; collect the eluate. Under reduced
pressure, evaporate the eluate to dryness. Dissolve the residue
in 2.0 mL of methanol R. Filter through a suitable membrane
filter (nominal pore size 0.45 μm).
Reference solution (b). Dissolve 3.0 mg of ginsenoside Rb1 CRS
in methanol R and dilute to 5.0 mL with the same solvent. p = percentage content of ginsenoside Rb1 in
Reference solution (c). Dissolve 3.0 mg of ginsenoside Rg2 R in
methanol R and dilute to 5.0 mL with the same solvent.
Reference solution (d). Dilute 1.0 mL of reference solution (b)
to 2.0 mL with reference solution (c).
Column :
– size : l = 0.125 m, Ø = 4.6 mm ;
– stationary phase : octadecylsilyl silica gel for
chromatography R (5 μm) ;
– temperature : 35 °C.
Mobile phase :
– mobile phase A : water R adjusted to pH 2 with phosphoric
acid R ;
– mobile phase B : acetonitrile R1 ;
Time Mobile phase A Mobile phase B
(min) (per cent V/V) (per cent V/V)
0-8 80 20
8 - 40 80 → 60 20 → 40
40 - 45 60 → 40 40 → 60
45 - 47 40 → 0 60 → 100
Flow rate : 1.0 mL/min.
Detection : spectrophotometer at 203 nm.
Injection : 20 μL.
Elution order : ginsenoside Rg1, ginsenoside Re,
ginsenoside Rf, ginsenoside Rb1, ginsenoside Rg2,
ginsenoside Rc, ginsenoside Rb2, ginsenoside Rd ; depending
on the operating conditions and the state of the column,
ginsenoside Rb1 may elute before or after ginsenoside Rg2.
Identification of peaks : use the chromatogram supplied
with ginseng dry extract HRS and the chromatogram
obtained with reference solution (a) to identify the peaks
due to ginsenosides Rg1, Re, Rf, Rc, Rb2 and Rd ; use the
chromatogram obtained with reference solution (b) to identify
the peak due to ginsenoside Rb1 ; use the chromatogram
obtained with reference solution (c) to identify the peak due
to ginsenoside Rg2.
Relative retention with reference to ginsenoside Rb1 (retention
time = about 33 min) : ginsenoside Rg1 = about 0.53 ;
ginsenoside Re = about 0.54 ; ginsenoside Rf = about 0.88 ;
ginsenoside Rg2 = about 0.98 ; ginsenoside Rc = about 1.04 ;
ginsenoside Rb2 = about 1.08 ; ginsenoside Rd = about 1.17.
System suitability : reference solution (d) :
– resolution : minimum 1.5 between the peaks due to
ginsenosides Rg2 and Rb1.
Calculate the percentage content of the sum of
ginsenosides Rb1, Rb2, Rc, Rd, Re, Rf, Rg1 and Rg2, expressed
as ginsenoside Rb1, using the following expression :
A1 ´ m 2 ´ p ´ 0.8
A 2 ´ m1
1458
EUROPEAN PHARMACOPOEIA 10.
A1 = sum of the areas of the peaks due to ginsenosides
Rb1, Rb2, Rc, Rd, Re, Rf, Rg1 and Rg2 in the
chromatogram obtained with the test solution ;
A2 = area of the peak due to ginsenoside Rb1 in
the chromatogram obtained with reference
solution (b);
m1 = mass of the extract to be examined used to prepare
the test solution, in grams ;
m2 = mass of ginsenoside Rb1 CRS used to prepare
reference solution (b), in grams ;
ginsenoside Rb1 CRS.
04/2014:1892
GOLDENROD
Solidaginis herba
DEFINITION
Whole or cut, dried, flowering aerial parts of Solidago gigantea
Aiton or Solidago canadensis L., their varieties or hybrids
and/or mixtures of these.
Content : minimum 2.5 per cent of flavonoids, expressed as
hyperoside (C21H20O12 ; Mr 464.4) (dried drug).
IDENTIFICATION
A. The stems are greenish-yellow or greenish-brown, partly
tinted reddish, roundish, more or less conspicuously
grooved, glabrous and smooth in the lower part, slightly or
densely pubescent in the upper part. They are solid with a
whitish pith.
The leaves are green, sessile, lanceolate, with a serrate
margin, 8-12 cm long and about 1-3 cm wide, the upper
surface is green and more or less glabrous, the lower
surface is greyish-green and pubescent, especially on the
veins. The inflorescence consists of a number of unilateral,
curved racemes which together form a pyramidal panicle
at the end of the stems.
Each capitulum has an involucre composed of
linear-lanceolate, imbricated yellowish-green bracts,
surrounding a single row of yellow ligulate florets about
the same length as the involucre ; yellow, radially arranged
tubular florets, as long as, or longer, than the ligulate
florets ; a brownish inferior ovary surmounted by a white
pappus of silky hairs.
B. Microscopic examination (2.8.23). The powder is green
or greyish-green. Examine under a microscope using
chloral hydrate solution R. The powder shows the following
diagnostic characters (Figure 1892.-1) : pappus bristles,
usually broken, consisting of multiseriate trichomes
composed of elongated cells with the tips free from
the surface and forming pointed projections over the
entire length [J] ; fragments of the mesophyll [F] with
vascular bundles [Fa], oil glands [Fb] and palisade
parenchyma [Fc] ; fragments of the leaf epidermises [E, H]
with sinuous or wavy-walled cells accompanied by palisade
parenchyma [Ea] (upper epidermis), with whip-like,
multicellular covering trichomes [Eb] and anomocytic
stomata (mainly on the lower epidermis) (2.8.3) [Ha] ;
multicellular uniseriate covering trichomes from the stems
and the margins of the leaves with up to 5 or 6 thick-walled
cells [C] ; in Solidago canadensis L., covering trichomes
with a terminal cell that may be bent at a right angle [G] ;
fragments of the style [A] with short papillae [Aa] and long,
See the information section on general monographs (cover pages)
EUROPEAN PHARMACOPOEIA 10.0
slender papillae with wrinkled walls [Ab] ; pollen grains
with 3 germinal pores and a spiny exine [B] ; a few rare
fragments of the ovary with twin covering trichomes [D].
Figure 1892.-1. – Illustration for identification test B of
powdered herbal drug of goldenrod
C. Thin-layer chromatography (2.2.27).
Test solution. To 0.75 g of the powdered herbal drug (355)
(2.9.12) add 5 mL of methanol R and boil in a water-bath
under a reflux condenser for 10 min. Cool and filter.
Reference solution. Dissolve 1.0 mg of chlorogenic acid R,
2.5 mg of quercitrin R and 2.5 mg of rutoside trihydrate R
in 10 mL of methanol R.
Plate : TLC silica gel plate R.
Mobile phase : anhydrous formic acid R, water R, methyl
ethyl ketone R, ethyl acetate R (6:6:18:30 V/V/V/V).
Application : 20 μL of the test solution and 10 μL of the
reference solution, as bands.
Development : over a path of 10 cm.
Drying : at 100-105 °C.
Detection : treat with a 10 g/L solution of diphenylboric acid
aminoethyl ester R in methanol R and then with a 50 g/L
solution of macrogol 400 R in methanol R. Allow to stand
for 30 min. Examine in ultraviolet light at 365 nm.
Results : see below the sequence of zones present in the
chromatograms obtained with the reference solution and
the test solution. Furthermore, other zones may be present
in the chromatogram obtained with the test solution.
General Notices (1) apply to all monographs and other texts
Goldenrod
Top of the plate
A bluish-green fluorescent zone
Quercitrin : a yellowish-brown A faint to intense yellowish-brown
fluorescent zone fluorescent zone (quercitrin)
_______ _______
A more or less intense
yellowish-brown zone
Chlorogenic acid : a light blue A light blue fluorescent zone
fluorescent zone (chlorogenic acid) and/or a
yellow fluorescent zone
Rutoside : an orange fluorescent A faint to intense yellowish-brown
zone fluorescent zone (rutoside)
_______ _______
Reference solution Test solution
TESTS
Foreign matter (2.8.2) : maximum 5 per cent of brownish
parts and maximum 2 per cent of other foreign matter.
Loss on drying (2.2.32): maximum 10 per cent, determined
on 0.500 g of the powdered herbal drug (355) (2.9.12) by
drying in an oven at 105 °C for 2 h.
Total ash (2.4.16) : maximum 7.0 per cent.
Ash insoluble in hydrochloric acid (2.8.1) : maximum 1.0 per
cent.
ASSAY
Stock solution. In a 100 mL round-bottomed flask, introduce
0.200 g of the powdered herbal drug (250) (2.9.12), add 1 mL
of a 5 g/L solution of hexamethylenetetramine R, 20 mL of
acetone R and 2 mL of hydrochloric acid R1. Boil the mixture
under a reflux condenser for 30 min. Filter the liquid through
a small plug of absorbent cotton into a 100 mL flask. Add the
absorbent cotton to the residue in the round-bottomed flask,
extract with 2 quantities, each of 20 mL of acetone R, each time
boiling under a reflux condenser for 10 min. Allow to cool.
Filter the combined acetone extracts through a filter paper
into a volumetric flask. Rinse the flask and the filter paper
and dilute to 100.0 mL with acetone R. Introduce 20.0 mL of
the solution into a separating funnel, add 20 mL of water R
and shake the mixture with 1 quantity of 15 mL and then with
3 quantities, each of 10 mL, of ethyl acetate R. Combine the
ethyl acetate extracts in a separating funnel, wash twice with
50 mL of water R and filter the extracts over 10 g of anhydrous
sodium sulfate R into a volumetric flask. Dilute to 50.0 mL
with ethyl acetate R, rinsing the separating funnel and the
sodium sulfate.
Test solution. To 10.0 mL of the stock solution add 1.0 mL of
aluminium chloride reagent R and dilute to 25.0 mL with a
5 per cent V/V solution of glacial acetic acid R in methanol R.
Compensation solution. Dilute 10.0 mL of the stock solution
to 25.0 mL with a 5 per cent V/V solution of glacial acetic
acid R in methanol R.
Measure the absorbance of the test solution (2.2.25) at 425 nm
after 30 min by comparison with the compensation solution.
Calculate the percentage content of flavonoids, expressed as
hyperoside, using the following expression :
A ´ 1.25
m
i.e. taking the value of the specific absorbance of hyperoside
to be 500.
A = absorbance measured at 425 nm ;
m
= mass of the herbal drug to be examined, in grams.
1459
Goldenrod, European EUROPEAN PHARMACOPOEIA 10.0

01/2013:1893 fibres [Lb] ; fragments of the epidermis of the petals with a

striated cuticle, through which run fine spiral vessels [S],
and bearing biseriate glandular trichomes (side view [P]) ;
spherical pollen grains, with 3 germinal pores and a spiny
exine [J] ; abundant pappus hairs and their fragments [C,
D], multiseriate with the marginal cells overlapping
GOLDENROD, EUROPEAN outwards ; fragments of parenchyma [Q], some showing
cells containing small, isolated cluster crystals of calcium
oxalate [Qa] ; fragments of bracts [R] with a finely striated
Solidaginis virgaureae herba cuticle, polygonal cells [Ra], bearing pennant-like covering
trichomes [Rb] and whose margin bears uniseriate,
multicellular covering trichomes [Rc].
DEFINITION
C. Thin-layer chromatography (2.2.27) as described in the test
Whole or fragmented, dried, flowering aerial parts of Solidago for Solidago gigantea Aiton and Solidago canadensis L.
virgaurea L.
Results : see below the sequence of the zones present in the
Content : minimum 0.5 per cent and maximum 1.5 per cent chromatograms obtained with the reference solution and
of flavonoids, expressed as hyperoside (C21H20O12 ; Mr 464.4) the test solution. Furthermore, other fluorescent zones
(dried drug). may be present in the chromatogram obtained with the
test solution.
IDENTIFICATION
Top of the plate
A. The stem is cylindrical, striated, the lower part often
A light blue fluorescent zone
reddish-violet, sometimes entirely glabrous or pubescent
with short, bent, apically directed hairs. The basal leaves Quercitrin : an orange fluorescent
are obovate or oblanceolate, with a serrate margin, and zone
_______ _______
taper at the base into a long, winged petiole ; the cauline
leaves are alternate, smaller than the basal leaves and more Chlorogenic acid : a light blue A light blue fluorescent zone
elliptical in outline, with an entire or slightly toothed fluorescent zone (chlorogenic acid)
margin ; they are sessile or with only a short petiole. Both Rutoside : an orange fluorescent An orange fluorescent zone
surfaces of the leaves are glabrous or only slightly pubescent zone (rutoside)
with a prominent reticulate venation on the lower surface. _______ _______
The capitula form a tightly packed panicle. At the base of
the pedicels there are 2 small, linear bracts with scarious Reference solution Test solution
margins. The involucre consists of 2-4 rows of loosely
arranged, imbricate bracts, each bract greenish-yellow with
a smooth and shiny inner surface, the outer surface hairy or
glabrous, with a scarious margin. Each capitulum contains
6-12 widely separated female ray florets, about twice as
long as the bracts, and about 10-30 hermaphrodite, tubular
florets. All florets are yellow. The brown, inferior ovary
tapers towards the base and has a ribbed surface, covered
with scattered hairs ; it is surmounted by a whitish pappus
composed of smooth or rough, bristly hairs.
B. Microscopic examination (2.8.23). The powder is light
green. Examine under a microscope using chloral hydrate
solution R. The powder shows the following diagnostic
characters (Figures 1893.-1 and 1893.-2) : fragments of
the upper epidermis of the leaf (surface view [B, H, M]),
covered by a distinctly striated cuticle, composed of
polygonal cells with straight, beaded, thickened walls [Ba,
Ma], uniseriate, multicellular covering trichomes [Ha]
or rounded, thick-walled, covering trichome scars with a
pitted lumen [Mb], and a few anomocytic stomata (2.8.3)
[Bb] sometimes accompanied by underlying palisade
parenchyma [Bc] ; fragments of the lower epidermis of
the leaf (surface view [A, K, N]) covered by a slightly
striated cuticle composed of cells with sinuous walls in
the area of the lamina [Aa] or with more rigid walls near
the veins [N], numerous anomocytic stomata (2.8.3) [Ab],
occasional glandular trichomes with a unicellular stalk and
a unicellular head [Ka, Na], covering trichomes some of
which are pennant-like [Ac, F], uniseriate, multicellular,
with 1-3 thin-walled basal cells [Fa], a flagella-like distal
cell [Fb], and an enlarged, more or less rounded cell [Fc]
between them, others are uniseriate, multicellular (up
to about 10 cells), with thick, finely wrinkled walls and
a rigid conical distal cell (side view [E]) ; rare fragments
from the ovary [G] bearing paired, covering trichomes
with a distinctly pitted central wall and a bifid apex
(surface view [Ga], side view [Gb]) ; vascular tissue from Figure 1893.-1. – Illustration for identification test B of
the stems [L] composed of vessels [La] and groups of powdered herbal drug of European goldenrod

1460 See the information section on general monographs (cover pages)