Pharmacopoeia Article
Eleutherococcus senticosus is gathered in fall, thoroughly cleaned of
soil, cut into pieces, and dried rootstocks and roots of the wild
eleutherococcus shrubs of the ginseng family (Araliaceae).
Appearance Indications
Pieces of rootstocks and roots, one-piece or split lengthwise, 8 cm long, up to
4 cm thick, ligneous, solid, straight or bent, sometimes branched. Bark is thin
and tight against the wood. At the surface, rootstocks are smooth or rugulose
with axillary buds and traces of died off stalks and broken off roots.
Roots' surface is smoother with light-colored transverse eminences.
Fracture is long-stapled and light-yellow or cream in color.
At the surface, rootstocks are light-brown, while roots are darker.
The odor is delicate and aromatic. The taste is slightly pungent.
Mill Stock.
Pieces of rootstocks and roots of different shapes, light-yellow or cream in
color with brown inclusions, run through a 7mm sieve (in compliance with the
State Standard (GOST) 214-83).
Microscopy
On examination of the root transverse section under a microscope, one can see
the periderm with 5 to 7 rows of brown cork; among the large cells of phloem
parenchyma are located secretory ducts and bast fibers. The secretory ducts with
4 to 5 epithelial cells are numerous, small, about 18 µm in diameter (the
diameter of the ducts is the same along the whole length of bark), with lumens
filled with brown contents.
Bast fibers with thick lignified walls are formed in groups or singly. In the
parenchyma cells are seen numerous druses of calcium oxalate; starch grains are
found only in parenchyma cells surrounding the secretory ducts and in the cells
of medullary rays (it sets the plant apart from other members of the Araliaceae,
the starch grains of which fill all the cells of the parenchyma bark).
The wood is wide, annular-vessel, but annular location of vessels is not always
present. Tyloses are found in the vessels. Medullar rays are polystichous,
straight in wood and bent in bark. Cells of medullar rays are filled with starch
grains. Rootstock differs from root by availability of larger secretory ducts
(about 25µm in diameter) and medulla.
Qualitative Reactions
0.5 grams of mill stock (see Section: Quantitative Determination below) is put
into a 25ml cone flask, then add 10ml of hot water, heat on a water bath for 5
minutes and filter. To 1ml of extraction should be added several drops of 1%
solution of oxide chloride iron; green coloration (polyphenols) appear.
Ultraviolet spectrum of Solution B (see Section: Quantitative Determination
below) in the area of from 225nm to 325nm must has maximum absorption at the
(278±2nm) wavelength.
20ml of Solution A is put into a round-bottom flask for distillation and
evaporated to 3ml volume under vacuum on a rotary evaporator.
0.030ml of received solution is applied on the starting line of Silufol* plate
UV-254 (CSFR) of 7.5 by 15cm in size and chromatographed by means of rising
method in the system of solvents: chloroform - methyl alcohol - water (71:33:7).
When the solvents front reaches the plate end, it should be taken out of the
chamber and dried in air for 10 minutes. The plate should be looked through UV
light at 254nm wavelength. On the chromatogram appears one spot of (dark) blue
color, having Rf about 0.45 (eleutherozide B). When scanning the chromatogram
through UV light at 366nm wavelength, there must be found not less than 3 spots
with different degrees of fluorescence and Rf values of approximately 0.5; 0.6
or o.95.
* transliterated from Russian
Numerical indicators
Undivided stock.
Sums of eleutherozides in terms of eleutherozide B is not less than 0.30%;
moisture content: not more than 14%; total ash: not more than 8%; stalk remains,
including those separated during analysis: not more than 1.5%; rootstocks and
roots got brown at fracture: not more than 3%; organic admixtures: not more than
1%; and mineral admixtures: not more than 1%.
Mill stock.
Sums of eleutherozides in terms of eleutherozide B: not less than 0.30%;
moisture content: not more than 14%; total ash: not more than 8%; particles that
cannot pass through the 7mm sieve according to State Standard (GOST)
214-83: not more than 10%; particles passed through the 0.5mm sieve according to
Specifications 14-4-1374-86: not more than 20%; organic admixtures: not more
than 1%; mineral admixtures: not more than 1%.
Quantitative determination
An analytical sample of stock should be pulverized down to the particle size
that passes through the 2mm sieve according to State Standard (GOST) 214-83.
About 1 gram (precise weight) of pulverized stock should be put into a 100ml
cone flask to carry out fractional extraction sequentially by 20ml two times by
70% ethyl alcohol and two times by 95% alcohol. Each extraction should be done
in a magnetic mixer heated at <50°C for 1 hour. The extract should be
filtered through a paper filter into a 100ml round-bottom flask for
distillation. Alcohol should be distilled to dryness under vacuum in a rotor
evaporator.
To the remains in the flask should be added 10ml of water and 10ml of carbon
tetrachloride. The flask contents should be thoroughly mixed and quantitatively
transferred to a 100ml separating funnel. The flask should be rinsed twice by
carbon tetrachloride by 5ml and rinse content should be added to the separating
funnel contents. Then into the flask should be added 10ml of the mixture:
chloroform and 95% alcohol (5:1) and then left for 10 minutes.
In the separating funnel, the water phase should be rinsed by thrice extraction
by carbon tetrachloride by 10ml.
After separation of carbon tetrachloride, to the separating funnel contents
should be added 20ml (including 10ml from the distillation flask) of the
mixture: chloroform and 95% alcohol (5:1) and then extract eleutherozides for 5
minutes. The lower layer should be filtered through a paper filter with 2 grams
of waterless sodium sulfate into a 100ml measuring flask.
Extraction of eleutherozides in the separating flask should be repeated four
times more sequentially 15ml, 15ml, 10ml, and 10ml of the same mixture,
collecting them into the same flask. Flask solution volume should be brought to
the mark with the mixture: chloroform and 95% alcohol (5:1, Solution A).
20ml of Solution A should be put into a 50ml measuring flask and the solution
volume should be brought to the mark with the mixture: chloroform and 95%
alcohol (5:1, Solution B). Optical density of Solution B should be measured by a
spectrometer with 278nm wavelength in a cuvette with a layer 10mm thick.
The mixture: chloroform and 95% alcohol (5:1) should be used as a reference
solution.
The content of the sum of eleutherozides in the terms of eleutherozide B and
absolutely dry stock in percentage terms (X) should be computed from the
formula:
Packing
In accordance with State Standard (GOST) 6077-80, solid stock should be
packed in bags according to GOST 18225-72 or GOST 19317-73 not
more than 30kg net weight or in fabric bales according to GOST 19298-73
not more than 40kg net weight.
Mill stock should be packed in bags in accordance with GOST 18225-72 or GOST
19317-73 not more than 30kg net weight.
Marking
In accordance with GOST 6077-80 and GOST 14192-77.
Transportation
In accordance with GOST 6077-80.
Storage
In accordance with GOST 6077-80.
Shelf life:
3 years.
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