Salvia miltiorrhiza root and rhizome EUROPEAN PHARMACOPOEIA 10.

ASSAY
In a 500 mL round-bottomed flask, place 30.0 g of the tincture
and add 100 mL of water R. Distil, using a descending
condenser, into a separating funnel which has been marked
beforehand at 50 mL. Stop the distillation process as soon as
the distillate reaches the 50 mL mark. Rinse the condenser
with 10 mL of pentane R. Dissolve in the distillate sufficient
sodium chloride R to produce a saturated solution. Shake
with 3 quantities, each of 20 mL, of pentane R. Dry the
combined pentane layers, including the pentane from rinsing
the condenser, over anhydrous sodium sulfate R and filter
through a plug of absorbent cotton into a weighed 100 mL
round-bottomed flask. Wash the sodium sulfate several times
with small quantities of pentane R. Remove the pentane
carefully at a temperature not exceeding 40 °C. Dry the
residue in a desiccator over diphosphorus pentoxide R and hard
paraffin at atmospheric pressure and at room temperature for
2 h. Weigh the residue (essential oil).

04/2017:2663

SALVIA MILTIORRHIZA ROOT AND

RHIZOME
Salviae miltiorrhizae radix et rhizoma
DEFINITION
Dried, whole or fragmented rhizome and root of Salvia
miltiorrhiza Bunge, collected in spring or autumn.
Content :
– salvianolic acid B (C36H30O16 ; Mr 719) : minimum 3.0 per
cent (dried drug);
– tanshinone IIA (C19H18O3 ; Mr 294.3) : minimum 0.12 per
cent (dried drug).
IDENTIFICATION
A. The rhizome is short and thick, sometimes with stem
remnants at the apex. The roots are numerous, about
10-20 cm long and 0.3-1 cm in diameter, cylindrical and
slightly curved ; some are branched, with secondary roots
and rootlets. The outer surface is reddish-brown or dark
reddish-brown, marked with longitudinal striations. The
bark of old roots comes off usually as purplish-brown
scales. The texture is hard and fragile. The fracture is soft,
fissured or slightly even and dense, with a reddish-brown
outer part and a greyish-yellow or purplish-brown wood,
showing bundles of yellowish-white vessels, arranged
radially.
Cultivars are relatively stout, about 0.5-1.5 cm in diameter.
The outer surface is brownish-red, longitudinally wrinkled.
The bark adheres closely to the wood and is difficult to
remove. The texture is compact ; the fracture is relatively
even.
B. Microscopic examination (2.8.23). The powder is
brownish-red. Examine under a microscope using chloral
hydrate solution R. The powder shows the following
diagnostic characters (Figure 2663.-1) : fragments of cork
consisting of subrectangular or polygonal cells, up to
150 μm in diameter, containing yellowish-brown pigment
(surface view [A], transverse section [E]); fragments
of parenchyma (longitudinal section [C]) consisting of
polygonal or elongated cells, often septate [Ca] ; fragments
of parenchyma (transverse section [B]) with rounded cells ;
xylem fibres usually in bundles [F], long and fusiform, with
walls showing pits shaped as crosses or oblique slits ; very
numerous reticulate or pitted vessels, 3-120 μm in diameter,
free, in bundles (longitudinal section [D], transverse
section [G]) or sometimes accompanying the fibres [Fa].
Figure 2663.-1. – Illustration for identification test B of
powdered herbal drug of salvia miltiorrhiza root and rhizome
C. Thin-layer chromatography (2.2.27).
Test solution. To 1 g of the powdered herbal drug (355)
(2.9.12) add 40 mL of methanol R. Sonicate for 15 min.
Filter. Evaporate the filtrate to 1 mL.
Reference solution. Dissolve 2 mg of salvianolic acid B R
and 2 mg of tanshinone IIA R in 1 mL of methanol R.
Plate : TLC silica gel F254 plate R (2-10 μm).
Mobile phase : methanol R, anhydrous formic acid R,
toluene R, methylene chloride R, ethyl acetate R
(5:20:20:30:40 V/V/V/V/V).
Application : 5 μL as bands of 8 mm.
Development : over a path of 6 cm.
Drying : in air.
Detection A : examine in daylight.
Results A : 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 upper third and middle part of the
chromatogram obtained with the test solution.
Top of the plate
Tanshinone II A : a prominent red A prominent red zone
zone (tanshinone IIA)
An orange zone
_______ _______
A faint brownish-green zone
Salvianolic acid B : a faint grey A faint grey zone (salvianolic
zone acid B)
_______ _______
Reference solution Test solution
Detection B : examine in ultraviolet light at 254 nm.

1610 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 10.0 Sanguisorba root

Results B : see below the sequence of zones present in Relative retention with reference to tanshinone IIA (retention
the chromatograms obtained with the reference solution time = about 33 min) : rosmarinic acid = about 0.3 ; salvianolic
and the test solution. Furthermore, other faint zones acid B = about 0.4.
may be present in the upper third and middle part of the System suitability :
chromatogram obtained with the test solution.
– resolution : minimum 5.0 between the peaks due to
Top of the plate rosmarinic acid and salvianolic acid B in the chromatogram
Tanshinone IIA : a prominent A prominent quenching zone obtained with reference solution (c) ;
quenching zone (tanshinone IIA) – symmetry factor : maximum 2.0 for the peak due to
A quenching zone salvianolic acid B in the chromatogram obtained with
_______ _______ reference solution (b).
A quenching zone
Calculate the percentage content of tanshinone IIA using the
following expression :
Salvianolic acid B : a prominent A prominent quenching zone
quenching zone (salvianolic acid B) A1 ´ m 2 ´ p1
_______ _______
A 2 ´ m1 ´ 5

A1 = area of the peak due to tanshinone IIA in the

Reference solution Test solution chromatogram obtained with the test solution ;
A2 = area of the peak due to tanshinone IIA in
TESTS the chromatogram obtained with reference
Loss on drying (2.2.32) : maximum 10.0 per cent, determined solution (a) ;
on 1.000 g of the powdered herbal drug (355) (2.9.12) by m1 = mass of the herbal drug to be examined used to
drying in an oven at 105 °C for 2 h. prepare the test solution, in grams ;
Total ash (2.4.16) : maximum 10.0 per cent. m2 = mass of tanshinone IIA CRS used to prepare
Ash insoluble in hydrochloric acid (2.8.1): maximum 3.0 per reference solution (a), in grams ;
cent. p1 = percentage content of tanshinone IIA in
ASSAY tanshinone IIA CRS.
Liquid chromatography (2.2.29). Protect the solutions from Calculate the percentage content of salvianolic acid B using
light. the following expression :
Test solution. Disperse 0.30 g of the powdered herbal drug A3 ´ m3 ´ p2 ´ 2
(355) (2.9.12) in 50.0 mL of a 70 per cent V/V solution of
methanol R. Sonicate for 1 h. Filter through a membrane filter A4 ´ m1
(nominal pore size 0.45 μm).
A3 = area of the peak due to salvianolic acid B in the
Reference solution (a). Dissolve 5.0 mg of tanshinone IIA CRS
in methanol R and dilute to 50.0 mL with the same solvent. chromatogram obtained with the test solution ;
Dilute 2.0 mL of the solution to 10.0 mL with methanol R. A4 = area of the peak due to salvianolic acid B in
Reference solution (b). Dissolve 5.0 mg of salvianolic the chromatogram obtained with reference
acid B CRS in methanol R and dilute to 25.0 mL with the same solution (b);
solvent. m1 = mass of the herbal drug to be examined used to
Reference solution (c). Dissolve 1 mg of rosmarinic acid R in prepare the test solution, in grams ;
methanol R, add 5 mL of reference solution (b) and dilute to m3 = mass of salvianolic acid B CRS used to prepare
10.0 mL with methanol R. reference solution (b), in grams ;
Column : p2 = percentage content of salvianolic acid B in
– size : l = 0.25 m, Ø = 4.6 mm ; salvianolic acid B CRS.
– stationary phase : end-capped octadecylsilyl silica gel for
chromatography R (5 μm).
Mobile phase :
– mobile phase A : 0.1 per cent V/V solution of anhydrous 04/2008:2385
formic acid R ;
– mobile phase B : acetonitrile for chromatography R ;
Time Mobile phase A Mobile phase B
(min) (per cent V/V) (per cent V/V)
0 - 10 79 → 71 21 → 29
SANGUISORBA ROOT
10 - 15 71 → 65 29 → 35
Sanguisorbae radix
15 - 25 65 → 28 35 → 72
DEFINITION
25 - 37 28 → 0 72 → 100
Whole or fragmented, dried underground parts of Sanguisorba
Flow rate : 1.0 mL/min. officinalis L. without rootlets.
Detection : spectrophotometer at 280 nm. Content : minimum 5.0 per cent of tannins, expressed as
Injection : 10 μL. pyrogallol (C6H6O3 ; Mr 126.1) (dried drug).
Identification of peaks : use the chromatogram obtained with
reference solution (a) to identify the peak due to tanshinone IIA CHARACTERS
and the chromatogram obtained with reference solution (b) to The adventitious roots are about 5-25 cm long and up to 2 cm
identify the peak due to salvianolic acid B. in diameter.

General Notices (1) apply to all monographs and other texts 1611