h a n g e Vi h a n g e Vi

XC e XC e
F- w F- w
PD

PD
er

er
!

!
W

W
O

O
N

N
y

y
bu

bu
to

to
k

k
lic

lic
C

C
w

w
m

m
w w
w

w
o

o
.d o .c .d o .c
c u -tr a c k c u -tr a c k

16 Drugs Containing Triterpenes

This drug group Includes Cimicifugae rhizoma (Tetracyclic triterpene glycosides,

actein, cimifugoside) and Ononidis radix (Triterpene n-onoccrin = onocol).

16.1 Preparation of Extracts

Powdered drug (1 g) is extracted with 10 ml methanol for 15 min 011 a water bath; 30 III of Cimidfugae
the filtrate is used for TLC. rhizoma

Powdered drug (l g) is extracted with chloroform for) h under reflux; 20 ul of the filtrate Ononidis
Is used for TLC of terpenes. radix
Powdered drug (I g) is extracted with 10 mlmethanol for 15 min on a water bath; 20 III is
used for TLC of flavonoid glycosides and terpenes.

16.2 Thin-Layer Chromatography

Caffeic and ferulic acid; ononin, rutin, chlorogenic acid, hyperoside as a 0.1% solution in Reference
methanol; 10 ftl is used for TLC. solutions

Silica gel 60 F",-precoated plates (Merck, Germany) Adsorbent

Ethyl formiate-toluene-formic acid (50:50: 15) -t Cimicifugae rhizorns Chromatography

Toluene-chloroform-ethanol (40:40: 10) -t Ononidis radix solvents
Ethyl acetate-glacial acetic acid-formic acid-water -t polar compounds
(100:11:11:26) ~} ononin (isoflavone)

16.3 Detection

• UV-254nm caffeic acid, its derivatives and isoflavones show

• UV-365nm
• Anisaldehyde-sulphuric
acid reagent (AS No.3)
(sec Appendix A)
quenching
caffeic acid, its derivatives and isoflavones
fluoresce blue
The sprayed TLC is heated for 6 min at 100"C;
evaluation in vis.: tritcrpenes blue-violet
(Cimicifugae rhizoma) and red to red-violet
(Ononidis radix)
 h a n g e Vi h a n g e Vi
XC e XC e
F- w F- w
PD

PD
er

er
!

!
W

W
O

O
N

N
y

y
bu

bu
to

to
k

k
lic

lic
C

C
w

w
m

m
w w
w

w
o

o
.d o .c .d o .c
c u -tr a c k c u -tr a c k

336

16.4 Drug List

Drug/plant source . Main constituents

Family/pharmacopeia

Fig. I Cimicifugae rhizoma •• Tetracyclic triterpene glyeosides:

Cimicifuga, black cohosh
Cimicifuga racemosa (L.) NUTT
(syn, Actaea racernosa L.)
Ranunculaceae
MD, Japan, China (other
Cimicifuga species)
actein (acetyl-acteol xyloside),
cimicifugoside (cimicigenol xyloside)
•• Isoflavone: formononetin
•• Caffeic and isoferulic acid
•• 15%-20% resins (cimicifugin)

Fig.2 Ononidis radix •• Triterpenes: u-onocerin (= onocol)

Restharrow root •• Isoflavones; ononin (= forrnononetin-
Ononis spinosa L. 7-glueoside), fOflllononetin-7-(6"-
Fabaceae O-maloyl)-glncoside, biochanin A-7-
OAB,MD glucoside
•• 0.02%-1 % essential oil: anethole, carven,
menthol

16.5 Formulae

Cimlfugoside R = Xylose
Cimicigenol R~H
 h a n g e Vi h a n g e Vi
XC e XC e
F- w F- w
PD

PD
er

er
!

!
W

W
O

O
N

N
y

y
bu

bu
to

to
k

k
lic

lic
C

C
w

w
m

m
w w
w

w
o

o
.d o .c .d o .c
c u -tr a c k c u -tr a c k

16 Drugs Ccntainiug Tril'etpenes 337

RD

Actein R ~ Xylose
Acetyl-acteol R~H

o-Onocerln (~Onocol)

Ononin R ~ Glucose
Formononetin R~H
 h a n g e Vi h a n g e Vi
XC e XC e
F- w F- w
PD

PD
er

er
!

!
W

W
O

O
N

N
y

y
bu

bu
to

to
k

k
lic

lic
C

C
w

w
m

m
w w
w

w
o

o
.d o .c .d o .c
c u -tr a c k c u -tr a c k

338

16.6 Chromatograms

Cimicifugae rhizoma

Drug sample j Cimicifugae rhizoma (methanolic extract, 10 ul)

Reference '1'1 caffeic acid (+ methylester of caffeic acid)
compound '1'2 formononetin
Solvent system Fig. I ethyl forrniatc-toluenc-formic acid (50:50:15)
Detection A UV-254nm
II Natural product polyethylene reagent (NST/PEG No. 28) --> UV-365nm
C Anisaldchydc-sulphuric acid reagent (AS No.3) --> vis.

j1ig.IA In UV-254 nm Cimidfugae rhizoma (l) shows two prominent quenching zones at R,
0.55-0.65 due to caffeic acid (TI) and the isoflavone formononetin (1'2). The zones at
It, 0.05-0.1 are phenol carboxylic acids.
n After treatment with the NST/PEG reagent, the main zones develop a bright blue
fluorescence in UV-365 nrn.
c Treatment with AS reagent reveals the violet zones of the trlterpene glycosides in the R,
range 0.3-0.55. The prominent zones arc due to actein and clmifugoside.

Ononidis radix

Drug sample 1 Onon idis radix (rnethanolic extract, 201'.1)

Reference 1'1 ononin
compound 1'2 rutin (R, -_.0.4) ~ chlorogenic acid (R, -~ 0.5) ~ hyperoside (R, -~ 0.6)
Solvent system Pig. 2 A ethyl acetate-glacial acetic add-formic
acid-water (100:11:11:26) system I
B+ C toluene-chloroform-ethanol (40:40:10) --> system 2
Detection A UV-254nm
B UV-365nm
C Anisaldchydc-sulphuric acid reagent (AS No.3) ----7 vis

Fig.2A Ononidis radix (I). The characteristic isoflavone fonnononetin-z-Ovglucosidc (ononinl

TJ) and its -6" -rnalonate are found as quenching zones in the R, range 0.65-0.75 in
system l , The tcrpcnes move with the solvent front.
B In the lipophilic solvent system 2 the green-blue fluorescent ononin (Tl) remains close
to the start; additional blue fluorescent zones ate found in the R, range 0.2-0.15. The
tcrpenes migrate to the lower R, range, detectable after treatment with the AS reagent.
c Treatment with AS reagent reveals five violet-red zones (R, 0.05-0.55) with onocerin as
the major terpene zone at R, -'. 0.1 (vis).
 h a n g e Vi h a n g e Vi
XC e XC e
F- w F- w
PD

PD
er

er
!

!
W

W
O

O
N

N
y

y
bu

bu
to

to
k

k
lic

lic
C

C
w

w
m

m
w w
w

w
o

o
.d o .c .d o .c
c u -tr a c k c u -tr a c k

16 Drllgs Containing Triterpenes 339

-F
Rf

-0.5

-s
T1 12 11 12 Fig. I

-F
Rf

11 12 T1 Fig. 2