Challenges in the development of quality control monographs for Chinese herbs

Challenges in the development of quality Challenges in the development of quality control monographs for Chinese herbscontrol monographs for Chinese herbs

Univ.Univ.--Prof. Dr. Rudolf BauerProf. Dr. Rudolf Bauer

Ins t i tu t e of Pharmaceu t i c a l

Sc iencesKar l-Franzens-Univers i t y

Graz

AOAC Europe International Workshop, 10.-12. October

2011, Erlangen -

Nuremberg

QualityQuality

ControlControl

of of BotanicalsBotanicals, TCM, , TCM, HerbalHerbal

Food Food Supplements and Supplements and HerbalHerbal

Medicinal ProductsMedicinal Products

••

China China exportsexports

200,000 200,000 tonstons

rawraw

herbsherbs

annuallyannually••

outputoutput

valuevalue

in 2005: ca. in 2005: ca. €€

11 11 billionbillion

••

expectedexpected

total TCM total TCM marketmarket

in China in 2015: in China in 2015: €€

18.8 18.8 billionbillion

http://www.hkc22.com/ChineseMedicine.html

Pharmaceutical quality is the basis for reproducible efficacy and safety

Pharmaceutical quality is the basis Pharmaceutical quality is the basis for reproducible efficacy and safetyfor reproducible efficacy and safety

Quality

Effic

acy

Safe

ty

RationalRational UseUse

Quality control is a mustQuality control is a mustQuality control is a must

Development

of modern monographs

for identification

and quality

control

of TCM herbs

DevelopmentDevelopment of modern of modern

monographsmonographs for for identificationidentification

and and qualityquality controlcontrol

of TCM of TCM herbsherbs

A4-0075/97

8. Incorporating non-conventional medical remedies to the European PharmacopoeiaThe European Pharmacopoeia, as drawn up by the Council of Europe, needs to be opened up to other pharmacopoeiae particularly the medicinal plants used in Chinese medicine.

Actual

Status of Work

of theTCM-Working

Party

Ca. 75

Monographs on the

workplan

Working

Party for the

Elaboration

of Monographs of Chinese Herbs for the

European Pharmacopoeia

WorkingWorking

Party for Party for thethe

EElaborationlaboration

of Monographs of of Monographs of Chinese Herbs for Chinese Herbs for thethe

European European PharmacopoeiaPharmacopoeia

Chairman Prof. Dr. Gerhard FRANZ (D)Specialists:A

Prof. Dr. Rudolf BAUER

A Mag. Erich STOEGER

A Doz. Dr. Reinhard LAENGERB Prof. Dr. Pierre DUEZCH

Dr. Albert BLARER

CH

Dr. Thomas LEHMANNCH

Dr. Eike REICH

D Dr. Uwe Michael GASSER

D Dr. Kim WUTHOLD

F

Prof. Isabelle FOURASTEF

Mr. Robert SOUSSAIN

I

Prof. Dr. Anna Rita BiliaTR

Prof. Dr. Kemal Husnü

Can BASER

NL Dr. Mei WANGPL Prof. Dr. Kazimierz GLOWNIAKUK Mr. M. WahleyAUS Prof. Kelvin CHAN (observer

status)

Members

and Observers of the European Pharmacopoeia

MembersMembers and Observers of and Observers of thethe

European European PharmacopoeiaPharmacopoeia

36 Member

States

+ EU22 Observer States

+ WHO

Aspects

of pharmaceutical

quality control

of herbal

medicinal

products

AspectsAspects of of pharmaceuticalpharmaceutical

qualityquality controlcontrol

of of herbalherbal medicinalmedicinal

productsproducts•

Clear

botanical

definition

•

Tests for identity•

Tests for purity–

Adulterations

–

Foreign

materials•

Heavy metals

•

Pesticides•

Mycotoxins

•

Fumigation agents–

Microbiological

contamination

–

Residual solvents•

Assay

for constituents of known therapeutic

activity or for (active) markers

•

Identification

/ Lack of authentic reference

material (HRS, CRS)

•

Insufficient

knowledge

on impact

of processing

•

Contaminants

and toxic constituents

•

Decision

on relevant marker compounds

for the

assay

Major challenges

in the

quality

control

of Chinese herbs

Major Major challengeschallenges

in in thethe

qualityquality

controlcontrol

of of Chinese Chinese herbsherbs

©2010 EDQM, Council of Europe, All rights reserved

Problems Problems withwith nomenclaturenomenclature

•

Many products have more than one Pinyin name e.g.:

Epimedii

Herba: yinyanghuo 淫羊藿, xianlingpi

仙灵脾

•

The same Pinyin name is assigned for different

species

e.g.:

Clematis armandii

小木通 -

xiao

mutong

Akebia

quinata

五叶木通 -

wuye

mutong

Akebia

trifoliata

三叶木通 -

sanye

mutong

•

Similarity between Chinese names e.g. Guan mutong

关木通 / Chuan

mutong

川木通

Hsu, H.Y., Oriental Healing Arts Int. Bull. 12, 387-408 (1987)

Missing Missing harmonizationharmonization

of of nomenclaturenomenclature in China and in China and neighboringneighboring

countriescountries

China Mainland

Taiwan

Hong Kong

Korea

Japan

Pugongying Taraxacum

officinale

T. formosum

Elephantopus

scaber

T. platycarpum

T. japonicum(Taraxaci

herba)

Lactuca

chinensis

(Compositae)

T. platycarpum(Compositae)

T. officinale

Bianxu Polygonum

aviculare

Euphorbia

thymifolia

Belamcanda

chinensis

P. aviculare(Polygoni

herba)

(Polygonaceae)

(Euphorbiaceae)

(Iridaceae)

Xixiancao S. orientalis

Anisomeles

ovata

Anisomeles

indica

S. glabrescens

S. pubescens(Siegesbeckiae

S. pubescens

(Labiatae)

(Compositae)herba)

S. glabrescens

Duhuo Angelica pubescens

A. pubescens

A. pubescens

Aralia

cont. Aralia

cordata(Ang. pub. radix)

(Umbelliferae)

(Araliaceae)

Guanzhong Dryopteris

Sphaeropteris

Blechnum

orientale

Dryopt. crass.

Cyrtomium(Dryopt. crass.

crassirhizoma

lepifera

(Polypodiaceae)

fortuneirhiz.)

(Aspidiaceae)

(Cyatheaceae)

(Polypodiaceae)

Different Different latinlatin namesnames in in useusePinyinPinyin Chinese Chinese PharmacopoeiaPharmacopoeia PorkertPorkertDuhuoDuhuo AngelicaeAngelicae pubescentispubescentis RadixRadix HeracleiHeraclei Radix Radix NiubangziNiubangzi ArctiiArctii FructusFructus BardanaeBardanae FructusFructusChuanxiongChuanxiong ChuanxiongChuanxiong RhizomaRhizoma LigusticiLigustici RhizomaRhizomaShanyaoShanyao DioscoreaeDioscoreae RadixRadix BatatatisBatatatis RhizomaRhizomaBaizirenBaiziren PlatycladiPlatycladi SemenSemen BiotaeBiotae SemenSemenHuoxiangHuoxiang PogostemonisPogostemonis HerbaHerba AgastachidisAgastachidis HerbaHerbaFangfengFangfeng SaposhnikoviaeSaposhnikoviae RadixRadix LedebouriellaeLedebouriellae RadixRadix

etc.etc.

HPLC Fingerpr in t

Analys is of Angel i c ae

pubescens

rad i x

獨活) and Adul teran ts

HPLC HPLC Fingerpr in tFingerpr in t

Analys is of Analys is of Angel i c aeAngel i c ae

pubescenspubescens

rad i xrad i x

獨活獨活) and ) and Adul teran tsAdul teran ts

Angelica Angelica dahuricadahurica

Angelica Angelica apaensisapaensis

HeracleumHeracleum moellendorffiimoellendorffii

10 20 30Time (min)

020

0m

AU

10 20 30Time (min)

020

040

0m

AU

10 20 30Time (min)

020

040

0m

AU11 0

000

Angelica Angelica pubescenspubescens

10 20 30Time (min)

020

040

0m

AU

10 20 30Time (min)

020

040

0m

AU

10 20 30Time (min)

0

200

400

0

20

0 4

00

mA

Um

AU

AraliaAralia cordatacordata

HeracleumHeracleum candicanscandicans

Zschocke

S., Liu J.-H., Stuppner H., Bauer R., Phytochemical Analysis 9, 283-290 (1998)

Fructus Piperis longi – Bibo 荜拨FructusFructus PiperisPiperis longilongi –– BiboBibo 荜拨荜拨

Stöhr, J., Bauer, R., Wagner, H., Xiao P.G., Chen J.M.: Chinese Drug Monographs and Analysis, 2007

Not Not acceptedaccepted::Piper Piper retrofractumretrofractum VahlVahlsyn. syn. Piper Piper officinarumofficinarum((MiqMiq.) C. DC..) C. DC. ((IndiaIndia: ": "BariBarioror Large Large PeepalPeepal")")

Piper longum var. (Round Peepal)

Piper longum Piper Piper retrofractumretrofractum

The

only

pecies

accepted by

Ch.P.:

Piper longum L.

Discrimination

of Piper longum and Piper retrofractum by

TLC

DiscriminationDiscrimination

of of Piper Piper longumlongum and and Piper Piper retrofractumretrofractum byby

TLCTLC

Stöhr, J., Bauer, R., Wagner, H., Xiao Peigen, Chen Jianmin: Chinese Drug Monographs and Analysis, (2007)

R = Piperin1 = P. retrofractum2 = P. retrofractum3 = P. longum4 = P. longum5 = P. longum6 = P. longum var.7 = P. retrofractum8 = P. retrofractum

R 1 2 3 4 5 6 7 8 R 1 2 3 4 5 6 7 8

R 1 2 3 4 5 6 7 8 R 1 2 3 4 5 6 7 8

Sorbent: silica

60F254Solvent: toluene-EtOAc-formic

acid

80:15:10

Anise aldehyde/sulfuric

acid

/

VIS

UV 254 nm

On the

Chinese market

we

found mainly

Piper retrofractum

AstragaliAstragali RadixRadix, , HuangHuang qiqi 黄芪黄芪•

Huang

qi

is

derived

from

Astragalus mongholicus BUNGE •

Other

taxa

such as A. hoantchy, A.

dshimensis, A. lehmannianus, A. sieversianus, A. chrysopterus, A. aksuensis, A. tribulifolius, A. floridulus, A. lepsensis, A. ernestii, A. laxmannii , A. penduliflorus and even Hedysarum polybotrys are

often

used as adulterants [Wagner, H. et al. (1997) Radix

Astragali

(Huang

Qi). In: Chinese Drug Monographs and Analysis. Verlag für Ganzheitliche Medizin Dr. E. Wühr

GmbH]

ITS ITS SequenceSequence

Analysis of Analysis of AstragaliAstragali

radixradix, , HHuuáángng qqíí

ITS1 5.8S ITS2

A. tribulifoliusA. austrosibiricusA. hoantchyA. dshimensisA. chinensisA. sieversianusA. lehmannianusA. lepsensisA. aksuensisA. ernestiiA. propinquusA. mongholicus TGGTGCGCAA

GTGTACGCT TGGCGCGCAATGGCGCGCAATGGCGCGCAATGGCGCGCAATGGCGCGCAATGGCGCGCAATGGCGCTCAATGGCGTGCAATGGCGCGCAAGGG- - - - - - - -GGACTCACAG

GTGTAGGCTGTGTAGGCTGTGTAGGCT

GTGTAGGCTGTGTAGGCTGTGTAGGCCGTGTAGGCTGTGTAGGCTGTGCATGCTGTGCGGGCA

GTGTAGGCT

Species

specific

mutations: C

T G

C

Heubl,

G.; Wölkart,

K.; Wenzig,

E. M.; Heydel,

B.; Bauer,

R., Planta Med. 73, 1022 (2007)

黃芪

Pyrrolizidine alkaloids are a threat to human health and safety

PyrrolizidinePyrrolizidine alkaloidsalkaloids areare a a threatthreat to human to human healthhealth and and safetysafety

•

International Programme on Chemical Safety

(IPCS). 1989. Pyrrolizidine

Alkaloids Health and Safety

Guide. Health and

Safety

Criteria

Guide 26. WHO: Geneva.•

In 1992, the

Federal Health Department of Germany has

restricted

“the

manufacture

and use

of pharmaceuticals containing

pyrrolizidine

alkaloids

with

a unsaturated

necine

skeleton”. The

herbal

plants

“may

be

sold

and used

only

if

daily external

exposure

to no more

than

100 μg

pyrrolizidine

alkaloids

and internal

exposure

to no more

than

1 μg

per day for no more

than

six

weeks

a year”.

Fu, P.P., Yang, Y.C., Xia, Q., Chou, M.C., Cui, Y.Y., Lin G.: Pyrrolizidine

alkaloids

- Tumorigenic

components

in Chinese herbal

medicines

and dietary

supplements,

Journal of Food and Drug Analysis, 10(4), 198-211 (2002)

•

Compositae

family: in plants

of the

Senecioneae

subtribe

(24 genera, the

genus

Senecio

is

prevelant) and the

Eupatorieae

subtribe

(mainly

in the

genera

Eupatorium and Ageratum); •

Boraginaceae

family: in virtually

all plants

of this

family; and

•

Fabaceae

family

(Leguminosae): in the

subtribe Crotalariaceae, mainly

in the

genus

Crotalaria, but

also in the

genera

Chromolaena and Lotononis.

Ligularia dentataChiyetuowu

Lithospermum erythrorhizonZicao

Senecio scandensQianliguang

Sources

of pyrrolizidine

alkaloidsSourcesSources of of pyrrolizidinepyrrolizidine

alkaloidsalkaloids

PyrrolizidinePyrrolizidine alkaloidsalkaloids in in ZicaoZicao

Ruan

Zicao

软紫草Arnebia euchroma

PA content

ca. 0.0006%, mainly

angeloylretronecin

Zicao

紫草Lithospermum erythrorhizon

PA content

ca. 0.02%, mainly intermedin

and myoscorpin

A Test for pyrrolizidine

alkaloides

is

currently developed

for the

European Pharmacopoeia.

A Test for A Test for pyrrolizidinepyrrolizidine

alkaloidesalkaloides

isis

currentlycurrently developeddeveloped

for for thethe

European European PharmacopoeiaPharmacopoeia..

•

Stir-baking•

Scalding

•

Calcining•

Carbonizing

•

Steaming•

Boiling

•

Stewing•

Blanching in boiling water

MethodsMethods

of of ProcessingProcessing

accordingaccording

to Chinese to Chinese PharmacopoeiaPharmacopoeia

•

Processing with wine•

Processing with vinegar

•

Processing with salt-water•

Levigating

•

Stir-baking with ginger juice

•

Stir-baking with honey•

Frost-like powder

•

Levigating

Processes

need

to be

standardized, effects

need

to be

studied, and end points

need

to be

defined!

The processing chart (a) and processing method of Radix Aconiti

Lateralis

Preparata

(b 1. Cutting; 2. Washing; 3. Stir-frying; 4. Drying; 5. Burying) recorded in the Ming Dynasty.

Z.Z. Zhao et al., Planta Medica 2010; 76(17): 1975-1986

AconiteAconite

rootsroots

in TCMin TCM Radix

Aconiti

(Chuanwu 川烏) Aconitum carmichaeli

Radix

Aconiti

lateralis

(Nifuzi

泥附子) Radix

Aconiti

kusnezoffii

(Caowu

草烏)

Aconitum carmichaeli

R1

R2

LD50Aconitine

C2

H5

OH

0.22 mg/kg Hypaconitine

CH3

H

0.27 mg/kg Mesaconitine

CH3

OH

0.50 mg/kg

R1

R2

Csupor D, Wenzig

EM, Zupkó

I, Wölkart K, Hohmann J, Bauer R. J. Chromatogr. A 1216(11): 2079-86 (2009)

Pattern of alkaloids

in unprocessed roots

of Aconitum carmichaeli

Pattern of Pattern of alkaloidsalkaloids in in unprocessedunprocessed

rootsroots of of AconitumAconitum carmichaelicarmichaeli

Mes

acon

itine

Hyp

acon

itine

Aco

nitin

e

10-O

H-M

esac

oniti

ne

Aco

nine

Deo

xyac

oniti

ne

14-B

enzo

ylac

onin

e

R1

R2

Aconitine

C2

H5

OHHypaconitine

CH3

HMesaconitine

CH3

OH

R1

R2

The

effect

of processing

on the

alkaloid composition

of roots

of Aconitum carmichaeli

TheThe

effecteffect

of of processingprocessing

on on thethe

alkaloidalkaloid compositioncomposition

of of rootsroots

of of AconitumAconitum carmichaelicarmichaeli

14-B

enzo

ylac

onin

e

Aco

nine

Aco

nitin

eAconite

roots

soaked

in water

for 1 hour, then

boiled

for 5 hoursProcessing

of pure aconitine

14-B

enzo

ylm

esac

onin

e

Mes

acon

ine

Csupor D, Wenzig

EM, Zupkó

I, Wölkart K, Hohmann J, Bauer R. J. Chromatogr. A 1216(11): 2079-86 (2009)

Code HPLC (%) Titration (%)Zhicaowu

–

Aconiti

kusnezoffi

praeparataB3 not detectable 0,065B4 0,007 0,013B6 not detectable 0,194Zhichuanwu

(Shanxi) –

Aconiti

praeparata

(radix)D1 0,054 0,207D2 not detectable 0,120Aconiti carmichaeli

radix

praeparata/Aconiti radix

praeparataE1 0,013 0,062E2 0,162 0,433Shanghai market –

Aconiti radix

praeparata/Aconiti radix

lateralis

praeparataF2 –

2003 not detectable 0,045F3 –

2004 not detectable 0,103F4 -

2005 not detectable 0,123F5 –

2008 0,125 0,342F6 –

2009 not detectable 0,070German

market –

Aconiti radix

praeparata/Aconiti radix

lateralis

praeparataG1 –

2004 0,003 0,113G2 –

2004 0,026 0,129G4 –

2007 0,043 0,097G5 –

2007 0,011 0,116

Csupor D, Borcsa B, Heydel B, Hohmann J, Zupkó I, Ma Y, Widowitz U, Bauer RPlanta Medica 76(12), 1332 (2010)

Content of toxic alkaloids in commercial aconite rootsContent of toxic alkaloids in commercial aconite roots

Determination of Determination of thethe contentcontent

of of thethe

„„relevantrelevant““ constituentsconstituents

Chinese Angelica RootAngelicae

sinensis

radix

当归Chinese Angelica is the dried root of Angelica sinensis (Oliv.) Diels (Fam. Umbelliferae). The drug is collected in late autumn, removed from rootlet and soil, slightly dried and tied up in small bundles, placed on a shelf and smoke dried.

Content: not less than 0.05 per cent of trans-ferulic

acid (C10

H10

O4, Mr 194.184), calculated with reference to the dried drug.

1,92, 86484

2,11, 458092

2,56, 4024882,91, 959897

13,41, 390698

14,48, 45363

0 5 10 15 20 25 30 35 40 45

Retention Time (min)

0,00

0,02

0,04

0,06

0,08

0,10Intensity (AU)

tran

s-Fe

rulic

acid

cis-

Feru

licac

id

Fingerprint

of constituents

of Danggui

当归FingerprintFingerprint

of of constituentsconstituents

of of DangguiDanggui

当归当归

Zschocke, S., Liu, J.-H., Stuppner, H., Bauer, R., Phytochemical Analysis 9, 283-290 (1998)

SenkyunolideO

O

Z-LigustilideO

O

Butylphthalide

O

O

0 10 20 30Time (min)

O

HO

OH

Angelicachromene

Riligustilide

O

OO

O

Levistolide A

O

O

OO

Tokinolide B

O

O

O

O

Coniferylferulate

HO

O

O

OH

OCH3

OCH3

E-B

utyl

iden

phth

alid

eE-

Ligu

stili

de

Z-B

utyl

iden

phth

alid

e

Falcarindiol

OH

OH

Lino

leic

acid

γ−Li

nole

nic

acid

Assay

of Danggui

当归in the

Hong Kong Monograph

AssayAssay

of of DangguiDanggui

当归当归in in thethe

Hong Kong Monograph Hong Kong Monograph

Z-LigustilideO

O

Ferulic

acid

Limits The sample contains not less than 0.60% of Z-ligustilide

(C12H14O2), calculated

with reference to the dried

substance.

Salvianolic

acid

BTanshinone

IIa

Rosmarinic

acid

SalviaeSalviae miltiorrhizaemiltiorrhizae radix radix ((DanshenDanshen

丹参丹参))

Tanshinone

IIA Salvianolic

acid B Rosmarinic

acidCP 2010 ≥

0,2 % ≥

3,0 %

HK Monograph ≥

0,12 % ≥

4,4 % ≥

0,17 %BP 2010 ≥

0,12 % ≥

3,0 % ≥

0,17 %

Pharmeuropa

23.3 ≥

0,2 % ≥

3,0 %

HPLC of HPLC of Salvia Salvia miltiorrhizamiltiorrhiza ((DanshenDanshen

丹参丹参):):Simultaneous determination Simultaneous determination

of hydrophilic and of hydrophilic and lipophiliclipophilic

compounds compounds

1,87

2,43

2,53 3,09

10,37

12,29

13,60

15,25

29,07

29,84

31,49

32,37 33,44

0 5 10 15 20 25 30 35 40

Retention Time (min)

0,0

0,1

0,2

0,3

0,4

Intensity (AU)

Rosm

arinic

acid

Salvianolic

acid

B

Tanshinon

IIA

Cao

J, Wei

YJ, Qi

LW, Li P, Qian ZM, Luo HW, Chen J, Zhao: J. Biomed Chromatogr. 2008; 22(2): 164-72

Assay of Assay of Salvia Salvia miltiorrhizamiltiorrhiza ((DanshenDanshen

丹参丹参))

Ph.PRC: MeOH

1 h refluxHong Kong: MeOH/DCM

8:2 30 min US

Cao

et. al.: 70 % MeOH

1 h US

Assay of Assay of Salvia Salvia miltiorrhizamiltiorrhiza ((DanshenDanshen

丹参丹参))

(60 % MeOH) 

(75 % MeOH) 

(75 % MeOH) 

Determination of Determination of salvianolicsalvianolic

acid B acid B in in SalviaeSalviae miltiorrhizaemiltiorrhizae radix radix ((DanshenDanshen

丹参丹参) )

Determination of Determination of tanshinonetanshinone

IIA IIA in in SalviaeSalviae miltiorrhizaemiltiorrhizae radix radix ((DanshenDanshen

丹参丹参) )

Suggested Specification:Suggested Specification:Tanshinone

IIA Salvianolic

acid B Rosmarinic

acid

≥

0,12 % ≥

3,0 %

Summary and ConclusionsSummarySummary and and ConclusionsConclusions•

In order to guarantee safety and efficacy, Chinese herbal drugs must undergo serious quality control, which should be based on pharmacopoeia monographs.

•

The methods should include simple but significant tests for identity and purity, and an assay for pharmaceutically relevant compounds.

•

The complexity of constituents should be considered by using polar and non-polar markers.

•

Harmonization of pharmacopoeia monographs should be a common goal.

Astragali radix, Huangqi 黄芪AstragaliAstragali radixradix, , HuangqiHuangqi 黄芪黄芪PhEur

7.0 / CP 2010:

Assay by HPLC: min. 0,04 % Astragaloside

IV;

Astragalus Astragalus mongholicusmongholicus Fisch.Fisch.

ContentContent

of of AstragalosideAstragaloside

IV (%) IV (%) in in variousvarious

batchesbatches

of of AstragaliAstragali

radixradix, ,

cultivatedcultivated

in Germanyin Germany

ExtractionExtraction

methodmethod

of of thethe European European PharmacopoeiaPharmacopoeia

7.07.0

••

Samples are Samples are extracedextraced

using using SoxhletSoxhlet

ExtractionExtraction••

Instrumental parameters:Instrumental parameters:

»»

4.0 g 4.0 g powderedpowdered

plant plant matrialmatrial

was was maceratedmacerated

withwith 40 ml 40 ml methanolmethanol

overover

nightnight

»»

80 ml 80 ml methanolmethanol

was was addedadded

and and extractedextracted

under under reflux at about 150 reflux at about 150 °°C for 4 hoursC for 4 hours

»»

Evaporated and reEvaporated and re--dissolved in 10 ml waterdissolved in 10 ml water»»

Washed with Washed with nn--butanolbutanol

(4 times with 40 ml)(4 times with 40 ml)

»»

Washed with Ammonia Washed with Ammonia RR [72 % Ammonia ([72 % Ammonia (≥≥ 25%) in water] (2 times with 40 ml)25%) in water] (2 times with 40 ml)

»»

Concentrated to dryness, reConcentrated to dryness, re--dissolved in 5 ml dissolved in 5 ml waterwater

»»

Applied to RP 18 SPE column and Applied to RP 18 SPE column and eluatedeluated

with with 25 ml ethanol 70 %25 ml ethanol 70 %

»»

Evaporated and reEvaporated and re--dissolved in 5.0 ml methanoldissolved in 5.0 ml methanol

HPLCHPLC--ELSD ELSD chromatogramschromatograms

of of AstragaliAstragali RadixRadix

extractextract

min0 10 20 30 40 50 60

mV

0

250

500

750

1000

1250

1500

1750

2000

ELS1 A, Signal Voltage (D:\DATA\KARIN\ASTRAGALUS 2011\060511\AST 6A LAUT AB2.D)

Area: 5

8982.2

34.

923

min0 10 20 30 40 50 60

mV

0

500

1000

1500

2000

2500

ELS1 A, Signal Voltage (D:\DATA\KARIN\ASTRAGALUS 2011\060511\AST 6B NUR KOCHEN2.D)

Area: 7

24.207

34.

878

Area: 1

1429.9

38.

084

Area

: 45900.3

46.

049

Area: 2

345.32

50.

137

WithoutWithout ammoniaammonia

UsingUsing

PhEurPhEur samplesample

preparationpreparation

AstragalosideAstragaloside

IVIV

AGSAGS--II

AGSAGS--IIII

Monschein M, Ardjomand-Woelkart

K, Rieder J, Wolf I, Kunert O, Bauer R, in preparation

ExtractionExtraction

of AGSof AGS--IV, AGSIV, AGS--II, AGSII, AGS--I, MI, M--AGS IAGS I usingusing

different different solventssolvents

(Extraction

without

hydrolysis)

Monschein M, Ardjomand-Woelkart

K, Rieder J, Wolf I, Kunert O, Bauer R, in preparation

Actual

Status of Work

of theTCM-Working

Party

•

21 Monographs adopted

by

the

European Pharmacopoeia

Commission

•

Drafts

of 22 Monographs in PHARMEUROPA•

Monographs for

General Methods:

• Chapter

on Processing: published• Test for Aristolochic

acids: published

• Test for Pyrrolizidine

Alkaloids: pending

Elaboration

of Monographs for Chinese herbs

for the

European Pharmacopoeia

ElaborationElaboration

of Monographs for Chinese of Monographs for Chinese herbsherbs

for for thethe

European European PharmacopoeiaPharmacopoeia

Monographs published

in the

European PharmacopoeiaMonographs Monographs publishedpublished

in in thethe

European European PharmacopoeiaPharmacopoeiaBistortae

rhizoma

(Quan

shen

拳参) Ph. Eur. 6.0

Notoginseng

radix (San qi

三七) Ph. Eur. 6.0Sanguisorbae

radix (Di yu

地榆) Ph. Eur. 6.1

Schisandrae

fructus

(Wu wei

zi

五味子)

Ph. Eur. 6.3Carthami

flos

(Hong hua

红花) Ph. Eur. 6.4

Ephedrae

herba

(Ma huang

麻黄) Ph. Eur. 6.7Stephaniae

tetrandrae

radix (Han Fangji

防己)

Ph. Eur. 7.0

Astragali

mongholici

radix (Huang qi

黄芪) Ph. Eur. 7.0Scutellariae

baicalensis

radix (Huang qin

黄芩) Ph. Eur. 7.1

Sophorae

japonicae

flos

immaturus

(Huai

hua

槐花) Ph. Eur. 7.2Acanthopanacis

gracilistyli

cortex (Wu jia

pi 五加皮) Ph. Eur. 7.3

Angelicae

dahuricae

radix (Bai

zhi

白芷)

Ph. Eur. 7.3Angelicae

pubescentis

radix (Du huo

独活)

Ph. Eur. 7.3

Isatidis

radix (Ban lan

gen 板蓝根)

Ph. Eur. 7.3Puerariae

lobatae

radix (Ge

gen 葛根) Ph. Eur. 7.3

Puerariae

thomsonii

radix (Fen ge

粉葛) Ph. Eur. 7.3

Monographs already

adopted

by the

European Pharmacopoeia

Commission

Monographs Monographs alreadyalready

adoptedadopted

byby thethe

European European PharmacopoeiaPharmacopoeia

CommissionCommission

Angelicae

sinensis

radix (Dang gui

当归) Atractylodis

macrocephalae

rhizoma

(Bai

zhu

白术)

Atractylodis

rhizoma

(Cang

zhu

苍术)Poria

(Fu

ling

茯苓)

Drynariae

rhizoma

(Gu

sui bu

骨碎补)

Monographs Monographs publishedpublished

in in PharmeuropaPharmeuropaArnebia

radix (Zi

cao

紫草) 19.1

Eucommia

ulmoidis

bark (Du zhong

杜仲) 19.3 Indigo naturalis

(Qing dai

青黛) 20.1

Acori

calami

rhizoma

(Shui

chang

pu

水菖蒲) 21.1Acori

tatarinowii

rhizoma

(Shi chang

pu

石菖蒲) 21.1

Coicis

semen (Yi yi

ren

薏苡仁) 21.3Aucklandiae

radix

(Mu

xiang

木香) 22.4

Citri

reticulatae

epicarpium

et mesocarpium

(Chen pi 陈皮) 22.4Polygoni

multiflori

radix (He shou

wu

何首乌) 22.4

Polygoni

multiflori

radix immutata

(Shi he shou

wu) 22.4Acebiae

caulis (Mutong

木通) 23.1

Ecliptae

prostatae

herba

(Mohanlian

墨旱蓮) 23.1Lycii

fructus

(Gouqizi

枸杞子) 23.1

Magnoliae

officinalis

caulis cortex (Houpo

厚樸) 23.1Paeoniae

radix alba (Baishao

白芍) 23.1

Paeoniae

radix rubra

(Chishao

赤芍) 23.1Sinomenii

acuti

caulis (Qing feng

teng

青风藤) 23.1

Ligustici

sinensis

radix et rhizome (Gaoben

藁本) 23.2Fraxini

cortex (Qinp

秦皮) 23.3

Clematidis

caulis (Chuanmutong

川木通) 23.3Magnoliae

officinalis

flos

(Xin Yi 辛夷) 23.3

Paeoniae

suffruticosae

cortex (## 牡丹皮) 23.3Salviae

miltiorhizae

radix et rhizome (Danshen

丹参) 23.3

IncreasedIncreased

useuse

of of drieddried

aqueousaqueous

extractsextracts ((„„granulesgranules““))

ConstituentsConstituents??ExcipientsExcipients??

- Front

- Start

Rhizom a

e t Radi x

Ligus t i c i , Gao

Ben 藳本RhizomaRhizoma

e t e t Radi xRadi x

Ligus t i c iL igus t i c i , , GaoGao

Ben Ben 藳本

UV 366 nm UV 254 nm Anis aldehyde, Sulf. Acid, VIS

D T D T D T D. Peinhopf, R. Bauer, 2008

Decoction

(D)

Ethanolic

extract

(T)

SpecificSpecific

qualityquality controlcontrol

methodsmethods

for for

drieddried

aqueousaqueous extractsextracts

((„„granulesgranules““) )

areare

neededneeded!!

AcknowledgementsAcknowledgementsAcknowledgementsIng. Barbara HeydelMag. Doris PeinhopfMag. Ute WidowitzMag. Isabella WolfDr. Karin Ardjomand-WölkartDr. Olaf KunertDr. Liu JianghuaDr. Marlene MonscheinDr. Eva-Maria Pferschy-WenzigDr. Jochen

Stöhr

Dr. Sibylle

Zschocke

Prof. Dr. Ulrich BommeDr. Heidi HeubergerDr. Johann Rieder Prof. Dr. Günther Heubl

Dr. Dezsö

CsuporProf. Dr. Ingrid HohmannProf. Dr. Hermann StuppnerProf. Dr. Xiao Pei-Gen

Thank you very much!

Welcome to Graz! 格拉茨欢迎你

13th

International Congress of the

Society for Ethnopharmacology

in Graz, September 2. -

6. 2012“Ethnopharmacology along the Silk Road”